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LYMPHATIC VASCULATURE
DEVELOPMENT
Guillermo Oliver
Although the process of blood vasculature formation has been well documented, little is
known about lymphatic vasculature development, despite its importance in normal and
pathological conditions. The lack of specific lymphatic markers has hampered progress
in this field. However, the recent identification of genes that participate in the formation of
the lymphatic vasculature denotes the beginning of a new era in which better diagnoses
and therapeutic treatment(s) of lymphatic disorders could become a reachable goal.
LYMPHANGIOGENESIS Lymphatic vessels were first described in the seventeenth agent, afferent lymphatic vessels are the route through
The growth of lymphatic vessels. century by G. Aselli1. However, until a few years ago, the which, after antigen uptake, dendritic cells migrate to
lymphatic system was not considered an interesting topic the lymph nodes and lymphoid organs. Recent studies
LYMPHOEDEMA
Occlusion of lymphatic
among most scientists. Despite the importance of the indicate that spindle cells and cells lining the irregular
drainage followed by the lymphatic system and the belief that metastatic tumour spaces in Kaposi’s sarcoma have a lymphatic endothelial
abnormal accumulation of cells are disseminated through the lymphatic vasculature, origin9,10. An increased understanding of normal
interstitial fluid and swelling modern anatomy textbooks generally include only a lymphatic development should allow us to address
in the affected body part.
minimal description of the development of this system. pathological lymphatic conditions that lead to inflam-
LYMPHATIC METASTASIS
Interest in basic lymphatic research has radically mation, autoimmunity and cancer, and to improve the
Invasion of detaching primary increased in the past few years, in part, thanks to the iden- clinical treatment of primary and secondary forms of
tumour cells through the tification of lymphatic-specific markers and growth lymphoedema. Here, I discuss a working model for
lymphatic system to lymph factors, the debate about tumour LYMPHANGIOGENESIS, and normal lymphatic vasculature development that
nodes. After re-entering of the
lymphatic vessels into the blood
the efforts of patients suffering from LYMPHOEDEMA and focuses mainly on the early aspects, and that summarizes
vasculature, tumour cells will their families, who have attracted public and govern- findings from different laboratories through the charac-
also spread to distant organs. mental attention by creating the Lymphatic Research terization of newly identified lymphatic markers and
Foundation (see further information for website). available mouse mutant lines. In this model, blood
Although few life-threatening diseases (BOX 1) result venous endothelial cells are the ground condition from
from malfunction of the lymphatic system, failure of which a lymphatic endothelial cell phenotype will be
lymph transport promotes lymphoedema — a disfig- progressively acquired by the stepwise expression of
uring, disabling and occasionally life-threatening different gene products.
disorder for which present treatment is limited and
ineffective2. Malignant tumours can directly activate Historical considerations
lymphangiogenesis and LYMPHATIC METASTASIS3–8; so, During embryonic development, blood vessels originate
Department of Genetics, understanding the basic biology that is involved in from mesodermally derived endothelial cell precursors
St Jude Children’s Research lymphatic development is vital. Metastasis of most (vasculogenesis) (FIG. 1). These vessels grow and remodel
Hospital, 332 N. Lauderdale, cancers mainly occurs through the lymphatic system, into the mature network by endothelial sprouting and
Memphis, Tennessee 38105, and the extent of lymph-node involvement is a useful splitting (angiogenesis). The lymphatic vasculature
USA.
e-mail: guillermo.oliver@
prognostic indicator. The lymphatic system is also appears after the blood vasculature forms, which was
stjude.org essential for the immune response to infectious agents, the first indication that lymphatics might have a blood
doi:10.1038/nri1258 for example, during exposure to an inflammatory vasculature origin.
Box 1 | Roles of the lymphatic system in health and disease thymus, all of which have key roles in the immune
response to infectious agents.
In health The lymphatic system develops in parallel, but sec-
• Drains lymph fluid from the extracellular spaces ondary to the development of the blood vascular system,
• Absorbs lipids from the intestinal tract and lymphatic vessels are not normally present in avascu-
• Maintains the fluid homeostasis lar structures such as the epidermis, hair, nails, cartilage
• Transports white blood cells and antigen-presenting cells to lymphoid organs and cornea, nor are they present in some vascularized
organs such as the retina and probably the brain. From an
Disease evolutionary point of view, specialized lymph hearts first
• Congenital and acquired defects of the lymphatic system appeared in amphibians and reptiles11; later, lymph nodes
• Lymphoedema: imbalance between lymph formation and adsorption into lymphatic appeared in birds, and the final refinement of the
vessels lymphatic vasculature occurred in mammals2.
