Letters

https://doi.org/10.1038/s41591-019-0673-2

Undulating changes in human plasma proteome

profiles across the lifespan
Benoit Lehallier   1,2,3*, David Gate1,2,3,4, Nicholas Schaum5, Tibor Nanasi1,2,3,6, Song Eun Lee1,2,3,4,
Hanadie Yousef1,2,3,4, Patricia Moran Losada1,2,3, Daniela Berdnik1,2,3,4, Andreas Keller   7,
Joe Verghese8,9, Sanish Sathyan8,9, Claudio Franceschi10,11, Sofiya Milman8,12, Nir Barzilai8,12
and Tony Wyss-Coray   1,2,3,4*

Aging is a predominant risk factor for several chronic diseases
that limit healthspan1. Mechanisms of aging are thus increas-
ingly recognized as potential therapeutic targets. Blood from
young mice reverses aspects of aging and disease across
multiple tissues2–10, which supports a hypothesis that age-
related molecular changes in blood could provide new insights
into age-related disease biology. We measured 2,925 plasma
proteins from 4,263 young adults to nonagenarians (18–95
years old) and developed a new bioinformatics approach that
uncovered marked non-linear alterations in the human plasma
proteome with age. Waves of changes in the proteome in the
fourth, seventh and eighth decades of life reflected distinct
biological pathways and revealed differential associations
with the genome and proteome of age-related diseases and
phenotypic traits. This new approach to the study of aging led
to the identification of unexpected signatures and pathways
that might offer potential targets for age-related diseases.
Aging underlies declining organ function and is the primary
risk factor for several diseases1. Thus, a deeper understanding of
aging is likely to provide insights into mechanisms of disease and
to facilitate the development of new antiaging therapeutics. A grow-
ing number of investigators have applied genomic, transcriptomic
and proteomic assays (collectively referred to as ‘omics’) to stud-
ies of aging11. Human genetic studies have uncovered relatively
few modifiers of aging, yet other omics modalities, which measure
more dynamic gene modifications or products, have provided valu-
able insights. For example, the transcriptome varies greatly during
aging across tissues and organisms12, pointing to evolutionarily
conserved, fundamental roles of developmental and inflammatory
pathways13. The protein composition of cells, bodily fluids and tis-
sues changes similarly with age and provides insights into complex
biological processes, as proteins are often direct regulators of cel-
lular pathways. In particular, blood, which contains proteins from
nearly every cell and tissue, has been analyzed to discover biomark-
ers and gain insights into disease biology. Accordingly, organismal
aging results in proteomic changes in blood that reflect aspects of
aging of different cell types and tissues.
Perhaps the strongest evidence that blood can be used to study
aging comes from experiments employing heterochronic parabio-
sis, which is a surgically induced state that connects the circulatory
systems of young and old mice. These studies show that multiple
tissues, including muscle, liver, heart, pancreas, kidney, bone and
brain, can be rejuvenated in old mice2–10. Plasma (the soluble fraction
of blood) from old mice is sufficient to accelerate brain aging after
infusion into young mice9, and young plasma can reverse aspects
of brain aging10,14. Together, these studies support the notion that
the plasma proteome harbors key regulators of aging. Identifying
such protein signatures may help in understanding mechanisms
of organismal aging. However, plasma proteomic changes with age
have not been thoroughly exploited and require new tools to derive
insights into the biology of aging. In this study, we carried out a
deep proteomic analysis of plasma from young adults to nonage-
narians. Using new analysis tools, we discovered changes in protein
expression across the lifespan and linked these changes to biological
pathways and disease.
Results
Linear modeling links the plasma proteome to functional aging
and identifies a conserved aging signature. We analyzed plasma
isolated from EDTA-treated blood acquired by venipuncture from
4,263 healthy individuals aged 18–95 years from the INTERVAL15
and LonGenity16 cohorts (Fig. 1a and Extended Data Fig. 1).
Currently, one of the most advanced tools for the measurement of
plasma proteins is the single‐stranded oligonucleotides known as
aptamers17,18, which bind to targets with high affinity and specific-
ity. To generate a proteomic dataset of the human lifespan, we used
the SomaScan aptamer technology, which is capable of quantifying
thousands of proteins (Supplementary Tables 1 and 2) with high
precision within and between runs19 (Supplementary Table 3). The
INTERVAL and LonGenity datasets analyzed here can be interro-
gated with an interactive web interface (https://twc-stanford.shin-
yapps.io/aging_plasma_proteome/).
Because females have a longer average lifespan than males20,
we assessed whether sex and aging proteomes are interconnected

1
Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA, USA. 2Wu Tsai Neurosciences Institute, Stanford University,
Stanford, CA, USA. 3Paul F. Glenn Center for the Biology of Aging, Stanford University, Stanford, CA, USA. 4Department of Veterans Affairs, VA Palo
Alto Health Care System, Palo Alto, CA, USA. 5Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA, USA.
6
Institute of Cognitive Neuroscience and Psychology, Hungarian Academy of Sciences Research Centre for Natural Sciences, Budapest, Hungary.
7
Clinical Bioinformatics, Saarland University, Saarbrücken, Germany. 8Institute for Aging Research, Department of Medicine, Albert Einstein College
of Medicine, Bronx, NY, USA. 9Department of Neurology, Albert Einstein College of Medicine, Bronx, NY, USA. 10Department of Experimental, Diagnostic
and Specialty Medicine, University of Bologna, Bologna, Italy. 11Department of Applied Mathematics, National Research Lobachevsky State University of
Nizhny Novgorod, Nizhny Novgorod, Russia. 12Department of Genetics, Albert Einstein College of Medicine, Bronx, NY, USA. *e-mail: lehallib@stanford.edu;
twc@stanford.edu

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Letters NATurE MEDicinE

(Fig. 1b–d). The proteins most strongly changed with sex included
the well-known follicle-stimulating hormone (CGA FSHB),
human chorionic gonadotropin (CGA CGB) and prostate-specific
antigen (KLK3). With age, the most prominent overall changes
with respect to fold change and statistical significance included
sclerostin (SOST), ADP ribosylation factor interacting protein 2
(ARFIP2) and growth differentiation factor 15 (GDF15), in addi-
tion to several proteins that differed with sex, such as CGA FSHB.
The proteins most strongly associated with age also changed sig-
nificantly with sex (Fig. 1d): 895 of the 1,379 proteins altered
with age were significantly different between the sexes (q < 0.05;
Supplementary Table 4). These results are aligned with several stud-
ies that demonstrated that males and females age differently21. To
determine whether these findings are representative of the general
population, we compared changes identified in this study with
findings from four independent cohorts from the US and Europe
(n = 171; age range, 21–107 years; Extended Data Fig. 1d) and with
an independent study22. Although these independent cohorts used
an older version of the SomaScan assay measuring only a subset of
the current proteins (1,305 proteins; Supplementary Table 2), we
observed high consistency of the aging and sex proteomes across
cohorts (Extended Data Fig. 2).
To establish the biological relevance of these changes, we que-
ried the Gene Ontology (GO), Kyoto Encyclopedia of Genes and
Genomes (KEGG) and Reactome databases and measured enrich-
ment of proteins in pathways using sliding enrichment pathway
analysis (SEPA) (Supplementary Tables 5 and 6). The heat maps
produced by SEPA first illustrated the relationship between the top
100 proteins and the biological pathways they represent; second,
the heat maps emphasized how a restricted list of top aging-related
proteins revealed biological pathways that would have escaped
common pathway mining modalities (Fig. 1e). SEPA indicated that
incremental lists of proteins are needed to determine the biologi-
cal functions of sex-related proteins and pointed to expected dif-
ferences in hormonal metabolism and activity. Conversely, an
extensive list of aging proteins contained enrichment for blood-
related pathways, such as heparin and glycosaminoglycan binding,
as recently reported22.
To determine whether the plasma proteome can predict biologi-
cal age and serve as a ‘proteomic clock’, we used 2,817 randomly
selected individuals to fine-tune a predictive model that was tested
on the remaining 1,446 individuals (Fig. 1f). We identified a sex-
independent plasma proteomic clock consisting of 373 proteins
(Supplementary Table 7), which was highly accurate in predict-
ing age in the discovery, validation and four independent cohorts
(r = 0.93–0.97; Fig. 1g and Extended Data Fig. 3a,b). Remarkably,
individuals who were predicted to be younger than their chrono-
logical age performed better on cognitive and physical tests (Fig. 1h
and Supplementary Table 8). Although a reduced model compris-
ing only nine proteins predicted age with good accuracy (Extended
Data Fig. 3c and Supplementary Table 7), a combination of different
sets of proteins may be required to model changes in a large set of
clinical and functional parameters (Extended Data Fig. 3d).
As most biological pathways that change with age are evolution-
arily conserved23, we next identified aging-related proteins conserved
between mice and humans. We analyzed mouse plasma (n = 110;
age, 1–30 months) using SomaScan, which reliably measures hun-
dreds of non-human proteins and has proven useful in mouse stud-
ies6,24 (Fig. 1i and Supplementary Table 9). In mice, 172 proteins
changed with age (out of 1,305 proteins measured; Supplementary
Table 10; q < 0.05), and 46 proteins overlapped with human aging-
related proteins (Fig. 1j). Remarkably, many of these proteins were
also modulated by heterochronic parabiosis: young mice exposed
to old plasma (young heterochronic mice) showed a relatively older
plasma signature, whereas aged mice exposed to young plasma
(old heterochronic mice) showed a younger signature (Fig. 1k).
Altogether, standard linear modeling of the plasma proteome dur-
ing the human lifespan revealed established aging pathways, possibly
indicating accelerated and decelerated aging in humans and mice.
Intriguingly, changes to the conserved aging-related proteins did not
occur simultaneously (Fig. 1l). Thus, the chronology of aging in the
plasma proteome requires further investigation.
Clustering protein trajectories reveals undulation of the aging
plasma proteome. Although standard linear modeling showed
prominent changes in plasma protein composition, the undulating
behavior of the 46 conserved proteins (Fig. 1l), and, more globally,
the 2,925 plasma proteins as a group when they were visualized as
z-scored changes across the lifespan, was striking (Fig. 2a,b). These
undulating patterns were detected in independent human cohorts
and in mice (Extended Data Fig. 4), suggesting that they are robust
and conserved.
To reduce the complexity of the proteome, we grouped proteins
with similar trajectories using unsupervised hierarchical cluster-
ing (Fig. 2c) and identified eight clusters of protein trajectories
changing with age, which ranged in size from 8 to 1,415 proteins
(Supplementary Table 11). In addition to linear patterns (clusters 1
and 5), several non-linear trajectories were evident, including step-
wise, logarithmic and exponential trajectories (clusters 2, 3, 4, 6, 7
and 8) (Fig. 2d). Notably, these cluster trajectories were similarly
detectable in independent cohorts (Extended Data Fig. 5). Of the
eight clusters analyzed, six were enriched for specific biological path-
ways (q < 0.05; Extended Data Fig. 6 and Supplementary Table 12),
suggesting distinct, yet orchestrated, changes in biological pro-
cesses during the lifespan. For example, proteins present in blood
microparticles consistently decreased with age (cluster 5), and other
blood-related pathways, such as heparin and glycosaminoglycan
binding, increased in a two-step manner (cluster 4), whereas levels
of proteins involved in axon guidance and EPH–ephrin signaling
remained constant until age 60 before rising exponentially (cluster 6)
(Fig. 2d). Altogether, most plasma proteome changes across the
lifespan were non-linear.

Fig. 1 | Linear modeling links the plasma proteome to functional aging and identifies a conserved aging signature. a, Schematic representation of analysis
of the plasma proteome. b,c, Volcano plots representing changes of the plasma proteome (n = 4,263) with sex (b) and age (c). Linear models (l.m.),
adjusted for age, sex and subcohort, were tested using the F-test. d, Relative percentage of variance explained by age and sex. Values for each plasma
protein are connected by edges. e, Pathways associated with sex and age identified by SEPA (n = 4,263). Proteins upregulated and downregulated were
analyzed separately. The top ten pathways per condition are represented. Enrichment was tested using Fisher’s exact test (GO) and the hypergeometric
test (Reactome and KEGG). f, Schematic representation of biological age modeling using the plasma proteome. g, Prediction of age in the validation cohort
(n = 1,446) using 373 plasma proteins. The Pearson correlation coefficient between chronological and predicted age is given. h, Association between
delta age (difference between predicted age and chronological age) and functional readouts in old. Top associations in both the discovery and validation
datasets are represented. i, Schematic representation of the comparison between the human and mouse aging proteomes. j, Conserved markers of
aging. Both human and mouse aging effects are signed by the beta age of the corresponding linear analysis. Forty-six plasma proteins change in the same
direction in mice and humans (red dots) and define a conserved aging signature. k, Alteration of the conserved aging signature by parabiosis. Normed
principal-component analysis was used to characterize changes of the conserved aging signature when mice were exposed to young or old blood.
l, Age-related changes of the conserved aging signature. Plasma protein levels were z scored, and aging trajectories were estimated by LOESS.

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NATurE MEDicinE Letters

a b Sex proteome c Aging proteome

4,263 participants (18–95 years) 350 CGA.FSHB
DEFB104A
SPINT3 KLK3 350 SOST CGA.FSHB
LEP
300 CGA.LHB 300

–log10 (P value age)

IGFBP6

–log10 (P value sex)

CGA.CGB ARFIP2
GDF15
250 CRISP2 250 MLN
IGDCC4
200 NUDT16L1
VIT CLIC5 200 RET
PTN CGA.LHB
LHB SLITRK4
MB IL1RL1 MSMP SCARF2 CGA.CGB
150 IGFBP3 150
100 100
2,925 Male
plasma 50 50
proteins 0 0
Female
–0.4 –0.2 0.0 0.2 0.4 0.6 –0.005 0.000 0.005 0.010 0.015
Age
Up in Up Down Up
female Sex effect (beta lm) in male with age Age effect (beta lm) with age

d e Aging vs sex pathways

Aging vs sex effects Aging Sex
10 10010 100
R-HSA-373076-Class A/1 (rhodopsin-like receptors)
Percentage of variance

R-HSA-500792-GPCR ligand binding

0.6 hsa04080-neuroactive ligand–receptor interaction
R-HSA-388396-GPCR downstream signaling
(partial Eta-2)

GO:0005179-hormone activity
GO:1901681-sulfur compound binding
0.4 GO:0008201-heparin binding
GO:0005539-glycosaminoglycan binding
GO:0044449-contractile fiber part
GO:0030016-myofibril
0.2 GO:0043292-contractile fiber
GO:0030017-sarcomere
hsa01100-metabolic pathways
GO:0005578-proteinaceous extracellular matrix
0.0 GO:0031012-extracellular matrix
R-HSA-392499-metabolism of proteins
Age Sex R-HSA-948021-transport to the Golgi and subsequent modification
R-HSA-975576-n-glycan antennae elongation in the medial/trans-Golgi
GO:0005179-hormone activity
Signed –log10 (FDR)
R-HSA-2980736-peptide hormone metabolism
–3 +3

f g Prediction of biological age h Associations with ∆age

100
Validation dataset (n = 1,446) 4 Trail making test B time
validation (signed effect)
Association with ∆age

Pearson: 0.97
Predicted age (years)

80
∆ age 2
Discovery Validation
(n = 2,817) (n = 1,446) 60
0
y

Aging
∆ age
clock 40 –2

20 –4 Physical grip
Clinical and
functional 20 40 60 80 100 –4 –2 0 2 4
readouts x age (years)
Chronological Association with ∆age
discovery (signed effect)

i j Conserved aging proteome k

10 INSR GDF15 Alteration of the conserved aging
Mouse aging (signed effect)

CGA TSHB signature by parabiosis

IL18BP
5 CCL7
MRC1
Human aging proteome CHRDL1 26 proteins Young–iso
0
Young–het

KLK7
Conserved –5 GDF11.MSTN
PC1 (33%)
aging markers MSTN Old–het
MIA
Mouse aging proteome –10 EPHB6
Old–iso
20 proteins

–200 –100 0 100 200 300

PC2 (15%)
Human aging (signed effect)

l Non-uniform changes of the conserved aging signature

Young

Old
Young

Old
STIP1
PLCG1
UBB
WNK3
SPOCK2
IGF1
GDF11.MSTN
MSTN
ISLR2
KLK7
SGTA
BOC
HAPLN1
TYK2
EDA
TNFAIP6
RTN4R
LSAMP
MIA
EPHB6
GDF15
TNFRSF1A
CHRDL1
INSR
CGA.TSHB
CAST
PPBP
GHRL
CD36
CCL5
KRT18
BMP7
SNX4
GCG
CCL7
CYP3A4
HMOX2
INHBA.INHBB
IL18BP
TNFSF15
MRC1
CHI3L1

ALCAM
PYY
EDA2R
MB

z score
–0.5 +0.5

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 a

d
Protein levels (z score Protein levels (z score) Protein levels (z score)

1846
–2
–1
0
1
2

2
–1
0
1
2
–2
–1
0
1
2
20

20
20

NA
n = 12

40
40

n = 1,002
40
Letters

60
60

Top pathway(s)
Top pathway(s)

Cluster 2
Cluster 7

Extracellular space

Age (years)
Age (years)
60

80
80
Age (years)
Protein profiles

listed in Supplementary Table 12.

100
100
80
Protein levels (z score) Protein levels (z score)

–2
–1
0
1
2
–2
–1
0
1
2
b

2,925 plasma proteins

20

20
20
n = 60

40
40

n = 1,415

60
60

Top pathway(s)
Top pathway(s)
Axon guidance/
40

Cluster 1
Cluster 6

Age (years)
Age (years)

Cellular localization
and binding functions
EPH–ephrin signaling

80
80

(Fig. 3c, Extended Data Fig. 7a and Supplementary Table 13). These
crests disappeared when the ages of individuals were permutated
the software tool differential expression-sliding window analysis
ers waves of aging-related proteins. To quantitatively understand

aging (Fig. 3b). Summing the number of differentially expressed

identified hundreds of proteins changing in waves throughout
we detected changes at particular stages of life and determined
(DE-SWAN) (Fig. 3a). This algorithm analyzes protein levels within
the proteomic changes occurring throughout life, we developed

(e.g., 35–45 years compared to 45–55 years), while sliding the win-

Quantification of proteomic changes across the lifespan uncov-

proteins at each age uncovered three crests at ages 34, 60 and 78

controlling for the effect of confounding factors). This approach
the sequential effects of aging on the plasma proteome (while also
dow in increments of 1 year from young to old. Using DE-SWAN,
a window of 20 years and compares two groups in parcels of 10 years
100
100
60
Age (years)

Protein levels (z score) Protein levels (z score)

Protein trajectories

–2
–1
0
1
2
–2
–1
0
1
2
Cluster trajectories
80

20
20
–0.5

n = 117

40
40

n = 204
95
z score

Top pathway(s)

60
60
Top pathway(s)

Cluster 5
Cluster 3

Blood microparticle

Age (years)
Age (years)
+0.5

glycosaminoglycan)

