00566nas a2200157   4500008004100000022001400041245010100055210006900156260001600225100002100241700002300262700001900285700002500304700002100329856005800350       2024                        eng d  a1464-793100aThe relationship between the secondary vascular system and the lymphatic vascular system in fish0 arelationship between the secondary vascular system and the lymph  cApr-06-20261 aPanara, Virginia1 aVaraliová, Zuzana1 aWilting, Jörg1 aKoltowska, Katarzyna1 aJeltsch, Michael  uhttps://onlinelibrary.wiley.com/doi/10.1111/brv.1311400926nas a2200253   4500008004100000022001400041245019700055210006900252260001600321300001800337490000700355100002600362700002500388700002400413700002300437700002500460700002300485700002100508700002400529700002100553700002200574700002400596856005200620       2024                        eng d  a1944-824400aStudy of the Synergistic Immunomodulatory and Antifibrotic Effects of Dual-Loaded Budesonide and Serpine1 siRNA Lipid–Polymer Nanoparticles Targeting Macrophage Dysregulation in Tendinopathy0 aStudy of the Synergistic Immunomodulatory and Antifibrotic Effec  cMay-04-2025  a18643 - 186570 v161 aLópez-Cerdá, Sandra1 aMolinaro, Giuseppina1 aParejaTello, Rubén1 aCorreia, Alexandra1 aKünig, Sarojinidevi1 aSteinberger, Peter1 aJeltsch, Michael1 aHirvonen, Jouni, T.1 aBarreto, Goncalo1 aStöckl, Johannes1 aSantos, Hélder, A.  uhttps://pubs.acs.org/doi/10.1021/acsami.4c0236300523nas a2200157   4500008004100000022001400041245008300055210006900138260001600207300001400223490000700237100002200244700002100266700002100287856005700308       2023                        eng d  a0969-697000aExpansion and collapse of VEGF diversity in major clades of the animal kingdom0 aExpansion and collapse of VEGF diversity in major clades of the   cJan-08-2023  a437 - 4610 v261 aRauniyar, Khushbu1 aBokharaie, Honey1 aJeltsch, Michael  uhttps://link.springer.com/10.1007/s10456-023-09874-900408nas a2200097   4500008004100000245008000041210006900121260005900190100002200249856003900271       2023                        eng d00aVEGF-C: The evolutionary origin, activation, and potential as a drug target0 aVEGFC The evolutionary origin activation and potential as a drug  aHelsinki. FinlandbUniversity of Helsinkic2023/06/09/1 aRauniyar, Khushbu  uhttp://hdl.handle.net/10138/35792300451nas a2200157   4500008004100000245003400041210003200075260001600107490000700123100002200130700002500152700001900177700002100196700002100217856005500238       2022                        eng d00aBioactive VEGF-C from E. coli0 aBioactive VEGFC from E coli  cJan-12-20220 v121 aRauniyar, Khushbu1 aAkhondzadeh, Soheila1 aGąciarz, Anna1 aKünnapuu, Jaana1 aJeltsch, Michael  uhttps://www.nature.com/articles/s41598-022-22960-000402nas a2200133   4500008004100000245006400041210006200105260001600167300001400183490000600197100002100203700001800224856002600242       2022                        eng d00aLymphatic-to-blood vessel transdifferentiation in zebrafish0 aLymphatictoblood vessel transdifferentiation in zebrafish  cJan-06-2022  a539 - 5410 v11 aJeltsch, Michael1 aAlitalo, Kari  uhttps://rdcu.be/cOjJ000294nas a2200097   4500008004100000245003500041210003400076260002200110100002100132856004300153       2021                        eng d00aDrug-induced lymphangiogenesis0 aDruginduced lymphangiogenesis  bJuzoc2021/09/04/1 aJeltsch, Michael  uhttps://doi.org/10.5281/zenodo.603430700459nas a2200145   4500008004100000245006500041210006400106260001600170300001000186490000700196100002100203700002100224700002100245856004700266       2021                        eng d00aKLK3 in the Regulation of Angiogenesis—Tumorigenic or Not?0 aKLK3 in the Regulation of Angiogenesis—Tumorigenic or Not  cJan-12-2021  a135450 v221 aKoistinen, Hannu1 aKünnapuu, Jaana1 aJeltsch, Michael  uhttps://www.mdpi.com/1422-0067/22/24/1354500414nas a2200133   4500008004100000022001400041245005100055210005100106260001600157490000700173100002100180700002100201856005800222       2021                        eng d  a1945-087700aOutside in and brakes off for lymphatic growth0 aOutside in and brakes off for lymphatic growth  cOct-08-20210 v141 aKünnapuu, Jaana1 aJeltsch, Michael  uhttps://www.science.org/doi/10.1126/scisignal.abj505800539nas a2200145   4500008004100000245014500041210006900186260001600255300000800271490000700279100002100286700002100307700002100328856004400349       