• Lymphangitis: inflammation of lymphatic vessels In 1902, Florence Sabin 12,13 proposed the most
widely accepted model of lymphatic vasculature
• Lymphangioma: benign neoplasm of lymphatic vessels
development. Sabin injected dye into pig embryos
• Lymphangiosarcoma: malignant neoplasm of lymphatic vessels (FIG. 3) , and her results culminated in the proposal
• Role in metastasis: migration of tumour cells that endothelial cells bud from veins to form primary
lymphatic sacs. From these sacs, budding cells then
centrifugally sprout towards the periphery, forming
PERICYTES The lymphatic system (FIG. 2) is composed of a vascu- capillaries that surround tissues and organs. In this
Smooth-muscle-like cells that lar network of blind-ended, thin-walled capillaries and model, the two jugular lymph sacs develop near the
cover the outer surface of the larger vessels that drain protein-rich interstitial fluid junction of the subclavian and anterior cardinal veins
endothelial cells of blood vessels.
(lymph) from the extracellular spaces within organs into (FIG. 3d). From these sacs, lymphatic capillaries spread
WOLFFIAN BODIES larger collecting ducts. In contrast to blood capillaries, to the head, neck, arms and thorax. Later, the remaining
A series of tubules that lymphatic capillaries lack fenestrations, a continuous lymph sacs originate from the mesonephric vein and
constitute the mesonephros. basal membrane and PERICYTES; instead, these vessels are veins in the dorsomedial edge of the WOLFFIAN BODIES.
lined with a continuous, single layer of overlapping The retroperitoneal lymph sac forms near the primi-
endothelial cells that form loose intercellular junctions. tive inferior vena cava and mesonephric veins, and
These characteristics make the lymphatic capillaries lymphatics spread to the abdominal viscera and
highly permeable to large macromolecules, pathogens diaphragm. The thoracic duct, which carries most
and migrating cells. lymph back to the blood vascular system, originates in
Lymph returns to the venous circulation through the cisterna chyli — a dilated sac that lies anterior to
the thoracic duct and the larger lymphatic collecting the uppermost lumbar vertebrae — which forms near
vessels, which contain a muscular layer and an adventitial the wolffian bodies. The posterior lymph sacs occur
layer. The lymphatic system is crucial for maintaining near the junctions of the primitive iliac veins and the
the colloid osmotic volume (or pressure) and for posterior cardinal veins; from these sacs, lymphatics
absorbing lipids from the intestinal tract. The lym- spread to the abdominal wall, pelvic region and legs
phatic system also includes lymphoid organs such as (FIG. 3d) (for a detailed anatomical description of the
the lymph nodes, tonsils, Peyer’s patches, spleen and lymphatic system in mice see REF. 14).
Vasculogenesis Angiogenesis
Endothelial
Artery
cell
Basement Vein
membrane
Mesoderm Haemangioblasts Tube formation Pruning and remodelling
cells
Figure 1 | Blood vascular development. Mesodermally derived haemangioblasts give rise to angioblasts, the precursors of
blood and vascular endothelial cells that will form the lining of the blood vessels (vasculogenesis). The remodelling of the primary
capillary network and the formation of new blood vessels from pre-existing vessels by endothelial sprouting and splitting
(angiogenesis) leads to the formation of mature arteries and veins. Figure reproduced, with permission, from REF. 68 © Sinauer
Associates, Inc. (2003).
state, and after expression of Prox1 these cells adopt a of the anterior cardinal vein results from a localized
lymphatic cell-type phenotype7,8,17. This proposal was inductive signalling mechanism or from heterogeneity
recently corroborated by experiments in which over- in the venous endothelial cells.
expression of Prox1 was sufficient to reprogramme cul- According to these results, venous endothelial cells
tured blood vascular endothelial cells into LECs; Prox1 could be considered the source of LEC progenitors,
induced LEC-specific gene expression while repressing which, after Prox1 expression, become biased (commit-
the expression of some blood vascular endothelial cell ted) to the lymphatic specification pathway. However,
markers25,26. In addition, Prox1 is expressed by LECs in we do not know whether these venous LEC progenitors
adult and tumour tissues7. Therefore, Prox1 is a reliable, are lymphangioblast cells. Once the expression of
specific marker of embryonic and adult LECs in normal Prox1 is initiated by the venous endothelial cells, the
and pathological conditions. expression of some receptor–ligand signalling system
In summary, Prox1 seems to be required and suffi- might be required to guide the subsequent polarized
cient to bias a subpopulation of competent venous budding of LECs towards the receptor–ligand source,
endothelial cells (lymphatic progenitors) into the lym- thereby ensuring the precise location of the primary
phatic programme. However, we do not yet know lymph sacs (FIG. 3d). The source and identity of the
whether the restricted expression of Prox1 on one side Prox1-inducing factor in the veins remains unknown.