80
80
c

Height
Protein binding (heparin,
0
5
10
15

100
100

WFDC2.11388.75.3
FSTL3.3438.10.2
NPPB.7655.11.3 GDF15.4374.45.2
RNASE1.7211.2.3
SVEP1.11109.56.3
SVEP1.11178.21.3 MMP12.4496.60.2
SCARF2.8956.96.3
CCDC80.3234.23.2
CHRDL1.3362.61.2
SUMF1.6941.11.3 PTN.3045.72.2
HLA.DMA.10639.1.3
FIGF.13098.93.3
ERLEC1.8957.72.3
CST8.10572.65.3
CLMP.10440.26.3
ZHX3.10036.201.3
EPHA10.6036.78.3 CTSZ.4971.1.1
CLSTN3.6291.55.3
PTHLH.2962.50.2
MANSC4.9578.263.3 CSMD2.9971.5.3
LGALS9.9197.4.3
CXCL16.2436.49.4
EDN2.12574.36.3
OAF.6414.8.3
REN.3396.54.2
IGLON5.6478.2.3
GNLY.3195.50.2
FAM3B.9177.6.3
TREML4.11139.4.3
SLITRK4.7139.14.3
PARVA.13434.172.3CTBS.6115.40.3
SLAMF1.7953.20.3MB.3042.7.2
MFAP4.5636.10.3
DCTN2.5879.51.3
CDNF.4962.52.1
DNAJB12.8006.12.3TTN.11352.42.3
RBFOX2.11462.8.3
S100A13.7223.60.3
CREB3L4.11308.8.3
PCDHAC2.9361.7.3
NPDC1.10424.31.3
PAPPA.4148.49.2 MXRA7.8005.1.3
UBE2J2.8802.24.3
POSTN.3457.57.1
FKBP7.9288.7.3
CECR1.6077.63.3
PVRL2.6245.4.3
NFASC.7179.69.3
SEMG2.6373.54.3 FAS.5392.73.2FAS.9459.7.3
LST1.9531.24.3
DLK1.6496.60.3 MXRA8.10521.10.3
PPY.4588.1.2
GREM2.5598.3.3
TPPP2.12800.5.3
CD163.5028.59.1 NPW.9986.14.3
SHISA3.7057.18.3
ASPN.6451.64.3
CD34.9023.9.3
CA3.3799.11.2
CBR1.12381.26.3
CSRP3.9171.11.3LHB.8376.25.4
SCGB1C1.5960.49.3
PLXNB2.9216.100.3 HAVCR1.9021.1.3
LAG3.9950.229.3
CCL3.3040.59.1
EDA2R.3083.71.1
SFRP1.3221.54.1
F8.13499.30.3THBS3.8982.65.3
KIR2DS2.10428.1.3
POMC.9204.33.3
IL15RA.14054.17.3
SPON1.4297.62.3
DPT.4979.34.2 MFAP2.9294.45.3
RSPO4.8464.31.3
TREM1.9266.1.3
TNFRSF11B.8304.50.3
LTBP4.13133.73.3
CHIT1.3600.2.3
REG4.11102.22.3
ADAMTS5.3168.8.2
CAPN2.14684.17.3
NTN1.6649.51.3
CHI3L1.11104.13.3
VEGFA.2597.8.3
ACYP2.12812.25.3
CXCL10.4141.79.1
IGFBP6.14088.38.3
MRC1.2637.77.2
CD55.5069.9.3
TIMP4.6462.12.3
HAVCR2.5134.52.2
RNASE4.5644.60.3
KCNE5.8908.14.3
RANBP3.14037.18.3
KRT20.12975.11.3
NPPA.5443.62.2
TNFSF15.2968.61.1
C1QTNF1.6304.8.3
FLRT2.13122.19.3
PLAUR.2652.15.1
NRXN3.5111.15.3
ESAM.7841.84.3 IL18BP.3073.51.2
RSPO3.13094.75.3
NPC2.6259.60.3
STC1.4930.21.1
DPY30.13943.38.3
VIT.6234.74.3
DCBLD2.9338.2.3
TBCA.12501.10.3
CXCL9.9188.119.3 PCOLCE.11237.49.3
IGFLR1.7244.16.3
CD59.11514.196.3
NEGR1.7050.5.3
APOF.12370.30.3
EPHA2.4834.61.2
UNC5C.5139.32.3 LMAN2.9468.8.3
RNASE6.5646.20.3
CHGB.8235.48.3
GAS1.5463.22.3
TWSG1.9234.8.3
UNC5B.7776.20.3
COL18A1.2201.17.6
DSC2.13126.52.3
TNFRSF1A.2654.19.1 ROR2.7861.9.3
TXNDC5.11212.7.3
COL28A1.10702.1.3
TMED10.6506.54.3 COL6A3.11196.31.3
TNFRSF1B.3152.57.1
SMOC1.13118.5.3
TNFRSF1B.8368.102.3 PTGDS.10514.5.3NBL1.2944.66.2
EPHB6.5078.82.3
PIANP.9599.6.3
EPHB2.8225.86.3
THBS2.3339.33.1
TGFBR3.3009.3.2
MMP2.4160.49.1DLK2.9359.9.3
MATN2.3325.2.2
FSTL1.13112.179.3 RGMB.3331.8.1
EFNA5.2615.60.2
KLK11.2831.29.1
TNFRSF21.5404.53.3
NELL2.6022.57.3
CCL14.2900.53.3
EFNB1.13104.32.3
PI3.4982.54.1
NEGR1.13109.82.3
DLL1.5349.69.3
LCN2.2836.68.2
SFN.4829.43.6
CD48.3292.75.1
ANGPT2.2602.2.2 EFNA2.14124.6.3
LYZ.4920.10.1 B4GALT1.13381.49.3
XXYLT1.6375.75.3
ROR1.2590.69.4
TFF2.9191.8.3
PCDH10.9018.38.3
NMB.9321.400.3
EFNA4.2614.28.2
TNFRSF19.5131.15.3
SECTM1.13093.6.3
PENK.9076.25.3
CTSV.3364.76.2
KDR.3651.50.5 RET.3220.40.2
NAGPA.11208.15.3
ADAMTS13.3175.51.5
GHR.2948.58.2 EGFR.2677.1.1
SERPINF2.3024.18.2
ATF6.11277.23.3
IGLL1.6485.59.3 CDH3.2643.57.2 APOL1.11510.31.3
HMGCS1.13496.19.3
NAB1.13933.276.3
RUFY1.11425.31.3
TNXB.5698.60.3
SLC35G2.13501.10.3
LMAN2.7638.30.3 KIT.2475.1.3
AHSG.3581.53.3
CBLN4.5688.65.3 KLK7.3378.49.2
ENPP5.6556.5.3
SELL.4831.4.2
MSMP.8080.24.3 CHAD.13460.4.3
COL11A2.11278.4.3 IGDCC4.9793.145.3
SPOCK2.5491.12.3 CDON.4541.49.2
CAT.3488.64.2
NELL1.6544.33.3
F10.3077.66.2
F10.4878.3.1
ALDOC.9876.20.3
FCN2.3313.21.2
FAM151A.7856.51.3
ERBB3.2617.56.35
DEFB1.6629.3.3
CA6.3352.80.3
IL1R2.14133.93.3
ITIH1.7955.195.3
SIGLEC12.8352.26.3
ASAH2.3212.30.3
CA10.13666.222.3
F2.5316.54.3
BPGM.12020.39.3 AIMP1.2714.78.2
WNK3.5493.17.3
UBB.6641.60.3
PMEL.6472.40.3
SMIM9.8888.33.3
SERPINA4.3449.58.2
PAICS.9841.197.3
GDF11.MSTN.2765.4.3
NDST1.6927.7.3
IL19.3035.80.2
SLC5A5.12826.5.3
NAALAD2.7986.98.3
PIGR.3216.2.2
SOD2.5008.51.1
SERPINC1.3344.60.4
MENT.5744.12.3
MFAP5.6440.31.3
IGFBP3.2571.12.3
PLG.3710.49.2
SGTA.3868.8.1
HBEGF.14094.29.3
APOM.10445.20.3
TLL1.6383.90.3
HPGDS.12549.33.3
REXO2.13590.1.3
QSOX2.8397.147.3 AK1.5012.67.1
RFESD.13603.7.3
TMOD1.12595.11.3
BLVRA.11382.5.3
MDH1.3853.56.1
S100A6.13090.17.3
LRIG3.3322.52.2
IL31.10455.196.3
TOP1.2876.74.2
ENTPD5.4437.56.3
ST3GAL6.6947.4.3
GPC3.4842.62.2
DNAJB1.3852.19.2
PPP2R1A.12621.55.3
AMIGO1.9979.13.3
FAM177A1.8039.41.3
INSIG1.10663.42.3
STMN4.6267.51.3
DIMT1.12694.28.3
ACPL2.6079.59.3 EGF.5509.7.3
NTN4.3327.27.1
SERPINA6.4785.30.3
MAP3K7.TAB1.5259.2.3 PSD2.9118.7.3
KIRREL2.9917.16.3
HMGCS2.13704.5.3
ETHE1.3847.56.2
NLRP1.11661.11.3
H1FX.12709.63.3
CXADR.11204.80.3
FAM213A.13423.94.3 FGF8.4394.71.2
GSTA3.4993.16.1
TMEM132C.11128.29.3 GP5.7185.29.3
GZMB.4133.54.2
CSDE1.12735.39.3
CYCS.2942.50.2
NMES1.6406.3.3 IST1.12434.25.3
STIP1.5489.18.3
PIWIL1.12793.4.3
ACE2.2805.6.2
CFB.4129.72.1
RRM1.11360.39.3
NXPH1.4562.1.2
CST7.3302.58.1
PCDHB2.10748.216.3 CNP.6609.22.3
CDC42.9840.2.3
FGR.3810.50.2
ARHGEF10.9061.3.3
QSOX1.6070.11.3
COPS2.14029.42.3
PLCG1.4563.61.2
SLC4A8.12798.46.3
NSDHL.8038.41.3
NRAC.13464.8.3
SIRT2.5030.52.1
ERAP2.8960.3.3
HCAR2.13495.48.3
CFP.2960.66.2
EFNB3.7785.1.3
IGSF11.10700.10.3
GLO1.9883.29.3
G0S2.8931.124.3
ARTN.2939.10.2
GPC5.4991.12.1 ATP1B2.7218.87.3
SELPLG.11266.8.3
CCDC126.6388.21.3
MET.2837.3.2
C1RL.9348.1.3
AGER.4125.52.2
C5orf63.13378.80.3
KLK15.6491.59.3
TNR.11302.237.3
NCAM2.6507.16.3
FAM19A4.6511.17.3
MOB1A.12426.19.3
RAD51L3.12554.10.3 GPC1.8697.38.3
FCRL1.5728.60.3
RB1.5024.67.1
SLC14A1.13430.50.3
LRRC37A2.8897.3.3
SERAC1.8985.13.3
IL36G.9117.4.3
TEK.3773.15.4
MFGE8.4455.89.2
RGS19.12713.365.3
ITGB7.11205.10.3
FUT9.6991.24.3
RNMTL1.9584.105.3
TLR4.LY96.3647.49.4
PRLH.6543.182.3
LRRK2.10990.21.3 H6PD.7161.25.3
EXTL2.6528.95.3 MATN4.7083.74.3
PSAPL1.8814.33.3MBL2.3000.66.1
BOC.4328.2.2
CPA2.9276.7.3
IL22RA2.5087.5.3
NDUFB11.7747.47.3
ADAMTS3.8845.2.3
CD63.9190.7.3
IL23R.5088.175.3
LIFR.5837.49.3
C2orf66.5677.15.3
TYMSOS.8263.64.3
ZNF334.12763.69.3
Protein levels (z score) Protein levels (z score) PMEPA1.6936.7.3
SLC6A14.13053.6.3
CACNA2D3.8885.6.3

LAMA1.LAMB1.LAMC1.2728.62.2
CAPN3.12385.4.3
MAPKAPK5.8382.47.3
HERC5.12934.1.3
EIF3J.13497.34.3
BST1.4535.50.2
TLR1.11149.3.3
PARK2.13013.41.3
SPINT1.2828.82.2
TMEM132A.7871.16.3
TF.4162.54.2
CCL21.2516.57.3
MASP1.3605.77.4
CLIC5.12475.48.3
GEM.12817.1.3
FKBP6.12529.32.3
IGF1.2952.75.2
LPO.4801.13.3

TPST1.7928.183.3
FCRL4.8973.23.3
COL1A1.11140.56.3
BCAN.3461.58.1
TMEM132B.8890.9.3
CTSF.9212.22.3
ITGAV.ITGB5.4917.62.1
HRSP12.14636.25.3
MYBPC1.7648.9.3
TFPI.3336.50.1
CCL11.5301.7.3
CGB2.6213.10.3
CGA.LHB.2953.31.2
CGA.CGB.4914.10.1
CST6.14711.27.3
CGA.FSHB.3032.11.2

GLTPD2.7948.129.3
SCUBE1.8989.40.3
PTPRU.8337.65.3
SSR1.8106.15.3
C1QTNF3.7251.64.3
SIGLEC7.2742.68.2
SOST.13101.60.3
MLN.5631.83.3
PRSS2.5034.79.1
CDSN.7085.81.3
ARFIP2.12630.8.3

BDNF.2421.7.3
CD36.2973.15.2
C5orf38.6378.2.3 MUSK.11547.84.3
TFF1.9185.15.3
CCL17.3519.3.2
ASIP.5676.54.3 CTRB1.5671.1.3
MAN1A2.9077.10.3
ORC6L.12389.4.3
ARSK.8269.327.3
CHST11.7779.86.3
STAT1.10370.21.3
ILF3.12759.47.3
RMDN1.7096.30.3
CHST9.11646.4.3
DCTPP1.4314.12.2
PTPN11.3397.7.4
GLCE.7808.5.3
GGT2.6334.9.3
TP53I11.13022.20.3
SEC13.14689.3.3
ARHGAP30.12807.89.3
PTH.5954.62.3
GFRA1.3314.74.2
IL5RA.13686.2.3
GFRAL.6920.1.3
CCL7.4886.3.1
FAM20A.6433.57.3
ITIH5.8233.2.3
ADSS.12644.63.3
S100A4.14116.129.3 AFP.5792.8.2
SYT11.7089.42.3
NPTX2.6521.35.3
SURF1.8009.121.3
COCH.7227.75.3
SOCS3.11440.58.3 KLK3.8468.19.3
GPC6.5350.14.2
CRYZL1.9207.60.3
KIAA0040.14603.51.3
SELP.4154.57.2
EIF2B1.10080.9.3
DKK4.3365.7.2 DCN.2666.53.2
FAM174A.6597.24.3
PRSS1.3049.61.2
AKT2.14685.17.3
ADAM11.6586.19.3
ASPH.6998.106.3
CD3E.8069.85.3
MMRN2.11895.21.3
TXNRD1.13967.14.3
PPP1R3B.12768.3.3
HMGB2.6913.189.3 CDY1.7097.8.3
CFC1.3294.55.2
SCARF2.9925.56.3
PABPC3.10447.18.3
CRP.4337.49.2
TWF1.12871.10.3
ADAMTS4.2809.25.2
ARL1.12392.30.3
EDC4.13066.42.3
PLAU.4158.54.2
EHMT2.5843.60.3
TOM1L1.13652.2.3
FCGR2B.3310.62.1
UGT1A6.7891.45.3 GRPEL1.7113.1.3
UBXN4.9997.12.3
BGN.3284.75.1
PLBD1.6315.58.3
ARL3.12571.14.3
MTHFS.14107.1.3
SULF2.8305.18.3
ARFGAP2.11664.32.3
HS3ST3A1.8268.98.3
SORBS3.12976.49.3
GMFG.13062.4.3
KIR3DL2.5096.51.3
PAIP1.12430.78.3
THPO.5947.90.3
PIANP.14114.18.3 CXCL13.3487.32.2 LRRC15.6557.50.3
STAMBPL1.12401.3.3TFF3.8323.163.3
TNFRSF17.2665.26.2
GFRA2.2515.14.3
PLA2G2A.2692.74.2
EPHA5.3806.55.1
EFNB2.14131.37.3
CRABP2.11696.7.3
JAG1.5092.51.3
AIFM1.13424.51.3
DCP1A.14008.22.3
PGRMC1.7863.50.3
TINAGL1.11192.168.3
IL22.2778.10.2
LILRA5.7787.25.3
NCR1.8360.169.3
INA.11436.6.3
LAYN.2635.61.2
PDCD1LG2.3004.67.2
IL6ST.2620.4.2
TNFRSF4.3730.81.2
GRN.4992.49.1
ANP32B.4194.26.3
MMP10.8479.4.3
GUCA2B.6223.5.3
SLURP1.6401.73.3
CHST15.4469.78.2
RFNG.7203.125.3
ANGPTL1.11142.11.3
GALNT1.7090.17.3
JTB.9038.12.3
PUF60.10575.31.3
DNAJB9.11214.40.3
VCAM1.2967.8.1
GPNMB.8289.8.3 IL16.2774.10.3
JAM2.2997.8.1
IL12B.IL23A.10365.132.3
ALCAM.5451.1.3
RETN.3046.31.1
CD300C.5066.134.3 CNTFR.14101.2.3
EPYC.9278.9.3
FJX1.7921.65.3
CCL23.3028.36.2
RPN1.6458.6.3
DLL4.3305.6.1
ICAM5.8245.27.3
ICOS.14084.191.3
CCL23.2913.1.2
EVI2B.13028.2.3
FAIM3.6574.11.3 ADPGK.6221.1.3
SCARA5.10419.1.3
QPCTL.8866.53.3
COL15A1.8974.172.3
LSAMP.2999.6.2
MANSC1.9557.5.3
MIA.2687.2.1
TFF3.4721.54.2 ESM1.3805.16.2
CD93.14136.234.3
WFIKKN2.3235.50.2 C9.3060.43.2
NAGK.3894.15.2
CALCOCO2.12534.10.3
FAM19A5.5609.92.3 BCAM.2816.50.2
GALNT16.8923.94.3
EPHA1.3431.54.2
SIRPB1.6247.9.3
ASGR1.5452.71.3
FBLN1.6470.19.3
TNNI3.5441.67.3
RELT.14112.40.3
CD300A.5630.48.3
KAL1.6603.18.3
TPSB2.3403.1.2
YWHAQ.7625.27.3
ARHGAP1.11955.1.3
MDM1.7898.29.3
PCYOX1L.5599.88.3
FAM3D.13102.1.3
ENTPD1.7999.23.3 MANEA.8014.359.3
PYY.3727.35.1
NDUFV2.7748.11.3
FABP1.11516.7.3
SCP2D1.14175.78.3IL9.5834.18.3
ARHGEF25.13519.112.3
SPARCL1.4467.49.2
TMEM190.10442.1.3
STX8.10903.50.3
FAM20B.7198.197.3
PLXNA1.9005.16.3
TRA2B.12373.73.3
EFNB3.2514.65.3
ADAMTS6.6441.62.3
HIST3H2A.14144.3.3
CILP.5717.2.3
NEO1.8900.28.3
CGA.TSHB.3521.16.2
RBM39.4284.18.3
CYR61.6264.9.3
CNTN2.3296.92.2
LAMC2.9580.5.3
STX2.7738.299.3
TMPO.8265.225.3
FGA.FGB.FGG.4907.56.1
KLRC3.7795.14.3 MAGI2.14066.49.3
AMY1A.7918.114.3
IGFBP7.3320.49.2
CHCHD10.11270.17.3SDF2L1.6990.44.3
APLP2.10627.87.3
PRKAR1B.12479.50.3
GPNMB.8606.39.3
GABARAPL1.12661.44.3 MCFD2.10476.23.3
FRZB.13740.51.3
SPATA9.7809.22.3
LMOD1.12504.26.3
CEACAM1.8031.11.3
ST6GALNAC6.7228.2.3 RIPK2.8993.151.3
NoneX.13231.90.3
PEX14.8300.82.3
CIRBP.12724.81.3 CXCL11.3038.9.2
RBM3.12747.89.3 SPINK6.5731.1.3
MYOM2.13534.20.3
CHGA.8476.11.3
SEMA3E.5363.51.3
GSTP1.4911.49.2
SGCB.7034.4.3 TGFBI.3283.21.1
SRSF6.11573.3.3
DIABLO.3122.6.2
FUT5.4549.78.2
TAX1BP3.12498.12.3
PTPN4.14254.27.3
MSMB.10620.21.3
CCL16.4913.78.1
PSG5.9314.9.3
TAC1.9337.43.3
NUDT9.9482.110.3
CANT1.6480.1.3
CBFB.10048.7.3
FCGR3B.3311.27.1
IGFL4.6353.60.3 BID.5798.3.3
AMBN.6522.57.3
UBE2G2.9199.6.3
STAR.10085.25.3
PSG3.6444.15.3 TESC.12831.21.3
DOCK9.14002.18.3
UPK3B.8286.44.3
VSIG1.8367.142.3 LILRB2.5091.28.3 CHRDL2.6086.15.3
PDGFRB.3459.49.2
CDH2.3797.1.1
CDC42BPA.12706.2.3
GSTT2B.11273.176.3
LIPK.6413.79.3
TMEM167B.13421.17.3
LPAL2.9246.1.3
FOXC2.14051.54.3
PTX3.6447.73.3
SCMH1.12604.16.3
FIBCD1.9378.6.3
TSNARE1.7069.9.4
PPAPDC1A.13548.53.3
YIPF6.9984.12.3
KLHL13.12463.7.3
–2
–1
0
1
2
–2
–1
0
1
2

