@article {623,
	title = {Key molecules in lymphatic development, function, and identification},
	journal = {Annals of Anatomy - Anatomischer Anzeiger},
	volume = {219},
	year = {2018},
	month = {2018/09/01/},
	pages = {25 - 34},
	type = {review},
	chapter = {25},
	abstract = {While both blood and lymphatic vessels transport fluids and thus share many similarities, they also show functional and structural differences, which can be used to differentiate them. Specific visualization of lymphatic vessels has historically been and still is a pivot point in lymphatic research. Many of the proteins that are investigated by molecular biologists in lymphatic research have been defined as marker molecules, i.e. to visualize and distinguish lymphatic endothelial cells (LECs) from other cell types, most notably from blood vascular endothelial cells (BECs) and cells of the hematopoietic lineage.Among the factors that drive the developmental differentiation of lymphatic structures from venous endothelium, Prospero homeobox protein 1 (PROX1) is the master transcriptional regulator. PROX1 maintains lymphatic identity also in the adult organism and thus is a universal LEC marker. Vascular endothelial growth factor receptor-3 (VEGFR-3) is the major tyrosine kinase receptor that drives LEC proliferation and migration. The major activator for VEGFR-3 is vascular endothelial growth factor-C (VEGF-C). However, before VEGF-C can signal, it needs to be proteolytically activated by an extracellular protein complex comprised of Collagen and calcium binding EGF domains 1 (CCBE1) protein and the protease A disintegrin and metallopeptidase with thrombospondin type 1 motif 3 (ADAMTS3).This minireview attempts to give an overview of these and a few other central proteins that scientific inquiry has linked specifically to the lymphatic vasculature. It is limited in scope to a brief description of their main functions, properties and developmental roles.},
	isbn = {09409602},
	doi = {10.1016/j.aanat.2018.05.003},
	url = {http://linkinghub.elsevier.com/retrieve/pii/S0940960218300712}
}
@proceedings {511,
	title = {From molecular biology to a causal treatment of lymphatic system disorders},
	year = {2014},
	month = {10/2014},
	publisher = {Deutsche Gesellschaft f{\"u}r Lymphologie},
	address = {Halle (Saale), Germany},
	abstract = {<a href="http://www.dglymph.de/fileadmin/global/pdfs/LymphForsch_02-14.pdf">Original Conference Abstracts (in German)</a>},
	url = {https://jeltsch.org/abstract-Halle2014}
}
@article {44,
	title = {VEGF-C/VEGFR-3 Signaling Regulates Inflammatory Response in Development of Obliterative Airway Disease},
	journal = {Journal of Heart and Lung Transplantation},
	volume = {30},
	year = {2011},
	month = {2011//},
	pages = {S118 - S118},
	author = {Krebs, R and Tikkanen, JM and Ropponen, JO and Jeltsch, M and Jokinen, JJ and Yl{\"a}-Herttuala, S and Koskinen, PK and Nyk{\"a}nen, AI and Lemstr{\"o}m, KB}
}
@article {21,
	title = {Adenoviral expression of vascular endothelial growth factor-C induces lymphangiogenesis in the skin},
	journal = {Circ Res},
	volume = {88},
	year = {2001},
	month = {2001/Mar/},
	pages = {623 - 9},
	abstract = {The growth of blood and lymphatic vasculature is mediated in part by secreted polypeptides of the vascular endothelial growth factor (VEGF) family. The prototype VEGF binds VEGF receptor (VEGFR)-1 and VEGFR-2 and is angiogenic, whereas VEGF-C, which binds to VEGFR-2 and VEGFR-3, is either angiogenic or lymphangiogenic in different assays. We used an adenoviral gene transfer approach to compare the effects of these growth factors in adult mice. Recombinant adenoviruses encoding human VEGF-C or VEGF were injected subcutaneously into C57Bl6 mice or into the ears of nude mice. Immunohistochemical analysis showed that VEGF-C upregulated VEGFR-2 and VEGFR-3 expression and VEGF upregulated VEGFR-2 expression at 4 days after injection. After 2 weeks, histochemical and immunohistochemical analysis, including staining for the lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), the vascular endothelial marker platelet-endothelial cell adhesion molecule-1 (PECAM-1), and the proliferating cell nuclear antigen (PCNA) revealed that VEGF-C induced mainly lymphangiogenesis in contrast to VEGF, which induced only angiogenesis. These results have significant implications in the planning of gene therapy using these growth factors.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/11282897},
