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examining
altered
Glycobiology
in Cancer

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Glycosylation and the Glycobiology Biomarkers Unraveling Glycobiology Glycobiology Under
tumor microenvironment for Cancer Therapeutics the Looking Glass
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Glycosylation
and the Tumor
microenvironment

impacts growth factor receptor activation well-known to aid immune evasion by

covalent attaching of gly and stabilizes transcription factors involved recruiting immunosuppressive cell types
cans to proteins during or in cell cycle progression (2). Finally, and driving the adoption of tolerogenic
after translation. It is integlycosylation plays a critical role in phenotypes (2). Glycosylation
Glycosylation refers to the
general to many programmed cell death by determining dysregulation results in the presentation
processes, including protein folding and ligand-receptor sensitivity. For example, of abnormal protein structures at the cell surface.
stability, and researchers have linked decreased fucosylation is linked to increased These structures are only mildly
abnormal glycosylation to numerous resistance to apoptosis in colon cancer antigenic and rarely immunogenic, thus
diseases (1). Altered glycosylation is cells (3), while N-glycan sialylation blocks contributing to immune evasion (7).
common in cancer that correlates with Fas-mediated apoptosis (4). Furthermore, Glycans also interact with lectin
oncogenesis and disease progression glycosyltransferases activate survival receptors expressed by immune cells,
(1). The full extent of glycoprotein signals such as focal adhesion kinase to such as sialic acid-bind ing
alterations in cancer is presently unknown, prevent cell death (5). Not all gly coproteins immunoglobulin-like lectins (Siglecs)
but researchers believe that gly can have are pro-proliferative; researchers have (8), to drive anti-inflammatory
the potential to serve as diagnostic identified proteins such as decorin or mechanisms (2). Finally, sialoglycans
biomarkers and therapeutic targets for cancerMGAT3
(1). which can reduce tumor growth interact to induce an antigen-specific
(2,6). Nonetheless, crosstalk between tolerogenic program, which enhances
cancer cells and the tumor microenvironment regulatory T cells at the expense of inflammatory
(TME) generally creates a pro-proliferative glycosylation profile (2).

How Glycobiology
Glycans are known to participate in
numerousCancer
Affects fundamental biological processes Mass spectrometry (MS) is commonly
that are affected by cancer, including Seeing
used Glycobiology
to characterize glycoconjugates,
inflammation, immune surveillance, cell Glycobiology and the Tumor
The TME, largely through the creation of a as the technique can provide structural
adhesion, and cellular metabolism. As Microenvironment
hypoxic environment, plays a major role in information with high sensitivity (9).
such, aberrant glycation contributes to driving cancer cell metabolism. Hypoxic However, MS has limited applicability
tumor progression by enhancing tumor induction of HIF-1ÿ elevates O- for in vivo contexts, so researchers are
proliferation, invasion, metastasis, and GlcNAcylation by funneling glucose into turning towards immunohistochemistry
angiogenesis (1,2). Glyco sylation is a the hexosamine biosynthetic pathway and flow cytometry to analyze
highly diverse process, drawing upon a glycobiology
(HBP), which is linked with enhanced tumor progression (1).within spatial and cellular contexts (1
large pool of glycoconjugate macro Hypoxia also induces the dynamic These techniques employ a number of
molecules, including O-glycans, N-glycans, glycosylation of glucose-6-phosphate different probes, including lectins, anti
glycosphingolipids, and glycoproteins (1), dehydrogenase and phosphofructokinase bodies, and aptamers, with lectins—
and potentially occurring at multiple sites 1. Both of these mechanisms contribute to carbohydrate-binding proteins ubiquitous
on a given protein. Unsurprisingly, cancer- increased cancer cell proliferation (2). in nature—being the most popular (11).
induced glycoproteomic profiles are heterogeneous andelevated
Finally, often case-specific.
O-Glc NAcylation in cancer Developments in probe technology have
cells indirectly stabilizes HIF-1ÿ, resulting led to the creation and expansion of gly
can(2).
in further promotion of additional O-GlcNAcylation arrays, capable of probing hundreds
of glycans simultaneously (12). Advances
The glycome regulates cancer cell like these will continue to aid researchers
Glycobiology
proliferation and Proliferation
in multiple ways. First, in their attempt to better understand the
heavily gly cosylated proteoglycans complexities of the glycome in cancer.
Glycobiology
produced by can cer-associated fibroblasts The and is essential for
immune response
the Immune
limitingalso
promote growth in nearby tumor cells. Glycosylation System
tumor progression, and the TME is Please see references on page 7.