USE1.8057.78.3
DUSP16.14631.22.3
DEFB103A.5679.16.3
RNF122.11160.56.3
HSD17B14.13972.4.3
IL7R.5089.11.3
HNF4A.10041.3.3 CDH15.11215.6.3
RARRES3.10961.15.3
HLA.DQA2.7757.5.3
UBTD2.12875.28.3ZP4.7766.25.3
ITGA5.6932.42.3
AP4M1.10076.1.3
PDE9A.5201.50.4
ART4.6576.1.3
TLN2.14082.56.3
RAET1E.7800.85.3
CUL4B.13743.56.3
SLAMF8.8994.65.3
COG8.9543.131.3
ACLY.12700.9.3
NPPB.3723.1.2
LEMD1.8040.9.3
PAXIP1.12894.3.3
CSF2RB.11137.43.3
VAV3.9830.109.3
SCGB1D2.6508.68.3
JPH1.6940.18.3
ADAM23.7049.2.3
LILRA5.8766.29.3
FGFR3.3809.1.2
FAM171B.8786.6.3
FN1.3435.53.2
INSL4.6410.26.3
GRAMD1C.8336.267.3 WBP1L.9532.5.3
LYSMD3.10563.13.3
FGF23.3807.1.2
CD274.5060.62.3
KMT2C.11402.17.3
WISP3.5927.4.3
TOR1AIP2.10553.8.3
WFDC10A.13429.3.3
DCUN1D5.8760.10.3 XG.8044.90.3
DPYSL5.12683.156.3
RNF34.13386.248.3
SAT1.9859.180.3
CNTNAP1.14617.7.3
PVR.8064.125.3
AMH.4923.79.1
PSG9.9335.28.3
ITSN1.14070.56.3
ERO1L.7060.2.3
IFNA10.14128.121.3
CLU.4542.24.2
ANKRD27.12445.50.3
MAN1A1.11291.73.3
RBM19.11468.15.3
NEURL1.13604.27.3LY9.3324.51.1
C6orf226.8078.15.3
NIPAL4.12864.9.3
WFDC5.6969.14.3
LMCD1.9530.6.3
FOXL2.11375.49.3
SEMA4C.7923.41.3
LCN8.5643.2.3
CD58.10938.13.3
LAT2.5613.75.3
NEURL4.13468.5.3
BACH1.12451.62.3
PECAM1.2695.25.1
ANKRD46.7851.30.3
EDA.14024.196.3
PLBD2.6536.54.3
ICAM3.2649.77.2
CLEC2B.7786.83.3
ELL.11459.81.3
CRB1.12012.33.3
ZNF264.9993.11.3
CRABP1.11967.23.3
ASTL.7993.23.3
CD33.3166.92.1
PAPOLG.8343.224.3
EDAR.2977.7.2
MMP16.9719.145.3
DCAF5.11283.13.3
TJP1.12001.7.3
ARPC3.13573.5.3
NMT1.5196.7.3
HDAC8.2859.69.2
MYO6.9894.13.3
CD38.11513.92.3
COMMD7.2823.7.1
B4GALT5.10425.3.3
FAM189A2.5719.66.3
RFXAP.12726.3.3
MZF1.14662.6.3
A4GALT.8759.29.3
PDZK1IP1.8260.13.3
ZNF843.8321.27.3
FANCL.10063.10.3
CCNC.7817.36.3
SPTLC2.7267.2.3
MNX1.9576.58.3
SLITRK3.10565.19.3
BTNL9.7950.142.3
IL1R1.2991.9.2
CASC4.8838.10.3
ST8SIA2.7920.30.3
HS3ST3B1.6986.17.3
SEC61B.7878.2.3
NEFH.9900.36.3
CLEC4D.7752.31.3
CGREF1.6257.56.3
CRISP2.9282.12.3
GNGT2.10917.40.3
NUDC.8887.21.3
YES1.2878.66.2
PLOD3.10612.18.3
SLCO5A1.11669.39.3
NTMT1.12454.105.3
RNF149.9773.15.3
NQO1.9837.60.3
SOCS7.11657.86.3
CYTH2.12533.135.3
CHCHD2.8015.144.3
ENDOU.5656.53.3
SARS2.12348.46.3
PRRG1.9008.6.3
RNF13.8087.250.3
ZNRF4.9467.24.3
S100A11.14011.17.3
CD70.5807.77.3
ARMC5.8785.1.3 HGF.2681.23.2
IL1RN.5353.89.2
ERH.11614.29.3
UBQLN4.12720.71.3
YWHAB.YWHAE.YWHAG.YWHAH.YWHAQ.YWHAZ.SFN.4707.50.2
CHST3.7189.55.3 TMUB2.9226.6.3
CSF2.4697.59.2
FAM19A2.6430.36.3
BRDT.12748.6.3
LRRC74A.8389.8.3
CBR3.14091.42.3
FAF2.9738.7.3
RBP7.14208.3.3
HRASLS2.7822.11.3
IL36B.14149.9.3
ADAMTS1.3174.2.1
PRNP.6545.58.3
CSF3R.2719.3.4
CD109.3290.50.2
DEFB119.13455.10.3
NLGN4X.5357.60.3
IL1RL1.4234.8.2
GHDC.9730.22.3
TMEM119.11110.4.3
RQCD1.8975.26.3
CCDC167.7797.11.3 HINT2.5612.16.3
MSRA.7137.8.3
PQLC2L.7939.1.3
GLP1R.13085.18.3
FOXJ2.14204.55.3
DEFB108B.5611.56.3 IL5.11071.1.3
PTH1R.13470.43.3
LRIT2.11716.28.3
FGFR2.3808.76.2
PSG7.5651.50.3
TXNDC11.8769.30.3
FLT3LG.14093.10.3
TSSK2.7873.32.3
DCUN1D3.13553.4.3
GLT8D1.8955.60.3
TLR4.11101.18.3
C1orf115.8366.19.3
CLTC.13711.10.3
NR1D1.5236.2.3
MRPL32.7982.10.3
TEX29.10557.6.3
HAMP.3504.58.2
ANGPTL1.9092.33.3
CA2.4970.55.1
ITM2B.8086.49.3
SMR3A.7838.27.3
EVPL.14019.73.3
KIRREL3.4557.61.2
LY6G6C.6256.9.3
TAX1BP1.12721.4.3
AKT1.AKT2.AKT3.3392.68.2
YWHAZ.5858.6.5
GTF2I.13609.11.3
STOM.8261.51.3
CAST.3026.5.2
PATE1.8386.11.3
SIGLEC9.3007.7.2
SEC61G.8872.1.3
C17orf89.9030.56.3
CD96.9735.44.3
STX6.10945.11.3
SPINK2.13405.61.3
FLRT3.9128.34.3
FBXL4.11416.23.3
ULBP1.3081.70.2
RNPC3.12814.17.3
LOXL2.6504.65.3
LRFN2.7200.4.3
MAPK14.5007.1.1
SYT17.9110.2.3
APMAP.10605.22.3
LYG1.9243.10.3
CHIC2.10914.4.3
TNMD.6578.29.3
C1QL4.7132.55.3
NGRN.7153.66.3
FAM96A.8787.21.3
FXR1.13076.4.3
KIN.13476.16.3
PLG.4150.75.2
CSNK1G2.12653.13.3
FBXL5.12846.3.3
U2AF2.13577.25.3
PRKG1.13067.5.3
SPRED1.8318.13.3
GFRA3.2505.49.3
EDF1.12415.122.3
HIST1H1C.2987.37.3
MTFR1.9095.5.3
SHC4.11692.21.3
ARFIP1.13488.3.3
MAPKAPK2.3820.68.2
FATE1.7740.33.3
RMI1.13926.1.3
KIAA1324L.8363.18.3
ZNF10.11567.23.3
FAF2.8761.7.3
CNEP1R1.13532.25.3
NRG4.14139.16.3
MATK.3823.9.1
RMDN3.9290.8.3
HECW1.12669.30.3
MAPKAPK3.3822.54.2
RNASE10.5602.62.3
PSG8.6238.55.3
C1orf198.8035.6.3
STAB1.14599.18.3
GCNT4.10842.7.3
CCNB1.5347.59.3
PVRL4.5734.13.3
LIF.7790.21.3
ABCC6.8935.22.3
KIAA0319.10013.34.3
PMS2.11312.40.3
HTATIP2.10630.5.3
GLIPR1.9265.10.3
IFIT2.9853.3.3
LCORL.4304.18.2
IL10RB.2631.50.2
C18orf32.8236.8.3
HBG1.10665.30.3
GRAMD1C.8842.16.3
DEAF1.6369.82.3
CHST1.7803.4.3
IL10.2773.50.2
LCMT1.4237.70.3
SERF1A.7842.52.3
DNAJC27.12799.65.3
GALNT10.7003.4.3
RBBP6.9887.40.3
RHPN2.11439.88.3
ANXA10.13605.16.3
ACBD6.10075.75.3
IL13.3072.4.2
RAD1.12670.15.3
SNX7.14245.195.3
NHEJ1.11351.233.3
RABEPK.13599.15.3
IMPDH2.5250.53.3
CKAP2.5345.51.3
ARHGAP36.6289.78.3
CLECL1.14308.192.3
CD300E.10798.4.3
C9orf89.7778.104.3
FGFR4.4988.49.2
PAPPA2.5756.66.3
NTRK1.3477.63.2
TRH.5659.11.3
VSIG2.8018.43.3
TMED4.9319.59.3
ASH2L.12832.10.3
CANX.8834.58.3
MMP16.5268.49.3
HRK.8008.28.3
DYNLT3.12867.40.3
OSTN.7265.32.3
CD300LB.10713.151.3
RFFL.14186.13.3
UGT1A8.8899.75.3
NDUFB8.9800.20.3
SPINK8.7962.11.3
PAPLN.11254.13.3
ADAM9.3795.6.2
APCDD1.6599.5.3
IGFBPL1.7815.49.3
UBE2D4.13475.10.3
OSM.14063.17.3
SPACA3.8076.4.3
BECN1.13032.1.3
RNASE13.6424.2.3
PTK2B.8918.64.3
ITGAL.11617.1.3
IL1B.3037.62.1
PNLIPRP1.6627.25.3
EIF4G3.13991.47.3
CALR.5264.65.3
EFHA2.6912.6.3
WBP1.10943.36.3
H2AFZ.4163.5.2
CFL1.4203.50.2
ANGPTL4.3796.79.2
MED1.3892.21.2
TENM3.11107.25.3
C10orf54.14123.34.3
PIK3CA.PIK3R1.3390.72.2
RAB14.14283.12.3
IL12RB2.3815.14.1
NCL.13655.34.3
MCEE.9388.18.3
PDGFRA.10366.11.3
PRRG1.8306.54.3
COTL1.4905.63.1
ANGPTL7.6371.50.3
GADD45GIP1.9302.90.3
GAL.13389.8.3
ADAM29.13549.15.3
TTC17.8800.14.3
CREB3L1.11198.37.3
CASS4.12855.16.3 GDF5.2752.62.2
B3GALT6.7981.230.3
P4HA1.11645.9.3
TNS4.9927.96.3
FKBP14.9340.17.3
CREBBP.13614.6.3
TGFBR2.5133.17.3
LILRA6.7059.14.3 CGA.14056.4.3
SEMA3C.6448.36.3
RAB27B.13596.3.3
TNF.5936.53.3
AOC2.6434.18.3
ISG15.14151.4.3
CLEC10A.10955.4.3
GJA1.9937.7.3
BPIFA1.6473.55.3
PSMA4.14099.20.3
DNAJC15.7197.2.3
C19orf18.9445.44.3
QRFP.6463.59.3
B3GNT6.7082.2.3
PCDHA7.8758.2.3
DHX38.13645.14.3
SPACA7.6466.7.3
KCNN1.13539.131.3
MRPL52.7123.25.3
SLC27A2.9045.3.3
RNF148.7742.11.3
HEPACAM2.9116.28.3 CD84.8770.136.3
CD3G.7924.7.3
GALNT11.8700.325.3 NGF.5801.72.3
SERPINI2.7117.21.3
RAB39B.12403.30.3
GNRH1.5627.53.3
C12orf49.9387.13.3
SPAG11A.5726.49.3
PCDHGC5.7983.1.3
TIGAR.12476.50.3
INSL6.5754.76.3
BRD1.11607.15.3
CEP57.9067.152.3
AHSA1.11633.89.3
IL4R.3055.54.2
COL6A5.11155.16.3 BNIP3.7045.4.3
SBSN.5724.58.3
CHST6.4429.51.2
CCL26.9168.31.3
GCH1.11185.145.3
CPXM1.6255.74.3
CADM4.8619.12.3
PPCDC.13996.16.3
BIRC3.4973.18.1
TP63.10040.63.3
ZNF774.12760.34.3
HSD17B2.8979.1.3
PYDC1.12835.101.3
CLEC4E.7077.9.4
OTUB2.12493.42.3
DSC3.4981.6.1
STMN2.10900.272.3
PDRG1.12593.33.3
STMN3.8019.73.3
KIAA1324.9097.5.3
HMGB3.12775.6.3
SUSD3.5752.63.3
HTATIP2.7753.21.3
SWAP70.13552.7.3
PRKCA.2644.11.2
ELMO1.12764.3.3 RIC3.7125.4.3
KLK4.2833.20.1
PLA2G16.7865.126.3
UBL4A.11490.42.3
DNAJC19.4545.53.3
GSTK1.13474.40.3
IL18R1.14079.14.3
NAB2.13680.3.3
RPLP2.10949.59.3
GAPDHS.8004.15.3
LAIR1.11284.24.3 CKB.3800.71.2
EHD4.11421.10.3
CTCF.14624.51.3
IL24.3321.2.2
ANXA9.13588.11.3
EID3.8079.39.3
CACYBP.12432.23.3
STAU1.12471.47.3
SMYD2.12636.113.3
MEPE.3209.69.1
APBB2.12753.6.3
PRSS37.5653.23.3
PLEKHA1.12459.13.3
CCL3L1.2783.18.2 G6B.8659.68.3
IMPDH1.5229.90.3
SEMA3B.5667.3.3
WSCD2.6274.15.3
GUCA1A.10008.43.3
ELL2.11494.4.3
SUMO3.14623.26.3
UFC1.3405.6.2
UNC13A.14052.26.3
B3GNT1.8259.25.3
FGF7.4487.1.1
AKT2.5360.9.2
RAD23B.12522.6.3
ISCU.7201.5.3
20
20

GGA3.11683.19.3
C11orf68.8307.47.3
ARPP19.4963.19.1
CSK.3363.31.4
ARHGDIB.9846.32.3 APOE.2937.10.2
EIF4EBP2.4184.43.3 VCY.6295.67.3
NCMAP.14614.41.3
FGF6.4130.71.1
FAM3B.5618.50.3
PUS1.11201.19.3
RFK.13059.33.3
NTS.7857.22.3
KIAA2013.6538.90.3
DGCR2.8055.33.3
COL26A1.9539.25.3
IFNA5.6210.100.3
CRIM1.8699.43.3
TMCO5A.6907.17.3
ZNRF3.14122.132.3
VWC2L.7995.16.3 A1CF.12423.38.3
CRH.5614.44.3
RELL1.13399.33.3
AMELX.8578.45.3
SLAMF7.5487.7.3
IFNLR1.7192.37.3
AXIN2.8429.16.3
CHRD.13438.115.3
CDHR3.9463.26.3
CRYGD.12366.16.3
SF3B4.13449.25.3
PGM1.9173.21.3
TSR3.7018.10.3
DEFB110.8340.9.3
ZNF75D.11596.47.3
NGFR.8949.3.3
MGAT4C.7208.60.3
ARF3.12578.13.3
CERS5.13494.6.3
BATF3.8858.21.3
XRCC4.9886.28.3
CALY.10671.67.3
N6AMT1.11096.57.3
NCF2.10047.12.3
IFNA7.14129.1.3
POMC.4890.10.1
PDILT.7092.7.3
FAM163B.10880.38.3
CDKN3.14178.18.3
GCKR.5223.59.3
ULK3.12437.18.3
C8orf33.9613.16.3
HDLBP.13570.43.3
TMPRSS15.3189.61.2
SIGLEC11.8913.22.3
FAM174B.9054.8.3
DOK2.14246.50.3
KIAA1024.9075.121.3
ODC1.13689.2.3
GSKIP.12849.25.3
PGK2.13936.24.3
LGALS1.8046.9.3
MAPK8.3825.18.2
FAM209B.7066.199.4
CD300LF.5623.11.3
KCT2.6365.62.3
PLEKHA7.12731.12.3 IL11.4493.92.1
CHST14.7262.191.3
CEP57.9905.8.3
WFDC3.6384.19.3
VAPB.7181.17.3
SMAP1.11649.3.3
TBCE.11211.7.3
TOLLIP.13963.7.3
EIF1AX.9850.38.3
SMAD4.12022.12.3
GRB2.5464.52.3
LPCAT2.11175.45.3
SH3BP2.7769.29.3
CSNK1D.11289.31.3
P2RX6.7233.73.3
PAFAH1B2.2642.4.1
NCAM1.7746.230.3
CRKL.9877.28.3
ASAH1.5748.20.3
EFNB2.8772.5.3
HPGD.4995.16.1
CDHR1.8372.29.3
MYC.10362.35.3
SHC1.5272.55.2
PRKAA1.PRKAB1.PRKAG1.5183.53.3 THPO.8059.1.3
IL17A.9170.24.3
C6orf89.10885.36.3
TGFB2.4156.74.1
PRB4.12590.67.3
PLA2G2C.8850.5.3
ELAVL1.11592.1.3
MRAP2.10889.2.3
ARF6.12425.104.3
ATP1A1.11993.227.3
VAMP5.8290.1.3
C2orf82.9439.454.3
PRDM1.14197.2.3
SHC2.14074.2.3
CD226.5062.60.3
MMP17.2838.53.1
TMEM237.12856.14.3
MZT1.8072.19.3
KCNAB2.10015.119.3
COLGALT2.7234.12.3 MFNG.5605.77.3
B4GALT3.6921.24.3
FLVCR1.9073.35.3 PGF.3078.1.2
CLMP.9585.80.3
ARHGAP5.14748.31.3
MRAP.7895.108.3
DPP7.8346.9.3
NETO1.5639.49.3
PRDX4.7789.182.3
POLI.13536.56.3
TMEM9.9249.17.3
SNX4.3903.49.2
NA

RBBP5.13631.1.3
HS3ST4.8998.15.3
DTX1.11430.49.3
ACAP2.12343.14.3
PPM1A.12619.14.3
KRT5.11177.16.3
DHX8.11601.26.3
IER3IP1.6412.26.3
C17orf67.9070.1.3NGFR.8374.5.3
CLPS.5749.53.3
LILRB1.5090.49.2
TFPI2.9233.71.3
ADGRF1.11243.90.3
CCL27.2192.63.10
LGALS7.9196.8.3
NUDT12.13947.371.3
GNAI3.12650.43.3
ANP32A.13073.14.3
RHOG.12540.25.3
ASCC1.10647.18.3
LEPR.5400.52.3
ITPRIPL1.9221.6.3
RBM23.11590.5.3
ELK3.5707.55.3
GGA1.13594.158.3
UGP2.13939.14.3
TOR1AIP1.9039.47.3
HNRNPM.12783.29.3
LONP1.6398.12.3
PROC.3758.68.3
NFU1.7770.25.3
TGFB3.3520.58.1
TGFB1.2333.72.1
LDHB.3890.8.2
ST6GAL1.6035.2.3
THSD7A.11174.8.3
OBP2A.6526.77.3
GCNT1.7016.12.3
ANGPT4.2500.2.3
DHFR.9823.2.3
TPT1.3872.2.1
PSME3.5204.13.3
PAK4.13719.19.3
CLN5.8874.53.3
LILRB4.6453.70.3
TAC3.7847.66.3
TNFRSF14.5352.11.3 STX7.8274.64.3
DEFB135.6411.58.3 GZMA.3440.7.2
CD4.3143.3.1
IGSF8.6984.6.3
OCRL.10011.65.3
TMED2.10761.5.3
SLC14A2.12818.159.3 SST.5957.30.3
CHP1.12458.79.3
TEAD4.12516.13.3
RAP1GDS1.14106.46.3FIGF.14705.1.3
BMP10.3587.53.2
GNPTG.10666.7.3 VTI1B.8963.8.3
GOLM1.8983.7.3
C10orf35.8220.15.3LRP1.10699.52.3
LTA4H.3204.2.2
PDGFRL.9713.67.3
MICB.5102.55.3
IFNGR1.5825.49.3
TNFSF8.3421.54.2 FURIN.6276.16.3
CRELD1.7628.40.3IL17RA.2992.59.2
SLAMF6.5128.53.3
SEMA4D.5737.61.3
CNTNAP2.6965.19.3
CRTAM.7968.15.3
KLK8.2834.54.1
UNC5A.7975.97.3
CADM3.3630.27.4
LY86.3623.84.4
RTN4R.5105.2.3
NTNG1.5637.81.3
PRKCSH.5687.5.3
PTPRS.6049.64.3
CXCL12.3516.60.2
PIM1.5359.65.3
RNF165.11561.32.3
CSF2RA.10438.19.3
KPNA2.2860.19.2
PTEN.3831.21.1
CD79A.7796.10.3
PSG6.6456.17.3
LMAN2L.8013.9.3
PLD3.10948.14.3
PKD2.13745.10.3
NR4A1.8089.173.3
GH1.8462.18.3
PC.13990.1.3
SCARB2.5100.53.3
TRAT1.7224.11.3
KITLG.9377.25.3
HMOX2.2622.18.1
PRKCZ.2645.54.1
CXCL8.3447.64.2
ST8SIA4.7038.45.3 SCIN.12684.5.3
SKIL.14670.1.3
CLEC14A.10461.57.3
IL6.4673.13.2
GCNT2.7143.9.3 BCL2A1.3413.50.2
MADCAM1.11258.41.3 SIRPB2.5669.26.3
FLNA.12906.137.3
VIMP.11286.78.3
BMP6.8459.10.3
GRID2.12758.47.3
PCDHA4.7157.22.3
UCMA.10977.55.3
SRSF7.12987.12.3
ITGA1.ITGB1.3503.4.2 NADK.13624.17.3
ACP5.3232.28.2GCG.4891.50.1
MDK.2911.27.2
n=8

