Download Possible Roles of Tumor-associated Carbohydrate Antigens1

[CANCER RESEARCH 56, 2237-2244,
May 15. 1996]
Perspectives in Cancer Research
Possible Roles of Tumor-associated
Carbohydrate Antigens1
Minoru Fukuda2
The Glycobiology Program, La Julia Cancer Research Center. The Burnham Institute, La Jolla, California 92037
the analysis into the roles of carbohydrate-dependent
Abstract
Recent progress in the studies on the roles of carbohydrates has
brought about critical discoveries, which allow us to have working hy
potheses for understanding the roles of tumor-associated carbohydrate
metastasis. Rather than collect a vast amount of knowledge on
changes of cell surface carbohydrates in tumor cells, I have selected
these three topics. By doing this, I hope that I will present clear
examples that can be a model for working on other systems. Readers
are encouraged to read other review articles to see other carbohydrate
changes reported in tumor cells (6-9).
antigens. In this report, I focus my description on three different aspects
of this progress. I discuss: («) the immunologie ¡ilresponse to oligosaccharides aberrantly expressed under pathological conditions; (In possible
roles of carbohydrate-dependent
cell adhesion during tumor metastasis;
and (c) the roles of carbohydrates in modulating the functions of proteins
that attach those carbohydrates. These three areas in the roles of carbo
hydrates will likely be the target for continuous research efforts to reveal
the roles of tumor-associated carbohydrates.
Expression of Branched Hexasaccharides Attached to
Leukosialin (CD43) during Development and Malignancy
Introduction
Analysis of cell surface glycoproteins of human leukocytes and
leukemic cell lines revealed that the structures of O-glycans aie cell
type-specific. Erythrocytes express a tetrasaccharide, NeuNAc «2—
»3
Galßl^>3(NeuNAc a.2—
>6) GalNAc, whereas granulocytes exptess a
hexasaccharide,
NeuNAc «2^3 Galßl^»3 (NeuNAc a2^>3
Galßl^4 GlcNAc ßl^6) GalNAc (10). The large mr.jorily of Tlymphocytes in peripheral blood express the same tetn.sacc'-iaride as
Cell surface glycoproteins play essential roles in maintaining the
function as well as the structure of cells. The importance of cell
surface carbohydrates is particularly evident during the course of
development and differentiation, as shown by sequential expression
and disappearance of cell surface carbohydrates specific to each cell
lineage or maturation stage in a given cell lineage (1-4). At the end,
mature cells acquire a vast array of different functions and cell surface
structures unique to each cell type.
In the past decade, a large amount of knowledge has been accu
mulated on the carbohydrate structures present in glycoproteins and
glycolipids. These studies revealed that those carbohydrates can be
classified into several groups, for instance those attached to proteins
through aspartylglycosylamine linkage, Af-glycans, and those through
O-glycosidic linkage, O-glycans. There appears to be a tremendous
erythrocytes express. T-lymphocytes are activated when enga /lag in
immune response. Those activated T-lymphocytes now acr/j/e the
hexasaccharide (11). This conversion is caused by an incrw.. •¿
i core
2 GnT, which forms a critical branch. In resting T-lymphocy .¿s,the
same enzyme is negligibly present, thus forming instead tre tetnsaccharide (Fig. 1). The expression of core 2 GnT suppresses fhf. expres
sion of the tetrasaccharide, because this enzyme competes "Atti a-2,6sialyltransferase at the same C-6 position of /V-acetylgalactosamine.
In T-lymphocytes, these O-glycans are attached to 1 ul'oJalin
heterogeneity among oligosaccharides belonging to the same group,
but yet those of apparent complexity can be deduced into the common
core structure and the variation in the side chains attached to the core
structure (5).
One of the most important findings in the past decade is that these
variations of oligosaccharides are well controlled during development
and differentiation. Thus, oligosaccharides are very often character
istic of different cell types. Under pathological conditions such as
tumor cells, changes in carbohydrate structures can be almost always
observed (6).
In this report, I would like to present three examples that exemplify
the roles of those oligosaccharides present under pathological condi
tions. I will thus first describe the changes in O-linked oligosaccharide
structures associated with immunodeficiencies
such as WiskottAldrich syndrome and AIDS. In this section, I will discuss immunological response to the altered oligosaccharides and its role in patho
logical processes. In the second part, I will concentrate my description
on roles of cell surface carbohydrates as ligands and their possible
roles in tumor metastasis. In particular, I will summarize the roles of
carbohydrates in cell adhesion during inflammation and then extend
Received 1/22/96; accepted 3/15/96.
1 Supported by Grants R37CA33000,
RO1CA33895,
RO1CA48737.
adhesion during
metastasis. In the third part, I will discuss the roles of carbohydrates
as a modulator of protein function. In particular, I will focus ;;n the
roles of polysialic acid in the N-CAM3 during development and i mor
(CD43, sialophorin). It has been shown that leukosialin is involved in
cell-cell interaction: (a) the adhesion of T-lymphocytes to HeLa cells
through LFA-1 binding to ICAM-1 was inhibited by expression of
CD43 in HeLa cells. This inhibitory activity was abolished, however,
when HeLa cells expressing leukosialin was treated with sialidase
(12); (¿?)
similarly, leukosialin was found to be concenVT;d in the
cleavage furrow during cell division (13). Such hig'ìlyi ,;>trictive
distribution of leukosialin likely provides repulsion jy v.rative
charges due to sialic acid and thus facilitates the dis;, a -.' r. of fwo
dividing cells; (c) leukosialin is shed from neutropiii*
•¿â€¢
/ the
spreading and adhesion of activated neutrophils (¡4.
-- .tioi!,
activated T lymphocytes also shed leukosialin (15). S'K
'I
; of
leukosialin appears to be required for cell adhesion, si
'..c ; alai
leukosialin works as an anti-adhesive molecule. In faci, :
. fr tm
:i;oof leukosialin is present in the plasma, and its carbo
-v sition is consistent in that the leukosialin was derive i
cytes and lymphocytes (16).
More recent studies suggest that mucin-type O-gly ;.
leukosialin play a critical role in thymocyte developrie
cells of T lymphocytes, which originated from the boi.'?
and POI
AI33189 and a grant from the Mizutani Foundation for Glycoscience.
2 To whom requests for reprints should be addressed, at The Glycobiology Program, La
Jolla Cancer Research Center, The Burnham Institute, 10901 North Torrey Pines Road. La
Jolla, CA 92037. Phone: (619) 455-6480; Fax: (619) 646-3193.
3 The abbreviations used are: N-CAM. neural cell adhesion i-i.
W-acetylneuramic acid; GalNAc, W-acetylgalactosamine; GlcNAc. A a
core 2 GnT. core 2 ß-l,6-A'-acetylglucosaminyltransferase; ICAM, i
molecule; Le, Lewis; PSGL. P-selectin glycoprotein ligand-1; PST, |
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ROLES OF TUMOR-ASSOCIATED
CARBOHYDRATE
ANTIGENS
GalNAcal -»Servir
B1->3Gallransterase
Galßl->3GalNAcoc1->Ser/Thr
(core 1)
ß1
->6GlcNAc transferase
(Core 2 GnT)
oc2—>3
NeuNAc Iransferase
Galßl->3GalNAca1~>Ser/Trir
(core 2)
NeuNAco2-»3Galß1
->3GalNAcce1-»Ser/Thr
o2—>6
NeuNAc transferase
ß1-»4Gallransferase
NeuNAco2
^6
NeuNAco2-»3Galß1
->3GalNAca1^->Ser/Thr
Fig. 1. The biosynthetic pathways of O-glycans (Ai and polyyV-acetyllactosaminyl O-glycans (B). A. it has been shown that the
tetrasaccharide (bottom left) is formed by sequential action of
aï—»3
sialyltransferase, followed by ct2—»6
sialyltransferase.