2021                        eng d00aProteolytic Cleavages in the VEGF Family: Generating Diversity among Angiogenic VEGFs, Essential for the Activation of Lymphangiogenic VEGFs0 aProteolytic Cleavages in the VEGF Family Generating Diversity am  cJan-02-2021  a1670 v101 aKünnapuu, Jaana1 aBokharaie, Honey1 aJeltsch, Michael  uhttps://www.mdpi.com/2079-7737/10/2/16700566nas a2200205   4500008004100000022001400041245003800055210003500093260001600128300001400144490000800158100002100166700002000187700001800207700001900225700001700244700002100261700002200282856005600304       2020                        eng d  a2242-328100aLymphatics and the eye. [Finnish]0 aLymphatics and the eye Finnish  c2020/02/10/  a1777-17880 v1361 aGucciardo, Erika1 aLehti, Timo, A.1 aKorhonen, Ani1 aSalvén, Petri1 aLehti, Kaisa1 aJeltsch, Michael1 aLoukovaara, Sirpa  uhttps://www.duodecimlehti.fi/lehti/2020/16/duo1573903948nas a2200109   4500008004100000245009000041210006900131260005900200520351100259100001803770856005003788       2020                        eng d00aMechanism of VEGF-C Activation and Effect on Lymphatic Vessel Growth and Regeneration0 aMechanism of VEGFC Activation and Effect on Lymphatic Vessel Gro  aHelsinki, FinlandbUniversity of Helsinkic2020/05/29/3 aLymphangiogenesis, the growth of the lymphatic vasculature, is a crucial process during embryonic development, and - if compromised by genetic damage - can lead to hereditary lymphedema. Although the molecular mechanisms that regulate the growth, development, and maintenance of the lymphatic vasculature have been researched with increasing intensity over the last 25 years, the therapeutic regeneration of lymphatic vessels is still a work in progress in the treatment of conditions such as lymphedema. Vascular endothelial growth factor-C (VEGF-C) is the primary growth factor responsible for the growth and development of the lymphatic vasculature. VEGF-C is activated by a complex process, which is indispensable for its ability to induce lymphangiogenesis via its primary receptor, VEGFR-3. The understanding of this process is a key factor for the development of VEGF-C as a drug target. The goal in my studies has been to increase our insights into VEGF-C activation at the molecular level, to identify its regulatory factors, and to establish its role in the lymphangiogenic process.Absence of the collagen- and calcium-binding EGF domains 1 (CCBE1) protein interrupts the lymphangiogenic process at about the same developmental stage when VEGF-C is first required. We utilized cell-based assays and adeno-associated viral-based gene transduction to investigate the role of CCBE1 on VEGF-C activation. In study I, we identified A disintegrin and metalloprotease with thrombospondin motifs-3 (ADAMTS3) as a protease that cleaves and activates VEGF-C, resulting in the major mature form of VEGF-C. We showed that CCBE1 acts as a cofactor in this process by enhancing the ability of ADAMTS3 to activate VEGF-C. Correspondingly, CCBE1 augmented the lymphangiogenic potential of VEGF-C in vivo.  The presence of N- and C- terminal domains and their proteolytic cleavage characterize both CCBE1 and VEGF-C. In study II, we investigated the role of these domains for VEGF-C activation and the lymphangiogenic process. Our study demonstrated a requirement for the C-terminal domain of VEGF-C for the robust activation of VEGF-C both in vitro and in vivo. Moreover, we identified that the N- and C-terminal domains of CCBE1 have independent roles in the process of VEGF-C activation. The C-terminal domain accelerates the proteolytic cleavage, while the N-terminal domain aids in the assembly of the VEGF-C/ADAMTS3/CCBE1 cleavage complex by mobilizing VEGF-C to the endothelial cell surface. In study III, we searched for additional proteases that can cleave VEGF-C. We identified kallikrein-related peptidase 3 (KLK3) in seminal plasma and cathepsin D in saliva as proteases that cleave and activate VEGF-C. In human seminal plasma, we found substantial amounts of VEGF-C, which became activated concurrently with the semen liquefaction process. The newly identified VEGF-C cleavage sites are conserved in VEGF-D and we found that KLK3 and cathepsin D were able to activate VEGF-D as well. We also found that cleaved forms of VEGF-C and VEGF-D differ in their abilities to activate VEGFR-2 and VEGFR-3. When