	author = {Enholm, B and Karpanen, T and Jeltsch, M and Kubo, H and Stenback, F and Prevo, R and Jackson, D G and Yla-Herttuala, S and Alitalo, K}
}
@article {22,
	title = {Signalling via vascular endothelial growth factor receptor-3 is sufficient for lymphangiogenesis in transgenic mice},
	journal = {EMBO J},
	volume = {20},
	year = {2001},
	month = {2001/Mar/},
	pages = {1223 - 31},
	abstract = {Vascular endothelial growth factor receptor-3 (VEGFR-3) has an essential role in the development of embryonic blood vessels; however, after midgestation its expression becomes restricted mainly to the developing lymphatic vessels. The VEGFR-3 ligand VEGF-C stimulates lymphangiogenesis in transgenic mice and in chick chorioallantoic membrane. As VEGF-C also binds VEGFR-2, which is expressed in lymphatic endothelia, it is not clear which receptors are responsible for the lymphangiogenic effects of VEGF-C. VEGF-D, which binds to the same receptors, has been reported to induce angiogenesis, but its lymphangiogenic potential is not known. In order to define the lymphangiogenic signalling pathway we have created transgenic mice overexpressing a VEGFR-3-specific mutant of VEGF-C (VEGF-C156S) or VEGF-D in epidermal keratinocytes under the keratin 14 promoter. Both transgenes induced the growth of lymphatic vessels in the skin, whereas the blood vessel architecture was not affected. Evidence was also obtained that these growth factors act in a paracrine manner in vivo. These results demonstrate that stimulation of the VEGFR-3 signal transduction pathway is sufficient to induce specifically lymphangiogenesis in vivo.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/11250889},
	author = {Veikkola, T and Jussila, L and Makinen, T and Karpanen, T and Jeltsch, M and Petrova, T V and Kubo, H and Thurston, G and McDonald, D M and Achen, M G and Stacker, S A and Alitalo, K}
}
@article {23,
	title = {Vascular endothelial growth factor-C-mediated lymphangiogenesis promotes tumour metastasis},
	journal = {EMBO J},
	volume = {20},
	year = {2001},
	month = {2001/Feb/},
	pages = {672 - 82},
	abstract = {Metastasis is a frequent and lethal complication of cancer. Vascular endothelial growth factor-C (VEGF-C) is a recently described lymphangiogenic factor. Increased expression of VEGF-C in primary tumours correlates with dissemination of tumour cells to regional lymph nodes. However, a direct role for VEGF-C in tumour lymphangiogenesis and subsequent metastasis has yet to be demonstrated. Here we report the establishment of transgenic mice in which VEGF-C expression, driven by the rat insulin promoter (Rip), is targeted to beta-cells of the endocrine pancreas. In contrast to wild-type mice, which lack peri-insular lymphatics, RipVEGF-C transgenics develop an extensive network of lymphatics around the islets of Langerhans. These mice were crossed with Rip1Tag2 mice, which develop pancreatic beta-cell tumours that are neither lymphangiogenic nor metastatic. Double-transgenic mice formed tumours surrounded by well developed lymphatics, which frequently contained tumour cell masses of beta-cell origin. These mice frequently developed pancreatic lymph node metastases. Our findings demonstrate that VEGF-C-induced lymphangiogenesis mediates tumour cell dissemination and the formation of lymph node metastases.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/11179212},
	author = {Mandriota, S J and Jussila, L and Jeltsch, M and Compagni, A and Baetens, D and Prevo, R and Banerji, S and Huarte, J and Montesano, R and Jackson, D G and Orci, L and Alitalo, K and Christofori, G and Pepper, M S}
}
@article {20,
	title = {Intravascular adenovirus-mediated VEGF-C gene transfer reduces neointima formation in balloon-denuded rabbit aorta},
	journal = {Circulation},
	volume = {102},
	year = {2000},
	month = {2000/Oct/},
	pages = {2262 - 8},