When ßl—»6
W-acetylglucosaminyltransferase,
core 2 GnT. is
present, the branched hexasaccharide (bottom right) is formed
(based on Refs. 10, 11, and 22). B, poly-W-acetyllactosaminyl
chain can be extended from the GlcNAcßl—*6linkage formed in
core 2. Sialylated
poly-/V-acetyllactosaminyl
extension or
W-acelyllactosamine can be further modified by al—»3fucosyltransferase. forming sialyl Lex termini. The core 2 branch is
emphasized by a box (based on Fukuda ei al. ( 10), Piller et al.(\\
and Maemura and Fukuda (47)].
a2-»3NeuNActransferasefsi
NeuNAco2->3Galß1-»4
NeuNAcot2-»3Galß1
->3GalNAcoc1-»SerAThr
B
Galßl->3GalNAca1->Ser;Thr
),
jj
1
B1-)3GlcNAc transferase
B1->4Gal transferase
(Galßl->4GlcN Acß1
->3)nGalß1
->4
NeuNAco2->3(Galß1-H.4GlcNAcß1
->3) Galßl->4
NeuNAca2->3Galß1->3GalN Acal -»Berühr
NeuNAco2->3(Galß1->4GlcN Acß1
->3) „¿Galßl
-»4
3
T
NeuNAco2-»3Galß1
->3GalNAca1 -»Ser/Thr
Fuc
into the thymus for further differentiation and maturation. T-lymphocyte progenitor cells first enter into the periphery of the thymus
cortex. The cortical thymocytes (T-lymphocyte precursor cells) thus
represent more immature T lymphocytes; then the cortical thymocytes
enter into the medulla of the thymus, and they represent more matured
T lymphocytes, medullary thymocytes.
It was discovered that the cortical thymocytes are strongly stained
by a monoclonal antibody, T305, which preferentially reacts with the
branched hexasaccharides, NeuNAc a2—>3 Galßl—¿>3
(NeuNAc
a2-»3 Galßl->4 GlcNAcßl-»6) GalNAc attached to leukosialin
undesirable thymocytes apparently takes place in the medullary thy
mus. These results thus suggest a very provocative possibility that the
selection for further maturation may be dependent on the turning off
of core 2 GnT. Ectopie expression of core 2 GnT in the medullary
thymus and its knock-out in the thymus should yield critical informa
tion to test this hypothesis.
It was also shown that adhesion of thymocytes to thymus epithe
lium can be inhibited by T305 antibody ( 17). It has been demonstrated
that interaction between thymocyte-thymus epithelium plays an es
sential role in thymocyte maturation (19). These results combined
together indicate that the interaction between the hexasaccharides on
thymocytes and thymus epithelia plays a critical role(s) during thy
mocyte maturation in the cortical thymus. Once thymocytes are rel
atively matured, they no longer require the hexasaccharides for at
tachment to thymus epithelia. It is most likely that the interaction
between cortical thymocytes and thymus epithelia is mediated through
another interaction, and such an interaction is likely hindered by the
presence of the hexasaccharides in leukosialin.
(17). In contrast, medullary thymocytes are negligibly stained by the
same antibody. Consistent with this finding, the mRNA of core 2
GnT, which forms the core 2 branches, is abundantly present in
cortical thymocytes whereas it is not detectable in medullary thymo
cytes (17). These results clearly indicate that the O-glycans attached
to leukosialin undergo a profound change from the hexasaccharides
and larger ones to the tetrasaccharides once thymocytes mature from
the cortical thymus to the medullary thymus.
It is known that the majority of active thymocytes die before they
enter into the medullary thymus. This process is thought to be a Production of Autoantibodies against the Branched
typical example of apoptosis. The most recent studies showed that the
Oligosaccharides under Pathological Conditions
selection in cortical thymus is mainly due to the selection for further
In the previous section, I have described the presence of the
maturation, and those selected for not being processed for further
maturation die in the cortical thymus (18). In contrast, the deletion of hexasaccharides in immature cortical thymocytes and its absence in
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ROLES OF TUMOR-ASSOCIATED
relatively matured medullary thymocytes as well as resting T lym
phocytes in the peripheral blood. Once activated, T lymphocytes
acquire again the hexasaccharides. This is a typical example in which
proliferating matured cells show a phenotype similar to their imma
ture precursor cells. This similarity can be extended further to the
property displayed by T lymphocytes under pathological conditions:
(a) It was shown that leukemic cells derived from T lymphocytes in
the peripheral blood are characterized by having a significant amount
of the branched hexasaccharides (20, 21). This increase in the hexas
accharides is more prominent in those patients with acute lymphocytic
leukemia than those with chronic lymphocytic leukemia (20).
(b) The increase of the hexasaccharides in the peripheral blood is
observed in patients with the Wiskott-Aldrich syndrome (22, 23). The
Wiskott-Aldrich syndrome is inherited as an X-linked recessive trait,
most likely due to a defect(s) in a single locus of the X chromosome
(24). The most recent study identified a gene, which is defective in the
syndrome (25). The putative protein product of this gene has the
sequence enriched with prolines, which is characteristic for SH3
binding, and a nuclear localization signal. In fact, a mouse homologue
of the Wiskott-Aldrich gene, named N-3AP1, was cloned by virtue of
its binding to the SH3 domains of the Nek oncogene.4 Nck protein is
phosphorylated in response to activation of various cytoplasmic and
receptor tyrosine kinases, and Nck has been shown to bind, via its
SH2 domain, to activated platelet growth factor and epidermal growth
factor receptors (for an example, see Ref. 26). These results indicate
that the signal pathways by Nck may be regulated by its binding to the
Wiskott-Aldrich gene product. It will be exciting to determine the
roles of the Wiskott-Aldrich gene product in the signal pathways.
This syndrome is characterized by severe eczema, thrombocytopenia, and susceptibility to opportunistic infection. Patients with this
syndrome fail to respond to polysaccharide antigens, rendering them
susceptible to bacterial infection. The second critical problem is that
T-cell number and function decline quickly with age. T-cell-dependent areas of lymph nodes and spleen becomes progressively depleted
(24). This raised the possibility that one of the defects in this syn
drome may be the defect in the thymus. Before the advent of better
therapy such as bone marrow transplantation, the median life span of
these patients was found to be 5.7 years. The cause of death is
infection (57% of the total deaths), hemorrhage (27%), and malignant
tumors (5%), which are mostly leukemia and Hodgkin's disease (24).
From the early stages of the studies, it was discovered that leukosialin behaves abnormally in leukocytes of patients having the
Wiskott-Aldrich syndrome. Initially, the studies appeared to suggest
that leukosialin is missing in leukocytes from the patients with this
syndrome (27). However, it turned out that leukosialin is present but
with a high molecular weight form that is the same as that found in
activated T lymphocytes (22, 23).
It was demonstrated that a substantially increased amount of the
branched hexasaccharides are present in leukosialin derived from
patients with the Wiskott-Aldrich syndrome. As described above, the
hexasaccharides are present in the immature cortical thymocytes but
absent in the mature resting T lymphocytes. The expression of the
hexasaccharides thus appear to reflect that lymphocytes from the
patients with the Wiskott-Aldrich syndrome are functionally imma
ture as well. These results strongly suggest that the conversion of the
hexasaccharide to the tetrasaccharide is critical for the maturation of
T lymphocytes and its failure might induce premature release of
immature T lymphocytes from the thymus.
Continuous expression of the hexasaccharide is due to the increased
amount of core 2 GnT. It is thus tempting to speculate that the
4 W. Li, personal communication
(see also Refs. 91 and 92).
CARBOHYDRATE
ANTIGENS
transcription of core 2 GnT is suppressed by the product of the
wild-type gene, which is mutated in the Wiskott-Aldrich syndrome.
Because of the mutation in the Wiskott-Aldrich syndrome, such gene
product may not suppress the core 2 GnT gene expression any longer.
During the analysis of lymphocytes from various patients, it was
discovered that the peripheral lymphocytes from AIDS patients also
contained an increased amount of the hexasaccharide (20). When two
different specimens were analyzed, the patient with more full-blown
AIDS symptoms showed an increased amount of the hexasaccharide.
The patients who were HIV-infected but asymptomatic in AIDS did
not show an increase of the hexasaccharide.
Interestingly, the sera from HIV-infected individuals were found to
contain antibodies reactive with leukosialin, and the sera reacted with
leukosialin in thymocytes but not with peripheral blood lymphocytes
(28). When peripheral T lymphocytes are partially digested by neuraminidase, the reactivity with the AIDS patients' sera was increased.