Unidentified inductive
Arteries Versus Veins
Veins Veins Unknown signal in surrounding
factor tissue
Prox1-negative Prox1
endothelial cells
c E10.5–E11.5 d LEC differentiation
Lymphatic specification
VEGFC
Nrp2
Lymphangiogenesis
Figure 4 | Model for differentiation of the lymphatic vasculature. The proposed four-step model for lymphatic vasculature
formation is summarized in this scheme. a | Lymphatic endothelial cell (LEC) competence is the autonomous ability of venous
endothelial cells to respond to a specific, inductive signal; this stage is characterized, for example, by the appearance or
disappearance of a receptor or by different expression levels of certain transcription factors. Competence is not a passive state,
but an actively acquired condition that occurs only in venous endothelial cells; arterial endothelial cells should not respond, even if
exposed to the inducer. b | LEC bias, or an intermediate state of bias towards LEC determination, is eventually lost in other venous
endothelial cells that are not exposed to the inductive signals. Expression of prospero-related homeobox 1 (Prox1) might be crucial
for this step. c | LEC specification occurs when biased venous endothelial cells differentiate independently from the regional
context; for example, the anterior cardinal vein should give rise to LECs, even if transplanted to another part of the embryo.
d | Finally, lymphatic vessel differentiation and maturation occurs in a stepwise manner leading to synthesis of all of the main
lymphatic vessel components. BEC, blood vascular endothelial cell; D6, β-chemokine receptor D6; E, embryonic day; Lyve1,
lymphatic vessel endothelial hyaluronan receptor 1; Nrp2, neuropilin 2; SLC, secondary lymphoid chemokine; VEGFC, vascular
endothelial growth factor C; VEGFR3, vascular endothelial growth-factor receptor 3.
Specification of lymphatic endothelial cells. Specification final lymphatic vascular differentiation. During
is the stage at which cells will differentiate into the this stage, the expression of more lymphatic markers
desired phenotype, even if isolated and cultured begins whereas the expression of some blood vascular
alone18. Lymphatic bias and specification are closely markers (for example, CD34 and laminin) progres-
linked. Specification of LECs is the first step towards sively decreases7. A clear morphological indication of
lymphatic specification is the budding of immature undetectable in those budding cells7,8,17 and primary
LECs from mouse veins on E10.5 (FIGS 5b, 6). The initi- lymph sacs appear along the embryo at around this
ation of LEC budding is independent of Prox1 func- stage (FIGS 3d, 5c).
tion7,8 and is the first morphological indication that The expression of Prox1 might maintain VEGFR3
lymphangiogenesis has begun. At E10.5, both Prox1 expression by LECs during lymphangiogenesis.
and Lyve1 are expressed by venous endothelial cells Unfortunately, functional inactivation of VEGFR3
and budding LECs (FIGS 4, 5, 6). causes cardiovascular failure and defects in the remodel-
In mice, LECs that bud from the anterior cardinal ling of the primary vascular networks; therefore, these
vein can be observed as late as E12.5, when the primary embryos die before the lymphatic vessels emerge31.
lymph sacs become apparent. A typical characteristic of Nonsense mutations in VEGFR3 have been identified in
the lymphatic capillaries is their lack of a continuous patients with hereditary lymphoedema37. Together,
basement membrane. From around E10.5–11.0, these findings show that VEGFR3 regulates the develop-
expression of extracellular matrix components laminin ment of the blood vascular and the lymphatic vascular
or collagen IV starts to decrease in the budding LECs7. systems38.
Neuropilin 2 (Nrp2) (TABLE 1) mediates axonal Further support for the importance of the VEGFR3-
guidance during neuronal development27. Nrp pro- signalling pathway during lymphatic development is
teins also bind various vascular endothelial growth provided by studies of its ligand VEGFC (TABLE 1),
factors (VEGFs) and have a role in VEGF–VEGFR which was the first lymphangiogenic growth factor to
signalling28. At around E10.0, uniform Nrp2 expres- be identified39. Although no functional inactivation of
sion is detected in the anterior cardinal vein, in the this ligand has been reported, much evidence supports
budding LECs and in the surrounding mesoderm29 an important role for this growth factor during lym-
(N. Harvey and G.O., unpublished observations). phatic development. VEGFC was identified as a potent
Nrp2–/– mice show an absence or severe reduction in inducer of lymphatic sprouting40.
the number of small lymphatic vessels and capillaries VEGFC is mainly expressed in mesenchyme of areas
— a phenotype that is associated with reduced DNA where embryonic lymphatics are developing39.