EXAMINING ALTERED GLYCOBIOLOGY IN CANCER

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Glycobiology
Biomarkers

yes. For example, the FDA approved yltransferases such as ST6GAL1 are
copteomic changes that the glycoprotein antigens CA-125 (aka abnormally upregulated in cancers.
develop during oncogene mucin 16), CA 19-9 (aka sialyl-Lew This phenomenon may have prognostic
sis and tumor progression isA), and CA 15-3 (derived from mucin application for colorectal cancer and is
The myriad glycomic
provide andwith
researchers gly 1) as serum markers for ovarian, pan linked to tumorigenesis in ovarian and
opportunities to identify novel creatic, and breast cancer, respectively. pancreatic cancer cells (5).
biomarkers. These can be diagnostic, Similarly, kallikrein-3 (better known as
such as the identification of glycosylation prostate-specific antigen) is a gly
moieties, sites, or patterns that are coprotein-modulating enzyme that is
specific to certain cancer types or widely used to screen for prostate
indicative of certain degrees of cancer (1,2). These, along with many Biomarkers
Exploring howascancer alters
progression. They can also potentially others, are readily detectable in glycosylation
Therapeuticmechanisms
Targets has the
serve as therapeutic targets, with patients' serum and have become added benefit of highlighting potential
researchers seeking to correct or routine elements of medical practice (3). markers that can be targeted to correct
counterbalance aberrant oncogenic or cancer-in duced modulations. Tumor-
cancer-promoting signaling or pathway modulations. associated carbohydrate antigens
(TACAs) are cell surface structures
resulting from cancer-induced alterations
The work done exploring to glycosylation, making them a prime
Cancer can affect the glycome in a how cancer alters target for potential cancer vaccines
Where to
number Look?generating many
of ways, (6,7). Researchers are also looking at
glycosylation mechanisms
different types of biomarkers. Individual cancer cell-specific gly cans as binding
have the added benefit
modu lations at the epitope, glycan targets for vectors carrying
moiety, or glycosylation-site level may of highlighting potential chemotherapeutics to improve specificity
indicate the presence of cancer (1,2). markers that can be and efficacy, with promising in vitro and in vivo re
These potential biomarkers are likely In the same vein, targeted
targeted to correct cancer-
differentiated not just by their presence attenuation of glycosylation using small
induced modulations.
or absence, but by relative changes in molecule inhibitors is a major avenue
either abundance or modification profile Beyond this, researchers are also of investigation, with several candidates
between cancer and healthy cells (2). looking at the proteins that modulate currently in clinical trials. The fact that
As such, multi-glyco protein panels offer key gly cosylation mechanisms—namely various cell types can easily take up
better specificity and sensitivity for gly cosyltransferases and glycosidases (4). these small molecules, combined with
disease identification, with researchers O-linked-N-acetylglucosamine the ubiquity of aberrant glycosylation
moving towards global N and O-glycan transferase (OGT) and O-GlcNAcase mechanisms across multiple cancers,
analysis for more comprehensive (OGA) add and remove O-linked-N- means that these agents can potentially
acetylglucosamine
disease (and biomarker) screening and discovery (1). (O-GlcNAc) treat multiple diseases. As a prime
moieties, respectively. Elevated O- example of this, galectin inhibitors are
GlcNAcylation stemming from increased currently part of clinical trials for
OGT expression is found in lung, colon, melanoma, head, neck, colorectal, lung,
and breast cancer. Indeed, a breast, and prostate cancers (10).
Biomarkers have
Researchers for Disease
made promising combination of increased OGT and
Diagnosis
strides and Prognosis
in identifying glycomic and decreased OGA expression is a Please see references on page 7.
glycopro teomic biomarkers for cancer diagno
possible prognostic marker for breast and prostate cancer (4). Similarly, sial