Leukosialin in thymocytes carry not only the hexasaccharides but
also the pentasaccharides, Galßl—»3
(NeuNAc al—»3Galßl—»4
GlcNAcj31^6)
GalNAc or NeuNAc a2^3
Gal (Galßl^4
GlcNAcßl—>6)GalNAc, which are partially sialylated oligosaccharides (20). In fact, the amount of Galßl-*3 GalNAc a-2,3-sialyltransferase transcript is more abundant in medullary thymocytes than
cortex thymocytes (29).
Taken together, these results strongly suggest that AIDS patients
produce antibodies against leukosialin, responding to the increased
amount of the partially sialylated oligosaccharides present on leuko
sialin under such pathological conditions.
It has been shown that antibodies to leukosialin can induce bio
chemical and functional changes in T lymphocytes in vitro (for an
example, see Ref. 30). It is thus tempting to speculate that human
anti-leukosialin antibodies in vivo may have similar effects on thy
mocytes, resulting in inappropriate activation of thymocytes during
the process of maturation. It is presumed that the thymus is required
for normal replenishment of CD4+ lymphocytes. Thymic dysfunction
thus would be expected to prevent the replenishment of mature CD4+
cells killed by HIV-1.
Complementary to the above discoveries, it was found that the
HIV-1-infected leukemic T-cell line CEM produced leukosialin that
was undersialylated. Such hyposialylation was found to be associated
with an impairment of leukosialin-mediated homotypic aggregation
(31).
Although there has been no report on autoantibodies in the WiskottAldrich syndrome, patients with the Wiskott-Aldrich syndrome may
produce similar autoantibodies. If that is the case, those antibodies
will likely induce the depletion of thymocytes, which is one of the
major symptoms in this syndrome. The presence of autoantibodies in
both syndromes could heighten the possibility that these autoantibod
ies are involved in the pathogenesis of these diseases.
Roles of CD43 (Leukosialin) Revealed by Its Overexpression
Gene Knock-out in Mice
or
The above described studies mostly address the roles of glycans
attached to leukosialin. To directly determine the roles of leukosialin
expression, overexpression of leukosialin in B-cell lineage and knock
out of the leukosialin gene were carried out in mice.
As described above, leukosialin is present in early B-cell progeni
tors but absent during maturation. Once B lymphocytes differentiate
into plasma cells, leukosialin is expressed again. By fusing with the
immunoglobulin heavy chain enhancer, leukosialin was continuously
expressed in B lymphocytes (32). Those B lymphocytes obtained in
the transgenic mice survive longer in vitro and show less sensitivity to
apoptosis. These results suggest that leukosialin may deliver a signal
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ROLES OF TUMOR-ASSOCIATED
CARBOHYDRATE
ANTIGENS
^¿g^lylylyl,,,^^,^!,^!.^^!^
Fig. 2. Schematic representation of neutrophil and monocyte adhesion to endothelial cells during inflammation. Adhesion of leukocytes is initiated by binding of their carbohydrates
(SLe") to E-selectin and P-selectin. Such binding ("rolling effect") leads into firm attachment to endothelial cells by integrin/ICAM interaction and then establishing vascular
extravasation through plateletyendothelial cell adhesion molecule 1 (PECAM-1) homotypic interaction. This whole process can be inhibited by administration of sialyl Le*-containing
glycoproteins. Tumor cells, in particular carcinoma cells, most likely use this mechanism to adhere to endothelial cells at metastatic sites [from Fukuda (5); see also Springer (36)].
that reduces the number of B lymphocytes that would have normally
undergone programmed cell death.
When the leukosialin gene was knocked out in mice, the knockout
mice developed normal hematopoiesis. However, T lymphocytes from
leukosialin-deficient mice showed an accelerated kinetics of T-cell
activation than T lymphocytes from wild-type mice (33). This in
creased T-cell activation was found to be due to an increase in the
proliferation ability of the leukosialin-deficient cells rather than in the
number of lymphocytes in the allo-specific precursor pool. To deter
mine how these proliferative abilities were achieved, the adhesion of
thymocytes and splenic T lymphocytes were investigated. Thymocytes from leukosialin-deficient mice showed substantially increased
homotypic adhesion compared with thymocytes from wild-type litter
mates. Binding of leukosialin-deficient splenic T lymphocytes was
nearly 100% greater than binding of leukosialin-containing cells to
fibronectin and 50% greater to ICAM-1.
When mice were infected with vaccinia virus, leukosialin-deficient
mice showed considerably greater CTL activity. Moreover, infectious
virus was recovered from lungs and gonads, which was not observed
from wild-type mice. These results suggest that defects involving
tissue localization of effector cells or post-lysis viral clearance by
phagocytes might occur in the absence of leukosialin.
These results, taken together, strongly suggest that leukosialin acts
as an antiadhesive property that retards T-lymphocyte interactions. Its
carbohydrate moiety, composed of a significant amount of sialic acid,
clearly contributes to this function. In this context, the presence of
partially sialylated oligosaccharides in leukosialin from the immunodeficient patients may have more significance than they act as immunogenes to produce autoantibodies. Moreover, it is possible that the
complexity of those oligosaccharides influences the antiadhesive
property of leukosialin and possibly its susceptibility to proteases.
In summary, I have described the roles of mucin-type O-glycans
attached to leukosialin and leukosialin itself in immunodeficient syn
dromes. This field may be one of the few fields where studies have
been extensively carried out on both a protein and the glycosylation in
a particular glycoprotein. Because of recent isolation of genes that
might determine the expression of core 2 GnT, further studies will be
expected to provide more insightful and exciting results that would
reveal the roles of this interesting molecule. Such studies are expected
to shed light on possible roles of autoantibodies formed against
tumor-associated carbohydrate antigens.
Oligosaccharides
in Inflammation and Tumor Metastasis
In the past several years, significant progress was made in under
standing the roles of oligosaccharides in inflammation. This series of
developments was initiated by structural studies on the cell surface
carbohydrates of neutrophils. These early studies demonstrated that
neutrophils contain Le\ GaljSl —¿*4(Fucal^>3)GlcNAc—>R and sia
lyl Le", NeuNAca2-*3 Galßl^4(Fucal^»3)GlcNAc^R
(34, 35),
but not blood group ABO antigens, which are the major oligosaccha
rides present in erythrocytes.
In 1989, the cDNAs for three different adhesion molecules, now
collectively called "selectin," that bind to neutrophils and monocytes
or endothelial cells were cloned. Among them, E- and P-selectin
appear in the onset of inflammation, and binding of these selectins to
neutrophils and monocytes results in the recruitment of these leuko
cytes into inflammatory sites (reviewed in Ref. 36).
Since all of these selectins were found to contain a carbohydratebinding motif observed in animal lectins, it was natural to test if
selectins bind to carbohydrates. These studies revealed that E- and
P-selectin binds to sialyl Le", present as a minor component in
neutrophils (37-39). This binding of E-and P-selectins slows down the
movement of neutrophils along the endothelial wall, which then
allows leukocytes to have enough access to chemokines. Once chemokines are bound to their receptors on leukocytes, a signal is
presumably transmitted through G-proteins, which activates integrins
on the surfaces of leukocytes, establishing a firm attachment to
endothelia. This firm attachment then leads into extravasation, possi
bly through homophilic interaction of platelet/endothelial cell adhe
sion molecule 1 (Fig. 2). It is possible that sialyl Le" and E-selectin
interaction also plays a role in cell-cell interaction during develop
ment. In fact, it has been shown that the interaction of E-selectin and
sialyl Le" is involved in capillary tube formation from bovine endo
thelial cells (40).
Tumor cells, in particular carcinoma and leukemic cells, are enriched
with sialyl Le" structures. Carcinoma cells are also enriched with sialyl
Lea structure NeuNAcc*2^3Galßl^3(Fucal->4)GlcNAc^R,
which
is the isomer of sialyl Lex (41, 42). Sialyl Lea was found to be a ligand
for E-selectin, which is as potent as sialyl Le" (43, 44). Considering a
similar carbohydrate binding specificity between E- and P-selectin, Pselectin might bind to sialyl Lea as well.