their N-termini were progressively shortened, the ability of VEGF-D to bind to and activate VEGFR-3 was decreased, while VEGF-C lost preferentially its ability to bind to and activate VEGFR-2. These findings contribute to the existing knowledge on the mechanisms of VEGF-C activation and the functional consequences thereof, and provide new opportunities to target VEGF-C for therapeutic purposes.1 aJha, Sawan, K  uhttps://helda.helsinki.fi/handle/10138/31471401961nas a2200337   4500008004100000020001400041245005600055210005300111260001600164300001100180490000600191520106000197653001901257653001601276653003401292653001301326653002201339653001001361653001101371653001101382100001801393700002201411700002001433700002001453700002101473700002101494700002401515700001801539700002101557856004501578       2019                        eng d  a2050-084X00aKLK3/PSA and cathepsin D activate VEGF-C and VEGF-D0 aKLK3PSA and cathepsin D activate VEGFC and VEGFD  c2019/05/17/  ae444780 v83 aVascular endothelial growth factor-C (VEGF-C) acts primarily on endothelial cells, but also on non-vascular targets, e.g. in the CNS and immune system. Here we describe a novel, unique VEGF-C form in the human reproductive system produced via cleavage by kallikrein-related peptidase 3 (KLK3), aka prostate-specific antigen (PSA). KLK3 activated VEGF-C specifically and efficiently through cleavage at a novel N-terminal site. We detected VEGF-C in seminal plasma, and sperm liquefaction occurred concurrently with VEGF-C activation, which was enhanced by collagen and calcium binding EGF domains 1 (CCBE1). After plasmin and ADAMTS3, KLK3 is the third protease shown to activate VEGF-C. Since differently activated VEGF-Cs are characterized by successively shorter N-terminal helices, we created an even shorter hypothetical form, which showed preferential binding to VEGFR-3. Using mass spectrometric analysis of the isolated VEGF-C-cleaving activity from human saliva, we identified cathepsin D as a protease that can activate VEGF-C as well as VEGF-D.10acancer biology10aCathepsin D10akallikrein-related peptidases10aKLK3/PSA10aLymphangiogenesis10amouse10aVEGF-C10aVEGF-D1 aJha, Sawan, K1 aRauniyar, Khushbu1 aChronowska, Ewa1 aMattonet, Kenny1 aMaina, Eunice, W1 aKoistinen, Hannu1 aStenman, Ulf-Håkan1 aAlitalo, Kari1 aJeltsch, Michael  uhttps://elifesciences.org/articles/4447802718nas a2200205   4500008004100000245007100041210006600112260001600178300001200194490000700206520212700213653002202340653001602362653001602378653001102394100002002405700002302425700002102448856004302469       2019                        eng d00aThe Proteolytic Activation of Vascular Endothelial Growth Factor-C0 aProteolytic Activation of Vascular Endothelial Growth FactorC  c2019/12/18/  a88 - 980 v233 aThe enzymatic cleavage of the protein backbone (proteolysis) is integral to many biological processes, e.g. for the break­down of proteins in the digestive system. Specific proteolytic cleavages are also used to turn on or off the activity of proteins. For example, the lymphangiogenic vascular endothelial growth factor-­C (VEGF-­C) is synthesized as a precursor molecule that must be converted to a mature form by the enzymatic removal of C-­ and N-­terminal propeptides before it can bind and activate its receptors. The constitutive C-­terminal cleavage is mediated by proprotein convertases such as furin. The subsequent ac­tivating cleavage can be mediated by at least four different proteases: by plasmin, ADAMTS3, prostate­-specific antigen (PSA) and cathepsin D. Processing by different proteases results in distinct forms of "ma­ture" VEGF­-C, that differ in their affinity and their receptor activation potential. This processing is tightly regulated by the CCBE1 protein. CCBE1 regulates the acti­vating cleavage of VEGF­C by ADAMTS3 and PSA, but not by plasmin. During embryonic development of the lymphatic system, VEGF-­C is activated primarily by the ADAMTS3 protease. In contrast, it is believed that plasmin is responsible for wound healing lymphangiogenesis and PSA for tumor-­associated pathological lym­phangiogenesis. Cathepsin D has also been implicated in tumor lymphangiogenesis. In addition, cathepsin D in saliva might activate latent VEGF­C upon wound licking, thereby accelerating wound healing. The molecular details of proteolytic activation of VEGF­-C are only