synthesis in the LECs29. Recombinant VEGFC promotes lymphangiogenesis in
VEGFR3 (also known as Fms-like tyrosine kinase 4, the chick chorioallantoic membrane41. Overexpression
Flt4) (TABLE 1) was one of the first lymphatic markers of VEGFC in the skin of transgenic mice results in
associated with lymphatic differentiation30. VEGFR3 hyperplasia of cutaneous lymphatic vessels42, and over-
binds the secreted glycoproteins VEGFC and VEGFD. expression of VEGFC through the use of a recombinant
Early during mouse development, VEGFR3 is widely adenovirus promotes lymphangiogenesis in adult mouse
expressed by both the blood and lymphatic endothelia, skin43. VEGFC gene transfer to the skin of mice with
including the anterior cardinal vein30–32 (FIG. 4). This lymphoedema induces regeneration of the cutaneous
expression is subsequently downregulated in blood vas- lymphatic vessel network44. In addition, VEGFC stim-
culature and becomes mostly restricted to the lymphatic ulates blood vascular angiogenesis in the mouse
endothelium during late development (FIG. 6c) and cornea in vivo — a result that is probably due to an
adulthood7,30. The pattern of expression of VEGFR3 interaction with VEGFR2, which is expressed by blood
and its ligand VEGFC (see later) in developing mice has vessels45. VEGFC also stimulates the migration and
provided additional support for Sabin’s model. proliferation of endothelial cells46 and promotes
The number and distribution of budding LECs tumour lymphangiogenesis5. VEGFD, another
increases markedly between E10.5 and E12.5. From VEGFR3 ligand, also stimulates lymphangiogenesis in
around E11.0, expression of the cell-surface mucin-type the skin of transgenic mice47. Both VEGFC and
glycoprotein T1α/podoplanin33–35 also starts to be VEGFD stimulate angiogenesis and lymphangiogene-
detected in the budding LECs (FIG. 4). In addition, sis, depending on whether they are bound to VEGFR2
podoplanin-deficient mice have lymphatic defects asso- (mainly an angiogenic receptor) or VEGFR3 (mainly a
ciated with diminished lymphatic transport, congenital lymphangiogenic receptor)48.
lymphoedema and dilation of lymphatic vessels35.
At E12.5, the number of budding Prox1+Lyve1+ Differentiation and maturation of lymphatic vessels and
Nrp2+podoplanin+VEGFR3+ LECs adjacent to the lymphangiogenesis. The final stage — differentiation —
anterior cardinal vein has clearly increased7; expres- results from an induction event promoted by a nearby
sion of secondary lymphoid chemokine (SLC) is now structure18. At around E14.5, the lymphatic vasculature
also evident in these cells7, but expression of blood has spread throughout the developing embryo (FIG. 5d) by
vascular markers, such as CD34 and laminin, is not budding and sprouting from the primary lymph sacs,
seen7,8,17 (FIG. 4); levels of VEGFR3 expression by blood and the immature LECs are in the process of terminal
vascular endothelial cells are now reduced7,30,36. So, the differentiation. As differentiation and lymphangiogenesis
maintenance of high levels of VEGFR3 and the progress, additional lymphatic markers are expressed by
expression of podoplanin and SLC by the budding the forming lymph vessels and capillaries.
Prox1+Lyve1+Nrp2+ immature lymphatic endothelial As proposed earlier, it is probable that the differenti-
cells indicates that, at E12.5, the budding venous ation of the lymphatic vasculature is a stepwise process
endothelial cells are now specified into the lymphatic that occurs over several days17. Therefore, the detection,
programme. Blood vascular markers are mostly as well as the level of expression, of some of the markers
determine precisely the mechanisms that are involved in unanswered; are new lymphatic vessels and capillaries
lymphatic vessel formation in adults during tissue formed only by the proliferation of endothelial cells
regeneration, wound healing and tumour formation. from pre-existing lymphatics, or can they also be
These studies will help to determine the functional role formed from endothelial precursors, lymphangioblast
of peritumoral and intratumoral lymphatics during cells or by endothelial budding from veins? Are lym-
tumour progression, and could eventually lead to the phangioblast cells present in both mammalian
generation of anti-lymphangiogenic therapy. In addi- embryos and adults? What signalling mechanisms
tion, the generation of other animal models of lym- guide the budding of LECs in embryos? What mole-
phatic disorders will increase our understanding of the cules and mechanisms are involved in the separation
molecular mechanisms that underlie the normal and of the lymphatic and blood vessels, and which ones
pathological formation of the lymphatic vessels and maintain their close proximity?
facilitate the development of gene therapy for patients Information gathered during this new molecular
suffering from lymphatic disorders. era, combined with previously gained knowledge
Although great progress has been made during the suggests that gene therapy to promote or inhibit
past few years towards understanding the biology of lymphatic growth should become available in the
the lymphatic system, important questions remain near future.