	abstract = {BACKGROUND: Gene transfer to the vessel wall may provide new possibilities for the treatment of vascular disorders, such as postangioplasty restenosis. In this study, we analyzed the effects of adenovirus-mediated vascular endothelial growth factor (VEGF)-C gene transfer on neointima formation after endothelial denudation in rabbits. For comparison, a second group was treated with VEGF-A adenovirus and a third group with lacZ adenovirus. Clinical-grade adenoviruses were used for the study. METHODS AND RESULTS: Aortas of cholesterol-fed New Zealand White rabbits were balloon-denuded, and gene transfer was performed 3 days later. Animals were euthanized 2 and 4 weeks after the gene transfer, and intima/media ratio (I/M), histology, and cell proliferation were analyzed. Two weeks after the gene transfer, I/M in the lacZ-transfected control group was 0. 57+/-0.04. VEGF-C gene transfer reduced I/M to 0.38+/-0.02 (P:<0.05 versus lacZ group). I/M in VEGF-A-treated animals was 0.49+/-0.17 (P:=NS). The tendency that both VEGF groups had smaller I/M persisted at the 4-week time point, when the lacZ group had an I/M of 0.73+/-0.16, the VEGF-C group 0.44+/-0.14, and the VEGF-A group 0. 63+/-0.21 (P:=NS). Expression of VEGF receptors 1, 2, and 3 was detected in the vessel wall by immunocytochemistry and in situ hybridization. As an additional control, the effect of adenovirus on cell proliferation was analyzed by performing gene transfer to intact aorta without endothelial denudation. No differences were seen in smooth muscle cell proliferation or I/M between lacZ adenovirus and 0.9\% saline-treated animals. CONCLUSIONS: Adenovirus-mediated VEGF-C gene transfer may be useful for the treatment of postangioplasty restenosis and vessel wall thickening after vascular manipulations.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/11056103},
	author = {Hiltunen, M O and Laitinen, M and Turunen, M P and Jeltsch, M and Hartikainen, J and Rissanen, T T and Laukkanen, J and Niemi, M and Kossila, M and H{\"a}kkinen, T P and Kivel{\"a}, A and Enholm, B and Mansukoski, H and Turunen, A M and Alitalo, K and Yl{\"a}-Herttuala, S}
}
@article {19,
	title = {Current biology of VEGF-B and VEGF-C},
	journal = {Curr Opin Biotechnol},
	volume = {10},
	year = {1999},
	month = {1999/Dec/},
	pages = {528 - 35},
	abstract = {Endothelial growth factors and their receptors may provide important therapeutic tools for the treatment of pathological conditions characterised by defective or aberrant angiogenesis. Vascular endothelial growth factor (VEGF) is pivotal for vasculogenesis and for angiogenesis in normal and pathological conditions. VEGF-B and VEGF-C provide this gene family with additional functions, for example, VEGF-C also regulates lymphangiogenesis.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/10600689},
	author = {Olofsson, B and Jeltsch, M and Eriksson, U and Alitalo, K}
}
@article {16,
	title = {Vascular endothelial growth factor D (VEGF-D) is a ligand for the tyrosine kinases VEGF receptor 2 (Flk1) and VEGF receptor 3 (Flt4)},
	journal = {Proc Natl Acad Sci U S A},
	volume = {95},
	year = {1998},
	month = {1998/Jan/},
	pages = {548 - 53},
	abstract = {We have identified a member of the VEGF family by computer-based homology searching and have designated it VEGF-D. VEGF-D is most closely related to VEGF-C by virtue of the presence of N- and C-terminal extensions that are not found in other VEGF family members. In adult human tissues, VEGF-D mRNA is most abundant in heart, lung, skeletal muscle, colon, and small intestine. Analyses of VEGF-D receptor specificity revealed that VEGF-D is a ligand for both VEGF receptors (VEGFRs) VEGFR-2 (Flk1) and VEGFR-3 (Flt4) and can activate these receptors. However. VEGF-D does not bind to VEGFR-1. Expression of a truncated derivative of VEGF-D demonstrated that the receptor-binding capacities reside in the portion of the molecule that is most closely related in primary structure to other VEGF family members and that corresponds to the mature form of VEGF-C. In addition, VEGF-D is a mitogen for endothelial cells. The structural and functional similarities between VEGF-D and VEGF-C define a subfamily of the VEGFs.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/9435229},
	author = {Achen, M G and Jeltsch, M and Kukk, E and M{\"a}kinen, T and Vitali, A and Wilks, A F and Alitalo, K and Stacker, S A}
}
@article {15,
	title = {Vascular endothelial growth factor (VEGF)-C synergizes with basic fibroblast growth factor and VEGF in the induction of angiogenesis in vitro and alters endothelial cell extracellular proteolytic activity},