recently extensively explored, and we likely do not know yet all activating proteases. It appears that the activity of VEGF­-C is regulated for different specific functions by different proteinases. Although VEGF­-C clearly plays a pivotal role for tumor progression and metastasis in experimental animal studies, the rele­vance of most correlative studies on the role of VEGF­-C in human cancers is quite limited until now, also due to the lack of methods to differentiate between inactive and active forms.10aLymphangiogenesis10aproteinases10aproteolysis10aVEGF-C1 aLackner, Marcel1 aSchmotz, Constanze1 aJeltsch, Michael  uhttps://doi.org/10.5281/zenodo.362926300395nas a2200097   4500008004100000245007600041210006900117260004400186100002100230856004600251       2017                        eng d00aWhat you should know about VEGF-C when working with lymphatics [German]0 aWhat you should know about VEGFC when working with lymphatics Ge  aBad Soden (Frankfurt), Germanyc06/20171 aJeltsch, Michael  uhttps://jeltsch.org/Abstrakt-BadSoden201701999nas a2200217   4500008004100000245016300041210006900204260001200273300001300285490000700298520124700305653001901552653002201571653001501593653002701608653001101635653001101646100001801657700002101675856008501696       2013                        eng d00aDie lymphangiogenic growth factors VEGF-C and VEGF-D. Part 2: The role of VEGF-C and VEGF-D in diseases of the lymphatic system. [bilingual: English, German].0 aDie lymphangiogenic growth factors VEGFC and VEGFD Part 2 The ro  c11/2013  a96 - 1040 v173 aVEGF-C and VEGF-D are the two central signaling molecules that stimulate the develop- ment and growth of the lymphatic system. Both belong to the vascular endothelial growth factor (VEGF) protein family, which plays important roles in the growth of blood vessels (angiogenesis) and lymphatic vessels (lymphangiogenesis). In mammals, the VEGF family comprises five members: VEGF-A, PlGF, VEGF-B, VEGF-C and VEGF-D. The family was named after VEGF-A, the first member to be discovered. VEGF-C and VEGF-D form a subgroup within this family in terms of function and structure. Their distinctive biosynthesis differentiates them from the other VEGFs: they are produced as inactive precursors and need to be activated by proteolytic removal of their long N- and C-terminal propeptides. Unlike the other VEGFs, VEGF-C and VEGF-D are direct stimulators of lymphatic vessel growth. They exert their lymphangiogenic function via VEGF receptor 3, which is expressed in the adult organism almost exclusively on lymphatic endothelial cells. In this review, we provide an overview of the VEGF protein family and their receptors. We focus on the lymphangiogenic VEGF-C and VEGF-D, discussing their biosynthesis and their role in embryonic lymphangiogenesis.10agrowth factors10aLymphangiogenesis10alymphedema10alymphogenic metastasis10aVEGF-C10aVEGF-D1 aKrebs, Rainer1 aJeltsch, Michael  uhttp://jeltsch.org/sites/jeltsch.org/files/JeltschMichael_Lymphforsch2013_96.pdf01909nas a2200193   4500008004100000245014000041210006900181260001200250300001200262490000700274520124700281653001901528653002201547653001101569653001101580100001801591700002101609856008501630       2013                        eng d00aThe lymphangiogenic growth factors VEGF-C and VEGF-D. Part 1: Basic principles and embryonic development. [bilingual: English, German].0 alymphangiogenic growth factors VEGFC and VEGFD Part 1 Basic prin  c05/2013  a30 - 370 v173 aVEGF-C and VEGF-D are the two central signaling molecules that stimulate the develop- ment and growth of the lymphatic system. Both belong to the vascular endothelial growth factor (VEGF) protein family, which plays important roles in the growth of blood vessels (angiogenesis) and lymphatic vessels (lymphangiogenesis). In mammals, the VEGF family comprises five members: VEGF-A, PlGF, VEGF-B, VEGF-C and VEGF-D. The family was named after VEGF-A, the first member to be discovered. VEGF-C and VEGF-D form a subgroup within this family in terms of function and structure. Their distinctive biosynthesis differentiates them from the other VEGFs: they are produced as inactive precursors and need to be activated by proteolytic removal of their long N- and C-terminal propeptides. Unlike the other VEGFs, VEGF-C and VEGF-D are direct stimulators of lymphatic vessel growth. They exert their