Although E- and P-selectin bind to these oligosaccharides,
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these
ROLES OF TUMOR-ASSOCIATED
two selectins differ in the requirement for how these oligosaccharide
ligands are presented. It has been observed repeatedly that E-selectin
binds to cells as long as they express sialyl Le" structures. However,
P-selectin binds to sialyl Lex structures only when they are attached to
particular glycoproteins. One of these carrier glycoproteins, PSGL-1,
was cloned by the virtue of its support of high affinity binding to
P-selectin (45). The amino acid sequence of PSGL-1 was found to
contain a domain composed of tandem repeats that should attach a
large number of mucin-type O-glycans. Similarly, the high affinity
ligand for L-selectin is presented by two mucin-type glycoproteins,
CD34 and GlyCAM-1 (46). These results strongly suggest that P- and
L-selectins bind mucin-type O-glycans that are present as a cluster. It
is likely that the multiple presentation of ligands leads into high
affinity binding of P- and L-selectin. In contrast, E-selectin apparently
binds to the carbohydrate ligands, regardless of how they are pre
sented (37).
For the synthesis of the sialyl Le" structure in mucin-type oligosaccharides, the presence of core 2 GnT is essential (Fig. 1; Ref. 47).
Only those cells expressing core 2 GnT can present efficient ligands
for P- and L-selectin. In some instances, it may even be important to
have sialyl Le" in mucin-type oligosaccharides for E-selectin. For
example, activated T-lymphocytes adhere to E-selectin whereas rest
ing T lymphocytes do not (48). As described above, activated T
lymphocytes express core 2 GnT, thus allowing the expression of
sialyl Le" in mucin-type oligosaccharides present in activated T
lymphocytes. These results strongly suggest that the expression of the
branched hexasaccharide and its undersialylated forms widely play
roles in cell-cell interaction.
Since both tumor cells and neutrophils contain sialyl Lex on the cell
surface, it is an attractive hypothesis that tumor cells use the same
selectin-carbohydrate interaction during metastasis. When tumor cells
disseminate, blood-borne tumor cells must adhere to endothelial cells
at metastatic sites (49). Once tumor cells adhere to endothelial cells,
they then penetrate through endothelial cells, moving into subendothelial tissues. Once tumor cells grow beneath the endothelia, tumor
metastasis is established (see also Fig. 2). In fact, it was shown in
early studies that tumor cells bind to endothelial cells by E-selectin/
carbohydrate interaction (50).
It has been demonstrated that highly metastatic colonie carcinoma
cells bind more strongly to E-selectin expressed on activated, human
endothelial cells than low-metastatic counterparts (51). A similar
difference was found in the binding of the same colon carcinoma cells
to mouse endothelial cells that were activated (51). The latter findings
indicate that metastasis in nude mice mostly reflects tumor metastasis
in humans, at least at the level of E-selectin-mediated adhesion.
Poorly metastatic tumor cells expressing low levels of sialyl Le"
structure were genetically engineered to increase the amount of sialyl
Le". Resultant cells were found to adhere more strongly to E-selectins
(52). More importantly, it was discovered that the amount of sialyl
Le" on colonie carcinoma cells is directly correlated to the prognosis
CARBOHYDRATE
ANTIGENS
contains not only sialyl Le" but also sulfated tyrosine, which appears
to play critical roles for the binding of PSGL-1 to P-selectin (54-56).
Consistent with this result, it was reported that different kinds of
tumor cells bind more efficiently to P-selectin after the cells were
transfected to express PSGL-1 (57). These tumor cells apparently
contain sulfated glycans that inhibit sialyl Le" and sialyl Le"-dependent adhesion to P-selectin (58). However, a recent report indicates
that colonie carcinoma cells, the same cells used in the above studies
(51, 52), can adhere to P-selectin under flow conditions (59), which
should be closer to the actual situation when tumor cells adhere to
platelets. These colonie tumor cells were shown to lack PSGL-1 (59).
It is thus possible that tumor cells express a P-selectin ligand presenter(s) that is different from PSGL-1.
(b) In tumor cell binding to selectins, which /V-glycans or mucintype glycans primarily carry high-affinity ligands for selectins? In this
regard, it is interesting to note that colo 205 colonie carcinoma cells
express sialyl Le" in mucin-type glycoprotein leukosialin, which may
be involved in E- and P-selectin-mediated adhesion (60).
(c) How can the amount of sialyl Le" in tumor cells be correlated
to the presence of other carbohydrates present in tumor cells'? For
example, the presence of sialyl Tn, NeuNAca2—»6GalNAcal—»Ser/
Thr or Tn antigen GalNAcal—»Ser/Thr in mucin-type oligosaccha
rides likely indicates that the mucin-type O-glycans present in those
tumor cells are shorter than those that do not express sialyl Tn or Tn
antigen (61). If this is the case, those tumor cells likely contain less
amounts of sialyl Le" and sialyl Le" structures. It is then possible that
colonie carcinoma cells expressing sialyl Tn or Tn antigen is less
metastatic than those that express less of the antigen. Contrary to this,
it was reported that compared to patients with colonie tumors con
taining less amounts of sialyl Tn and Tn, those containing increased
amounts of these antigens showed a poorer prognosis (62). Further
studies are thus necessary to clarify this point. Those studies address
ing the above three points should provide a comprehensive under
standing on the roles of tumor-associated carbohydrate antigens dur
ing metastasis.
It is also necessary to mention here that carbohydrate-protein in
teraction may play a role beyond the scope of selectin-carbohydrate
interaction. In fact, it has been shown that galectin-1 on endothelial
cells, which belongs to a family of S-type lectin, is involved in
apoptosis during T-cell development (63). It is noteworthy that the
binding of galectin-1 to /V-glycans on thymocytes is critical in this
process. Mucin-type O-glycans are inhibiting this process (63). Con
sidering that the conversion of O-glycans from the hexasaccharide to
the tetrasaccharide in thymocyte development (17), it is likely that
bulkier O-glycans are more efficient in inhibiting the interaction
between galectin-1 and iV-glycans. These results suggest that carbo
hydrate-protein interaction in tumor metastasis may also use mecha
nisms other than selectin-carbohydrate interaction. Further studies
will be of great significance to reveal such interactions.
of the patients with these carcinomas (53). The patients with a higher
expression of cell surface sialyl Le" structures display much poorer
prognosis than those with lower expression of sialyl Lex, although
Modulation of the N-CAM by Polysialic Acid
other histological criteria were apparently similar.
These combined studies strongly suggest that one of the key factors
in metastatic spread is the abundance of sialyl Le" structure and
possibly sialyl Lea, the ligand for E- and P-selectin expressed in tumor
Polysialic acid is a developmentally regulated carbohydrate com
posed of a linear homopolymer of a-2,8-linked sialic acid residues.
Polysialic acid is mainly attached to the N-CAM. Although polysialylated N-CAM is abundant in embryonic tissues, the majority of
N-CAM in adult tissues lacks this unique glycan (64-66). The pres
cells. There are several points that need to be understood:
(a) It has not been determined whether tumor cells bind to Pselectin as well. This is a critical point since tumor cells aggregate
around platelets (49) and there is a possibility that this aggregation is
due to the interaction of P-selectin on platelets and carbohydrates on
tumor cells. In this regard, it is important to point out that PSGL-1
ence of this large negatively charged carbohydrate modulates the
adhesive property of N-CAM, and removal of polysialic acid in
creases binding between N-CAM-bearing liposomes and neuroblas
toma cells expressing N-CAM (67, 68). During embryonic develop
ment, the polysialylated, embryonic N-CAM is restricted to specific
cell types where cells are migrating (69, 70), and the removal of
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ROLES OF TUMOR-ASSOCIATED
polysialic acid from the N-CAM was reported to influence motor
neuron projections in the embryonic tissues (69). Other studies sug
gest that the polysialylated form of N-CAM not only exhibits antiadhesive properties on homophilic N-CAM interaction but also attenu
ates cell-cell interactions carried by other cell adhesion molecules (65,
71, 72). The recent studies of N-CAM knock-out mice demonstrated
a defect in spatial learning and memory, due to an anomaly of the
olfactory bulb and hippocampus (73 74), where polysialic acid is
continuously expressed also in adult brain (75, 76). These results
strongly suggest that polysialylation regulates the function of NCAM. Due to the presence of polysialic acid in these tissues under
going synaptic rearrangement and cell migration, polysialic acid is
implicated in reducing N-CAM adhesion and thus perhaps allowing
increased neurite outgrowth and cellular mobility.