NET1.14260.112.3
F3.4931.59.1
FAM150B.6284.7.3
EWSR1.12988.49.3
VAPA.7167.102.3
IL17RC.5468.67.3
EPO.5813.58.3
PSG4.5649.83.3
OLFML3.8660.5.3
TMPRSS11A.7231.37.3
GPX7.8345.27.3
VEGFC.3132.1.1
NXT1.9942.2.3
ARRDC3.12352.70.3
SPINT2.2843.13.2
HSD17B10.4217.49.3
CXCL14.5730.60.3
PABPC4.12603.87.3
ETNK1.8328.9.3
HDGFRP2.4553.65.3
IL13RA1.2633.52.2
FUT10.7156.2.3
SERPING1.4479.14.2
CELF2.7245.2.3
EPS15L1.4212.5.3
LIPN.8097.77.3
PDGFD.9341.1.3
RRM2B.8925.25.3
ABHD14A.5715.4.3
BAGE2.6294.11.3
NTRK3.2658.27.1
ROCK2.13654.1.3
CRK.4976.57.1
SEMA7A.7019.13.3
GLRX2.12486.8.3
TNFRSF6B.5070.76.3 SYT7.7121.2.3
ZNF18.11372.2.3
EIF1AD.13545.97.3
TBX22.11146.4.3
VPS24.12508.9.3
PSMA2.4280.47.2DNAJC10.8297.8.3
CELA3B.6357.83.3
RETSAT.8381.18.4
AKR1A1.4192.10.2
DNAJB11.7110.2.3
IFNA14.7180.114.3
NCR2.2734.49.4
TIRAP.9839.148.3
HTRA2.3317.33.1
GALNT3.6593.5.3
RAB27A.9504.19.3
LRP1.9182.3.3
FLRT3.13123.3.3
CCL19.4922.13.1
ANXA2.13700.10.3
LRIT3.11919.84.3
ANXA2.4961.17.1
PDCD5.12517.52.3
CSRP2.12968.2.3
LGALS2.3033.57.1
TENM4.11365.17.3
LILRB5.7015.8.3
RND1.13574.50.3
HP.3054.3.2
DPP10.7890.68.3
EIF3G.11454.87.3
ANAPC10.12345.4.3
GPHA2.6395.58.3 INSR.3448.13.2
TNS2.11667.29.3
NID1.3213.65.2
FCGR1A.3312.64.1
ISM1.8355.80.3
IGF1R.4232.19.2
RPN1.10490.3.3
STAT3.10346.5.3
ST3GAL2.6281.51.3
DEFB107A.6399.52.3
CCL13.4144.13.1
MRPL34.6933.20.3
ANAPC7.11690.47.3
IL15RA.3445.53.2
CISD2.8094.20.3 DRGX.8034.6.3
HERC1.12705.9.3
FGF5.3065.65.1
C19orf80.7183.102.3
ABL1.3341.33.4
ATAD1.6625.31.3
STX1B.6966.144.3
PFDN5.4271.75.2
NRG2.8060.7.3
CCL28.2890.59.2
FCRL6.6617.12.3
NT5C.12560.9.3
POMGNT2.6359.50.3 TAPBP.12378.71.3
BAG3.10078.5.3
RAB7B.12409.90.3
INHBA.INHBB.8467.9.3
TDGF1.5810.25.3
CYGB.11546.7.3CISD1.7745.3.3
UQCC3.8228.10.3
BCL2L2.13097.11.3
CXCL9.11593.21.3
MACROD1.13653.335.3 FBP1.7206.20.3
ADSSL1.13998.26.3 NOTUM.8252.2.3
APOB.2797.56.2 ASL.11241.8.3
C14orf93.6439.59.3
DEFB104A.5763.67.3 LILRA4.8299.66.3
CEL.9796.4.3
CCL25.2705.5.2
PDGFB.4149.8.2
PDGFA.4499.21.1 NSF.13992.12.3
QDPR.11257.1.3
C4A.C4B.4481.34.2
PLOD2.6923.1.3
PPBP.2790.54.2
PIP4K2A.12697.30.3
DNAJC17.14655.1.3
RPS27A.2846.24.2
CCL5.5480.49.3
APCS.2474.54.5
IGBP1.12358.6.3
PRDX1.3855.56.1SELE.3470.1.2
SPINK9.8042.88.3
FAH.11424.4.3
UBE2N.3905.62.1
TPI1.4309.59.3 FGF20.2763.66.2
APP.3171.57.2
CUL3.10045.47.3
TKT.4306.4.2
CSNK2A1.CSNK2B.5225.50.3 RPIA.12333.87.3
AREG.2970.60.2
PDXP.3897.61.1
FGF16.4393.3.1
IL2RG.2634.2.2
RAD23A.10058.1.3
HDHD2.13472.35.3
NDUFS4.10584.7.3
MLEC.6285.71.3
HMBS.11530.37.3
UBE2V1.11626.7.3
OLA1.12659.13.3
XDH.11264.33.3
CENPW.8864.59.3
SEMA5A.13132.14.3
DNAJA4.9744.139.3
SERPINE1.2925.9.1 CELA2A.7140.1.3
PLA2G12B.9380.2.3
ADH4.8325.37.3
SERPINA10.13119.26.3
POR.2731.29.2
PCBD2.6899.37.3
SPARC.3043.49.2
CA1.4969.2.1
CARS.14098.28.3
PSMD11.13572.43.3
CFI.2567.5.6
ENPP7.4435.66.2
NSG2.13409.9.3
ST3GAL1.5657.28.3
MLYCD.11538.216.3
LRPAP1.3640.14.3
HNRNPDL.10852.114.3PCBD1.11313.100.3
CUZD1.7943.16.3
BAGE3.6442.6.3
SIRT5.12461.8.3
MCTS1.12488.9.3BMP7.2972.57.2
BAG4.12844.10.3
RDH16.12881.17.3
PSMD7.3898.5.2
MPP7.12732.13.3
LOC652493.6561.77.3 LCK.4560.34.2
CYTH4.12746.4.3
VPS29.14318.1.3
L1CAM.4246.40.2
S100A12.5852.6.3
PSME1.5918.5.3
PIR.13634.209.3
EIF5.2612.5.2
SIAE.9263.57.3
NR1D2.12885.42.3
TMEM2.8992.1.3
ENO2.10339.48.3
DPYSL3.12707.26.3
TBXAS1.8098.37.3
DNAJB2.11438.6.3
SNX8.6925.26.3
XPNPEP1.3481.87.1
ANXA1.4960.72.1
INPP5E.11370.20.3
CROT.13929.27.3
SNRPA.12678.66.3
MAPK3.2855.49.2 SYNE2.9789.52.3
MGAT4B.7141.21.3 INHBC.6408.2.3
CTSA.3179.51.2
ICAM4.6550.4.3
CTSH.8465.52.3
OAS1.10361.25.3
VCL.8750.46.3
MPG.12438.127.3
CETN2.13078.3.3
PLK1.3394.81.2
NRN1.8806.18.3
CXCL5.2979.8.2
RPS6KA6.12688.115.3
MSLN.3893.64.1
PDK2.12651.21.3
APBB1.12822.34.3
SUSD2.10021.1.3
FBXL4.9951.36.3
BCL2.3412.7.1
FUT8.8244.16.3
LAMTOR3.12490.92.3
MTAP.9910.9.3
TFRC.6895.1.3
COL9A2.9804.11.3
IRF2.12801.33.3
APEX1.9849.13.3
TMEM8B.12742.160.3
40
40
n = 107

RAP1GAP.13735.1.3
LRP1B.11275.94.3
EHBP1.12813.18.3
PPIL1.9884.8.3
ADAMTS15.4533.76.2
IFNW1.7196.21.3
UBE2L3.3874.8.1
RGS10.11634.32.3
KREMEN2.3202.28.2
EEF1B2.5882.34.2
LY6G6D.6469.62.3
CD1D.8749.194.3
ASIC4.6951.26.3
ARID3A.3875.62.1
FUT3.4548.4.2
SDHAF2.6420.4.3
GRB14.13628.58.3
SCGB2A1.5001.6.2
PSMA6.3860.7.2
SKP1.3902.21.2
CARTPT.5743.82.3
GMEB2.12804.5.3
CALB1.9918.23.3
RPS27A.4474.19.2
CTGF.2975.19.2
CA5A.8791.151.3
NETO2.10562.42.3
ING1.3888.8.1
HNRNPAB.8894.80.3
HK2.13130.150.3
DDX39B.9742.59.3
RPL30.12478.15.3
RSRP1.9050.170.3
CTRC.5626.20.3
WFDC13.9345.436.3
FTH1.FTL.5934.1.3
LCT.9017.58.3
ESD.4984.83.1
UBE2I.2877.3.1
ACP1.3858.5.1
GSTO1.12436.84.3
DKK1.3535.84.1
WISP1.13692.154.3
PRPSAP1.9478.69.3
CYB5A.11287.14.3
PPARA.12954.71.3
SPTLC1.7886.26.3
FHIT.9826.135.3
NHP2L1.13602.6.3
RPS7.3864.5.2
B4GALT7.7806.33.3
PPIH.12449.16.3
HINT1.5900.11.2
NAAA.3173.49.2
UROS.11248.43.3
SNCA.8458.111.3
TSN.12477.42.3
EIF5A.5888.29.3
GNPDA1.8909.77.3
RPE.12646.2.3
UBB.6651.74.3
GXYLT1.8229.1.3
ERP44.6064.4.3
EIF4B.14675.20.3
UGT2A1.8907.11.3
UAP1.13580.2.3
DEPP.7178.59.3 MAN1B1.7071.23.4MAP2K4.5242.37.3
CSH1.CSH2.13103.125.3
MBD4.3891.56.1
ADM.14115.34.3 PCDHB10.9963.19.3
LRPPRC.9046.46.3
SLC22A16.9969.8.3
PDXK.11098.1.3
CPNE1.5346.24.3
FBXO3.14628.72.3
MYZAP.9055.81.3
VRK1.12553.5.3
MAP2K1.2864.2.3
GNS.3616.3.5
NAMPT.5011.11.1
SLC5A8.13691.10.3 GHRL.8447.11.3
CDH7.7959.34.3
IL1RL2.2994.71.2
NPS.6390.18.3
SSB.13526.5.3
DEFB119.8315.5.3
ANGPT1.2811.27.1
RARRES1.8398.277.3 PIN4.12718.43.3
PTK6.3832.51.1
JAM3.2998.53.2
S100A9.5339.49.3
NPLOC4.12993.21.3 IDO1.9759.13.3
PDZD11.6219.14.3
SMR3B.8595.75.3
GABARAPL2.12494.99.3
MYSM1.11536.9.3
TNFSF4.2839.2.1
SIGLEC10.6048.4.3
SATB1.13511.29.3
DEFB121.5765.53.3 CDKN1B.3719.2.2
CSDC2.12754.14.3
HIF1A.13089.6.3
SPATA20.11117.2.3
PGK1.5020.50.1
CLEC2L.8242.9.3
IFNGR2.8818.13.3
COX6C.8903.1.3
NLRP4.12794.6.3
PROL1.6530.63.3
PTGFRN.12727.7.3
TMIE.7992.3.3
DBNL.4978.54.2
CNTN5.3299.29.2
SSMEM1.8298.8.3 SCG2.8377.87.4
PCBP1.11458.30.3
IGDCC3.11952.1.3
CSNK2A1.3427.63.2
RHOA.9855.10.3
CASP2.4904.7.1
TTMP.7963.36.3
SPG21.11122.97.3
NXF1.12453.161.3
PDIA5.5593.11.3
AGFG1.11681.8.3
UFM1.3836.51.2
SSU72.13566.2.3
PRKCB.5475.10.3
CXCL1.2985.35.1
TDP1.10073.22.3
LARGE.7935.26.3
RPS3A.5484.63.3

indicating the robustness of these age-related waves.

BIN1.9574.11.3
PTPN7.14688.6.3
TSG101.13044.5.3
GNPNAT1.13954.9.3
MSR1.11207.3.3
SEPHS1.14083.25.3
MPL.3473.78.2
SUN5.11260.47.3
COMMD1.12509.115.3
POGLUT1.6467.65.3
DAPK2.4355.13.1
TNFRSF11A.8316.36.3
POFUT1.5634.39.3
TPST2.8024.64.3
HMG20A.12712.9.3
PTP4A2.11699.16.3
LGALS4.2982.82.2
SMAD2.10364.6.3
DEFB123.9362.11.3
SMC3.14324.52.3
GPI.4272.46.2
ARHGAP25.11333.82.3
RPS4X.9758.17.3
ST8SIA6.6930.95.3
MAPK13.5006.71.1
MAPK1.3115.64.2
HNRNPA2B1.5351.52.3 PRCP.5722.78.3
ATG7.12627.97.3 HMGN1.9187.2.3
SPOCK3.9906.21.3 CD209.3029.52.2
ADGRF5.6409.57.3
PYGL.11441.11.3
CYP3A4.2943.5.2
ARHGEF2.12848.9.3
MRPL14.8021.59.3 INS.4883.56.2 FAP.5029.3.1
C1QC.14100.63.3
NME2.4249.64.2
YBX2.6372.7.3
CD74.6974.6.3
BAD.5870.23.2
NQO2.9754.33.3
ABO.9253.52.3
TPM1.5033.27.1
CLPSL2.7767.1.3
NSG1.7802.53.3
COX7A1.8390.25.3
PSMD1.8824.2.3
FSTL4.9350.3.3
SLC6A16.13056.18.3
BDP1.8929.7.3
GALNT7.10888.4.3
SETD2.12647.52.3
PPM1L.7187.3.3
UNC93B1.13487.24.3
LECT1.9928.125.3
PRR16.10621.26.3
VTA1.4209.60.2
UXS1.8258.22.3
NR3C2.12931.16.3
CSMD1.9598.23.3
FLRT1.4547.59.2
TGIF2.9847.21.3
LRRTM4.6572.10.3
SRXN1.14268.4.3
CDHR3.8222.49.3
ZNF382.14616.16.3
STAU2.12970.35.3
MMP14.5002.76.1
PURA.12537.88.3
IL2.3070.1.2
RECQL.11431.235.3
AMN.4322.28.3
C22orf15.7073.69.4
DDX58.12382.2.3
CCNB1IP1.9728.4.3 NRXN1.8971.9.3
TACSTD2.2619.72.2
IGHE.IGK.IGL.4135.84.2
PXDNL.11324.3.3
IL1RAP.14048.7.3
ERMAP.8631.13.3
MAP2K3.6151.18.3
ZNF180.12771.19.3
GRP.5897.58.3
PITPNA.9934.29.3
ULBP3.2747.3.2
LRRN1.11293.14.3
PF4V1.5663.18.3
LRRN3.10471.25.3
CSNK2A1.5224.20.3
ATAD2.13043.157.3
CRLF1.CLCF1.2607.54.2 HBZ.6919.3.3
C1orf226.7989.5.3
TMEM57.12367.52.3
LRRC3.6917.49.3
FGF1.3486.58.2
VPS4A.11476.43.3
PRKCI.3379.29.1
TMPRSS11D.6547.83.3
HNF1A.11193.27.3
CARHSP1.12808.103.3
PSMG3.12729.12.3
RBM28.11927.3.3
RGS3.12827.37.3 DMPK.9010.3.3
ASH1L.12622.96.3
VSTM2L.6549.60.3
IL27.EBI3.2829.19.2
DYNLL1.3881.49.2
RNF148.9955.40.3
PSMD5.10716.35.3
PROK1.2247.20.11
IL31RA.8273.84.3
MRE11A.11319.106.3
TSLP.3010.53.2
DHH.4389.2.1
KIF23.5228.25.2
CCL24.4128.27.2
SEZ6L2.3867.49.1
ARPP21.12860.7.3
RXFP1.14135.3.3GOT1.4912.17.1
CD40LG.3534.14.2
FOLH1.5478.50.2
CCL20.2468.62.3STX18.9037.1.3
IFNL1.4396.54.1
KLK5.3201.49.2
CFH.4159.130.1
UTS2R.13530.5.3
MINOS1.7956.11.3
KLRC3.11571.75.3
ACBD7.13563.259.3
CCL1.2770.51.2
MCF2L.13934.3.3
IL34.4556.10.2
RASA1.5481.16.3
HLA.DPB1.8755.202.3
PDLIM4.12387.7.3
NOVA1.12557.18.3
FCAR.4987.17.1
GOSR1.7805.52.3
PSD.13055.53.3
RAD51.2871.73.2IL37.2723.9.2
PAK3.3387.1.2
ZNF23.11565.58.3
VARS.13083.18.3
PPP2R3A.13665.35.3
STK17B.5249.31.3 DLG3.7897.75.3
SYT3.10452.24.3
GPR135.11465.4.3
C1GALT1C1.5735.54.3
UBE2T.12400.25.3
TMPRSS5.8002.27.3
AURKA.3091.70.2
ENTPD1.3182.38.2
RS1.6497.10.3
PVRL3.13557.3.3
RYK.5583.62.3
SEC11C.10658.28.3
ERO1LB.7994.41.3 SPR.9287.6.3
GLTP.12513.8.3
PDE6D.13491.40.3
STAP1.12510.3.3
IL17RD.3376.49.2
PCSK7.4459.68.2
KYNU.4559.64.2
JAG2.5093.47.3
NENF.9219.70.3
ZFP91.13651.54.3
IL3.4717.55.2
PDE3A.5254.69.3
LILRB3.11334.7.3
IL1RAPL2.5082.51.3
GRAP2.5265.12.3
ASAP2.13518.5.3
ST6GALNAC5.7927.16.3ITM2A.7765.15.3
SNX17.12845.18.3
DGCR6.9444.70.3
KIAA1324.10637.50.3
MAGEB10.11456.2.3
ERAP1.4964.67.1
ASNA1.13620.10.3
CD22.2891.1.3
CHST5.7020.13.3
DHX58.14012.17.3
CD86.5337.64.3
KCNIP3.10513.13.3
SSB.13625.19.3
APOBEC3G.13930.3.3
CEACAM21.7204.1.3
UBE2J1.6900.30.3
FAM24B.8775.61.3
LDLR.13129.40.3
TTL.13973.62.3
SCN2B.8353.15.3
SYT2.9577.26.3
PRMT3.12696.166.3
YWHAE.14157.21.3
UCK2.12515.45.3
GDF11.14587.16.3
DHX9.10527.22.3
TXNIP.11682.7.3
C2orf40.6362.6.3
C8A.C8B.C8G.2429.27.4
EMID1.13021.12.3
KCNG4.13525.17.3
NDUFB4.10677.9.3 CLEC4M.3030.3.2
FLT3.3437.80.3
SPSB1.13942.140.3 IMPAD1.9231.23.3
CAMK1D.3418.12.2
TSTA3.12657.2.3
NISCH.12738.43.3
CRYBB2.10000.28.3
Protein clusters