lymphangiogenic function via VEGF receptor 3, which is expressed in the adult organism almost exclusively on lymphatic endothelial cells. In this review, we provide an overview of the VEGF protein family and their receptors. We focus on the lymphangiogenic VEGF-C and VEGF-D, discussing their biosynthesis and their role in embryonic lymphangiogenesis.10agrowth factors10aLymphangiogenesis10aVEGF-C10aVEGF-D1 aKrebs, Rainer1 aJeltsch, Michael  uhttp://jeltsch.org/sites/jeltsch.org/files/JeltschMichael_Lymphforsch2013_30.pdf02103nas a2200241   4500008004100000245010100041210006900142260001400211300001400225490000800239520135400247100002601601700002101627700002101648700002001669700001501689700002101704700001901725700002601744700002401770700001801794856004901812       2011                        eng d00aStructural determinants of vascular endothelial growth factor-D receptor binding and specificity0 aStructural determinants of vascular endothelial growth factorD r  c2011/Feb/  a1507 - 150 v1173 aVascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. Two different proteolytic cleavage sites have been reported in the VEGF-D N-terminus. We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. Consistent with this, the affinity and the thermodynamic parameters for VEGFR-2 binding are very similar. In comparison with VEGF-C structures, however, the VEGF-D N-terminal helix was extended by 2 more turns because of a better resolution. Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. Thus, we define here a VEGFR-2-specific form of VEGF-D that is angiogenic but not lymphangiogenic. These results provide important new insights into VEGF-D structure and function.1 aLeppänen, Veli-Matti1 aJeltsch, Michael1 aAnisimov, Andrey1 aTvorogov, Denis1 aAho, Kukka1 aKalkkinen, Nisse1 aToivanen, Pyry1 aYlä-Herttuala, Seppo1 aBallmer-Hofer, Kurt1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/2114808500667nas a2200217   4500008004100000245010700041210006900148260001100217300001600228490000700244100001300251700001700264700001700281700001500298700001600313700002200329700001700351700001700368700001800385856004600403       2011                        eng d00aVEGF-C/VEGFR-3 Signaling Regulates Inflammatory Response in Development of Obliterative Airway Disease0 aVEGFCVEGFR3 Signaling Regulates Inflammatory Response in Develop  c2011//  aS118 - S1180 v301 aKrebs, R1 aTikkanen, JM1 aRopponen, JO1 aJeltsch, M1 aJokinen, JJ1 aYlä-Herttuala, S1 aKoskinen, PK1 aNykänen, AI1 aLemström, KB  uhttps://jeltsch.org/Krebs2011?language=hu01603nas a2200265   4500008004100000245011900041210006900160260001400229300001300243490000700256520077800263100002101041700002001062700002001082700002101102700002201123700002101145700002301166700002301189700001601212700002101228700002101249700001801270856004901288       2010                        eng d00aClaudin-like protein 24 interacts with the VEGFR-2 and VEGFR-3 pathways and regulates lymphatic vessel development0 aClaudinlike protein 24 interacts with the VEGFR2 and VEGFR3 path  c2010/May/  a875 - 800 v243 aThe Claudin-like protein of 24 kDa (CLP24) is a hypoxia-regulated transmembrane protein of unknown function. We show here that clp24 knockdown in Danio rerio and Xenopus laevis results in defective lymphatic development. Targeted disruption of Clp24 in mice led to enlarged lymphatic vessels having an abnormal smooth muscle cell coating. We also show that the Clp24(-/-) phenotype was further aggravated in the Vegfr2(+/LacZ) or Vegfr3(+/LacZ) backgrounds and that CLP24 interacts with vascular endothelial growth factor receptor-2 (VEGFR-2) and VEGFR-3 and attenuates the transcription factor CREB phosphorylation via these receptors. Our results indicate that CLP24 is a novel regulator of VEGFR-2 and VEGFR-3 signaling pathways and of normal lymphatic vessel structure.1 aSaharinen, Pipsa1 aHelotera, Hanna1 aMiettinen, Juho1 aNorrmen, Camilla1 aD'Amico, Gabriela1 aJeltsch, Michael1 aLangenberg, Tobias1 aVandevelde, Wouter1 aNy, Annelii1 aDewerchin, Mieke1 aCarmeliet, Peter1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/2043942801838nas a2200289   4500008004100000245014100041210006900182260001400251300001300265490000700278520091500285100002001200700002101220700001501241700002601256700002001282700002701302700002501329700002101354700002201375700002101397700002101418700002101439700002101460700001801481856004901499       