HISTOPATHOLOGY PRESENTATION.

2.10 CELLULAR AGING

BMS 3.1,

30TH MARCH 2023

DANIEL CHEBET 2020-08-00202.

MAYINJA LIVINGSTONE 2020-08-00216.

TWAKYIRE DERRICK 2019-08-08424

Definition of Cellular aging .
•Cellular aging is the progressive decline in
cellular function and viability caused by
genetic abnormalities and the accumulation
of cellular and molecular damage due to
the effects of exposure to exogenous
influences.
qShakespear characterized aging in his elegant
description of the seven ages of man beginning
from ;

i. Conception, this is a stage of reproduction following

fertilization of an ovum by a sperm.

ii.Differentiation, the state of development of

pluripotent cells into different cells having specialized
functions.
iii. Maturation of the organism, is a process an
organism goes through as it ages.

iv. Maturation of its cells, this is cellular developmental

process that is required for a specific cell to attain its
fully functional state.

v. Progressive loss of functional capacity,

vi. Senescence,(synonym is permanent cell
cycle arrest) is the impaired ability of the cells
to undergo replication and repair, and ultimately
lead to cell death culminating in

vii. Death of the individual, the irreversible

cessation of all biological functions that sustain
an organism.
ØT h e c o n s e q u e n c e s o f a g i n g a p p e a r a f t e r
reproductive age but however, aging is distinct from
mortality and disease although aged individuals are
more vulnerable to disease.

ØSurvival is longer in women compared to the

men the ratio being (3:2).
•The higher mortality rate in men is attributed to
violent causes and greater susceptibility to
cardiovascular disease, cancer, cirrhosis and
respiratory diseases,
for which cigarette smoking and alcohol consumption
are two most important contributory factors.
Hypotheses explaining cellular basis of aging.

1. Experimental cellular senescence. By in vitro

studies of tissue culture,
observations;
i. Cultured human fibroblasts replicate for up
to 50 population doublings and then the
culture dies out, thus there is reduced
functional capacity to proliferate with age.
ii. There is either loss of chromosome 1 or
deletion of its long arm, 1q.

iii.with every cell division, there is

progressive shortening of telomere present
at the tips of chromosomes which is
repaired in a normal cell by the presence
of RNA enzyme telomerase.
2. Genetic control in invertebrates.

•Clock (clk) genes responsible for controlling

the rate and time of aging have been identified
in lower invertebrates, e.g. clk-1 gene mutation
in the metazoan.
• Located on chromosome 4.

• Eg, ROR-A and ROR-B,(Retinoic acid Receptor – Related

Orphan receptor A and B) and Casein kinase-1
3. Diseases of accelerated aging.
ØAging under genetic control in human beings is
supported by the observation of high
concordance in lifespan of identical twins.
Such as;
ØProgeria,
An inheritable condition associated with signs of
accelerated aging process characterized by baldness, cataracts
and coronary artery disease.

§Caused by a gene mutation in Lamin A,(LMNA) gene,

located at position 1q22.

§Lamin A and C are supporting components of the nuclear

envelope.

• The genes function to maintain cell structural stability, chromosome organization, cell
motility, gene regulation, cell differentiation, DNA damage repair, telomere protection.
4. Oxidative stress hypothesis (free
radical-mediated injury).

•Aging is partly caused by progressive and

reversible molecular oxidative damage due
to persistent oxidative stress on human
cells.
•In normal cells, very small amounts (3%) of
total oxygen consumption by the cell is
converted into reactive oxygen species.

•The rate of generation of ROS is directly

correlated with the metabolic rate of the
organisms.
Hypotheses continued….
•in aging, there is low metabolic rate with
generation of toxic oxygen radicals which
fail to get eliminated causing their
accumulation and hence cell damage.

•the underlying mechanism is the oxidative

damage to the mitochondria.
Factors determining life expectancy of individuals.

1. Intrinsic genetic processes i.e. the genes

controlling response to endogenous and
exogenous factors initiating apoptosis in senility.

2. Environmental factors e.g. consumption and

inhalation of harmful substances, diet, role of
antioxidants etc.
3. Lifestyle of the individual such as diseases due
to alcoholism (e.g. cirrhosis, hepatocellular
carcinoma), smoking (e.g. bronchogenic carcinoma
and other respiratory diseases), drug addiction.