Although the expression of polysialic acid is diminished in the
majority of tissues in the adult, some tumors are known to re-express
polysialic acid; thus, polysialic acid represents an oncodevelopmental
antigen. The first example of this expression was discovered in
Wilms' tumor (77). Wilms' tumor is a primary renal tumor of child
hood and is characterized by the presence of structural elements,
which are found during embryonic development of the human kidney.
In particular, undifferentiated blastoma regions, which are composed
of small closely packed cells, express the highest amount of polysialic
acid. Regions showing epithelial differentiation, such as glomeruloid
structures and tubules, exhibited only a weak staining. The majority of
these polysialic acid glycans were found to be attached to N-CAM.
These results clearly establish that Wilms' tumors express polysialic
acid, which is suppressed in normal adult kidney.
Very recently, a cDNA encoding a PST was cloned (78, 79). The
predicted amino acid sequences of both human and hamster PST show
that the enzyme consists of 359 amino acid residues and has homology with other sialyltransferases belonging to the sialyltransferase
superfamily. It was also demonstrated that PST can form all a-2,8sialic acid residues necessary for polysialic acid formation (80).
Consistent with the above results on tissue distribution, mRNA of PST
is substantially present in fetal kidney but absent in adult kidney (78).
It will be thus of interest to test if the amount of the PST transcript is
elevated in Wilms' tumor. It will be also significant if Wr-1, the tumor
suppressor gene identified in Wilms' tumor (81), regulates the gene
expression of PST.
Small cell lung carcinoma is a highly malignant tumor showing
some neuroendocrine characteristics. It was found that human small
cell lung carcinomas, irrespective of their histológica! type, express
polysialic acid (82). Metastatic tumor cells, derived from small cell
lung carcinomas, also express polysialic acid. In contrast, polysialic
acid was not detected in the bronchial as well as gastrointestinal
carcinoids. In one of the cell lines derived from a human small cell
lung carcinoma (NCI-H69), a subline containing a negligible amount
of polysialic acid (E2) and that containing a significant amount of
polysialic acid (F3), respectively, were established (83). A most recent
study showed that the expression of polysialic acid in this small cell
lung carcinoma cell line is directed by sialyltransferase X, which is
highly homologous to PST (84). In the majority of other cells, it
appears that PST determines the expression of polysialic acid (78).
However, these results indicate that it is necessary to determine
whether PST or sialyltransferase X directs the expression of polysialic
acid in a given cell (see Refs. 80 and 84 for detailed discussion).
When these two carcinoma sublines, derived from NCI-H69 as
described above, are compared, F3 shows a much more retarded
reaggregation than E2, after cells were dissociated and let them
aggregate. This retarded reaggregation disappears after removal of
polysialic acid by endoneuraminidase.
More importantly, compared to the polysialic acid-negative clonal
CARBOHYDRATE
ANTIGENS
N-CAM + PSA
Fig. 3. Neurite outgrowth dependent on polysialic acid attached to N-CAM. Parent
HeLa cells. HeLa cells stably expressing N-CAM alone, and HeLa cells expressing
N-CAM and polysialic acid were cultured as monolayers. Neurons from embryonic
chicken were seeded on those HeLa cells and cultured for 15 h. The neuntes were
visualized by immunofluorescent staining for neurofilament. Neurons cultured on HeLa
cells expressing N-CAM and polysialic acid grew significantly longer neuntes, which
exhibited more branching. AAA. polysialic acid [based on Nakayama el al. (78)].
subline, (E2), polysialic acid-positive clonal subline (F3) forms much
more colonies in soft-agar or methyl cellulose. Most significantly, i.e.
metastasis took place when F3 subline was s.c. injected into nude
mice. In contrast, the polysialic acid-negative subline produced only a
negligible amount of metastasis, despite the fact that both E2 and F3
express comparable amounts of N-CAM (83).
These results strongly suggest that masking or weakening of NCAM homophilic or heterophilic interaction may allow those tumor
cells to detach more easily from primary tumor. They also may be able
to survive in the blood stream by preventing the attachment to endothelial cells.
Consistent with these discoveries, the amount of N-CAM itself is
reduced in some tumor métastases(85, 86). In these tumors, either
decreasing of N-CAM itself or weakening the adhesive capability of
N-CAM by polysialic acid results in increased metastasis.
When tumor cells migrate and adhere to certain tissues, polysialic
acid-containing N-CAMs appear to play roles. Natural killer cellderived lymphomas express polysialylated N-CAM, and their metastatic patterns are characterized by unusual tissue involvement, such
as the central nervous system, nasopharyx, gastrointestinal tract, skel
etal muscle, and kidney (87, 88). The recent studies showed that
mRNA of PST can be detected in all of these tissues (78). Moreover,
those tissues express polysialic acid detected by anti-polysialic acid
antibody. These results strongly suggest that lymphoma cells express
ing polysialic acid and N-CAM migrate to the tissues where N-CAM
and polysialic acid are expressed, achieving the binding of those
lymphoma cells at the metastatic sites. For successful attachment of
lymphoma cells, however, the amount of polysialic acid should be
moderate in both lymphomas and tissues where the attachment takes
place. If the polysialic acid content is too high, the detachment of
lymphoma cells can take place, but the attachment of those cells to
particular tissues may not be possible. Although there is no quantita
tive data of the polysialic acid content on lymphoma cells, it is
noteworthy that the heart, which expresses a very high amount of
polysialic acid (78), is not a preferential site for the attachment of
lymphoma cells.
It has been shown clearly that neurite outgrowth is facilitated much
better on substrate cells that express both N-CAM and polysialic acid
than those expressing N-CAM alone (Fig. 3; Ref. 78). The presence of
polysialic acid thus allows cells to migrate more efficiently, prevent
ing static adhesion (89). Such modulated cell movement may be also
critical when tumor cells eventually adhere to the sites where they
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ROLES OF TUMOR-ASSOCIATED
grow, after invading into subendothelial tissues and further migrating
into the metastatic sites. Further studies will be important to determine
the roles of polysialic acid by testing the metastatic capability of
tumor cells that express a different amount of polysialic acid by gene
manipulation. In this regard, it is also now possible to determine the
roles of polysialylated gangliosides by using a cDNA encoding the
newly identified ganglioside-specific polysialyltransferase (90).
In summary, I have described three different roles of tumor-asso
ciated carbohydrates in the pathogenesis of tumor formation. The first
section concerned immunogenicity toward oligosaccharides that are
aberrantly synthesized. The second section concerned the ligand func
tion of oligosaccharides in protein-carbohydrate interaction. The third
CARBOHYDRATE
ANTIGENS
19. Patel,
D. D.. and Haynes. B. F. Cell adhesion molecules involved in intrathymic T cell
development. Semin. Immunol.. 5: 282-292, 1993.
20. Saitoh. O.. Piller. F.. Fox, R. !.. and Fukuda. M. T-lymphocytic leukemia express
complex, branched O-linkcd oligosaccharides on a major sialoglycoprotein, leuko
sialin. Blood, 77: 1491-1499. 1991.
Brockhausen. I.. Kuhns, W., Schachter, H., Matta, K. L.. Sutherland. D. R., and
Baker. M. A. Biosynthesis of O-glycans in leukocytes from normal donors and from
patients with leukemia: increase in O-glycan core 2 UDP-GlcNAc:Galß3 GalNAc
a-R (GlcNAc to GalNAc ß-(l,6)-/V-acetylglucosaminyltransferase in leukemic cells.
Cancer Res., 51: 1257-1263, 1991.
22. Piller, F.. Le Deist. F., Weinberg. K. !.. Parkman, R., and Fukuda, M. Allered
O-glycan synthesis in lymphocytes from patients with Wiskott-Aldrich syndrome. J.
Exp. Med.. ¡73: 1501-1510, 1991.
23. Higgins. E. A., Siminovitch. K. A.. Zhuang. D.. Brockhausen. !.. and Dennis, J. W.
Aberrant O-linked oligosaccharide biosynthesis in lymphocytes and platelets from
patients with the Wiskotl-Aldrich syndrome. J. Biol. Chem., 266: 6280-6290. 1991.
Remold-O'Donnell. E.. and Rosen, F. S. Sialophorin (CD43) and the Wiskott-Aldrich
24.
section described the modulatory function of oligosaccharides at
tached to functional proteins. I believe that these three different
functions of oligosaccharides will be continuously revealed in further
studies on tumor-associated carbohydrate antigens.
syndrome. Immunodefic. Rev., 2: 151-173. 1990.