1. Aselli, G. De Lacteibus sive Lacteis Venis, Quarto Vasorum 18. Grainger, R. M. Embryonic lens induction: shedding light on 36. Partanen, T. A. et al. VEGF-C and VEGF-D expression
Mesarai corum Genere novo invento Mediolani, Milano vertebrate tissue determination. Trends Genet. 8, 349–355 in neuroendocrine cells and their receptor, VEGFR-3, in
(1627). (1992). fenestrated blood vessels in human tissues. FASEB J.
2. Witte, M. H. et al. Lymphangiogenesis and 19. Banerji, S. et al. LYVE-1, a new homologue of the CD44 14, 2087–2096 (2000).
lymphangiodysplasia: from molecular to clinical lymphology. glycoprotein, is a lymph-specific receptor for hyaluronan. 37. Karkkainen, M. J. et al. Missense mutations interfere with
Microsc. Res. Tech. 55, 122–145 (2001). J. Cell Biol. 144, 789–801 (1999). VEGFR-3 signalling in primary lymphoedema. Nature Genet.
3. Karpanen, T. et al. Vascular endothelial growth factor C 20. Prevo, R. et al. Mouse LYVE-1 is an endocytic receptor for 25, 153–159 (2000).
promotes tumor lymphangiogenesis and intralymphatic hyaluronan in lymphatic endothelium. J. Biol. Chem. 276, Identification that mutations in vascular endothelial
tumor growth. Cancer Res. 61, 1786–1790 (2001). 19420–19430 (2001). growth-factor receptor 3 (VEGFR3) are responsible for
4. Mandriota, S. J. et al. Vascular endothelial growth factor-C- 21. Mouta-Carreira, C. et al. LYVE-1 is not restricted to the certain forms of hereditary lymphoedema.
mediated lymphangiogenesis promotes tumour metastasis. lymph vessels: expression in normal liver blood sinusoids 38. Jussila, L. & Alitalo, K. Vascular growth factors and
EMBO J. 20, 672–682 (2001). and downregulation in human liver cancer and cirrhosis. lymphangiogenesis. Physiol. Rev. 82, 673–700 (2002).
5. Skobe, M. et al. Induction of tumor lymphangiogenesis by Cancer Res. 61, 8079–8084 (2001). 39. Kukk, E. et al. VEGF-C receptor binding and pattern of
VEGF-C promotes breast cancer metastasis. Nature Med. 22. Knudson, C. B. & Knudson, W. Hyaluronan-binding proteins expression with VEGFR-3 suggests a role in lymphatic
7, 192–198 (2001). in development, tissue homeostasis, and disease. vascular development. Development 122, 3829–3837
This paper presents important results about the role FASEB J. 7, 1233–1241 (1993). (1996).
of tumour lymphangiogenesis. 23. Jackson, D. G., Prevo, R., Clasper, S. & Banerji, S. LYVE-1, 40. Saaristo, A. et al. Lymphangiogenic gene therapy with
6. Stacker, S. A. et al. VEGF-D promotes the metastatic the lymphatic system and tumor lymphangiogenesis. Trends minimal blood vascular side effects. J. Exp. Med. 196,
spread of tumor cells via the lymphatics. Nature Med. 7, Immunol. 22, 317–321 (2001). 719–730 (2002).
186–191 (2001). 24. Wigle, J. T. & Oliver, G. Prox1 function is required for the 41. Oh, S. J. et al. VEGF and VEGF-C: specific induction of
7. Wigle, J. T. et al. An essential role for Prox1 in the induction development of the murine lymphatic system. Cell 98, angiogenesis and lymphangiogenesis in the differentiated
of the lymphatic endothelial cell phenotype. EMBO J. 21, 769–778 (1999). avian chorioallantoic membrane. Dev. Biol. 188, 96–109
1505–1513 (2002). The first report of a gene product, targeted (1997).