4. Age-related diseases e.g. atherosclerosis and

ischaemic heart disease, diabetes mellitus,
hypertension, osteoporosis, Alzheimer’s disease,
Parkinson’s disease etc.
 Organ changes in aging.
organ changes
1.Cardiovascular system. 1.Atherosclerosis,
arteriosclerosis with
calcification, aneurysms.
2.Nervous system. 2.Demyelination of nerves.

3.Musculoskeletal
system. 3.Degenerative bone
diseases, age related
muscular degeneration.
 organ changes
4.Eyes. 4.Deterioration of vision due to
cataract and vascular changes in
the retina.

5.Hearing. 5.Hearing disability due to senility

related to otosclerosis.

6.Reduced IgG response to antigens

6.Immune system. and frequent infections.
organ changes

7.Skin. 7.Laxity of skin due to loss of

elastic tissue.

8.Cancers. 8.About 80% of cancer occur

in the age range of 50 and 80
years.
 Hallmarks of aging.

1. Genomic instability,
Ø the increased tendency of DNA mutations due to defects
and other changes that occur during cell division due to
defects in DNA repair genes.

2. Telomere attrition.
ØThis is the progressive shortening of telomere.
3. Epigenetic alterations.
Øthis are alterations involved in DNA methylation
patterns and the post translational modification of
histones(i.e. acetylation, methylation, trimethylation,
deacetylation) and chromatin remodeling.
4. Loss of proteostasis.
Øthis is the reduced/ filed activity of mechanisms
involved in protein folding (by chaperones) and
degradation of proteins (by the proteosome or
lysosome).
5. Deregulated nutrient sensing.
Øthis is the reduced activity of the insulin, insulin-like
growth factor-1,(I,IGF-1) signaling leading to increased
activity of downstream effectors like the mTOR signaling
pathway.
6. Mitochondrial dysfunction.
Øthis is the reduced or altered functioning of the
mitochondria as a result of diminishing efficacy of the
respiratory chain with aging thus increasing proton
leakage and reducing ATP generation.
7. Cellular senescence.
ØThis is the reduced replicative capacity of cells resulting from telomere
shortening.

8. Stem cell exhaustion.

ØThe decline in the regenerative potential of tissues. for example,
hematopoiesis declines with age, resulting in a diminished production of
adaptive immune cells—a process termed immunosenescence—and in an
increased incidence of anemia and myeloid malignancies.
9. Altered intercellular communication.
ØThis is decreased ability to maintain neuro-hormonal signal transmission
resulting from increasing inflammatory reactions with aging.
MECHANISMS
OF CELLULAR
AGING
The mechanisms of cellular aging are;

1) DNA damage.
2) Replicative Senescence.
3) Defective protein homeostasis.
4) Deregulated nutrient sensing.
 Mechanism 1. DNA damage.

•A variety of exogenous (physical, chemical, and

biologic) agents and

•Endogenous factors such as Reactive Oxygen

Species threaten the integrity of nuclear and
mitochondrial DNA.
•Although most DNA damage is repaired by
DNA repair enzymes (e.g., DNA Glycosylases,
DNA Polymerases),some persist and
accumulate as cells age. These enzymes recognize
and correct damage in DNA.

•It is likely that this accumulating damage

explains why, like most cancers, the most
c ommon he ma tologic ma ligna nc i e s a r e
diseases of the aged.
DNA damage cont…….
vWerner syndrome,

vA rare inherited disorder marked by rapid aging that

begins in early adolescence or young adulthood and
with increased risk of cancer.

vSigns and symptoms include; shorter than average

height, thinning of arms and legs, unusual facial
changes like wrinkled skin.
ØGene involved is wrn gene located at position 8p11.2-
12.
Øthe defective gene product is a DNA helicase, a
protein involved in DNA replication and repair.

ØThis enzyme defect causes rapid accumulation of

chromosomal damage mimicking the injury that
normally accumulates during ageing.
DNA damage cont……

vGenetic instability in somatic cells,

ØThe increased tendency for DNA mutations

and other genetic changes to occur during
cell division.

Patients display some manifestations of

ageing at an increased rate such as;-
1. Bloom syndrome-

Øinherited disorder marked by shorter than

average height, narrow face, red skin rash
and increased risk of cancer especially
squamous cell carcinoma, leukemia, etc.

§Caused by a mutation in the BLM gene

located on chromosome 15q26.
2. Ataxia- telangiectasia,
Øa rare childhood neurological disorder characterized by;
unsteady walking, slurred speech, difficulty swallowing,
difficulty coordinating eye movements, etc.

§Is due to mutations of the ATM (Ataxia Telangiectasis

Mutated) genes located on chromosome 11q22.23.