25. Derry. J. M. J.. Ochs. H. D.. and Francke. U. Isolation of a novel gene mutated in
Wiskott-Aldrich syndrome [published erratum appears in Cell. 79: 1994]. Cell. 78:
635-644. 1994.
26. Lee, C. H., Li, W., Nishimura. R., Zhou. M.. Batzer. A. G.. Myers, M. G., Jr., White,
M. F., Schlessinger, J., and Skolnik, E. Y. Nek associates with (he SH2 domaindocking protein 1RS-1 in insulin-stimulated cells. Proc. Nati. Acad. Sci. USA, 90:
11713-11717. 1993.
Remold-O'Donnell, E., Kenney. D. M.. Parkman, R., Cairns, L., Savage, B.. and
Acknowledgments
I thank my colleagues and collaborators who played key roles in obtaining
the results summarized in this article; Drs. Jun Nakayama and Edgar Ong for
critical reading of the manuscript; and Susan Greaney for the arrangement of
the manuscript.
References
1. Hakomori. S. Glycosphingolipids in cellular interaction, differentiation, and oncogenesis. Annu. Rev. Biochem.. 50: 733-764, 1981.
2. Fukuda, M., and Fukuda. M. N. Cell surface glycoproteins and carbohydrate antigens
in development and differentiation of human erythroid cells. In: R. J. Ivatt, The
Biology of Glycoproteins (éd.),pp. 183-234. New York: Plenum Publishing Corp..
1984.
3. Feizi, T. Demonstration by monoclonal antibodies that carbohydrate structures of
glycoproteins and glycolipids are onco-developmental antigens. Nature (Lond.). 314:
53-57, 1985.
4. Jessell, T. M., Hyncs, M. A., and Dodd, J. Carbohydrates and carbohydrate-binding
proteins in the nervous system. Annu. Rev. Neurosci., 13: 227-255, 1990.
5. Fukuüa,M. Cell type-specific expression of cell surface carbohydrates. In: M. Fukuda
and O. Hindsgaul (eds.). Molecular Glycobiology. pp. 1-52. Oxford: Oxford University Press, 1994.
6. Hakomori, S. Aberrant glycosylation in cancer cell membranes as focused on glycolipids: overview and perspectives. Cancer Res.. 45: 2405-2414, 1985.
7. Dennis, J. W. Changes in glycosylation associated with malignant transformation and
tumor progression. In: M. Fukuda (ed.). Cell Surface Carbohydrates and Cell Development, pp. 161-194. London: CRC Press, 1992.
8. Fukuda. M. Cell surface glycoconjugates as onco-differentiation markers in hematopoietic cells. Biochem. Biophys. Acta, 780: 119-150, 1985.
9. Muramatsu. T. Carbohydrate signals in metastasis and prognosis of human carcinomas. Glycobiology. 3: 291-296. 1993.
10. Fukuda. M.. Carlsson. S. R., Klock, J. C., and Dell. A. Structures of O-linked oligosacchandes isolated from normal granulocytes, chronic myelogenous leukemia cells and
acute myelogenous leukemia cells. J. Biol. Chem., 26/: 12796-12806, 1986.
11. Piller, F., Piller, V., Fox, R. L, and Fukuda, M. Human T-lymphocyte activation is
associated with changes in O-glycan biosynthesis. J. Biol. Chem.. 263: 15146-15150,
1988.
12. Ardman, B., Sikorski, M. A., and Staunton, D. E. CD43 interferes with T-lymphocyte
adhesion. Proc. Nati. Acad. Sci. USA, 89: 5001-5005, 1992.
13. Yonemura. S., Nagafuchi, A., Sato, N., and Tsukita. S. Concentration of an integral
membrane protein. CD43 (leukosialin. sialophorin). in the cleavage furrow through
the interaction of its cytoplasmic domain with actin-based cytoskeletons. J. Cell Biol..
120: 437-449, 1993.
14. Nathan. C.. Xie, Q. W., Halbwachs-Mecarelli. L.. and Jin. W. W. Albumin inhibits
neutrophil spreading and hydrogen peroxide release by blocking the shedding of
CD43 (sialophorin. leukosialin). J. Cell Biol., /22: 243-256. 1993.
15. Bazil. V.. and Strominger. J. L. CD43. the major sialoglycoprotein of human leuko
cytes, is proteolvtically cleaved from the surface of stimulated lymphocytes and
granulocytes. Proc. Nati. Acad. Sci. USA, 90: 3792-3796. 1993.
16. Schmid. K.. Hediger. M. A.. Brossmer. R., Collins. J. H.. Haupt. H.. Marti. T.. Offner.
G. D., Schaller, J.. Takagaki, K.. Walsh, M. T., Schwick, H. G., Rosen. R. S., and
Remold-O'Donnell, E. Amino acid sequence of human plasma galactoglycoprotein:
identification with the extracellular region of CD43 (sialophorin). Proc. Nati. Acad.
Sci. USA, 89: 663-667. 1992.
17. Baum, L. G., Pang. M., Perillo, N. L., Wu, T., Delegeane. A.. Uittenbogaart. C. H.,
Fukuda. M., and Seilhamer, J. J. Human thymic epithelial cells express an endogenous lectin. galectin-l. which binds to core 2O-glycans on thymocytes and T lymphoblastoid cells. J. Exp. Med.. 181: 877-887, 1995.
18. Weissman, I. L. Developmental switches in the immune system. Cell, 76: 207-218,
1994.
27.
Rosen. F. S. Characterization of a human lymphocyte surface sialoglycoprotein that
is defective in Wiskott-Aldrich syndrome. J. Exp. Med., 15V: 1705-1723, 1984.
Ardman, B., Sikorski. M. A., Settles. M.. and Staunton. D. E. Human immunodefi
ciency virus type 1-infected individuals make autoantibodies that bind to CD43 on
normal thymic lymphocytes. J. Exp. Med., /72: 1151-1158, 1990.
29. Gillespie, W.. Paulson. J. C., Keim, S., Pang, M., and Baum, L. Regulation of
a2,3-sialyltransferase
expression correlates with conversion of peanut agglutinin
(PNA)* to PNA~ phenotype in developing thymocytes. J. Biol. Chem.. 26/Õ:380130.
31
32.
33.
34.
35.
36.
37.
38.
39.
40.
41.
3804. 1993.
Mentzer, S. J.. Remold-O'Donnell,
E.. Crimmins. M. A., Bierer, B. E., Rosen, F. S.,
and Burakoff. S. J. Sialophorin, a surface sialoglycoprotein defective in the WiskottAldrich syndrome, is involved in human T lymphocyte proliferation. J. Exp. Med.,
165: 1383-1392. 1987.
Lefebvre, J. C., Giordanengo, V., Limouse, M., Doglio, A., Cucchiarini, M.,
Monpoux. F.. Mariani, R.. and Peyron. J. F. Altered glycosylation of leukosialin.
CD43, in HIV-1-infected cells of the CEM line. J. Exp. Med., 180: 1609-1617, 1994.
Dragone, L. L., Barth, R. K., Sitar, K. L.. Disbrow, G. L., and Frelinger, J. G.
Disregulation of leukosialin (CD43, Ly48, sialophorin) expression in the B-cell
lineage of transgenic mice increases splenic B-cell number and survival. Proc. Nati.
Acad. Sci. USA, 92: 626-630, 1995.
Manjunath. N.. Correa. M.. Ardman. M., and Ardman, B. Negative regulation of
T-cell adhesion and activation by CD43. Nature (Lond.). 377: 535-538, 1995.
Fukuda. M.. Spooncer. E., Dates, J. E., Dell, A., and Klock, J. C. Structure of
sialylated fucosyl lactosaminoglycan isolated from human granulocytes. J. Biol.
Chem., 259: 10925-10935, 1984.
Mizoguchi. A., Takasaki, S.. Maeda. S.. and Kobata, A. Changes in asparagine-linked
sugar chains of human promyelocytic leukemic cells (HL-601 during monocytoid
differentiation and myeloid differentiation. Decrease of high-molecular-weight oli
gosaccharides in acidic fraction. J. Biol. Chem., 259: 11949-11957. 1984.