EXAMINING ALTERED GLYCOBIOLOGY IN CANCER

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Unraveling Glycobiology
for Cancer Therapeutics
traditional cancer therapeutic
I n techniques,
responseincluding
to the radiation,
drawbacks of
surgery,
and chemotherapy, sci
Entists are developing more tar geted
approaches such as small mole cule
inhibitors and immunotherapy for better
precision and efficacy. Naturally, scientists
are adapting these approaches for
glycobiology targets as well. How ever, this
process has been hampered by the inherent
complexity associated with glycosylation, as
well as difficulties in replicating glycobiology
ex vivo. New advances, particularly in high-
through put screening (HTS), have made it
easier to characterize the glycoproteome and
identify modulatory agents (1).
Adapting Drug Discovery for
Glycobiology
Today, cancer drug development typically
starts by screening a library of candidate
compounds for “hits” that potentially act on
the target of interest, a process that has been
greatly stream lined by automated HTS. To
that end, researchers have adapted HTS for
glycobiology, developing assays for individually proliferation, immune evasion, and metastasis,
targeting key glycosyltransfer ases (1).
Because a given glycosylation enzyme can
modulate multiple proteins, implementing
HTS for cellular systems is vital. Researchers
are making progress on that front, with a
2017 report documenting the first use of a
cellular model to find an inhibitor for the
transferase ppGalNAc-T3 (2).
EXAMINING ALTERED GLYCOBIOLOGY IN CANCER
Targeting Glycosylation
In addition to targeting key glycosylation
enzymes, researchers are testing the
feasibility of targeting specific glycan-protein
interactions that drive cancer progression.
Selectins and their glycan ligands, for
example, are heavily implicated in metastasis
(3), and selectin inhibition can limit or prevent
bone marrow metastasis in acute myeloid
leukemia (4). Similarly, sialic acid-binding
immu noglobulin-like lectins (Siglecs), a
receptor family that binds sialylated ligands,
are important immune homeostasis regulators
that facilitate immune evasion in cancer
scenarios (3). Siglec or Siglec ligand
antagonism—via desialylation or physical
impediment—represents an interesting
anticancer strategy that clinicians have
employed against melanoma and breast
cancer (3,5).
Glycobiology and Combination
Therapeutics
The glycome not only directly affects can
cer progression by modulating import ant
mechanisms associated with sur vival,
but can also impact the efficacy of existing
treatment strategies. Indeed, aberrant
glycosylation has been implicated in
chemotherapy resistance and poor tumor cell
collection by immunotherapeutic antibodies.
Researchers are therefore investigating
combination therapies that use glycobiology
modulators alongside tar- geted therapeutic
agents (1). For exam ple, conjugating a
sialidase to the anti HER2 monoclonal
antibody trastuzumab
resulted in increased natural killer cell
activation and elevated cytotoxicity against
tumor cells (6).
These combination strategies are also
important for better targeting, especially
when using nanoparticle vectors (7).
Researchers have used TKH2 monoclonal
antibody binding to the tumor cell-exclusive
sialyl-Tn antigen to selectively deliver the
chemotherapeutic agent cisplatin, resulting
in attenuated in vitro and in vivo tumor growth
(8). Researchers have seen similar results
when they treated sialyl-LewisA-expressing
gastric cancer cells with targeted nanoparticles
containing 5-fluorouracil and pacl itaxel (9).
Glycans as Vaccine Targets?
Finally, tumor-associated carbohydrate
antigens (TACAs) are interesting can didates
for anticancer vaccine development, owing
to their potential commonality across different
cancer types (3).
Antigenicity has posed a problem, especially
for monomeric vaccines, as carbohydrates
alone are not highly immunogenic (3,10). To
combat this, researchers are coupling TACAs
to other elements such as T-cell epitopes or
chemically modifying them in vitro (10).
Multivalent vaccines using agents that
simultaneously target multiple glycans may
also be a solution, as recent studies
investigating heptavalent and pentavalent
vaccines induced better immune responses
than prior monomeric attempts (3).
Please see references on page 7.
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Glycobiology Under the Looking Glass

How Cancer Changes Glycosylation

N-linked
In addition to expression level shifts, tumor cells glycan branching
sialylation
often display glycans not found in normal cells. The Structural truncations Be/Thr

most common changes include increased sialylation, Be/Thr

Fucosylation and O-linked Be/Thr
asn asn glycan branching
fucosylation, structural truncations, and N- and O- Be/Thr

linked glycan branching (1).