§The ATM gene encodes proteins involved in repairing

double strand breaks in DNA.
 Mechanism 2. Replicative senescence.

ØA terminally non-dividing state of cells after a

fixed number of divisions.

ØAging is associated with progressive replicative

senescence of cells.
Mechanisms involved are:

i. Telomere attrition.
vInvolves progressive shorte ning of
telomeres resulting in cell cycle arrest.

vTelomeres are repeated segments of DNA

found at the ends of chromosomes.
 vTelomeres protect chromosomes from
end fusion and degradation.

vWhen somatic cells replicate, the ends of

chromosomes are less protected and are
seen as broken DNA which signals cell
cycle arrest.
Telomere attrition conti…

vTelomerase activity is expressed in Germ cells and

present in low levels in stem cells, but absent in most
somatic tissues.

ØGerm cells are the reproductive cells of the body, (ova

and sperms).

ØStem cells- (embryonic, adult, induced pluripotent),

cells with unique ability to develop into specialized cell
types in the body.
ØSomatic cells include all body cells except
reproductive/ germ cells.

• As most somatic cells age, their telomeres

become shorter and exit the cell cycle hence
inability to generate new cells to replace
damaged ones.

ØTelomerase is an enzyme that adds telomeres to

ends of chromosomes thus preventing cell death.
 §In immortalized cancer cells, telomerase is
usually reactivated and telomere length is
stabilized allowing uncontrolled
proliferation.

vTelomere shortening is also associated with

premature development of diseases like;
1. Pulmonary fibrosis, lung disease resulting
from damage and scarring of lung tissue due to
failed regeneration of alveoli due to alveolar
stem cell dysfunction resulting from telomerase
shortening.

ØInflammation of lung stem cells influence local

signals like the TGF-β(transforming growth factor-
β) signaling in the lungs thus inducing pro-fibrotic
conditions resulting in fibrosis in a periphery to
center pattern.
2. Aplastic anemia, a condition in which the body
stops producing adequate new blood cells.

Øresults from bone marrow failure in patients with

mutations in the TERT or TERC genes.

ØWhen telomeres associate with the TERT (Telomerase

Reverse Transcriptase) and TERC(Telomerase RNA
Component) mutated genes,
• mutations cause low telomerase activity,
accelerated telomere shortening and diminished
proliferative capacity of hematopoietic progenitors.

• TERC genes are located on chromosome 3,

• TERT gene on chromosome5, at 5P15.33.

• Both genes provide instructions for making a

complement of telomerase enzyme.
 Replicative senescence conti…..
ii. Activation of tumor suppressor genes.

vActivation of certain tumor suppressor genes

particularly those encoded by the CDKN2A (Cyclin
Dependent Kinase Inhibitor 2A)locus also seem to be
involved in controlling replicative senescence.

vCDKN2A locus encodes two tumor suppressor

proteins, P14, P16.
vOthers include; P19, P21.
vexpression of one of which, known as P16 or
INK4a located on 9p21.3 chromosome, is
correlated with chronologic age in virtually all
humans and mouse tissues examined.

ØP16/INK4a involved in regulation of the cell

cycle.
Øp16 slows the G1 to S phase progression
during the cell cycle.
ØP16 binds directly to CDK4, CDK6 inhibiting their
activity, thus no phosphorylation of the Rb,
(Retinoblastoma protein) hence activation.

ØActivated Rb protein binds to E2F family of proteins

hence inhibiting its transcriptional activity, inhibiting
expression of transcription factors thus results into cell
cycle progression is irreversibly arrested allowing a
state of cellular senescence.
ØP14 binds directly to MDM2(Murine Double Minutes),
stabilizing it inhibits its binding with p53, hence p53 is not
bound and thus not degraded.

ØP53 mediated transcription activity is induced and increased

hence genes expressed cause cell cycle arrest.
Øsuch genes include p21, and p19 which are cyclin
dependent kinase inhibitors of CDK1, CDK2 and CDK4
respectively that mediate cell cycle arrest.
Mechanism 3: DEFECTIVE PROTEIN HOMEOSTASIS.
ØThis is reduced activity of mechanisms involved in
protein folding and degradation.

ØDefects in two mechanisms are involved;

1. Maintenance of proteins in their correctly folded
conformations (mediated by Chaperones).

2. Degradation of misfolded proteins by the autophagy-

lysosome system and ubiquitin-proteosome system.
 1. Maintenance of proteins in their correctly folded
conformations(mediated by chaperones).
ØChaperones are proteins belonging to a family of heat shock
proteins(HSPs).
ØThey direct and guide metabolic molecules to the sites of
metabolic activity, e.g. protein folding, disaggregation of
protein-protein complexes and transport of proteins into
various intracellular organelles.