Springer. T. A. Traffic signals for lymphocyte recirculation and leukocyte emigration:
the multistep paradigm. Cell, 76: 301-314, 1994.
Lowe, J. B.. Stoolman, L. M.. Nair, R. P., Larsen, R. D.. Berhend, T. L., and Marks,
R. M. ELAM-1-dependent cell adhesion to vascular endothelium determined by a
transfected human fucosyltransferase cDNA. Cell, 63: 475-484, 1990.
Walz, G.. Aniffo, A.. Kolanus. W.. Bevilacqua. M., and Seed, B. Recognition by
ELAM-1 of the sialyl-Lex determinant on myeloid and tumor cells. Science (Wash
ington DC), 250: 1132-1135, 1990.
Phillips, M. L.. Nudelman, E.. Gaeta, F. C. A., Perez, M., Singhai, A. K., Hakomori.
S-I-. and Paulson, J. C. ELAM-1 mediates cell adhesion by recognition of a carbo
hydrate ligand. sialyl Le". Science (Washington DC), 250: 1130-1132, 1990.
Nguyen, M., Strubel. N. A., and Bischoff. J. A role for sialyl Lewis-X/A glycocon
jugates in capillary morphogenesis [published erratum appears in Nature (Lond.I.
366: 368. 1993). Nature (Lond.). 365: 267-269. 1993.
Fukushima. K.. Hirota. M.. Terasaki. P. I.. Wakasaka, A-, Togashi. H., Ghia, D..
Suyama. N., Fukushi. Y.. Nudelman. E.. and Hakomori. S-I. Characterization of
sialosylated Lewis" as a new tumor-associated antigen. Cancer Res.. 44: 5279-5285.
1984.
42. Magnani. J. L.. Nilsson. B.. Brockhaus. M.. Zopf. D., Steplewski. Z.. Koprowski. H.,
and Ginsburg, V. A monoclonal antibody-defined antigen associated with gastroin
testinal cancer is a ganglioside containing sialylated lacto-JV-fucopenlaose II. J. Biol.
Chem., 257: 14365-14369, 1982.
43.
Berg, E. L., Robinson, M. K.. Mansson. O., Butcher. E. C., and Magnani. J. L. A
carbohydrate domain common to both sialyl LeÃ-a)and sialyl Le(X) is recognized by
the endothelial cell leukocyte adhesion molecule ELAM-1. J. Biol. Chem.. 266:
14869-14872, 1991.
44. Takada. A.. Ohmori. K., Takahashi. N.. Tsuyuoka, K., Yago, A., Zenita. K.. Hasegawa. A., and Kannagi, R. Adhesion of human cancer cells to vascular endothelium
mediated by a carbohydrate antigen, sialyl Lewis A. Biochem. Biophys. Res. Com
mun.. /79: 713-719. 1991.
45. Sako. D.. Chang, X-J.. Barone, K. M.. Vachino, G.. White, H. M., Shaw, G.,
2243
Downloaded from cancerres.aacrjournals.org on June 17, 2017. © 1996 American Association for Cancer Research.
ROLES OF TUMOR-ASSOCIATED
. Bean. K. M.. Ahem. T. J.. Furie. B.. Gumming. D. A., and Larsen.
>cloning of a functional glycoprotein ligand for P-selectin. Cell. 75:
Singer. M. S.. Henzel. W.. Hemmerich. S.. Renz. M.. Rosen, S. D..
i. Binding of L-selectin lo the vascular sialomucin CD34. Science
K'»,262.- 436-438. 1993.
;>nd Fukuda. M. Poly-W-acetyllactosaminyl O-glycans attached to
.. presence of sialyl Le* structures in O-glycans. J. Biol. Chem.. 267:
b ',992.
i i »
. :;ada, A., Yoneda. T.. Buma. Y.. Hirashima. K., Tsuyuoka, K.,
.,:id Kannagi, R. Differentiation-dependent expression of sialyl stage•¿
-j'O'iic antigen-1 and I-antigens on human lymphoid cells and its impliLjr.nhydrate-mediated
adhesion to vascular endothelium Blood. 81:
'3
I
Metastatic tumor cell interactions with endothelium. basement
M :•
M-: ¡issue.Curr. Opin. Cell Biol.. /: 1009-1019, 1989.
•¿â€¢.)
' vi'
I. P., Stengelin. S.. Gimbrone. M. A.. Jr.. and Seed. B. Endothelial
IT ..' '' trtli-e ion molecule I: an inducible receptor for neutrophils related to
*);•,>!
n; ; •¿â€¢
,-ulatory proteins and lectins. Science (Washington DCI. 243: 1160; 'f*
•¿â€¢;•
..
51. ?aiv^
-i., i 31.boi. S.. and Fukuda. M. Differential E-selectin-dependent adhesion
f.i .i' i •¿;/
" sublines of a human colon cancer with distinct metastatic potentials. J.
Biol. -...i-m., 269: 1425-1431, 1994.
52. Siva«: ,;. Lowe. J. B.. and Fukuda. M. E-Selectin-dependent adhesion efficiency of
colorie carcinoma cells is increased by genetic manipulation of their cell surface
lamp-1 expression levels. J. Biol. Chem., 26«.12675-12681, 1993.
53. Nak: non. S.. Kameyama. M.. Imaoka. S.. Furukawa. H.. Ishikawa. 0.. Sasaki. Y..
Kabu o. T., Iwanaga. T.. Matsushita. Y.. and Irimura. T. Increased expression of sialyl
Lewi^* antigen correlates with poor survival in patients with colorectal carcinoma:
clÃ-nico ittiologica! and immunohistochemical study. Cancer Res., 53: 3632-3637,
m\
54. Vii'k.n." P. P.. Moore, K. L., McEver, R. P.. and Cummings. R. D. Tyrosine sulfation
i/ I' sel. clin glycoprotein ligand-1 is required for high affinity binding to P-selectin.
J. Dio! "hem., 270: 22677-22680, 1995.
5f. ^iko. > , Comess, K. M.. Barone. K. M.. Camphausen. R. T.. Cumming. D. A., and
£haw G. D. A sulfated peptide segment at the amino terminus of PSGL-I is critical
: ,- P- el;i.Jn binding. Cell. 83: 323-331. 1995.
56. i1 mv i,l . and Seed, B. PSGL-1 recognition of P-selectin is controlled by a tyrosine
suli 't, -n ronsensus at the PSGL-1 amino terminus. Cell, S3: 333-343. 1995.
y!. K KM K. White. T.. Ito, K.. Fang, H.. Wang, S., and Hakomori. S. P-Selectin•¿
,*' ¿l adhesion of human cancer cells: requirement for co-expression of a
1 '..-;- ike" core protein and the glycosylation process for sialosyl-Lex or sialosylL,' ;. t. J Dn^ol., 6: 773-781, 1995.
58. Ht.i.; i.K,t- udelman, E. D., Stroud, M. R., Shiozawa, T.. and Hakomori. S. Selectin
CÕ'F- 14i: ('JD62 Padgem) binds to sialosyl-Lea and sialosyl-Le*. and sulfated glycans
modulate thi- binding. Biochem. Biophys. Res. Commun.. 181: 1223-1230. 1991.
59. GoetÃ-,I •¿.
I. Ding H.. and Luscinskas, F. W. A colon carcinoma cell line adheres to
P-sc'cc'- i jiider flow conditions independent of PSGL-1. Mol. Biol. Cell. 6: 410a.
19°i.
M). Baerkstrorr. D.. Znang. K.. Asker. N.. Riietschi, U.. Ek. M.. and Hansson. G. C.
Expression of the leukocyte-associated sialoglycoprotein CD43 by a colon carcinoma
celi line. J. B'ol. Chen., 270: 13688-13692. 1995.
61. Itzkcr.vitz. T H.. Yuan. M., Montgomery, C. K.. Kjeldsen. T.. Takahashi. H. K..
Bigbee. W. i .. and Kim, Y. S. Expression of Tn, sialosyl-Tn. and Tantigens in human
cole,'! c.mcer. Cancer Res., 49: 197-204, 1989.
62. Itzkfwitz, S. H., Bloom, E. J., Kokal. W. A.. Modin, G., Hakomori. S., and Kim. Y. S.
Sialos>i-Tn: a novel mucin antigen associated with prognosis in colorectal cancer
patients. Cancer (Phila.), 66: 1960-1966. 1990.