	journal = {J Cell Physiol},
	volume = {177},
	year = {1998},
	month = {1998/Dec/},
	pages = {439 - 52},
	abstract = {Vascular endothelial growth factor-C (VEGF-C) is a recently characterized member of the VEGF family of angiogenic polypeptides. We demonstrate here that VEGF-C is angiogenic in vitro when added to bovine aortic or lymphatic endothelial (BAE and BLE) cells but has little or no effect on bovine microvascular endothelial (BME) cells. As reported previously for VEGF, VEGF-C and basic fibroblast growth factor (bFGF) induced a synergistic in vitro angiogenic response in all three cells lines. Unexpectedly, VEGF and VEGF-C also synergized in the in vitro angiogenic response when assessed on BAE cells. Characterization of VEGF receptor (VEGFR) expression revealed that BME, BAE, and BLE cell lines express VEGFR-1 and -2, whereas of the three cell lines assessed, only BAE cells express VEGFR-3. We also demonstrate that VEGF-C increases plasminogen activator (PA) activity in the three bovine endothelial cell lines and that this is accompanied by a concomitant increase in PA inhibitor-1. Addition of alpha2-antiplasmin to BME cells co-treated with bFGF and VEGF-C partially inhibited collagen gel invasion. These results demonstrate, first, that by acting in concert with bFGF or VEGF, VEGF-C has a potent synergistic effect on the induction of angiogenesis in vitro and, second, that like VEGF and bFGF, VEGF-C is capable of altering endothelial cell extracellular proteolytic activity. These observations also highlight the notion of context, i.e., that the activity of an angiogenesis-regulating cytokine depends on the presence and concentration of other cytokines in the pericellular environment of the responding endothelial cell.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/9808152},
	author = {Pepper, M S and Mandriota, S J and Jeltsch, M and Kumar, V and Alitalo, K}
}
@article {13,
	title = {Genomic organization of human and mouse genes for vascular endothelial growth factor C},
	journal = {J Biol Chem},
	volume = {272},
	year = {1997},
	month = {1997/Oct/},
	pages = {25176 - 83},
	abstract = {We report here the cloning and characterization of human and mouse genes for vascular endothelial growth factor C (VEGF-C), a newly isolated member of the vascular endothelial growth factor/platelet-derived growth factor (VEGF/PDGF) family. Both VEGF-C genes comprise over 40 kilobase pairs of genomic DNA and consist of seven exons, all containing coding sequences. The VEGF homology domain of VEGF-C is encoded by exons 3 and 4. Exons 5 and 7 encode cysteine-rich motifs of the type C6C10CRC, and exon 6 encodes additional C10CXCXC motifs typical of a silk protein. A putative alternatively spliced rare RNA form lacking exon 4 was identified in human fibrosarcoma cells, and a major transcription start site was located in the human VEGF-C gene 523 base pairs upstream of the translation initiation codon. The upstream promoter sequences contain conserved putative binding sites for Sp-1, AP-2, and NF-kappaB transcription factors but no TATA box, and they show promoter activity when transfected into cells. The VEGF-C gene structure is thus assembled from exons encoding propeptides and distinct cysteine-rich domains in addition to the VEGF homology domain, and it shows both similarities and distinct differences in comparison with other members of the VEGF/PDGF gene family.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/9312130},
	author = {Chilov, D and Kukk, E and Taira, S and Jeltsch, M and Kaukonen, J and Palotie, A and Joukov, V and Alitalo, K}
}
@article {10,
	title = {Hyperplasia of lymphatic vessels in VEGF-C transgenic mice},
	journal = {Science (80- )},
	volume = {276},
	year = {1997},
	month = {1997/May/},
	pages = {1423 - 5},
	abstract = {No growth factors specific for the lymphatic vascular system have yet been described. Vascular endothelial growth factor (VEGF) regulates vascular permeability and angiogenesis, but does not promote lymphangiogenesis. Overexpression of VEGF-C, a ligand of the VEGF receptors VEGFR-3 and VEGFR-2, in the skin of transgenic mice resulted in lymphatic, but not vascular, endothelial proliferation and vessel enlargement. Thus, VEGF-C induces selective hyperplasia of the lymphatic vasculature, which is involved in the draining of interstitial fluid and in immune function, inflammation, and tumor metastasis. VEGF-C may play a role in disorders involving the lymphatic system and may be of potential use in therapeutic lymphangiogenesis.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/9162011},