Øchaperones are labelled HSPs because they are increased in

expression as a response to heat shock.
 ØStudies show that mutant mice deficient in
chaperones of heat shock protein family age
rapidly.

• Those that over-express such chaperones are long-

lived.

• They are of 5 major classes of chaperones based on

their observed molecular weights.

• HSP 60, HSP 70, HSP 90, hsp104 and other

small chaperones.
ØBoth normal folding and degradation of
misfolded proteins are impaired with aging.

ØAbnormal protein homeostasis can have many

effects on cell survival , replication and
functions.

•May lead to accumulation of misfolded

proteins which can trigger pathways of
apoptosis.
2. Degradation of misfolded/ unfolded proteins by
the autophagy-lysosome system and ubiquitin-
proteasome system.

ØThe autophagy-lysosome system involves encircling

unfolded proteins with a double membrane known as
autophagosome which is derived from the endoplasmic
reticulum.
The autophagosome fuses with the lysosome to form an
autophagolysosome which is followed by degradation of the
denatured proteins by lysosomal enzymes and the digested
materials are released for recycling of metabolites.

The ubiquitin-proteasome system,

Øinvolves polyubiquination of the misfolded proteins.
ØThe polyubiquitinated molecules are then funned into the
polymeric proteasome complex containing proteasomes that
execute protease activity by digesting proteins into small
fragments known as amino acids that can later be recycled.
 Mechanism 4: DEREGULATED NUTRIENT SENSING.

ØThis is reduced activity of the insulin, insulin-like growth

factor-1 signaling leading to increased activity of
downstream effectors like the mTOR signaling pathway.

vCaloric restriction increases lifespan in all eukaryotic species

in which it has been tested.

vThus eating less increases longevity.

vThere are two major neurohormonal circuits that regulate

metabolism; a defect in any increases aging.
a)Insulin and Insulin- like Growth Factor- 1,
(IGF-1) signaling pathway.

ØIGF-1 mimics intracellular signaling by insulin and

thereby informs the cells of the availability of glucose,
promoting an anabolic state as well as cell growth and
replication.
ØIGF-1 signaling has multiple downstream targets;
relevant to this discussion are two kinases: AKT and
its downstream target, mTOR (mammalian target of
rapamycin), which, as the name implies, is inhibited
by rapamycin.
b)Sirtuins.
vAre a family of NAD- dependent protein deacetylases.
vThere are seven types ; Sirt 1-sirt 7.
vDistributed in different cellular compartments,
Sirt 1,sirt 6, and Sirt 7 mainly reside in the nucleus, Sirt 1, Sirt 6
also in the cytoplasm.
Sirt 2 mainly in the cytoplasm, Sirt 3,4 and 5 are primarily found in
the mitochondria.
vSirtuins have non-redundant functions designed
to adapt bodily functions to various environmental
stresses including food deprivation and DNA
damage.

vAre thought to promote the expression of several

genes whose products increase longevity.
 Counteracting cellular aging.
vInvolves mechanisms that delay cell aging, increasing their life
span with proper functionality.

i. Caloric restriction. This refers to reduction in daily

calorie intake without malnutrition.

ØIt’s thought that caloric restriction increases

longevity by;
i. reducing the signaling intensity of the Insulin-
like Growth Factor-1 pathway.
ii. Increasing Sirtuins.
ØAn increase in Sirtuins, particularly sirtuin-6, serves
dual functions;
i. Contribute to metabolic adaptations of caloric
restriction by;
• shifting energy from growth to maintenance and
repair.
• Reducing the signaling intensity of insulin like
growth factor-1.

ii. Promote genetic integrity by activating DNA

repair genes such as;
• p53, (protein 53).
• BRCA1,2,(breast cancer 1 and 2 genes).
• MSH, (Muts Homologue 1, 2..etc ),
• XPC,(Xeroderma pigmentosum complementation
group C).
• BLM, (Bloom gene).
• ATM, (Ataxia Telangiectasia Mutated gene).
• FANCA(Fanconi Anemia complementation group A
genes). through deacylation.
Functions of Sirtuins.
i. Inhibit metabolic activity by deacetylation of
pyruvate dehydrogenase E1a subunit and this leads to
decreased PDH enzymatic activity.
ii. Reduce apoptosis by down regulating the expression
of C/EBP-Homologous Binding Protein(CHOP),
Caspases, and p53.
iii. Stimulate protein folding by removal of a 26 base
pair intron yielding the spliced XBP1(X-Box
Binding Protein-1) mRNA which is a potent
transcription activator which induces expression of
genes involved in protein folding.
iv. Increase insulin sensitivity and glucose
metabolism by
•repressing transcription of Protein Tyrosine Non-
receptor type 1(PTPN1) which is a negative
regulator of the insulin signalling transduction
cascade.
•Epigenetic modifications of enzymes involved in
insulin resistance such as