The first report that budding venous endothelial cells inactivation of which leads to embryos without any 42. Jeltsch, M. et al. Hyperplasia of lymphatic vessels in VEGF-C
are a default ground state and after expression of lymphatic vasculature. transgenic mice. Science 276, 1423–1425 (1997).
prospero-related homeobox 1 (Prox1) they acquire a 25. Hong, Y. K. et al. Prox1 is a master control gene in the 43. Enholm, B. et al. Adenoviral expression of vascular
lymphatic phenotype. program specifying lymphatic endothelial cell fate. endothelial growth factor-C induces lymphangiogenesis
8. Oliver, G. & Detmar, M. The rediscovery of the lymphatic Dev. Dyn. 225, 351–357 (2002). in the skin. Circ Res. 88, 623–629 (2001).
system: old and new insights into the development and 26. Petrova, T. V. et al. Lymphatic endothelial reprogramming of 44. Szuba, A. et al. Therapeutic lymphangiogenesis with human
biological function of the lymphatic vasculature. Genes Dev. vascular endothelial cells by the Prox-1 homeobox recombinant VEGF-C. FASEB J. 16, 1985–1987 (2002).
16, 773–783 (2002). transcription factor. EMBO J. 21, 4593–4599 (2002). 45. Cao, Y. et al. Vascular endothelial growth factor C induces
9. Jussila, L. et al. Lymphatic endothelium and Kaposi’s 27. Chen, H. et al. Neuropilin-2, a novel member of the angiogenesis in vivo. Proc. Natl Acad. Sci. USA 95,
sarcoma spindle cells detected by antibodies against the neuropilin family, is a high affinity receptor for the 14389–14394 (1998).
vascular endothelial growth factor receptor-3. Cancer Res. semaphorins Sema E and Sema IV but not Sema III. 46. Makinen, T. et al. Isolated lymphatic endothelial cells
58, 1599–1604 (1998). Neuron 19, 547–559 (1997). transduce growth, survival and migratory signals via the
10. Skobe, M. et al. Vascular endothelial growth factor-C 28. Soker, S. et al. Neuropilin-1 is expressed by endothelial and VEGF-C/D receptor VEGFR-3. EMBO J. 20, 4762–4773
(VEGF-C) and its receptors KDR and flt-4 are expressed in tumor cells as an isoform-specific receptor for vascular (2001).
AIDS-associated Kaposi’s sarcoma. J. Invest. Dermatol. endothelial growth factor. Cell 92, 735–745 (1998). 47. Veikkola, T et al. Signalling via vascular endothelial growth
113, 1047–1053 (1999). 29. Yuan, L. et al. Abnormal lymphatic vessel development in factor receptor-3 is sufficient for lymphangiogenesis in
11. Kampmeier, O. F. Evolution and Comparative Morphology neuropilin 2 mutant mice. Development 129, 4797–4806 transgenic mice. EMBO J. 20, 1223–1231 (2001).
of the Lymphatic System. (Springfield, Illinois, Charles C. (2002). 48. Alitalo, K. & Carmeliet, P. Molecular mechanisms of
Thomas, 1969). 30. Kaipainen, A. et al. Expression of the fms-like tyrosine kinase 4 lymphangiogenesis in health and disease. Cancer Cell.
12. Sabin, F. R. On the origin of the lymphatic system from the gene becomes restricted to lymphatic endothelium during 1, 219–227 (2002).
veins, and the development of the lymph hearts and development. Proc. Natl Acad. Sci. USA 92, 3566–3570 49. Gale, N. W. et al. Angiopoietin-2 is required for postnatal
thoracic duct in the pig. Am. J. Anat. 1, 367–389 (1995). angiogenesis and lymphatic patterning, and only the latter
(1902). The first identification of a gene product with role is rescued by angiopoietin-1. Dev. Cell. 3, 411–423
A remarkable, almost accurate, description of the restricted expression in the lymphatic system. (2002).
mechanisms involved in lymphatic vasculature 31. Dumont, D. J. et al. Cardiovascular failure in mouse 50. Sato, T. N. et al. Distinct roles of the receptor tyrosine
development carried out 100 years ago. embryos deficient in VEGF receptor-3. Science 282, kinases Tie-1 and Tie-2 in blood vessel formation.
13. Sabin, F. R. On the development of the superficial lymphatics 946–949 (1998). Nature 376, 70–74 (1995).
in the skin of the pig. Am. J. Anat. 3, 183–195 (1904). 32. Karkkainen, M. J. & Alitalo, K. Lymphatic endothelial 51. Suri, C. et al. Requisite role of angiopoietin-1, a ligand for the
14. van der Putte S. C. The early development of the lymphatic regulation, lymphoedema, and lymph node metastasis. TIE2 receptor, during embryonic angiogenesis.
system in mouse embryos. Acta Morphol. Neerl. Scand. 13, Semin. Cell Dev. Biol. 13, 9–18 (2002). Cell 87, 1171–1180 (1996).