ATP5B.4965.27.1
SIGIRR.8326.63.3
CAV2.9457.3.3
RLBP1.12936.38.3
FGF8.2443.10.4
PSG2.6405.74.3
ASGR2.9474.22.3
RNF43.14120.2.3
COL8A1.4807.13.3
AGR2.4959.2.1
MANBA.6382.17.3
RHOD.12442.4.3
COX4I2.7850.1.3
MAGEA10.13610.9.3
BIRC5.3472.40.2
GCK.12960.9.3
TYK2.5260.80.3
GALNT2.5764.4.3
JUN.10356.21.3
MCEMP1.6283.60.3
BGLAP.11067.13.3
PITPNB.12484.67.3
M6PR.10491.21.3
PCDHB4.10522.167.3 MMP9.2579.17.5
HSPA6.13672.3.3
PTPRH.11988.24.3
TFRC.8795.48.3
ADGRG5.4551.72.3 ISL1.11549.6.3
CLEC2D.7054.87.3 LTF.2780.35.2
ARSB.3172.28.2
C1QBP.4967.1.1
DCK.9836.20.3
MGA.11587.5.3
HSD17B1.4708.3.2
DGKB.12895.28.3
DEFB112.5689.1.3
HERC4.7860.9.3
CCPG1.8889.5.3
CLK2.11327.56.3
SERPINA12.6551.94.3
MT1F.10416.79.3
PRDM4.12779.30.3
CBS.10086.39.3
PTPN1.3005.5.2
CFAP45.6553.68.3
CSF2RB.10512.13.3
TMEM52.11186.12.3 ENG.4908.6.1
UBC.6647.55.3
SPINK13.10624.45.3
SERPINA9.7266.4.3
DYNAP.10692.48.3
ENOX2.13422.66.3
SAP30.12888.18.3
LEP.8484.24.3
MPST.12686.15.3
TREML1.9329.28.3
DEFB116.11144.10.3 C3.2755.8.2
TBP.2875.15.2
CD200R1L.8980.19.3
CD300LG.9909.4.3
CD200R1.5103.30.3
TNFRSF18.5526.53.3
KIAA1161.8068.43.3
GJD2.11678.105.3
CD5.14065.11.3
CD8B.9310.2.3
PRRG4.9579.59.3
MGAT1.10571.14.3
SUN3.8852.10.3
SEMA3A.3222.11.2
GSTA4.14645.253.3
RCAN1.13465.5.3
NTF3.4145.58.2
KCNF1.12834.3.3
TMEM70.8074.32.3
VTN.13125.45.3
B3GAT3.6897.38.3
WFIKKN1.3191.50.2
IQCF1.7991.54.3
NPFF.5617.41.3
GALP.9398.30.3
KPNA4.12698.72.3
RACGAP1.13587.10.3
FNDC4.13451.2.3
CBX5.4540.11.2
RELL2.6600.70.3
THYN1.12424.107.3
SCARA3.11292.13.3
PVRIG.6464.40.3
PCDH10.8780.2.3
KCNE3.8067.21.3
PEAR1.8892.14.3
SORT1.11300.32.3
NoneX.13230.174.3
PAPSS1.14007.22.3
MUC4.10659.49.3
PARP11.12882.7.3
CDC25B.12427.8.3
TUFT1.5690.49.3
MLL2.13623.4.3
ETS2.12350.86.3
SLC26A5.9127.4.3
CLEC2A.10953.14.3
APBB2.12761.12.3
AGGF1.8051.10.3
CYTIP.13612.7.3
MINPP1.5586.66.3
CLPB.11860.3.3
CRLF2.13694.24.3
DNER.11442.1.3
ZNF134.12787.47.3
C17orf78.6571.75.3
NAT1.12632.14.3
KLRC1.5629.58.3
B3GNT8.9297.12.3
CSAG1.5716.49.3
PARP1.10534.40.3
ZNF174.11486.26.3
RCVRN.14334.3.3
CNTF.3489.9.2
TMEM41B.10661.4.3
EPHB3.9220.7.3
IGSF9.10518.14.3
TNFRSF10A.4832.75.2 BMX.3414.40.2
60
60

DVL2.13575.40.3
SIT1.11194.6.3
GDA.12667.2.3 PTPRJ.8250.2.3
TMEM87B.13485.20.3
CTXN3.10467.58.3
ADH1B.9834.62.3
SLC9B2.9088.20.3
RAP2A.9885.41.3
AKR1C1.12618.50.3
RNASEH1.8470.213.3
ITM2A.10516.53.3
LYZL2.6377.54.3
CA9.3798.71.1
CNTN4.3298.52.2
TST.12663.1.3
PDK1.5227.60.3
ALOX15B.12422.143.3
COA3.7888.58.3
UST.8364.74.3
MTPAP.9028.5.3
MID2.14249.68.3
KIR2DL2.7773.20.3
CEACAM4.10910.6.3 TCN2.5584.21.3
DDX25.13984.23.3
KRT7.11383.41.3
LRRC4C.9369.174.3
FGFR3.13669.6.3
PAM.5620.13.3
SPOP.13727.44.3
TNFAIP3.14009.65.3
TNFSF18.2708.54.2
SIRPG.9241.40.3
RNF150.9774.59.3
DEFB128.6360.7.3
WDR1.10723.41.3
RGS8.11666.72.3
CHI3L2.9383.24.3
TEX29.13500.9.3
NPIPB3.9954.2.3
SEMA6A.7945.10.3
NUP210L.9606.4.3
RAB35.13514.121.3
PSMA1.7880.9.3 FNDC5.8041.5.3
RELB.11464.9.3
APBB1.14206.28.3
EDA.2826.53.2
TNFSF13B.3059.50.2
PCDHA4.10533.1.3
STK16.3471.49.1
SEPT10.12690.33.3
ICOSLG.5061.27.3
Top pathway(s)
Top pathway(s)

TRAF4.13041.47.3
RNF128.6510.56.3
HNRNPC.11429.80.3NDC80.12730.3.3
HTN1.10608.9.3
ERLIN1.8776.10.3
C1orf185.10667.78.3
CUL9.12991.49.3
COLGALT1.5638.23.3
ZNF415.12811.55.3
RIC3.11228.37.3
TYRO3.2611.72.2
DEFB134.10610.8.3
ARL11.12433.8.3
HAAO.5861.78.3
IL3RA.13744.37.3
FBP2.9867.23.3
PSG11.7846.44.3
AIFM1.9522.3.3
CASP10.5340.24.3
SPRY1.9512.24.3
DEFB106A.5664.57.3
ATP4B.9994.217.3
CNOT1.13482.14.3
OPALIN.7736.28.3
VPS4B.12668.7.3
GORAB.7247.1.3
GRB7.11281.6.3
CTSO.9264.11.3
SPHK2.4468.21.2
UBXN4.9970.7.3
BARD1.13977.28.3
FARS2.13941.82.3
FAXDC2.9074.6.3
AIF1.2849.49.1
ROBO3.5117.14.3
TMPRSS11B.10895.28.3
DPEP1.8794.13.3
ENTHD2.7947.19.3
B3GALTL.5727.35.3
PTH2.7257.18.3
KDELC2.8296.117.3
OSMR.10892.8.3
PDE4D.5255.22.3
IL21.7124.18.3
DNER.9769.48.3
MED4.14021.81.3
PPIE.5238.26.3
IL17D.4136.40.2
TMEM234.8107.12.3
POMC.2558.51.3
JPH3.9089.77.3
C7orf69.9544.24.3
CCNH.9848.22.3
LINGO3.10827.67.3
MRRF.12355.223.3
KCNE5.8756.41.3
CSGALNACT2.10772.21.3
ACVR1B.2806.49.2
LCK.3452.17.2
2,925 plasma proteins

GRP.8400.74.3
TMEM230.11542.11.3
ZNF566.12795.2.3
TK1.4301.58.2
GPLD1.11939.11.3
CMA1.3423.59.2
APOO.9373.405.3
SAYSD1.6594.64.3
MUL1.9315.16.3
DNAJC4.8016.19.3
MAPKAPK5.3821.28.1
KIF1C.9899.28.3
PLA2G2E.2447.7.4
DNAJC30.7866.11.3
EIF4A3.4997.19.1
ALDOA.5864.10.3
DEFB125.9486.13.3
B4GALT6.10832.24.3
NCOA2.14045.12.3
HGS.13644.30.3
FSTL5.7099.33.3
CRYAA.10087.10.3
CSNK2A2.CSNK2B.5226.36.3
Receptor activity

SAT2.12524.18.3
DUSP15.12838.28.3
ERCC1.12585.39.3
DEFB118.9306.7.3
ERVV.1.12531.5.3
RNGTT.12847.27.3
SCAMP5.13509.5.3
FCAMR.9568.289.3
TNFSF12.5939.42.3
FAM159B.10504.6.3
SENP7.12626.6.3GZMK.9545.156.3
SUSD1.9582.93.3
GPR107.12963.1.3
CXCL3.CXCL2.3148.49.1 ULBP2.3082.9.2
NCR3.3003.29.2
SPTAN1.5031.10.1
THRA.12527.50.3
LRRC19.7014.27.3
KLK5.14039.33.3
MUTYH.10030.8.3
C1D.7821.6.3
IL12RB1.2632.5.2
EVA1C.8877.22.3
FAM107A.2760.2.2
P4HA2.11348.132.3
KLRF1.5098.79.3
IFI16.12893.159.3
LAMA4.6577.64.3
LAT.10551.7.3
USP25.9215.117.3
TBK1.3400.49.2
EZR.9753.17.3
STAT6.10372.18.3
PDCL2.14192.31.3
CAMK2A.3350.53.2
FST.4132.27.2
NRG1.9178.30.3
USP21.12681.63.3
IL32.9051.13.3
Cluster 8
Cluster 4

SEMA6C.14597.5.3
RBL1.12879.5.3
MAN2B2.9251.28.3
DEFB136.9332.6.3
ZNF410.12843.6.3
PSMA1.3859.50.2
C16orf54.12394.53.3
ANK2.7624.19.3
CAPSL.13688.2.3
BOLA2.8404.102.3
S100A2.11969.5.3
CTNNA2.9872.23.3
ABLIM3.13578.98.3
CCDC51.12790.10.3
DNAJC18.8033.1.3 PTS.12014.19.3
FGF10.2441.2.4
TMEM167A.11112.18.3
PLA2G5.2449.1.4
CDHR5.9962.1.3
NOV.2737.22.2
WNT7A.4889.82.1
LRP1.8601.167.3
ADAM19.8948.13.3
PTPN9.12633.3.3
MFAP1.5606.24.3
RIPPLY1.9767.22.3
BNIP3L.7835.2.3
POMGNT1.8253.2.3
OSBPL11.12878.60.3
KLHL12.12695.62.3
C1orf210.8088.56.3
PANK3.12658.72.3
TEAD3.12854.3.3
NDNF.6604.59.3
CD177.13116.25.3
USB1.11328.9.3
OPHN1.12591.27.3
SYT8.7932.23.3
PHF3.11544.39.3
NME1.5909.51.3
LAD1.6407.63.3
ENAH.9757.29.3
HSP90B1.6393.63.3
FAAH2.8396.42.3 GNMT.14006.36.3
GDF9.3067.67.1
GAN.10054.3.3
KIR3DS1.5097.14.3
PCOLCE2.6081.52.3
MXI1.9035.2.3
NXPH3.6054.6.3
NRSN1.11654.77.3
ISOC1.9816.37.3
ASMTL.10089.7.3
CDK2.CCNA2.3357.67.2
MILR1.11173.29.3
CD207.3361.26.2
TGFB1I1.8777.5.3
SPOCK1.5490.53.3
KIR2DL5A.8000.17.3
DTX3L.11643.73.3
AMICA1.5094.62.3
NRBP1.12616.45.3
C21orf33.5981.6.3
GCA.12594.5.3
YTHDC1.8878.48.3
EMC1.11989.35.3
NBR1.7696.3.3
ALDH3B1.12940.35.3
NAPB.12655.30.3
PATE4.8065.245.3
APAF1.13583.19.3
VAMP4.7732.45.3
FAM107B.7801.30.3
XCL1.4143.74.2
PARP16.7881.244.3
KRT18.5354.11.3
PRIM1.13591.31.3
FGF4.4123.60.2
CALCA.10494.48.3
EDDM3B.7782.34.3
MSRB3.7824.88.3 ELF5.13457.33.3
COL20A1.8804.39.3
CHD7.14005.2.3
CASP3.3593.72.3
FAM172A.5615.62.3
BCAR3.12634.79.3
PSPN.2696.87.2
ARRB1.12643.4.3
LDHC.9828.86.3
MAP3K3.12990.39.3
PLA2R1.10916.44.3
GPCPD1.12786.61.3
GDNF.5822.22.3
CD47.6653.58.3
Age (years)
Age (years)

KIAA0319L.13698.28.3
CRISP3.3187.52.2
HGD.9832.33.3
PRKACA.3466.8.2
FABP5.4985.11.1
TNFRSF12A.5138.50.3
PDE7A.5178.5.3
ZBTB33.12785.49.3
STX10.12842.43.3
ENTPD6.8932.1.3
NAP1L2.13529.39.3
TTC17.9550.153.3IL25.4137.57.2
TTC9B.9386.42.3
TOPBP1.9947.22.3 RTBDN.8613.97.3
TLR2.3835.11.2
CBL.12016.60.3
SAP18.12830.4.3 FCGR2A.3309.2.2
TXN.8813.160.3
NFE2L1.11154.3.3
RUNX3.11138.16.3
KEL.7070.25.4
CBX7.13027.20.3
CREG1.9357.4.3
LRRC4B.11911.13.3
GABBR2.13948.50.3
HHLA2.14132.21.3
MAGEA3.12576.21.3
SEC22A.13713.164.3
DUSP6.12341.8.3
CTF1.2889.37.2
LAMP3.9355.26.3
LRTM2.8906.60.3
HSPA1L.12041.33.3
PPIB.4718.5.2
ATP2A3.13510.7.3
TMEM59L.9959.60.3
NCR3LG1.7854.38.3
PPIL2.14314.6.3
PRPF6.9581.4.3
CASC4.10613.33.3
PRLR.5114.65.3
KCNA10.13042.7.3
ARHGEF1.13976.9.3
RBM24.11380.84.3
PODXL.8792.17.3
HSP90AB1.5467.15.3
UQCRB.12957.62.3
SSRP1.5032.64.1
DES.12030.82.3
MYCT1.13541.1.3
RAPGEF5.9059.14.3
SLC6A9.11653.69.3
LCN10.13007.66.3
ICAM2.5486.73.3
PRH1.10502.15.3
CLC.11094.104.3
CD74.8748.45.3
MMP13.4925.54.2
BCAP29.11570.94.3
FAM134B.8556.5.3
OSM.2693.20.3
ZAP70.4476.22.2
NUDT8.7872.5.3
CDK5.CDK5R1.3358.51.2
BCL2L1.4423.77.2
CDK8.CCNC.3359.11.2
ST6GALNAC1.7867.154.3
LRMP.10704.91.3
RORB.12483.62.3
SPAST.8388.24.3
CDK2AP1.9450.18.3
KRT17.12923.51.3
CDC2.CCNB1.3422.4.2
PAK7.3388.58.2
ZAP70.3837.6.2
ENTPD3.4436.1.2
SETMAR.12462.20.3
MAD1L1.13618.15.3
KLRK1.14095.1.3
ZPLD1.5590.11.3
ADCYAP1.8285.64.3
PNKP.13657.2.3
ZFYVE27.9102.28.3 C6.4127.75.1
GBP6.7818.101.3
MICALL2.12891.1.3
POLR1C.12939.1.3
PIK3CG.3391.10.2
IGFL3.6961.14.3
SUV420H2.12452.32.3
AURKB.3346.72.2
DUSP4.10035.6.3
KCTD5.12473.48.3
KCNIP1.13650.11.3
CCM2.12347.29.3
SLC30A5.9594.30.3
GPD1L.12420.10.3
HK1.13131.5.3
GPR26.13540.1.3
CT55.9363.11.3 SEMG1.7115.5.3
MOCS3.14229.5.3
PTCHD3.13517.3.3
PIK3C2A.14028.22.3
DIRAS3.12406.119.3
BACH2.12756.3.3
SATB2.10081.17.3
JAKMIP3.9068.17.3
CAMK1.3592.4.3
ITM2C.10560.1.3
STC2.6231.46.3
METTL1.12514.16.3
RPRD1A.13515.8.3
NTF4.4146.58.2
RNF114.9957.9.3
APBB3.13589.10.3
COLEC10.6558.5.3
EXOSC3.12605.1.3
DLK1.8380.244.4
SYTL4.11563.51.3
TGFA.9358.3.3
MPP6.13490.1.3
GPLD1.9500.5.3
DEFB113.13374.4.3
LEPR.9566.103.3
HOGA1.10024.44.3
FEN1.12577.100.3
HMGB1.2524.56.3
IARS.12815.9.3
GZMH.3373.5.2
PMVK.12450.42.3
ADAM12.4420.7.2
FAM175B.8945.7.3
ITFG1.9347.13.3
CNDP2.3192.3.2
SMIM10.9379.248.3
RAB22A.12408.333.3
OAZ1.9893.27.3
SULT4A1.8357.43.3
DEFB115.8391.12.3 IRF6.9999.1.3
TPK1.12018.84.3
LGALS8.4909.68.1
EHF.13387.55.3
FCRL3.4440.15.2
IL17C.9255.5.3
KIF16B.11672.17.3
INSL3.5723.4.3
MYRF.8843.34.3
SAMHD1.11303.7.3
APOE.5312.49.3
RNF215.7758.217.3
LYPLAL1.12428.2.3
LDOC1.8378.3.4
ZNF276.14692.3.3
LTBR.2636.10.2
KLK12.3199.54.2CPA4.9267.2.3
MUC1.9176.3.3
C3orf18.6994.19.3
ECE1.8767.44.3
MST1R.2640.3.2
ADCK4.9794.17.3
UPK3BL.6633.43.3
NRXN3.9799.3.3
STAB2.3399.31.2
TPM2.4472.5.2
ST6GALNAC2.7823.22.3
CHEK1.2853.68.2PLD5.8081.55.3
DSG1.2976.58.2
INSL5.10462.14.3
GGH.9370.69.3
BRSK2.9790.28.3
LCTL.10890.135.3
IER3.9786.310.3
COX8A.10497.242.3
XRCC1.12535.2.3
BRPF1.11671.19.3
GRAP.12820.1.3
BCL10.8768.4.3
NAGS.11247.20.3
SNAPIN.6975.52.3
PLA2G10.2949.6.2
BET1L.10959.125.3
FAM19A3.7839.99.3
TBX3.13978.122.3
EMILIN3.8964.14.3
CLEC5A.8282.15.3
OSCAR.7116.31.3
DSCAM.9175.48.3
ERBB2.2616.23.18
SYNCRIP.4224.7.2
SIRPA.5430.66.3 CPZ.6493.9.3
AARS.12340.17.3
PSMD4.13568.30.3
ZDHHC14.11677.17.3 CTRL.9229.9.3
GFAP.3034.1.2
CD79B.6351.55.3
NPTX1.9256.78.3
MGAT2.6909.40.3
PCDHB1.9941.70.3
SIGLEC12.10037.98.3
DEFB105A.10962.46.3
CLEC12A.11187.11.3
HMG20B.11551.16.3
TNFSF9.2599.51.2
ST6GALNAC3.10705.14.3
EFCAB14.10830.5.3
ELK1.10006.25.3
COX5B.7887.57.3
ALDH3A1.11480.1.3
SLC25A18.5280.68.5
80
80