63. Perillo, N. L., Pace. K. E.. Seilhamer, J. J.. and Baum. L. G. Apoptosis of T cells
mediated by galectin-1. Nature (Lond.). 378: 736-739, 1995.
64. Edelman. G. M. Cell adhesion and the molecular processes of morphogenesis. Annu.
Rev. Biochem., 54: 135-169, 1985.
65. Rutishauser. U.. Acheson. A.. Hall, A. K., Mann. D. M.. and Sunshine. J. The neural
cell adhesion molecule (NCAM) as a regulator of cell-cell interactions. Science
(Washington DC). 240: 53-57. 1988.
66. Finne, J. Occurrence of unique polysialosyl carbohydrate units in glycoproteins of
developing brain. J. Biol. Chem.. 257: 11966-11970. 1982.
67. Sadoul, R.. Hirn. M.. Deagostini-Bazin. H.. Rougon. G.. and Goridis. C. Adult and
embryonic mouse neural cell adhesion molecules have different binding properties.
Nature (Lond.), 304: 347-349. 1983.
68. Hoffman, S., and Edelman. G. M. Kinetics of homophilic binding by embryonic and
adult forms of the neural cell adhesion molecule. Proc. Nati. Acad. Sci. USA. 80:
5762-5766, 1983.
69. Tang, J., Rutishauser. U.. and Landmesser. L. Polysialic acid regulated growth cone
ehavior during sorting of motor axons in the plexus region. Neuron. 13: 405-414.
"¿94.
CARBOHYDRATE
ANTIGENS
70. Predette. B.. Rutishauser. U., and Landmesser. L. Regulation and activity-dependence
of N-cadherin, NCAM ¡soforms. and polysialic acid on chick myotubes during
development. J. Cell Biol.. 123: 1867-1888. 1993.
71. Kadmon. G.. Kowitz. A.. Altevogt. P.. and Schachner. M. Functional cooperation
between the neural adhesion molecules LI and N-CAM is carbohydrate dependent. J.
Cell Biol.. 110: 209-218. 1990.
72. Bixby. J. L.. Pratt. R. S.. Lilien. J.. and Reichardt. L. F. Neunte outgrowth on muscle
cell surfaces involves extracellular matrix receptors as well as Ca2 * -dependent and
-independent cell adhesion molecules. Proc. Nati. Acad. Sci. USA, 84: 2555-2559,
1987.
73. Tomasiewicz. H., Ono. K.. Yee. D., Thompson. C.. Goridis. C.. Rutishauser, U.. and
Magnuson. T. Genetic deletion of a neural cell adhesion molecule variant (N-CAM180) produces distinct defects in the central nervous system. Neuron. //: 1163-1174,
1993.
74. Cremer. H.. Lange. R.. Christoph. A.. Plomann. M.. Vopper. G.. Roes. J.. Brown, R..
Baldwin, S.. Kraemer. P.. and Scheff. S. Inactivation of the N-CAM gene in mice
results in size reduction of the olfactory bulb and deficits in spatial learning. Nature
(Lond.), 367: 455-459, 1994.
75. Theodosis. D. T., Rougon, G.. and Poulain. D. A. Retention of embryonic features by
an adult neuronal system capable of plasticity: polysialylated neural cell adhesion
molecule in the hypothalamo-neurohypophysial
system. Proc. Nati. Acad. Sci. USA,
88: 5494-5498. 1991.
76. Key. B.. and Akeson. R. A. Delineation of olfactory pathways in the frog nervous
system by unique glycoconjugates and N-CAM glycoforms. Neuron, 6: 381-396.
1991.
77. Roth, J.. Zuber, C.. Wagner. P.. Taatjes, D. J., Weisgerber, C., Heitz, P. U., Rajewsky,
K.. and Bitter-Suermann. D. Reexpression of poly (sialic acid) units of the neural cell
adhesion molecule in Wilms tumor. Proc. Nati. Acad. Sci. USA. «5:2999-3003,
1988.
78. Nakayama. J.. Fukuda. M. N.. Predette. B.. Ranscht. B.. and Fukuda. M. Expression
cloning of a human polysialyltransferase that forms the polysialylated neural cell
adhesion molecule present in embryonic brain. Proc. Nati. Acad. Sci. USA. 92:
7031-7035. 1995.
79. Eckhardt. M.. Muhlenhoff. M., Bcthe. A.. Koopman. J.. Frosch. M., and GerardySchahn. R. Molecular characterization of eukaryotic polysialyltransferase-1. Nature
(Lond.). 373: 715-718. 1995.
80. Nakayama. J., and Fukuda. M. A human polysialyltransferase directs in vitro syn
thesis of polysialic acid. J. Biol. Chem.. 27/: 1829-1832. 1996.
81. Haber. D. A., and Housman. D. E. The genetics of Wilms' tumor. Adv. Cancer Res..
59: 41-68, 1992.
82. Komminoth. P., Roth, J.. Lackie. P. M.. Bitter-Suermann. D.. and Heitz. P. U.
Polysialic acid of the neural cell adhesion molecule distinguishes small cell lung
carcinoma from carcinoids. Am. J. Pathol., 139: 297-304, 1991.
83. Scheidegger, E. P.. Lackie. P. M.. Papay. J.. and Roth, J. In vitro and in vivo growth
of donai sublines of human small cell lung carcinoma is modulated by polysialic acid
of the neural cell adhesion molecule. Lab. Invest.. 70: 95-106. 1994.
84. Scheidegger, E. P.. Stemberg. L. R.. Roth. J.. and Lowe. J. B. A human STX cDNA
confers polysialic acid expression in mammalian cells. J. Biol. Chem.. 270: 2268522688, 1995.
85. Andersson, A. M., Moran, N.. Gaardsvoll. H.. Linnemann. D.. Bjerkvig, R.. Laerum.
O. D.. and Bock. E. Characterization of NCAM expression and function in BT4C and
BT4Cn glioma cells. Int. J. Cancer. 47: 124-129. 1991.
86. Linnemann, D., Raz. A., and Bock. E. Differential expression of cell adhesion
molecules in variants of K1735 melanoma cells differing in metastatic capacity. Int.
J. Cancer. 43: 709-712. 1989.
87. Kem. W. F.. Spier. C. M.. Hanneman. E. H., Miller, T. P.. Matzner. M., and Grogan.
T. M. Neural cell adhesion molecule-positive peripheral T-cell lymphoma: a rare
variant with a propensity for unusual sites of involvement. Blood, 79: 2432-2437.
1992.
88. Wong. K. F., Chan, J. K. C., Ng, C. S., Lee, K. C.. Tsang, W. Y. W., and Cheung.
M. M. C. CD56(NKH-1) positive hematolymphoid malignancies: an aggressive
neoplasm featuring frequent cutaneous/mucosal involvement cytoplasmic azurophilic
granule and angiocentricity. Hum. Pathoi.. 23: 798-804. 1992.
89. Livingston. B. D.. Jacobs, J. L., Glick. M. C., and Troy, F. A. Extended polysialic acid
chains (n greater than 55) in glycoproteins from human neuroblastoma cells. J. Biol.
Chem.. 263: 9443-9448. 1988.
90. Nakayama. J., Fukuda. M. N.. Hirabayashi, Y.. Kanamori. A.. Sasaki, K., Nishi. T.,
and Fukuda. M. Expression cloning of a human GT, synthase. GD, and GT1 are
synthesized by a single enzyme. J. Biol. Chem.. 27/: 3684-3691. 1996.
91. Rivero-Lezcano. O. M., Marcilla. A.. Sameshima, J. H.. and Robbins. K. C. WiskottAldrich syndrome protein physically associated with Nek through Src homology 3
domains. Mol. Cell. Biol., 15: 5725-5731. 1995.
92. Symons. M.. Derry, J. M., J.. Karlak, B.. Jiang, S.. Lemanieu. V.. McCormick, F..
Francke. U.. and Abo. A. Wiskott-Aldrich syndrome protein, a novel effector for the
GTPase CDC42Hs. is implicated in actin polymerization. Cell. 84: 723-734. 1996.
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Possible Roles of Tumor-associated Carbohydrate Antigens
Minoru Fukuda
Cancer Res 1996;56:2237-2244.
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