2010                        eng d00aEffective suppression of vascular network formation by combination of antibodies blocking VEGFR ligand binding and receptor dimerization0 aEffective suppression of vascular network formation by combinati  c2010/Dec/  a630 - 400 v183 aAntibodies that block vascular endothelial growth factor (VEGF) have become an integral part of antiangiogenic tumor therapy, and antibodies targeting other VEGFs and receptors (VEGFRs) are in clinical trials. Typically receptor-blocking antibodies are targeted to the VEGFR ligand-binding site. Here we describe a monoclonal antibody that inhibits VEGFR-3 homodimer and VEGFR-3/VEGFR-2 heterodimer formation, signal transduction, as well as ligand-induced migration and sprouting of microvascular endothelial cells. Importantly, we show that combined use of antibodies blocking ligand binding and receptor dimerization improves VEGFR inhibition and results in stronger inhibition of endothelial sprouting and vascular network formation in vivo. These results suggest that receptor dimerization inhibitors could be used to enhance antiangiogenic activity of antibodies blocking ligand binding in tumor therapy.1 aTvorogov, Denis1 aAnisimov, Andrey1 aZheng, Wei1 aLeppänen, Veli-Matti1 aTammela, Tuomas1 aLaurinavicius, Simonas1 aHolnthoner, Wolfgang1 aHeloterä, Hanna1 aHolopainen, Tanja1 aJeltsch, Michael1 aKalkkinen, Nisse1 aLankinen, Hilkka1 aOjala, Päivi, M1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/2113004302522nas a2200241   4500008004100000245008800041210006900129260001200198300001400210490000800224520178600232100002602018700002102044700002102065700002102086700002102107700002002128700002102148700002002169700002402189700001802213856004902231       2010                        eng d00aStructural determinants of growth factor binding and specificity by VEGF receptor 20 aStructural determinants of growth factor binding and specificity  c02/2010  a2425 - 300 v1073 aVascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of three receptor tyrosine kinases, VEGFR-1, -2, and -3. The extracellular domain of VEGF receptors consists of seven immunoglobulin homology domains, which, upon ligand binding, promote receptor dimerization. Dimerization initiates transmembrane signaling, which activates the intracellular tyrosine kinase domain of the receptor. VEGF-C stimulates lymphangiogenesis and contributes to pathological angiogenesis via VEGFR-3. However, proteolytically processed VEGF-C also stimulates VEGFR-2, the predominant transducer of signals required for physiological and pathological angiogenesis. Here we present the crystal structure of VEGF-C bound to the VEGFR-2 high-affinity-binding site, which consists of immunoglobulin homology domains D2 and D3. This structure reveals a symmetrical 22 complex, in which left-handed twisted receptor domains wrap around the 2-fold axis of VEGF-C. In the VEGFs, receptor specificity is determined by an N-terminal alpha helix and three peptide loops. Our structure shows that two of these loops in VEGF-C bind to VEGFR-2 subdomains D2 and D3, while one interacts primarily with D3. Additionally, the N-terminal helix of VEGF-C interacts with D2, and the groove separating the two VEGF-C monomers binds to the D2/D3 linker. VEGF-C, unlike VEGF-A, does not bind VEGFR-1. We therefore created VEGFR-1/VEGFR-2 chimeric proteins to further study receptor specificity. This biochemical analysis, together with our structural data, defined VEGFR-2 residues critical for the binding of VEGF-A and VEGF-C. Our results provide significant insights into the structural features that determine the high affinity and specificity of VEGF/VEGFR interactions.1 aLeppänen, Veli-Matti1 aProta, Andrea, E1 aJeltsch, Michael1 aAnisimov, Andrey1 aKalkkinen, Nisse1 aStrandin, Tomas1 aLankinen, Hilkka1 aGoldman, Adrian1 aBallmer-Hofer, Kurt1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/2014511602377nas a2200265   4500008004100000245014000041210006900181260001100250300001100261490000600272520152800278100001901806700002101825700002401846700002001870700002301890700002501913700002001938700002101958700001801979700001801997700002502015700002202040856004902062       2010                        eng d00aSuppressive effects of vascular endothelial growth factor-B on tumor growth in a mouse model of pancreatic