245–286 (1975). 33. Wetterwald, A. et al. Characterization and cloning of the 52. Abtahian, F. et al. Regulation of blood and lymphatic
15. Huntington, G. S. & McClure, C. F. W. The anatomy and E11 antigen, a marker expressed by rat osteoblasts and vascular separation by signaling proteins SLP-76 and Syk.
development of the jugular lymph sac in the domestic cat osteocytes. Bone 18, 25–32 (1996). Science 299, 247–251 (2003).
(Felis domestica). Am. J. Anat. 10, 177–311 (1910). 34. Breiteneder-Geleff, S. et al. Angiosarcomas express mixed Intriguing results providing indication of genes and
16. Schneider, M., Othman-Hassan, K., Christ, B. & Wilting, J. endothelial phenotypes of blood and lymphatic capillaries: mechanisms that are involved in the separation of
Lymphangioblasts in the avian wing bud. Dev. Dyn. 216, podoplanin as a specific marker for lymphatic endothelium. the blood and lymphatic vasculature.
311–319 (1999). Am. J. Pathol. 154, 385–394 (1999). 53. Paavonen, K., Puolakkainen, P., Jussila, L., Jahkola, T. &
17. Oliver, G. & Harvey, N. A stepwise model of the development 35. Schacht, V. et al. T1α/podoplanin deficiency disrupts normal Alitalo, K. Vascular endothelial growth factor receptor-3 in
of lymphatic vasculature. Ann. NY Acad. Sci. 979, 159–165 lymphatic vasculature formation and causes lymphedema. lymphangiogenesis in wound healing. Am. J. Pathol.
(2002). EMBO J. 22, 3546–3556 (2003). 156,1499–1504 (2000).
54. Milroy, W. F. An undescribed variety of hereditary oedema. 67. Lawson, N. D. & Weinstein, B. M. Arteries and veins: making 82. Sosa-Pineda, B., Wigle, J. T. & Oliver, G. Hepatocyte
NY Med. J. 56, 503 (1892). a difference with zebrafish. Nature Rev. Genet. 3, 674–682 migration during liver development requires Prox1. Nature
55. Meige, H. Dystophie oedematoeuse hereditaire. (2002). Genet. 25, 254–255 (2000).
Presse Med. 6, 341–343 (1898). 68. Gilbert, S. F. Developmental Biology. 7th edn (Sinauer 83. Dyer, M. A., Livesey, F. J., Cepko, C. L. & Oliver, G. Prox1
56. Irrthum, A. et al. Congenital hereditary lymphedema caused Associates Inc., Massachusetts, 2003). function controls progenitor cell proliferation and horizontal
by a mutation that inactivates VEGFR3 tyrosine kinase. 69. Szuba, A., Shin, W. S., Strauss, H. W. & Rockson S. The cell genesis in the mammalian retina. Nature Genet. 34,
Am. J. Hum. Genet. 67, 295–301 (2000). third circulation: radionuclide lymphoscintigraphy in the 53–58 (2003).
57. Ferrell, R. E. et al. Hereditary lymphedema: evidence for evaluation of lymphedema. J. Nucl. Med. 44, 43–57 (2003). 84. Gunn, M. D. A chemokine expressed in lymphoid high
linkage and genetic heterogeneity. Hum. Mol. Genet. 7, 70. Carlson, B. M. Human Embryology and Developmental endothelial venules promotes the adhesion and chemotaxis
2073–2078 (1998). Biology. 2nd edn (Mosby Inc., USA, 1999). of naive T lymphocytes. Proc. Natl Acad. Sci. USA 95,
58. Karkkainen, M. J. et al. A model for gene therapy of human 71. Stratmann, A., Risau, W. & Plate KH. Cell type-specific 258–263 (1998).
hereditary lymphedema. Proc. Natl Acad. Sci. USA 98, expression of angiopoietin-1 and angiopoietin-2 suggests 85. Gunn, M. D. et al. Mice lacking expression of secondary
12677–12682 (2001). a role in glioblastoma angiogenesis. Am. J. Pathol. lymphoid organ chemokine have defects in lymphocyte
An important demonstration that gene therapy could 153,1459–1466 (1998). homing and dendritic cell localization. J. Exp. Med. 189,
eventually be achieved in patients with lymphoedema. 72. Zagzag, D. et al. In situ expression of angiopoietins in 451–460 (1999).
59. Yoon, Y. S. et al. VEGF-C gene therapy augments astrocytomas identifies angiopoietin-2 as an early marker 86. Witmer, A. N. et al. VEGFR-3 in adult angiogenesis.
postnatal lymphangiogenesis and ameliorates secondary of tumor angiogenesis. Exp. Neurol. 159, 391–400 J. Pathol. 195, 490–497 (2001).
lymphedema. J. Clin. Invest. 111, 717–725 (2003). (1999). 87. Yoffey, J. M. & Courtice, F. C. Lymphatics, Lymph and the
60. Fang, J. et al. Mutations in FOXC2 (MFH-1), a forkhead 73. Nibbs, R. J. et al. The β-chemokine receptor D6 is Lymphomyeloid Complex. (Academic Press, London,
family transcription factor, are responsible for the hereditary expressed by lymphatic endothelium and a subset of 1970).
lymphedema–distichiasis syndrome. Am. J. Hum. Genet. vascular tumors. Am. J. Pathol. 158, 867–877 (2001).