PTPN2.3401.8.2
ARHGDIA.6454.38.3
TNFSF14.5355.69.3
REPIN1.13554.78.3
USP8.13450.49.3
DNAJB14.8053.16.3
SULT2A1.9829.91.3
APPL1.12825.18.3
CLEC6A.6911.103.3
LRCH4.11252.30.3
MTRF1L.11134.30.3
PEX5.5915.58.3
GPA33.6419.75.3
NRXN2.8876.51.3
IRF9.12439.67.3
BUB1.12033.3.3
EREG.4956.2.1
MAMDC2.6119.14.3
CUBN.12904.180.3
ZNF329.12803.9.3
TPO.3873.51.2
TBX5.11202.70.3
DNASE2B.6533.20.3
GJA8.12711.19.3
ITPKA.13473.55.3
NDRG4.7934.11.3
VSTM4.7242.14.3
RRBP1.9492.5.3
DERL1.13393.46.3
MEGF10.11168.3.3
MPDZ.14036.116.3
MMRN2.9723.105.3
KLK6.3450.4.2
SLC30A3.9081.39.3
ZNF358.10529.19.3
IFNA6.5714.88.3
EXOSC1.7930.3.3
CPVL.8891.7.3
LIPF.9353.8.3
SIGLEC8.7864.3.3
STIM1.9271.101.3
VCAN.9561.21.3
ST8SIA3.6929.10.3
TAC4.6468.37.3
CYP3A4.7879.12.3
SYT5.9099.19.3
ABHD12.7825.7.3
LYSMD4.9106.87.3
SMDT1.9497.3.3
CAND1.13937.75.3
CCDC90B.7792.58.3
SLC35B3.10660.33.3
TNFRSF13B.2704.74.1
PES1.4267.81.3
DGCR14.11356.19.3
RGS18.13982.33.3
CLINT1.11659.31.3
BRICD5.6612.90.3
POU2F1.11715.1.3
IDE.3197.70.2
GYPA.9389.12.3
GPR101.11371.1.3
IL4.2906.55.3
KIR3DL3.7944.1.3
ERP29.13728.19.3
IL2RA.3151.6.1
ADRBK1.3347.9.2
LYN.3381.24.2
LYN.3453.87.2
IL1A.4851.25.1
MRVI1.8255.34.3
LMNB1.3889.64.2
IFIT3.13642.90.3
IGDCC3.7118.24.3
MMGT1.7225.51.3
MMEL1.3627.71.4
PRKAA2.PRKAB2.PRKAG1.5245.40.5 VTI1A.7952.2.3
CALN1.10933.107.3
CTSG.2431.17.3
NMRAL1.13988.67.3
PELI2.12702.13.3
SEMA3G.5628.21.3
IFI16.13940.19.3
FAM171B.8061.102.3
NTRK2.4866.59.2
SYTL1.12892.10.3
COLEC11.4430.44.3
PILRA.8825.4.3
NLRP10.12821.6.3
PRC1.11591.43.3
ERCC4.9895.77.3
PCDH9.10558.26.3
EIF4E2.9745.20.3
MEF2C.8933.84.3
GABBR2.9930.48.3
SNAP29.12357.41.3
FAM173A.7148.42.3
AXIN2.12925.105.3
LRFN1.7910.41.3
KIAA1161.8093.13.3
SPINT3.7926.13.3
UMODL1.9114.84.3
PLS1.12737.12.3
TAPBPL.6364.7.3
DMKN.8535.102.3
MTMR1.11167.6.3
ACP2.9237.54.3
CCDC101.12788.6.3
IFNGR2.9180.6.3
CSPG4.8951.162.3
CDKN2B.9874.28.3
RNF219.8822.163.3
NAT14.11120.49.3
ARHGEF7.13932.45.3
ASPRV1.13023.8.3
TMEM40.12941.24.3
PCDH15.14228.1.3
FASLG.3052.8.2
and unsupervised hierarchical clustering was used to group plasma proteins with similar trajectories. c, Hierarchical clustering dendrogram. The eight

SPAG11B.5762.35.3
POLH.10022.207.3
HCK.3374.49.2
LRIT3.11534.6.3
CTSC.3178.5.2
IFNA8.6214.84.3
HMGCR.5230.99.3
ALKBH5.8793.13.3
TOX3.12717.65.3
PAX4.12398.15.3
SMCO2.7925.18.3
FCRL5.6103.70.3
IL12A.IL12B.10367.62.3
UBE2B.9865.40.3
GRIA4.10760.107.3
IL20.4138.25.2
TACSTD2.14034.22.3
CD44.9283.8.3
CAMK2B.3351.1.1
TOR1AIP2.10640.9.3
FIG4.6948.82.3
SV2A.12880.1.3
SNAI2.10014.31.3
IFNL2.4397.26.2
SMPDL3A.4771.10.3
DDC.3538.26.1
RIPK2.8970.9.3
ITM2C.9523.34.3
SLC41A2.12511.83.3
PGRMC2.8681.93.3
TMEM27.7816.23.3
NHLRC3.9087.8.3
CRISPLD2.5691.2.3
ZNF175.12716.3.3
MYZAP.9964.10.3 VAMP8.7064.2.4
LYPD3.13107.9.3
SNX1.8807.13.3
IFNAR1.9183.7.3
POLM.12851.5.3
LRRTM4.8646.61.3
NR5A2.12444.39.3
TFIP11.9043.7.3
RPS3.5026.66.1
DCLK3.7826.1.3
MFN1.11364.18.3
WFDC12.6037.6.3
VAMP3.7903.18.3
LGI3.8003.57.3
CCBL2.12682.5.3
HEG1.8947.268.3 AGRP.2813.11.2
SCG3.7957.2.3
PTPRD.9296.15.3 OMD.5358.3.3
NRCAM.5109.24.3COLEC12.5457.5.2
POFUT2.6042.52.3
IDUA.3169.70.2
HAPLN1.3196.6.2
ACAN.3280.49.2
CD200.5112.73.3
SF1.12777.11.3
FKBP2.9339.204.3
CDC42BPB.3629.60.4CDH12.10701.30.3
ABL2.5261.13.3
GZMM.5704.74.3
KRTDAP.5739.75.3
PRG3.9015.1.3
SEL1L2.9082.25.3
CDH15.5410.53.3
PTPRR.6361.49.3
KIAA1467.7892.132.3
APOL1.9506.10.3
RNF8.14663.44.3
IL27RA.5132.71.3
CYB5R3.7215.18.3
ROBO1.5740.17.3
HYAL1.8309.12.3
PSMB5.12580.7.3
HEPHL1.6354.13.3
LINGO1.6620.82.3
PGRMC2.10506.53.3
ACACB.12900.29.3
GP1BA.4990.87.1
PEAR1.8275.31.3
MPO.2580.83.2
C1QTNF5.7810.20.3
DUSP13.6525.17.3 TNFAIP6.5036.50.1
NOTCH1.5107.7.2
OBP2B.5680.54.3
DPEP2.8327.26.3
EFNA3.14153.8.3
SFRP2.7751.121.3
PTPRH.11222.62.3
LTA.LTB.3506.49.1
NR1H2.9016.12.3
PDE2A.5246.64.3
THSD1.5621.64.3
EMILIN3.8773.172.3
TMEM132D.13416.8.3 MSTN.14583.49.3
SLC26A11.13502.2.3
PI15.5745.64.3
TREML2.5736.1.3
SH2D1A.4567.82.2
TIE1.2844.53.2
B3GNT2.7980.72.3
NUDT16L1.12497.29.3
ISLR2.13124.20.3
LGMN.3622.33.4
DYRK3.4359.87.1
RCN2.10645.72.3
BPI.4126.22.1
MPZ.10615.18.3
SPINK5.8028.22.3
PCSK1.13388.57.3
PPT1.9244.27.3
PRSS22.4534.10.2
RGMA.5483.1.3
TNFRSF10D.14121.24.3
FRK.10034.16.3
KCNE2.10427.2.3
CADM1.3326.58.2
SERPINF1.9211.19.3
PGLYRP1.3329.14.2
PLXNC1.4564.2.2
LCN1.11708.2.3
RNASE2.8394.56.3
EEA1.14043.12.3
MRPL33.13453.2.3
LRTM1.9368.64.3
GNRH2.10708.3.3
SNAP25.13105.7.3
TNFRSF8.2605.49.2
SIGLEC14.8248.222.3 LTA.4703.87.2
LRP8.3323.37.1
TMEM132C.7173.141.3
CX3CL1.2827.23.2
B4GALT2.9595.11.3 KRTAP2.4.14615.46.3
MAZ.13436.54.3
AZGP1.9312.8.3
RAB26.6997.32.3
IL2RB.9343.16.3
CD2.7100.31.3
IGFBP1.2771.35.2
IBSP.3415.61.2
FCER2.3291.30.2
SLITRK5.4568.17.2CRHBP.6039.24.3
NRXN1.5110.84.3MRC2.3041.55.2
EVL.11656.110.3
DAG1.8369.102.3
CLEC11A.2966.65.2
NCAM1.4498.62.2
EMC4.13516.46.3
CBLN1.9313.27.3
PRTN3.13720.95.3
CTSS.3181.50.2
QPCT.7849.3.3
PRL.2585.2.5
HS6ST1.5465.32.3CDH5.2819.23.2
ART3.7970.315.3
DSG2.9484.75.3
CCL22.3508.78.3 SCO1.7853.19.3
HFE2.3332.57.1 LRRTM2.6904.14.3
PRELP.5675.6.3
SFTPC.5738.25.3
FAM163A.6260.14.3 TNC.4155.3.2
HS6ST2.13524.25.3
were z scored, and trajectories of the 2,925 plasma proteins were estimated by LOESS. b, Trajectories are represented in two dimensions by a heat map,

ROBO2.5116.62.2

are presented for each cluster. Pathway enrichment was tested using the GO, Reactome and KEGG databases. The top 20 pathways for each cluster are
clusters identified are represented by orange boxes. d, Protein trajectories of the eight identified clusters. Clusters are grouped by the similarity of global
Fig. 2 | Clustering of protein trajectories identifies linear and non-linear changes during aging. a, Protein trajectories during aging. Plasma protein levels

trajectories, with the thicker lines representing the average trajectory for each cluster. The number of proteins and the most significant enriched pathways
100
100

teins changing at age 34, 60 and 78 years was statistically significant

(MMP12), were significantly changed only at the last two crests,
as chordin-like protein 1 (CHRDL1) or matrix metallopeptidase 12

us to use SEPA to determine whether these waves reflected distinct

age were not identified by linear modeling (Fig. 3f). This prompted
nounced increase across the lifespan (Fig. 3a). Other proteins, such
but a few proteins were among the top ten differentially expressed
posed of different proteins (Fig. 3d and Supplementary Table 14),

(P < 0.05) but limited (Fig. 3e), and most proteins changing in old
reflecting their exponential increase with age. Overlap between pro-
in each crest, such as GDF15, which was consistent with its pro-
Intriguingly, the three age-related crests were largely com-
(Extended Data Fig. 7b) but were still detectable using different sta-

Data Fig. 7c, Supplementary Fig. 1 and Supplementary Table 13),

tistical models (e.g., smaller or larger sliding windows) (Extended

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NATurE MEDicinE
NATurE MEDicinE
biological processes. Strikingly, we observed a prominent shift
in multiple biological pathways with age (Fig. 3g). At young age
(34 years), we observed a downregulation of proteins involved in
structural pathways, such as the extracellular matrix. These changes
were reversed in middle and old age (60 and 78 years, respectively).
At age 60 years, we found a prominent role of hormonal activity,
binding functions and blood pathways. At age 78 years, key pro-
cesses still included blood pathways but also bone morphogenetic
protein signaling, which is involved in numerous cellular func-
tions25. Pathways changing with age by linear modeling overlapped
most strongly with the crests at age 34 and 60 years (Fig. 3g), indi-
cating that dramatic changes occurring in the elderly might be
masked in linear modeling by more subtle changes at earlier ages.
Altogether, these results showed that aging is a dynamic, non-linear
process characterized by waves of changes in plasma proteins that
reflect complex shifts in biological processes.
Proteins linked to age-related diseases are enriched in distinct
waves of aging. The plasma proteome is sensitive to the physi-
ological state of an individual but is also genetically influenced26.
To deconvolute complexity between the genome, proteome and
physiology, we asked whether the top aging-related proteins change
owing to genetic polymorphisms or whether they are among the
top predictors of disease or phenotypic traits. More specifically,
we sought to determine whether proteins that comprised the three
waves of aging were uniquely linked to the genome or proteome of
age-related diseases and traits (Fig. 4a). We used the ranked lists of
the top proteins identified by DE-SWAN at each of the three crests
(Fig. 3c and Supplementary Table 14) and summed the number of
proteins linked to the genome and proteome of specific diseases and
traits separately for each wave (i.e., the cumulative sum) (Fig. 4b–i
and Extended Data Fig. 8). First, we mined the genomic atlas of the
human plasma proteome26 (Fig. 4b) and discovered that the aging
proteome is also genetically determined (Fig. 4c and Extended
Data Fig. 8). However, the rank of proteins determined by trans-
association appeared more random with aging (Fig. 4c), suggesting
that other sources drive the aging plasma proteome. We then tested
whether the waves of aging proteins were differentially linked with
changes in cognitive and physical functions identified in Fig. 1h.
Interestingly, the proteome associated with these traits overlapped
with the proteome defining middle and old age, when these func-
tions decline the most (Fig. 4d,e). Finally, we used public datasets
and summary statistics from SomaScan proteomic studies focused
on age-related diseases, including Alzheimer’s disease (AD)27, Down
syndrome (DS)28 and cardiovascular disease (CVD)29. A plasma
proteomic study predicting body mass index (BMI)30 was used as a
control because weight gain varies widely with age (Supplementary
Fig. 2). As expected, the proteome linked to BMI was not selectively
enriched for proteins defining waves of aging (Fig. 4f). Conversely,
Letters
CVD-associated proteins were strongly enriched in waves of pro-
teins defining middle and old age compared to young age (Fig. 4g).
This enrichment corresponded to an increased incidence of CVD
after 55 years of age31. Finally, AD- and DS-associated proteins
overlapped with the top proteins defining middle and old age but
not those defining young age (Fig. 4h,i). The fact that the proteome
defining these two diseases also changed in old individuals of a sep-
arate disease-free cohort supports the notion of accelerated aging
in DS and AD32,33. Altogether, these results show that waves of pro-
teomic aging are differentially linked to the genomic and proteomic
traits of various diseases.
Discussion
Our analysis of the plasma proteome reveals complex, non-
linear changes over the human lifespan. Although linear analysis
provides information about the aging plasma proteome, modeling
non-linear protein trajectories is necessary to fully appreciate these
undulating changes.
It is well known that men and women age differently21, yet we
were surprised that two-thirds of proteins that changed with age
also changed with sex (895 of 1,379 proteins). This supports the
National Institutes of Health (NIH) policy on the inclusion of
women in clinical research and using sex as a biological variable
in experiments. Nevertheless, a unique proteomic clock can be
used to predict age in men and women, and deviations from this
plasma proteomic clock are correlated with changes in clinical and
functional parameters (Fig. 1f–h). This panel of 373 proteins can be
used to assess the relative health of an individual and to measure
healthspan, analogous to epigenetic clocks based on DNA meth-
ylation patterns34. More large-scale plasma proteomic studies
are required to establish the validity and utility of this clock and
whether specific protein subsets are more appropriate to reflect
particular clinical and functional parameters.
Blood is a sensitive marker of functional aging that also plays
an active role in aging. Several studies have shown that soluble
factors from young mouse blood reverse aspects of aging2–10,35.
Here we describe a 46-protein aging signature that was conserved
in humans and mice, containing known aging-related proteins
such as GDF1536 and IGF1–INSR37 but also less investigated ones
(Fig. 1l). This conserved signature may allow deeper investigation
of translational aging interventions in mice, such as heterochronic
parabiosis, which partially reverses age-related changes of these
proteins (Fig. 1k).
By deep mining the aging plasma proteome, we identified undu-
lating changes during the human lifespan. These changes were
the result of clusters of proteins moving in distinct patterns, cul-
minating in the emergence of three waves of aging. Unexpectedly,
we found that these clusters were often part of shared biological
pathways, particularly cellular signaling (Fig. 2). In addition,

Fig. 3 | Sliding window analysis distinguishes waves of aging plasma factors. a, DE-SWAN. DE-SWAN compares protein levels between groups of
individuals in parcels of 10 years (e.g., 30–40 years compared to 40–50 years). DE-SWAN identifies linear and non-linear changes during aging. Examples
are shown of DE-SWAN for three proteins and five age windows. Red and blue rectangles show the two parcels, and the red and blue lines represent the
mean within each parcel. DE-SWAN provides statistics for each age window and each plasma protein, allowing detailed analysis of plasma proteomic
changes during aging. b, Waves of aging plasma proteins characterized by DE-SWAN (n = 4,263). Within each window, –log10 (P values) and –log10
(q values) were estimated by linear modeling adjusted for age and sex, and significance was tested using the F-test. Local changes attributable to age
were signed on the basis of the corresponding beta age. c, Number of plasma proteins differentially expressed during aging. Three local peaks at the ages
of 34, 60 and 78 years were identified by DE-SWAN. d, Top ten plasma proteins identified by DE-SWAN at age 34, 60 and 78 years (n = 4,263). Linear
models adjusted for age, sex and subcohort were used, and significance was tested using the F-test. Blue and yellow represent local decrease and increase,
respectively. # and $ indicate different SOMAmers targeting the same protein. *q < 0.05, **q < 0.01, ***q < 0.001. e, Intersections between waves of aging
proteins (n = 4,263; q < 0.05). Linear models adjusted for age, sex and subcohort were used, and significance was tested using the F-test. f, Intersections
between linear modeling and the aging waves (n = 4,263; q < 0.05). Linear models adjusted for age, sex and subcohort were used, and significance was
tested using the F-test. g, Visualization of pathways significantly enriched for aging-related proteins identified by linear modeling and DE-SWAN at age
34, 60 and 78 years (n = 4,263). Proteins upregulated and downregulated were analyzed separately. The top ten pathways per condition are represented.
Enrichment was tested using Fisher’s exact test (GO) and the hypergeometric test (Reactome and KEGG).

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Letters NATurE MEDicinE

we provided biological relevance for the three main waves of aging- life (Fig. 3g). We conclude that linear modeling of aging based on
related proteins (Fig. 3), which are characterized by key biologi- omics data does not capture the complexity of biological aging
cal pathways with little overlap. In comparison, linear modeling across the lifespan. Thus, DE-SWAN will be invaluable for analyz-
failed to identify changes occurring late in the eighth decade of ing longitudinal datasets with linear and non-linear quantifiable

a Differential expression-sliding window analysis (DE-SWAN)

Plasma NME1
Detailed statistics
1

2,925 pasma
Protein trajectories

proteins
Fold
Protein levels (z score)

2
Individual protein trajectories
change

Plasma GDF15
1
20 40 60 80
0
1

2,925 pasma
–1

proteins
P /q values
Plasma ARFIP2

–2
20 40 60 80
Age (years) 20 40 60 80
Age (years)

Age (years) Age (years) Age (years) Age (years) Age (years)

b c d Top aging proteins

Waves of aging proteins
Peaks of aging proteins
1,400
* *** *** SVEP1$
** *** *** SVEP1#
# of significant proteins (q < 0.05)

1,200 * *** *** WFDC2

*** *** CHRDL1
** *** *** CCDC80
2,925 plasma proteins

1,000 *** *** *** SMOC1

*** *** PTGDS

800 ** *** *** FSTL3

*** *** *** RSPO4
** *** *** SCARF2
600 * *** *** PTN
*** *** MMP12
*** *** *** GDF15
400 * *** *** NPPB
*** ** *** SOST

200
*** ARFIP2
*** *** EPHB6
*** *** SCG3
0 * *** *** SERPINF2
*** ** OMD

20 40 60 80 *** *** MSMP

20 40 60 80 95 *** CHAD

Signed –log10 (FDR) Age (years) Beta age *** *** *** CDON
Age (years) *** COL11A2

–3 3 –0.03 0.03 *** *** *** RET

Age 34 Age 60 Age 78

e g Aging pathways Signed –log10

Overlap between waves of
aging proteins Lm Age 34 Age 60 Age 78 (false-discovery rate)
–3 +3
Young age R-HSA-375276-peptide ligand-binding receptors
R-HSA-418594-Gαi signalling events
R-HSA-388396-GPCR downstream signaling
GO:0005179-hormone activity
GO:0008201-heparin binding
GO:0005539-glycosaminoglycan binding
GO:1901681-sulfur compound binding
R-HSA-373076-class A/1 (Rhodopsin-like receptors)
R-HSA-500792-GPCR ligand binding
hsa04080-neuroactive ligand–receptor interaction
Middle age
GO:0044420-extracellular matrix component
GO:0005604-basement membrane
GO:0072562-blood microparticle
Old age GO:0071772-response to BMP
GO:0030509-BMP signaling pathway
GO:0071773-cellular response to BMP stimulus
f Overlap between aging waves GO:0030514-negative regulation of BMP signaling pathway
and linear modeling GO:0070848-response to growth factor
600 GO:0003723-RNA binding
GO:0017076-purine nucleotide binding
GO:0019199-transmembrane receptor protein kinase activity
400 GO:0004714-transmembrane receptor protein tyrosine kinase activity
Overlap