neuroendocrine tumorigenesis0 aSuppressive effects of vascular endothelial growth factorB on tu  c2010//  ae141090 v53 aBACKGROUND: The family of vascular endothelial growth factors (VEGF) contains key regulators of blood and lymph vessel development, including VEGF-A, -B, -C, -D, and placental growth factor. The role of VEGF-B during physiological or pathological angiogenesis has not yet been conclusively delineated. Herein, we investigate the function of VEGF-B by the generation of mouse models of cancer with transgenic expression of VEGF-B or homozygous deletion of Vegfb. METHODOLOGY/PRINCIPAL FINDINGS: Ectopic expression of VEGF-B in the insulin-producing β-cells of the pancreas did not alter the abundance or architecture of the islets of Langerhans. The vasculature from transgenic mice exhibited a dilated morphology, but was of similar density as that of wildtype mice. Unexpectedly, we found that transgenic expression of VEGF-B in the RIP1-Tag2 mouse model of pancreatic neuroendocrine tumorigenesis retarded tumor growth. Conversely, RIP1-Tag2 mice deficient for Vegfb presented with larger tumors. No differences in vascular density, perfusion or immune cell infiltration upon altered Vegfb gene dosage were noted. However, VEGF-B acted to increase blood vessel diameter both in normal pancreatic islets and in RIP1-Tag2 tumors. CONCLUSIONS/SIGNIFICANCE: Taken together, our results illustrate the differences in biological function between members of the VEGF family, and highlight the necessity of in-depth functional studies of VEGF-B to fully understand the effects of VEGFR-1 inhibitors currently used in the clinic.1 aAlbrecht, Imke1 aKopfstein, Lucie1 aStrittmatter, Karin1 aSchomber, Tibor1 aFalkevall, Annelie1 aHagberg, Carolina, E1 aLorentz, Pascal1 aJeltsch, Michael1 aAlitalo, Kari1 aEriksson, Ulf1 aChristofori, Gerhard1 aPietras, Kristian  uhttp://view.ncbi.nlm.nih.gov/pubmed/2112484102729nas a2200349   4500008004100000245015500041210006900196260001400265300001400279490000800293520163900301100001501940700002001955700002201975700002101997700002002018700002002038700002102058700001902079700002102098700002102119700002102140700002302161700002202184700002602206700001902232700001902251700002302270700001902293700001802312856004902330       2010                        eng d00aVascular endothelial growth factor-B acts as a coronary growth factor in transgenic rats without inducing angiogenesis, vascular leak, or inflammation0 aVascular endothelial growth factorB acts as a coronary growth fa  c2010/Oct/  a1725 - 330 v1223 aBACKGROUND: Vascular endothelial growth factor-B (VEGF-B) binds to VEGF receptor-1 and neuropilin-1 and is abundantly expressed in the heart, skeletal muscle, and brown fat. The biological function of VEGF-B is incompletely understood. METHODS AND RESULTS: Unlike placenta growth factor, which binds to the same receptors, adeno-associated viral delivery of VEGF-B to mouse skeletal or heart muscle induced very little angiogenesis, vascular permeability, or inflammation. As previously reported for the VEGF-B(167) isoform, transgenic mice and rats expressing both isoforms of VEGF-B in the myocardium developed cardiac hypertrophy yet maintained systolic function. Deletion of the VEGF receptor-1 tyrosine kinase domain or the arterial endothelial Bmx tyrosine kinase inhibited hypertrophy, whereas loss of VEGF-B interaction with neuropilin-1 had no effect. Surprisingly, in rats, the heart-specific VEGF-B transgene induced impressive growth of the epicardial coronary vessels and their branches, with large arteries also seen deep inside the subendocardial myocardium. However, VEGF-B, unlike other VEGF family members, did not induce significant capillary angiogenesis, increased permeability, or inflammatory cell recruitment. CONCLUSIONS: VEGF-B appears to be a coronary growth factor in rats but not in mice. The signals for the VEGF-B-induced cardiac hypertrophy are mediated at least in part via the endothelium. Because cardiomyocyte damage in myocardial ischemia begins in the subendocardial myocardium, the VEGF-B-induced increased arterial supply to this area could have therapeutic potential in ischemic heart disease.1 aBry, Maija1 aKivelä, Riikka1 aHolopainen, Tanja1 aAnisimov, Andrey1 aTammela, Tuomas1 aSoronen, Jarkko1 aSilvola, Johanna1 aSaraste, Antti1 aJeltsch, Michael1 aKorpisalo, Petra1 