67, 1382–1388 (2000). 74. Virata, M. L., Wagner, R. M., Parry, D. A. & Green, K. J. Acknowledgements
The authors identified forkhead box C2 (FOXC2) as Molecular structure of the human desmoplakin I and II amino I thank J. Wigle and N. Harvey, whose outstanding work in my
a gene responsible for the hereditary lymphoedema terminus. Proc. Natl Acad. Sci. USA 89, 544–548 (1992). laboratory has been instrumental in the progress of many of the
–distichiasis syndrome. 75. Schmelz, M., Moll, R., Kuhn, C. & Franke, W. W. Complexus ideas presented here, J. Morgan and S. Rockson for help with
61. Kriederman, B. M. et al. FOXC2 haploinsufficient mice are adhaerentes, a new group of desmoplakin-containing the figures, N. Harvey for critical reading of the manuscript and A.
a model for human autosomal dominant lymphedema– junctions in endothelial cells: II. Different types of lymphatic McArthur for scientific editing. This work was supported by the
distichiasis syndrome. Hum. Mol. Genet. 12, 1179–1185 vessels. Differentiation 57, 97–117 (1994). National Institutes of Health, Cancer Center Support from the
(2003). 76. Ebata, N. et al. Desmoplakin as a specific marker of National Cancer Institute and the American Lebanese Syrian
62. Irrthum, A. et al. Mutations in the transcription factor lymphatic vessels. Microvasc. Res. 61, 40–48 (2001). Associated Charities (ALSAC).
gene SOX18 underlie recessive and dominant forms of 77. Winnier, G. E., Hargett, L. & Hogan, B. L. The winged helix
hypotrichosis–lymphedema–telangiectasia. Am. J. Hum. transcription factor MFH1 is required for proliferation and Competing interests statement
Genet. 72, 1470–1478 (2003). patterning of paraxial mesoderm in the mouse embryo. The author declares that he has no competing financial interests.
63. Mandriota, S. J. et al. Vascular endothelial growth factor-C- Genes Dev. 11, 926–940 (1997).
mediated lymphangiogenesis promotes tumour metastasis. 78. Ayadi, A., Suelves, M., Dolle, P. & Wasylyk, B. Net, an Ets
EMBO J. 20, 672–682 (2001). ternary complex transcription factor, is expressed in sites of Online links
64. Beasley, N. J. P. et al. Intratumoral lymphangiogenesis and vasculogenesis, angiogenesis, and chondrogenesis during
lymph node metastasis in head and neck cancer. Cancer mouse development. Mech Dev. 102, 205–208 (2001). DATABASES
Res. 62, 1315–1320 (2002). 79. Ayadi, A. et al. Net-targeted mutant mice develop a vascular The following terms in this article are linked online to:
65. Padera, T. P. et al. Lymphatic metastasis in the absence of phenotype and upregulate egr-1. EMBO J. 20, 5139–5152 LocusLink: http://www.ncbi.nlm.nih.gov/LocusLink/
functional intratumor lymphatics. Science 296, 1883–1886 (2001). Ang1 | Ang2 | β-chemokine receptor D6 | desmoplakin | FOXC2 |
(2002). 80. Breiteneder-Geleff, S. et al. Podoplanin, novel 43-kd Lyve1 | Nrp2 | podoplanin | Prox1 | SLC | SOX18 | Tie2 | VEGFC |
References 64 and 65 provide antagonistic points of membrane protein of glomerular epithelial cells, is VEGFD | VEGFR2 | VEGFR3
view about the role of intratumoral lymphatics in downregulated in puromycin nephrosis. Am. J. Pathol.
tumour progression. 151, 1141–1152 (1997). FURTHER INFORMATION
66. Valtola, R. et al. VEGFR-3 and its ligand VEGF-C are 81. Wigle, J. T., Chowdhury, K., Gruss, P. & Oliver, G. Prox1 Lymphatic Research Foundation:
associated with angiogenesis in breast cancer. function is crucial for mouse lens-fibre elongation. Nature http://www.lymphatic research.org
Am. J. Pathol. 154, 1381–1390 (1999). Genet. 21, 318–322 (1999). Access to this interactive links box is free online.