Glycans Stimulate Glycosylation

cell proliferation Promotes Migration
Cancer-associated
and Metastasis
fibroblasts produce Selectins, as mediators of
growth-promoting cell-cell adhesion, figure
proteoglycans such prominently in mechanisms
as syndecan-1 and -2 of cancer cell migration,
that stimulate proliferation extravasation, and
in nearby cancer cells. metastasis. Selectin binding
Other secreted glycans by glycan ligands is a major
enhance growth factor promoter of metastasis in
binding and mitogenic both solid tumor and blood-
activity either by acting on based cancers (3).
receptors (largely receptor
tyrosine kinases) or
downstream transcription factors (2).

How Glycosylation Affects

metabolism
Hypoxic stress and subsequent HIF-1ÿ
activation significantly impact glycosylation, SLex

resulting in increased O-GlcNAcylation and

sialylation. This contributes to the stabilization e-selectin

of oncogenic transcription factors, promoting

oncogenesis and tumor aggression (2).

Glycobiology and Immune Evasion

Glycan-binding to cell surface lectins such as sialic acid
binding immunoglobulin-type lectins (Siglecs) interferes
with cancer-cell antigen recognition by immune cells—
particularly NK cells. Siglec binding also downregulates
immune cell activation proteins and upregulates
immunosuppressive cytokines (4).

references
1. Pinho S, Reis C. 2015. Glycosylation in Cancer: Mechanisms and Clinical Implications. Nature Reviews Cancer.
2. Peixoto A, et al. 2019. Protein Glycosylation and Tumor Microenvironment Alterations Driving Cancer Hallmarks. Frontiers in Oncology.
3. Smith BAH, Bertozzi CR. 2021. The Clinical Impact of Glycobiology: Targeting Selectins, Siglecs and Mammalian Glycans. Nature Reviews Drug Discovery.
4. Nardy AFFR, et al. 2016. The Sweet Side of Immune Evasion: Role of Glycans in the Mechanisms of Cancer Progression. Frontiers in Oncology.
Machine Translated by Google
references
1 | Glycosylation and the Tumor
microenvironment
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Mechanisms and Clinical Impli
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Death Receptor by ST6Gal-I Provides
Protection against Fas-Mediated Apoptosis in
Colon Carcinoma Cells. Journal of Biological
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5) LiC, et al. 2014. BCMab1, a Monoclonal
Antibody Against Aberrantly Glyco sylated
Integrin ÿ3ÿ1, has Potent Antitu mor Activity
of Bladder Cancer In Vivo.
Clinical CancerResearch.
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EXAMINING ALTERED GLYCOBIOLOGY IN CANCER
2 | Glycobiology Biomarkers
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Glycoproteins as Specific Biomarkers for
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Cancer Therapeutics
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Based Treatment Regimen for Enhanced
Efficacy and Sequen tial Administration of
Synergistic Drug
Combination in Pancreatic Cancer. day
Final of Pharmacology and Experimental
Therapeutics.
9) Fernandes E, et al. 2015. New Trends in Guided
Nanotherapies for Digestive Cancers: A
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controlled release.
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High-quality, purified lectins

and reagents for glycobiology
Lectins are the key to help you profile, characterize, and capture complex glycans in biological systems or
leverage functional assays to ask questions that were previously beyond reach.

Specifically, lectins can be used in glycobiology to:

• Separate oligosaccharides, even those with identical sugar compositions
• Discriminate between oligosaccharide structures • Isolate a specific
glycoconjugate, cell, or virus from a mixture

Bound to discover more

Explore more
at vectorlabs.com/glycobiology

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