	author = {Jeltsch, M and Kaipainen, A and Joukov, V and Meng, X and Lakso, M and Rauvala, H and Swartz, M and Fukumura, D and Jain, R K and Alitalo, K}
}
@article {14,
	title = {Proteolytic processing regulates receptor specificity and activity of VEGF-C},
	journal = {EMBO J},
	volume = {16},
	year = {1997},
	month = {1997/Jul/},
	pages = {3898 - 911},
	abstract = {The recently identified vascular endothelial growth factor C (VEGF-C) belongs to the platelet-derived growth factor (PDGF)/VEGF family of growth factors and is a ligand for the endothelial-specific receptor tyrosine kinases VEGFR-3 and VEGFR-2. The VEGF homology domain spans only about one-third of the cysteine-rich VEGF-C precursor. Here we have analysed the role of post-translational processing in VEGF-C secretion and function, as well as the structure of the mature VEGF-C. The stepwise proteolytic processing of VEGF-C generated several VEGF-C forms with increased activity towards VEGFR-3, but only the fully processed VEGF-C could activate VEGFR-2. Recombinant {\textquoteright}mature{\textquoteright} VEGF-C made in yeast bound VEGFR-3 (K[D] = 135 pM) and VEGFR-2 (K[D] = 410 pM) and activated these receptors. Like VEGF, mature VEGF-C increased vascular permeability, as well as the migration and proliferation of endothelial cells. Unlike other members of the PDGF/VEGF family, mature VEGF-C formed mostly non-covalent homodimers. These data implicate proteolytic processing as a regulator of VEGF-C activity, and reveal novel structure-function relationships in the PDGF/VEGF family.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/9233800},
	author = {Joukov, V and Sorsa, T and Kumar, V and Jeltsch, M and Claesson-Welsh, L and Cao, Y and Saksela, O and Kalkkinen, N and Alitalo, K}
}
@article {12,
	title = {Vascular endothelial growth factors VEGF-B and VEGF-C},
	journal = {J Cell Physiol},
	volume = {173},
	year = {1997},
	month = {1997/Nov/},
	pages = {211 - 5},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/9365524},
	author = {Joukov, V and Kaipainen, A and Jeltsch, M and Pajusola, K and Olofsson, B and Kumar, V and Eriksson, U and Alitalo, K}
}
@article {9,
	title = {VEGF-C receptor binding and pattern of expression with VEGFR-3 suggests a role in lymphatic vascular development},
	journal = {Development},
	volume = {122},
	year = {1996},
	month = {1996/Dec/},
	pages = {3829 - 37},
	abstract = {The vascular endothelial growth factor family has recently been expanded by the isolation of two new VEGF-related factors, VEGF-B and VEGF-C. The physiological functions of these factors are largely unknown. Here we report the cloning and characterization of mouse VEGF-C, which is produced as a disulfide-linked dimer of 415 amino acid residue polypeptides, sharing an 85\% identity with the human VEGF-C amino acid sequence. The recombinant mouse VEGF-C protein was secreted from transfected cells as VEGFR-3 (Flt4) binding polypeptides of 30-32x10(3) Mr and 22-23x10(3) Mr which preferentially stimulated the autophosphorylation of VEGFR-3 in comparison with VEGFR-2 (KDR). In in situ hybridization, mouse VEGF-C mRNA expression was detected in mesenchymal cells of postimplantation mouse embryos, particularly in the regions where the lymphatic vessels undergo sprouting from embryonic veins, such as the perimetanephric, axillary and jugular regions. In addition, the developing mesenterium, which is rich in lymphatic vessels, showed strong VEGF-C expression. VEGF-C was also highly expressed in adult mouse lung, heart and kidney, where VEGFR-3 was also prominent. The pattern of expression of VEGF-C in relation to its major receptor VEGFR-3 during the sprouting of the lymphatic endothelium in embryos suggests a paracrine mode of action and that one of the functions of VEGF-C may be in the regulation of angiogenesis of the lymphatic vasculature.},
	url = {http://view.ncbi.nlm.nih.gov/pubmed/9012504},
	author = {Kukk, E and Lymboussaki, A and Taira, S and Kaipainen, A and Jeltsch, M and Joukov, V and Alitalo, K}
}