GO:0009617-response to bacterium
R-HSA-3000178-ECM proteoglycans
200 GO:0005578-proteinaceous extracellular matrix
GO:0031012-extracellular matrix
R-HSA-1474244-extracellular matrix organization
0 R-HSA-3000171-non-integrin membrane–ECM interactions
Age 60
Age 34
R-HSA-2022090-assembly of collagen fibrils & other multimeric structures
Age 78 R-HSA-1474228-degradation of the extracellular matrix
Lm adj GO:0005578-proteinaceous extracellular matrix
1,000 500 0
# of proteins (q < 0.05)
GO:0031012-extracellular matrix

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NATurE MEDicinE Letters
a Relevance of the aging waves b Genome–proteome associations
c Trans-associations
and aging waves
CFH
NLRP12 1,200
VTN

Number of trans-associations
1,000

HRG
800
Proteins ranked by
600
P value at 34, 60 and 78 years
400
ABO NS P < 0.05
200 Age 34
Age 60
ZFPM2 Age 78
0 Random
Link with genome and proteome Gene
of disease and traits Protein 0 500 1,000 1,500 2,000 2,500 3,000
Proteins ranked in each wave

d e f Body mass index

Cognition and aging waves Handgrip and aging waves and aging waves
12 NS P < 0.05 20 NS P < 0.05 35
with trail making test B time (q < 0.05)

Age 34 Age 34

with body mass index (q < 0.05)

Number of proteins associated

Age 60

Number of proteins associated

Number of proteins associated

Age 60 30
10 Age 78 Age 78
with handgrip (q < 0.05)

Random Random
15
25
8
20
6 10
15
4
10
5
NS P < 0.05
2 5 Age 34
Age 60
Age 78
0 0 0 Random

0 20 40 60 80 100 0 20 40 60 80 100 0 20 40 60 80 100

Proteins rank in each wave Proteins rank in each wave Proteins rank in each wave

g Cardiovascular disease
h Alzheimer’s disease
i Down syndrome
and aging waves and aging waves and aging waves
12 NS P < 0.05 25 Chr. 21 NS P < 0.05
with cardiovascular disease (q < 0.05)

with Alzheimer’s disease (q < 0.05)

Age 34 Age 34
50
Number of proteins associated

Number of proteins associated

Number of proteins associated

Age 60 Age 60
with down syndrome (q < 0.05)

10 Age 78 Age 78
Random 20 Random
40
8
15
30
6
10
20 4
NS P < 0.05 5
10 Age 34 2
Age 60
Age 78
0 Random 0 0
0 20 40 60 80 100 0 20 40 60 80 100 0 20 40 60 80 100
Proteins rank in each wave Proteins rank in each wave Proteins rank in each wave

Fig. 4 | Waves of aging-related proteins are differentially linked to the genomes and proteomes of diseases and traits. a, Relevance of the aging waves.
Schematic representation of analysis. The proteins changing at 34, 60 and 78 years were ranked by P value and were associated with the genome and
proteome of diseases and traits. b, Association between the genome and the proteome. The network was created using the protein quantitative trait
locus associations identified by Sun et al.26. c, Enrichment for trans-association in the waves of aging-related proteins identified by DE-SWAN. Aging-
related proteins at age 34, 60 and 78 years were ranked on the basis of P value, and the cumulative number of trans-associations was enumerated. One-
sided permutation tests (1 × 105 permutations) were used to assess significance. d,e, Enrichment for proteins involved in cognitive (d) and physical (e)
performance in the waves of aging-related proteins. f–i, Enrichment for disease-associated proteins in the waves of aging proteins, including for BMI (f),
CVD (g), AD (h) and DS (i).

changes and for integrating non-linear changes in the analysis of with cognitive and physical impairments. These proteins also dis-
omics datasets. criminated patients from age-matched controls in AD, DS and CVD
Sources of variation of the plasma proteome can be diverse and (Fig. 4g–i), suggesting that the characteristic plasma proteins of
under genetic control26,38. Intriguingly, we observed that the rela- aging are amplified in these age-related diseases. Using an AD- and
tive importance of trans-associations decreased with aging (Fig. 4c), DS-free aging cohort, we provided evidence of accelerated aging for
which led us to investigate sources of variance with a focus on dis- these two diseases32,33. Further investigation of these proteins is war-
ease-associated proteomes. Proteins comprising the waves in mid- ranted to determine whether these associations indicate aging bio-
dle and old age differentially overlapped with proteins associated markers and/or causal mechanisms of disease. Nonetheless, these

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Letters
results suggest that variance within the aging plasma proteome
slowly transitions from hard coding factors (i.e., genomic) to soft
coding factors (e.g., diseases, environmental factors and resulting
changes in cognitive and physiological functions).
The undulating nature of the aging plasma proteome and its
interactions with diseases should be considered when develop-
ing proteomic signatures for diagnostic purposes. Indeed, disease
proteomes overlap significantly with the waves of aging proteins
(Supplementary Table 15). Accounting for heterogeneous and com-
plex changes to the plasma proteome during life will likely improve
the sensitivity and specificity of prognostic and diagnostic tests.
Moreover, these results are pertinent when considering the use of
blood or blood products to treat aging and age-related diseases39.
Specifically, identifying plasma proteins that promote or antagonize
aging at different stages of life could lead to more targeted therapeu-
tics and/or preventative therapies. Such reliable tests and treatments
are urgently needed for several diseases, and, in the future, we hope
to describe plasma proteome changes that predict subjects transi-
tioning to disease. Of particular interest are studies of AD, for which
blood-based biomarkers are unavailable, and clinical symptoms are
believed to occur up to two decades after disease onset.
Online content
Any methods, additional references, Nature Research reporting
summaries, source data, extended data, supplementary informa-
tion, acknowledgements, peer review information; details of author
contributions and competing interests; and statements of data and
code availability are available at https://doi.org/10.1038/s41591-
019-0673-2.
Received: 18 January 2019; Accepted: 30 October 2019;
Published online: 5 December 2019
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NATurE MEDicinE
Methods
Plasma proteomics measurements. The SomaScan platform was used to quantify
relative levels of protein involved in several processes, such as intercellular
signaling, extracellular proteolysis and metabolism. This platform was
established to identify biomarker signatures of diseases and conditions, including
cardiovascular risk29, cancer40 and neurodegenerative diseases27. The SomaScan
platform is based on modified single-stranded DNA aptamers (SOMAmer
reagents) binding to specific protein targets. Assay details were previously
described19. Different versions of the SomaScan assay were used in the LonGenity,
INTERVAL and four independent human cohorts. These versions contained 5,284,
4,034 and 1,305 aptamers, respectively.
Of the 4,034 aptamers measured in the INTERVAL cohort, 3,283 were
contained in the publicly available dataset (European Genoma–Phenome Archive
EGAS00001002555). Our study focused on 2,925 aptamers with identical SeqId
and SeqIdVersion in both INTERVAL and LonGenity cohorts (Supplementary
Table 1). Of the 2,925 aptamers, 888 were measured in the four independent
cohorts and in mice (Supplementary Table 2).
Human cohort characteristics. INTERVAL cohort. Participants in the INTERVAL
randomized controlled trial (ISRCTN24760606) were recruited with the active
collaboration of the National Health Service (NHS) Blood and Transplant (http://
www.nhsbt.nhs.uk), which has supported field work and other elements of the
trial. DNA extraction and genotyping were co-funded by the National Institute
for Health Research (NIHR), the NIHR BioResource (http://bioresource.nihr.
ac.uk/) and the NIHR Cambridge Biomedical Research Centre at the Cambridge
University Hospitals NHS Foundation Trust. The INTERVAL study was funded by
NHS Blood and Transplant (11-01-GEN). The academic coordinating center for
INTERVAL was supported by core funding from the NIHR Blood and Transplant
Research Unit in Donor Health and Genomics (NIHR BTRU-2014-10024), the
UK Medical Research Council (MR/L003120/1), the British Heart Foundation
(RG/13/13/30194) and the NIHR Cambridge Biomedical Research Centre at the
Cambridge University Hospitals NHS Foundation Trust. Proteomic assays were
funded by the academic coordinating center for INTERVAL and Merck Research
Laboratories (Merck & Co.). A complete list of the investigators and contributors
to the INTERVAL trial was previously reported15. The academic coordinating
center would like to thank blood donor center staff and blood donors for
participating in the INTERVAL trial. For more information, see the Nature
Research Reporting Summary.
Proteomics measurements from 3,301 human plasma samples (1,685 males
and 1,616 females) from two different subcohorts were used for this study.
Age ranged from 18 to 76 years with a median age of 45 years (first quartile = 31;
third quartile = 55). Sample selection, processing and preparation were
detailed previously26.
LonGenity cohort. LonGenity is an ongoing longitudinal study initiated in 2008
and designed to identify biological factors that contribute to healthy aging16. The
LonGenity study enrolls older adults of Ashkenazi Jewish descent with age 65–94
years at baseline. Approximately 50% of the cohort consists of offspring of parents
with exceptional longevity, defined as having at least one parent who survived
to 95 years of age. The other half of the cohort includes offspring of parents with
usual survival, defined as not having a parental history of exceptional longevity.
Proteomics measurements from 1,030 human plasma samples (457 males and 573
females) collected at baseline in LonGenity participants were used for this study.
Age ranged from 61 to 95 years with a median age of 74 years (first quartile = 69;
third quartile = 80). LonGenity participants are thoroughly characterized
demographically and phenotypically at annual visits that include collection
of medical history and physical and neurocognitive assessments. Sixty-eight
individuals without clinical and functional data were excluded from the analysis.
The LonGenity study was approved by the institutional review board (IRB) at the
Albert Einstein College of Medicine.
Four additional independent cohorts. One hundred seventy-one human plasma
samples (84 males and 87 females) were obtained from four different cohorts from
the US and Europe (VASeattle, PRIN06, PRIN09 and GEHA). Sample selection,
processing and preparation of the VASeattle cohort were detailed previously41.
Participants from the PRIN06, PRIN09 and GEHA42 cohorts were enrolled by
multiple Italian study centers. Participants were mainly of European ancestry. Age
ranged from 21 to 107 years with a median age of 70 years (first quartile = 58; third
quartile = 89). Written informed consent was obtained from each subject. The
IRB determined that our research did not meet the definition of human subject
research per Stanford’s Human Research Protection Program policy, and no IRB
approval was required for this study.
For these cohorts, all samples were stored at −80 °C, and 150 µl aliquots of
plasma were sent on dry ice to SomaLogic. Plasma samples were analyzed in
three different batches: 24 samples in 2015, 70 samples in 2016 and 77 samples in
2017. In addition to these 171 plasma samples, 12 additional aliquots from 4 of
these samples were measured in the different batches to estimate intra- and inter-
assay variability (Supplementary Table 3). Data for 1,305 SOMAmer probes were
obtained. No sample or probe data were excluded. HybNorm.plateScale.medNorm
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files provided by SomaLogic were bridged to data from the first batch of samples
using calibrators.
Normalization of INTERVAL and LonGenity datasets. Relative fluorescence
units (RFUs) of each plasma protein were log10transformed. We normalized the
levels of each protein within each subcohort on the basis of the average of the
subjects in the 60- to 70-year range. Supplementary Figure 3 shows representative
normalization examples. Note that this normalization is needed when fitting aging
trajectories (Fig. 2) but does not affect the results when ‘subcohort’ is included as a
covariate in the modeling.
The data from the four independent cohorts were log10 transformed and
bridged together using the SomaLogic procedure on the basis of calibrators.
However, the number of samples in the 60- to 70-year range was too small to
reliably bridge these data to the INTERVAL and LonGenity cohorts.
Linear changes in the aging plasma proteome. To determine the effect of age and
sex at the protein level, we used the following linear model:
Protein level  α þ β1 age þ β2 sex þ β3 subcohort þ ε
The type II sum of squares was calculated using the ANOVA function of the R
car package43. This sum of squares type tests for each main effect after the other
main effects. q values were estimated using the Benjamini–Hochberg approach44.
It should be noted that the age range differed between cohorts. If the adjustment
for cohort effect decreases the number of false positives, it could also alter the
true-positive rate. In the four independent cohorts, the ‘subcohort’ covariate also
accounted for batch effect, as samples from different cohorts were measured in
different batches (except for PRIN06 and GEHA, which were measured together).
To determine the relative proportion of variance explained by age and sex, we
calculated the partial Eta2 as follows:
Sum of squareseffect
Partial Eta2 ¼
ðSum of squareseffect þ Sum of squareserror Þ
Validation of the aging-related proteins. To provide confidence in the
reproducibility of the protein assays, we compared our findings with the associations
with age reported by Tanaka et al.22. To this end, we merged our results with those
from Tanaka et al. using ‘SomaId’. Of note, Tanaka et al. used the same version of the
SomaScan platform that we used for the four independent cohorts (1,305 proteins).
SEPA. To determine the biological meaning of a group of plasma proteins, we
ranked the top 100 proteins on the basis of the product of –log10(P values) and
beta age (or beta sex) and queried three of the most comprehensive biological
annotation and pathway databases: GO45, KEGG46 and Reactome47. Using these
databases, we tested enrichment for pathways in the top 10 to top 100 proteins
in increments of 1 protein. The 2,925 proteins measured in this study cover 90%
of the human GO, Reactome and KEGG terms containing more than eight genes
(Supplementary Figure 4).
To analyze each incremental list of proteins, we used the R topGO package48 for
GO analysis and the R clusterProfiler package49 for KEGG and Reactome analyses.
As input for SEPA, we used gene symbols provided by SomaLogic (Supplementary
Table 1). The 2,925 proteins measured by SomaScan served as the background set
of proteins against which to test for over-representation. Because several individual
proteins (33 of 2,925) were mapped to multiple gene symbols, we kept only the
first gene symbol provided by SomaLogic to prevent false-positive enrichment.
For KEGG and Reactome analysis, clusterProfiler requires EntrezID as input.
Therefore, we mapped gene symbols to EntrezID using the org.Hs.eg.db package50.
Again, to avoid false-positive enrichment, only the first EntrezID was used when
gene symbols were mapped to multiple EntrezID. q values were estimated using
the Benjamini–Hochberg approach44 for the different databases taken separately.
For GO analysis, q values were calculated for the three GO classes (molecular
function, cellular component and biological process) independently. To identify
the most biologically meaningful terms and pathways, we reported only those
with 20–500 proteins measured by the SomaScan assay. In addition, we focused on
pathways that were consistently highly significant (q < 0.05 for at least 20 different
incremental lists of proteins) and kept the top ten pathways per condition (e.g., for
each wave of aging proteins). Ranking was performed on the basis of the minimum
false-discovery rate across the incremental lists of proteins. SEPA can be viewed as
an extension of the gene set enrichment analysis approach51, with more control for
true and false positives.
Validation of the aging signature in mice. Male and virgin female C57BL/6JN
mice were shipped from the National Institute on Aging colony at Charles River
(housed at 67–73 °F) to the Veterinary Medical Unit (VMU; housed at 68–76 °F)
at the VA Palo Alto (VA). At both locations, mice were housed on a 12-h light/
dark cycle and provided food and water ad libitum. The diet at Charles River was
NIH-31; the diet at the VA VMU was Teklad 2918. Littermates were not recorded
or tracked. Mice that were 18 months old and younger were housed at the VA
VMU for no longer than 2 weeks before being killed; mice older than 18 months
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were housed at the VA VMU until they reached the experimental age. After
anaesthetization with 2.5% vol/vol Avertin, blood was drawn by cardiac puncture.
All animal care and procedures were carried out in accordance with institutional
guidelines approved by the VA Palo Alto Committee on Animal Research.
Heterochronic parabiosis was conducted as previously described3,10,52 with 3- and
18-month-old mice. Briefly, incisions in the flank were made through the skin and
peritoneal cavity of both sets of mice, and adjacent peritoneal cavities were sutured
together. Adjacent knee and elbow joints were then sutured together to facilitate
coordinated locomotion. Skin was then stapled together using surgical autoclips
(9 mm; Clay Adams), and mice were placed under heat lamps to recover from
anesthesia. Each individual mouse was injected subcutaneously with Baytril antibiotic
(5 μg g−1) and buprenorphine (0.05–0.1 μg ml−1 in phosphate-buffered saline) for
pain management and 0.9% (wt/vol) NaCl for hydration. Mice were monitored and
administrated drugs and saline over the next week as previously described.
EDTA-treated plasma was isolated by centrifugation at 1,000 g for 10 min at
4 °C before aliquotting and storing at −80 °C. A total of 110 plasma samples were
analyzed, and aliquots of 150 µl of plasma were sent on dry ice to SomaLogic.
Samples were sent in two different batches: 29 samples in 2016 and 81 samples in
2018. Data for 1,305 SOMAmer probes were obtained, and no sample or probe
data were excluded. RFUs of each plasma protein were log10 transformed.
The SomaScan assay was developed and validated for human fluids but has
been successfully used in mouse research6,24. To understand how similar mouse and
human sequences are, we downloaded all homologies between mouse and human
along with sequence identifiers for each species (HOM_MouseHumanSequence.
rpt) from Mouse Genome Informatics (http://www.informatics.jax.org/) as plain
text files. Then, the protein reference sequences for both organisms were extracted
from UniProt (https://www.uniprot.org/). On these matched sequence pairs, for
each protein we computed a global pairwise sequence alignment. The alignments
were calculated by using the R ‘Biostrings’ library53. The average identity was 0.85,
supporting the use of the SomaScan assay with mouse plasma.
To determine the effect of age and sex at the protein level, we used the 81 samples
from 1 month to 30 months. To this end, we fitted the following linear model:
Protein level  α þ β1 age þ β2 sex þ ε
The type II sum of squares was calculated, and q values were estimated using the
Benjamini–Hochberg approach.
To characterize the effects of young and old blood on the aging plasma
proteome, normed scaled principal-component analysis was performed using the
R ade4 package54.
Prediction of human biological age using the plasma proteome. To determine
whether the plasma proteome could predict biological age, we used glmnet55 and
fitted a LASSO model (alpha = 1; 100 lambda tested; ‘lamda.min’ as the shrinkage
variable was estimated after tenfold cross-validation). Input variables consisted of
z-scaled log10–transformed RFUs and sex information. Two-thirds (n = 2,817) of
the INTERVAL and LonGenity samples were used for training the model, and the
remaining 1,446 samples were used as a validation. In addition, the 171 samples
from the four independent cohorts were used to further assess the robustness of
the predictive model.
To estimate whether a subset of proteins in the the aging clock could provide
similar predictive results, we used a two-step approach that we described previously56.
One hundred models (100 lambda) including 0–373 proteins were created in step 1,
and we estimated the accuracy of each of these models on the discovery and
validation datasets, separately. Broken-stick regression was used to determine the best
compromise between the number of variables and prediction accuracy.
Associations between delta age and clinical and functional variables in old age.
We used the individuals from the LonGenity cohort to identify associations
between deviations from the proteomic clock (delta age = predicted age –
chronological age) and 334 clinical and functional variables (Supplementary
Table 8). To this end, we tested the following linear model:
Variable of interest  α þ β1 age þ β2 age þ β3 sex þ ε
For binary outcomes, logistic regression was used. This analysis was separately
performed in the discovery (n = 2,817) and validation (n = 1,446) cohorts. Type II
sum of squares were calculated using the ANOVA function of the R car package43.
Clustering of protein trajectories. To estimate protein trajectories during aging,
plasma protein levels were z scored, and locally estimated scatterplot smoothing
(LOESS) regression was fitted for each plasma factor. To group proteins with
similar trajectories, pairwise differences between LOESS estimates were calculated
on the basis of the Euclidian distance, and hierarchical clustering was performed
using the complete method. To understand the biological functions of each cluster,
we queried the Reactome, KEGG and GO databases, as described above.
DE-SWAN. To identify and quantify linear and non-linear changes of the plasma
proteome during aging, we developed the DE-SWAN approach. Considering
a vector l of k unique ages, we iteratively used lk as the center of a 20-year
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window and compared protein levels of individuals in parcels below and above lk
(i.e., ½lk � 10y; lk ½vslk ; lk þ 10y). To test for differential expression, we used the
following
I linear model:
Protein level  α þ β1 ageLow=High þ β2 sex þ ε
with age binarized according to the parcels. For each lk, q values were estimated
using the Benjamini–Hochberg correction. The type II sum of squares was
calculated using the ANOVA function of the R car package43.
To assess the robustness and relevance of DE-SWAN results, we tested multiple
parcel widths (5, 10, 15 and 20 years). In addition, we used multiple q-value
thresholds and compared these results with those obtained by chance. To this end,
we randomly permutated the phenotypes of the individuals and applied DE-SWAN
to this new dataset. To keep the data structure, age and sex were permuted
together. In addition, we analyzed the INTERVAL and LonGenity cohorts
separately (Supplementary Fig. 1). Finally, we tested the same linear model when
adjusting for subcohort. This led to a loss of statistical power when the age range of
the INTERVAL and LonGenity cohorts overlapped, but the three waves of aging-
related proteins remained and the ranks of the top proteins were nearly identical
(Supplementary Fig. 1). We used the model adjusted for subcohort when trying
to understand the waves of aging proteins (Figs. 3d–g and 4). The significance
levels of the intersections between aging plasma protein signatures identified by
linear modeling and DE-SWAN at different ages were determined using the R
SuperExactTest package57.
Relationships between the aging waves and the genome and the proteome of
diseases and traits. To quantify the overlap between proteins changing with age
at different stages of life and the genome and the proteome of diseases and traits,
we ranked DE-SWAN results on the basis of P values and created a k-ranked list of
aging-related proteins, Lk. To reflect the degree to which the genome or proteome
is linked to the waves of aging plasma proteins, we walked down Lk and counted
the number of proteins associated with the genome or specific proteome. When
different versions of the SomaScan platform were used, we walked down Lk until
reaching the top 100 proteins measured in both studies.
To identify specific genetic variants associated with the aging plasma proteome,
we mined the summary statistics generated by Sun et al.26., who found 1,927
associations with 1,104 plasma proteins. The Qgraph58 R package was used to
create a network between the genome and the 2,925 proteins analyzed in this study.
To determine whether the aging proteome was associated with the proteome
of clinical and functional variables, we used the individuals from the LonGenity
cohort and tested the following linear model for the top variables identified in
Supplementary Table 8:
Protein level  α þ β1 age þ β2 sex þ β3 variable of interest þ ε
The type II sum of squares was calculated using the ANOVA function of the
R car package43.
To determine whether the aging proteome was associated with disease
proteomes, we integrated data and results from previous proteomic studies using
the SomaScan platform. We re-analyzed one AD dataset publicly available by
AddNeuroMed27 and used summary statistics from published studies focused on
CVD29, DS28 and BMI30.
AddNeuroMed is a European multi-center study in which the AD proteome
was quantified in plasma samples from 681 control individuals and individuals
with mild cognitive impairment (MCI) and AD using a previous version of the
SomaScan assay. Files used were downloaded from the Synapse portal in March
2016 (syn5367752) and included measurements of 1,016 plasma proteins from 931
samples. We limited our analysis to the 645 samples available at visit 1 (191 control,
165 MCI and 289 AD). Raw data were log10 transformed. Four samples (two
control and two AD) were considered as outliers on the basis of visual inspection
of the results of a principal-component analysis and filtered out.
To identify plasma proteins associated with AD, we used linear models with
diagnosis, age, sex and center as covariates:
Protein level  α þ β1 diagnosis þ β2 age þ β3 sex þ β4 center þ ε
The type II sum of squares was calculated using the ANOVA function of the R
car package43.
To determine whether aging plasma proteins were involved in other disease
signatures, we identified three studies using the SomaScan platform in large
human cohorts providing detailed summary statistics. Carayol et al. mined the
plasma proteome to obtain new insights into the molecular mechanisms of obesity.
Of 1,129 proteins measured, they identified 192 plasma proteins significantly
associated with BMI (P < 0.05 after Bonferroni correction). Summary statistics we
used were obtained from Supplementary Data 1 of their publication30. Ganz et al.
derived and validated a nine-protein risk score to predict risk of cardiovascular
outcomes29. In addition to these 9 proteins, 191 other proteins were significantly
associated with cardiovascular risk (P < 0.05 after Bonferroni correction). Summary
statistics for these 200 proteins are available in their eTable 4, and the 1,130 proteins
measured in this study are listed in their eTable 1. Finally, Sullivan et al. used an
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extended version of the SomaScan platform to study DS and identified a large
number of dysregulated proteins28. We used the results for the discovery cohort
(sheet A of Supplementary File 1) in which 258 of 3,586 proteins were reported to
be associated with DS (P < 0.05 after Bonferroni correction).
Because detailed protein information was not available for all studies, we used
either gene symbols or unitprotID to merge disease proteomes characterized in
published studies with the aging proteome identified in this study. When multiple
P values were reported for the same gene symbol (or a combination of gene
symbols), only the most significant P value was retained.
Reporting Summary. Further information on research design is available in the
Nature Research Reporting Summary linked to this article.
Data availability
We created a searchable web interface to mine the human INTERVAL and
LonGenity datasets: https://twc-stanford.shinyapps.io/aging_plasma_proteome/.
The independent human cohorts and mouse protein data are available in
Supplementary Tables 16 and 17. The INTERVAL data are available through the
European Genome–Phenome Archive under accession EGAS00001002555.
Code availability
An R package for DE-SWAN is available in GitHub: http://lehallib.github.io/DEswan/.
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Acknowledgements
We thank the members of the Wyss-Coray laboratory for feedback and support. We
thank the clinical staff for human blood and plasma collection/coordination. We
thank A. Butterworth for his help in getting access to the INTERVAL proteomics data.
The AddNeuroMed data are from a public–private partnership supported by EFPIA
companies and the European Union Sixth Framework program priority FP6-2004-
LIFESCIHEALTH-5. Clinical leads responsible for data collection were I. Kłoszewska
(Lodz), S. Lovestone (London), P. Mecocci (Perugia), H. Soininen (Kuopio), M. Tsolaki
(Thessaloniki) and B. Vellas (Toulouse); imaging leads were A. Simmons (London),
L.O. Wahlund (Stockholm) and C. Spenger (Zurich); and bioinformatics leads were
R. Dobson (London) and S. Newhouse (London). This work was supported by National
Institutes of Health National Institute on Aging (NIA) F32 1F32AG055255 01A1 (D.G.),
Hungarian Brain Research Program Grant No. 2017-1.2.1-NKP-2017-00002 (T.N.), the
Fulbright Foreign Student Program (T.N.), the Cure Alzheimer’s Fund (T.W.-C.),
Nan Fung Life Sciences (T.W.-C.), the NOMIS Foundation (T.W.-C.), the Stanford Brain
Rejuvenation Project (an initiative of the Stanford Wu Tsai Neurosciences Institute),
the Paul F. Glenn Center for Aging Research (T.W.-C.), NIA R01 AG04503 and DP1
AG053015 (T.W.-C.) and the NIA-funded Stanford Alzheimer’s Disease Research
Center P50AG047366, NIA K23AG051148 (S.M.), R01AG061155 (S.M.), the American
Federation for Aging Research (S.M.), R01AG044829 (J.V. and N.B.), NIA R01AG057909
(N.B.), the Nathan Shock Center of Excellence for the Basic Biology of Aging
P30AG038072 (N.B.) and the Glenn Center for the Biology of Human Aging (N.B.).
Author contributions
B.L. and T.W.-C. planned the study. D.B., C.F., S.M., J.V., S.S. and N.B. provided human
plasma samples. N.S., S.E.L. and H.Y. performed the mouse experiments. B.L. analyzed the
data, with contributions from T.N. and A.K. P.M.L. developed the searchable web interface
(shiny app). B.L., D.G. and T.W.-C. wrote the manuscript. A.K., C.F., S.M., J.V., S.S., N.B.
and T.W.-C. supervised the study. All authors edited and reviewed the manuscript.
Competing interests
The authors declare no competing interests.
Additional information
Extended data is available for this paper at https://doi.org/10.1038/s41591-019-0673-2.
Supplementary information is available for this paper at https://doi.org/10.1038/
s41591-019-0673-2.
Correspondence and requests for materials should be addressed to B.L. or T.W.-C.
Peer review information Brett Bennedetti and Jennifer Sargent were the primary editors
on this article and managed its editorial process and peer review in collaboration with
the rest of the editorial team.
Reprints and permissions information is available at www.nature.com/reprints.
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Extended Data Fig. 1 | Sample demographics. Age (a, b), cohort (a, b) and sex distributions (c) of the 4,263 subjects from the INTERVAL and LonGenity
cohorts. (d) Age and cohort distributions of the 171 subjects from the 4 independent cohorts.