ØSUV39H1(a histone lysine methyl

transferase functioning as a protein coding
gene),
ØEZH2,(Enhancer of Zeste Homologue 2)
and
ØT2DM, (Type 2 diabetes melitus gene).
v. Mediate DNA repair by remodelling of the
chromatin structure which facilitates access and
recruitment of DNA repair proteins to the DNA damage
site thus modulating the repair activity.
What is the dietary source of Sirtuins?

•Optimistic wine lovers have been delighted to

hear that a constituent of Red Wine may
activate Sirtuins and thus increase life span.
Figure 1:Mechanisms that cause and counteract cellular
aging.
 Referrences.
1. Harsh-Mohan-Textbook of pathology,6th edition.
2. Robins and Cotran Pathologic Basis of Disease, 9th edition.
3. Lopez-Otin C, Blasco MA, Partridge L, et al: The
hallmarks of aging. Cell 153:1194, 2013. [A landmark
review that suggest nine hallmarks of aging and directions
for future research.]
4. www.ncbi.nlm.nih.gov, international journal of Molecular
Sciences
 CORRECTIONS.
1. What is type of cell death occurs in decreased telomerase
activity?
Ø Mitotic crisis.
2. Which hall mark of cancer corelates to increased telomerase
activity?
ØCancer immortality.
3. What are the mechanisms of cancer immortality?
i. evasion of senescence,
ii. evasion of mitotic crisis.
iii. Self renewal, i.e. they become stem cells.
4. What is the mechanism of oxidative mitochondrial
damage?
reactive oxygen species cause increased mitochondrial
permeability transition pores resulting in leakage of protons into
the cytosol thus disrupting the proton gradient responsible ATP
formation.

5. What is cirrhosis.
ØThis is a process of scarring involving nodular formation.
ØMorphological patterns of cirrhosis;
i. Macronodular,
ii. Micronodular and iii. Mixed.
Corrections conti……

6. What is ischaemic heart disease?

ØThis is a syndrome resulting from reduced blood supply to
the heart muscle marked by;
i. myocardial infarction,
ii. angina pectoris,
iii. chronic ischaemic disease, and
iv. sudden cardiac death.
7. What is the synonym for cancer immortality?
ØLimitless proliferative potential.

8. What kind of protein is P16?

ØIt’s a cyclin dependent kinase inhibitor.
9. What is INK4a in full?
ØInhibitor of cyclin dependent kinase-a.

10. Define Sirtuins?

ØSirtuins are signaling proteins belonging to a family of NAD-
dependent protein deacetylases involved in processes such as
DNA repair and metabolic regulation.

11. Insulin sensitivity; is defined as the ability of the target

tissues to respond to the effects of insulin.
12. How do Sirtuins increase insulin sensitivity?
•repressing transcription of Protein Tyrosine Non-
receptor type 1(PTPN1) which is a negative
regulator of the insulin signalling transduction
cascade.

13. How does red wine activate Sirtuins?

ØRed wine contains resveratrol,(3,4,5-trihydroxystilbene)
which is a natural polyphenol Sirtuin activating
compound(STAC).
ØResveratrol is a Sirtuin activating compound that binds
allosterically to Sirtuins causing a conformation change
modulating the affinity of Sirtuins for NAD+ and
protein substrates resulting into increasing activity.

14. How does a defect in autophagy lead to cellular

aging?
a defect in autophagy will lead to accumulation of
intracellular aggregates like broken down red blood cells,
unfolded proteins that will lead to apoptosis and thus
accelerating the ageing process.
References.
1. https://www.ucl.ac.uk/news/2020/apr/red-wine-
component-mimics-oestrogen-support-healthy-ageing.

2. https://www.ncbi.nlm.nih.gov, international journal of

Molecular Sciences.
3. Harsh-Mohan-Textbook of pathology,6th edition.
4. Robins and Cotran Pathologic Basis of Disease, 9th edition.