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Extended Data Fig. 2 | Comparing age and sex effects in independent cohorts. (a) Age and sex effects in the INTERVAL and LonGenity studies
(n = 4,263) were compared to age and sex effects in 4 independent cohorts analyzed together (n = 171) and to age effect from Tanaka et al. (n = 240,
2018). The aging plasma proteome was measured with the SomaScan assay in these cohorts and 888 proteins were measured in all studies (b) Scatter
plot representing the signed -log10(q value) of the sex effect in the INTERVAL/LonGenity cohorts (x axis, n = 4,263) vs the 4 independent cohorts (y-axis,
n = 171). Similar analysis for the age effect in the 4 independent cohorts (c, n = 171) and in Tanaka et al study (d, n = 240).

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Extended Data Fig. 3 | Deeper investigation of the aging proteomic clock. (a) Prediction of age in the 4 independent cohorts (n = 171) using the
proteomic clock. Only 141 proteins out of the 373 constituting the clock were measured in these samples. (b) Prediction of age in the discovery cohort
(n = 2,817) using the 373 plasma markers. (c) Feature reduction of the aging model in the Discovery and Validation cohorts to estimate whether a subset
of the aging signature can provide similar results to the 373 aging proteins. Dashed lines represent a broken stick model and indicate the best compromise
between number of variables and prediction accuracy. (d) Heatmap representing the associations between delta age and 334 clinical and functional
variables. For quantitative traits, linear models adjusted for delta age, age and sex were used and significance was tested using F-test. For binary outcomes,
binomial generalized linear models adjusted for delta age, age and sex were used and significance was tested using likelihood ratio chi-square test. As
in (c) the analysis was performed for the top 2 to top 373 variables predicting age. The non-uniformity in the heatmaps suggests that specific subsets of
proteins may best predict certain clinical and functional parameters.

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Extended Data Fig. 4 | Proteins and proteome undulations in independent human cohorts and in mouse. (a) Trajectories of 5 selected proteins based
on the INTERVAL and LonGenity cohorts (n = 4,263, left) and 4 independent human cohorts (n = 171, right). Trajectories were estimated using LOESS
regression. Undulation of the 1,305 plasma proteins measured in 4 independent cohorts (b, n = 171) and in mouse (c, n = 81). Plasma proteins levels were
z-scored and LOESS regression was fitted for each plasma factor.

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Extended Data Fig. 5 | Cluster trajectories in independent cohorts. Protein trajectories for the 8 clusters identified in the INTERVAL and LonGenity
cohorts (left column). Thicker lines represent the average trajectory for each cluster. Cluster trajectories for the subset of proteins measured in the 4
independent cohorts (middle column). Corresponding cluster trajectories in 4 independent cohorts (right column).

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Extended Data Fig. 6 | Pathways in clusters. Pathway enrichment was tested using GO, Reactome and KEGG databases (n = 4,263). Enrichment was
tested using Fisher’s exact test (GO) and hypergeometric test (Reactome and KEGG). The top 4 pathways for each cluster are shown. Pathway IDs and
number of plasma proteins associated are represented in the table.

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Extended Data Fig. 7 | DE-SWAN age effect for multiple q-values cutoffs, windows size and after phenotypes permutations. Different Q-value cutoffs
are represented in (a). Similar analysis with different after phenotype permutations (b) and different windows size in (c). The 3 local peaks identified at
age 34, 60 and 78 are indicated by colored vertical lines.

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Extended Data Fig. 8 | Cis-associations and aging waves. Enrichment for cis-association in the waves of aging proteins identified by DE-SWAN. Aging
proteins were ranked based on p-values at age 34, 60 and 78 and the cumulative number of cis-associations was counted. One-sided permutation tests
(1e + 5 permutations) were used to assess significance.

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 nature research | reporting summary
Tony Wyss-Coray (twc@stanford.edu)
Corresponding author(s): Benoit Lehallier (lehallib@stanford.edu)
Last updated by author(s): 10/22/2019

Reporting Summary
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Statistics
For all statistical analyses, confirm that the following items are present in the figure legend, table legend, main text, or Methods section.
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Only common tests should be described solely by name; describe more complex techniques in the Methods section.

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A description of any assumptions or corrections, such as tests of normality and adjustment for multiple comparisons
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AND variation (e.g. standard deviation) or associated estimates of uncertainty (e.g. confidence intervals)

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Give P values as exact values whenever suitable.

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Our web collection on statistics for biologists contains articles on many of the points above.

Software and code

Policy information about availability of computer code
Data collection No software was used for data collection

Data analysis R version 3.6.1 with packages: DEswan (0.0.0.9001), car (3.0-3), topGO (2.36), clusterProfiler(3.12.0) , org.Hs.eg.db (3.8.2) , NHANES
(2.1.0), ade4 (1.7-13), SuperExactTest (1.0.7), qgraph (1.6.3). Details and references can be found within text in the relevant Methods
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- Accession codes, unique identifiers, or web links for publicly available datasets
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October 2018

We created a searchable web interface to mine the human INTERVAL and LonGenity datasets (https://twc-stanford.shinyapps.io/aging_plasma_proteome/)
The Human independent cohorts and mouse protein data are available in Supplementary Tables 16 and 17. The INTERVAL data is available through the European
Genome-Phenome Archive (https://ega-archive.org/studies/EGAS00001002555).

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 nature research | reporting summary
Field-specific reporting
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Life sciences study design

All studies must disclose on these points even when the disclosure is negative.
Sample size Methods, "Human cohorts characteristics" and "Validation of the aging signature in mice" subsections.
Human cohorts. This is a discovery study, not focusing on specific effect-size. Power calculation are not applicable for our study but we are
well-powered (4000+ subjects) and we identified hundreds of highly significant changes after adjustment for multiple comparisons. Sample
sizes of the Human cohorts are listed in Extended Data 1)
Mouse cohorts. No statistical methods were used to predetermine sample size. Sample size was determined based on the number of animals
used in prior experiments conducted in the Wyss-Coray lab (Villeda et al., Nature 2011; Villeda et al., Nature Medicine 2014, Yousef et al.,
Nature Medicine 2019). Again, we are well powered (110 mice) and we identified hundreds of highly significant changes. Sample sizes of the
mouse cohorts are listed in Supplementary Table 9.

Data exclusions Methods, "Human cohorts characteristics" and "Validation of the aging signature in mice" subsections.
For the 4 independent cohorts and the mouse data. "Data for 1305 SOMAmer probes were obtained and no sample or probe data were
excluded"
For the Interval cohort, we used the dataset publicly available without excluding samples / proteins
For the LonGenity cohort. "Sixty-eight subjects without clinical and functional data were excluded from the analysis." By excluding these
samples, the same subjects (and not different subsets) were used in the whole paper, making the interpretation of the results more
straightforward.
For the Addneuromed data, no exclusion criteria were pre-established but "we filtered out 4 addneuromed samples based on visual
inspection of the results of a Principal Component Analysis (PCA)".

Replication All attempts at replication were successful (See Extended Data 2, 3a, 4).

Randomization Human cohorts. Within each subcohort, the age was balanced and age distribution between cohorts overlapped to assess cohort and batch
effect. Sex was balanced.
Mouse cohorts. Number of male mice per group was balanced. Female mice were available only at 3m, 12, 18 and 21 months.

Blinding Somalogic measured proteins levels without any information about the samples.

Reporting for specific materials, systems and methods

We require information from authors about some types of materials, experimental systems and methods used in many studies. Here, indicate whether each material,
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Materials & experimental systems Methods

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Antibodies ChIP-seq
Eukaryotic cell lines Flow cytometry
Palaeontology MRI-based neuroimaging
Animals and other organisms
Human research participants
Clinical data

Animals and other organisms

October 2018

Policy information about studies involving animals; ARRIVE guidelines recommended for reporting animal research
Laboratory animals Methods, "Validation of the aging signature in mice" subsection. A total of 110 male and virgin female C57BL/6JN mice were
used. Mouse groups are summarized in ST9.
In the aging cohort, 6 1 months old (mo), 10 3mo, 6 6mo, 6 9mo, 10 12mo, 6 15mo, 10 18mo, 10 21 mo, 5 24 mo, 6 27mo and 6
30mo were used.
In the parabiosis cohort, 11 4mo and 18 19mo were used.

Wild animals This study did not involve wild animals

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 Field-collected samples This study did not involve samples collected from the field.

nature research | reporting summary

Ethics oversight Methods, "Validation of the aging signature in mice" subsection. All animal care and procedures were carried out in accordance
with institutional guidelines approved by the VA Palo Alto Committee on Animal Research
Note that full information on the approval of the study protocol must also be provided in the manuscript.

Human research participants

Policy information about studies involving human research participants
Population characteristics Participants were healthy blood donors from the Interval, LonGenity, VAseattle, GEHA, PRIN06 and PRIN09 cohorts. For Interval,
age ranged from 18 to 76 years with a median age of 45 (1st Quartile=31, 3rd Quartile=55). For LonGenity, age ranged from 61
to 95 years with a median age of 74 (1st Quartile=69, 3rd Quartile=80). For the 4 independent cohorts (combined). Age ranged
from 21 to 107 years with a median of 70 years (1st Quartile=58, 3rd Quartile=89).

Recruitment Cohorts characteristics are summarized in Extended data 1

Participants in the INTERVAL randomized controlled trial were recruited with the active collaboration of the National Health
Service Blood and Transplant England (www.nhsbt.nhs.uk), which has supported field work and other elements of the trial.
As described by Sun et al. (Nature, 558, pages73–79, 2018), 50,000 participants were enrolled in the randomized trial of varying
blood donation intervals. People with a history of major diseases (e.g. myocardial infarction, stroke, cancer, HIV, and hepatitis B
or C) or with recent illness or infection were excluded . For proteomics measurements, subjects were randomly selected.
LonGenity is an ongoing longitudinal study initiated in 2008, designed to identify biological factors that contribute to healthy
aging. The LonGenity study enrolls older adults of Ashkenazi Jewish descent, age 65-94 years at baseline. Approximately 50% of
the cohort consists of offspring of parents with exceptional longevity (OPEL), defined by having at least one parent that survived
to 95 years of age. Subjects were randomly selected within each cohort.
Samples from the 4 independent cohort were obtained from cohorts in US (VAseattle) and Europe (GEHA, PRIN06 and PRIN09).
GEHA cohort (Franceschi et al. Ann N Y Acad Sci 2007) and VAseattle samples (Britschgi, M., et al. Mol Cell Proteomics 2011)
used were described previously. Subjects from the PRIN06 and PRIN09 cohorts were enrolled by multiple Italian study centers.
Subjects were randomly selected within each cohort.

Ethics oversight All participants from the Interval cohort gave informed consent before joining the study and the National Research Ethics Service
approved this study (11/EE/0538)
The LonGenity study was approved by the Institutional Review Board (IRB) at the Albert Einstein College of Medicine.
The IRB has determined that our research using VAseattle, GEHA, PRIN06 and PRIN09 cohorts does not meet the definition of
human subject research per STANFORD's HRPP policy because
1) we are not obtaining or receiving private individually identifiable information
2) data or specimens were not collected specifically for this study
3) no direct intervention or interaction.
For these reasons, this part of the study did not require approval from the IRB
Note that full information on the approval of the study protocol must also be provided in the manuscript.

Clinical data
Policy information about clinical studies
All manuscripts should comply with the ICMJE guidelines for publication of clinical research and a completed CONSORT checklist must be included with all submissions.

Clinical trial registration ISRCTN24760606

Study protocol Study protocol is described by Moore et al (Trials. 17;15:363, 2014, https://www.ncbi.nlm.nih.gov/pubmed/25230735)

Data collection According to Moore et al (Trials. 17;15:363, 2014), INTERVAL is a randomised trial of whole blood donors enrolled from all 25
static centres of NHS Blood and Transplant. Recruitment of about 50,000 male and female donors started in June 2012 and was
completed in June 2014.

Outcomes According to Moore et al (Trials. 17;15:363, 2014), the primary outcome is the number of blood donations made. Multiple
secondary outcome were investigated. The most important are (i) donor quality of life (assessed using the Short Form Health
Survey) and (ii) the number of 'deferrals' due to low haemoglobin (and other factors), iron status, cognitive function, physical
activity, and donor attitudes.
Several papers published the results of this clinical trial
2016 results in: https://www.ncbi.nlm.nih.gov/pubmed/27645285
2017 results in: https://www.ncbi.nlm.nih.gov/pubmed/28941948
October 2018

2019 extension study results in: https://www.ncbi.nlm.nih.gov/pubmed/31383583