aCarmeliet, Peter1 aLemström, Karl, B1 aShibuya, Masabumi1 aYlä-Herttuala, Seppo1 aAlhonen, Leena1 aMervaala, Eero1 aAndersson, Leif, C1 aKnuuti, Juhani1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/2093797402406nas a2200229   4500008004100000245009500041210006900136260001400205300001400219490000800233520168800241100002101929700002201950700002101972700002001993700002302013700002602036700002102062700002602083700001802109856004902127       2009                        eng d00aActivated forms of VEGF-C and VEGF-D provide improved vascular function in skeletal muscle0 aActivated forms of VEGFC and VEGFD provide improved vascular fun  c2009/Jun/  a1302 - 120 v1043 aThe therapeutic potential of vascular endothelial growth factor (VEGF)-C and VEGF-D in skeletal muscle has been of considerable interest as these factors have both angiogenic and lymphangiogenic activities. Previous studies have mainly used adenoviral gene delivery for short-term expression of VEGF-C and VEGF-D in pig, rabbit, and mouse skeletal muscles. Here we have used the activated mature forms of VEGF-C and VEGF-D expressed via recombinant adeno-associated virus (rAAV), which provides stable, long-lasting transgene expression in various tissues including skeletal muscle. Mouse tibialis anterior muscle was transduced with rAAV encoding human or mouse VEGF-C or VEGF-D. Two weeks later, immunohistochemical analysis showed increased numbers of both blood and lymph vessels, and Doppler ultrasound analysis indicated increased blood vessel perfusion. The lymphatic vessels further increased at the 4-week time point were functional, as shown by FITC-lectin uptake and transport. Furthermore, receptor activation and arteriogenic activity were increased by an alanine substitution mutant of human VEGF-C (C137A) having an increased dimer stability and by a chimeric CAC growth factor that contained the VEGF receptor-binding domain flanked by VEGF-C propeptides, but only the latter promoted significantly more blood vessel perfusion when compared to the other growth factors studied. We conclude that long-term expression of VEGF-C and VEGF-D in skeletal muscle results in the generation of new functional blood and lymphatic vessels. The therapeutic value of intramuscular lymph vessels in draining tissue edema and lymphedema can now be evaluated using this model system.1 aAnisimov, Andrey1 aAlitalo, Annamari1 aKorpisalo, Petra1 aSoronen, Jarkko1 aKaijalainen, Seppo1 aLeppänen, Veli-Matti1 aJeltsch, Michael1 aYlä-Herttuala, Seppo1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1944383502437nas a2200325   4500008004100000245014100041210006900182260001400251300001400265490000800279520142200287100002001709700001501729700002101744700003101765700002001796700002001816700002001836700002001856700001901876700002101895700002001916700002301936700001901959700002201978700002602000700001802026700001802044856004902062       2008                        eng d00aOverexpression of vascular endothelial growth factor-B in mouse heart alters cardiac lipid metabolism and induces myocardial hypertrophy0 aOverexpression of vascular endothelial growth factorB in mouse h  c2008/Oct/  a1018 - 260 v1033 aVascular endothelial growth factor (VEGF)-B is poorly angiogenic but prominently expressed in metabolically highly active tissues, including the heart. We produced mice expressing a cardiac-specific VEGF-B transgene via the alpha-myosin heavy chain promoter. Surprisingly, the hearts of the VEGF-B transgenic mice showed concentric cardiac hypertrophy without significant changes in heart function. The cardiac hypertrophy was attributable to an increased size of the cardiomyocytes. Blood capillary size was increased, whereas the number of blood vessels per cell nucleus remained unchanged. Despite the cardiac hypertrophy, the transgenic mice had lower heart rate and blood pressure than their littermates, and they responded similarly to angiotensin II-induced hypertension, confirming that the hypertrophy does not compromise heart function. Interestingly, the isolated transgenic hearts had less cardiomyocyte damage after ischemia. Significantly increased ceramide and decreased triglyceride levels were found in the transgenic hearts. This was associated with structural changes and eventual lysis of mitochondria, resulting in accumulation of intracellular vacuoles in cardiomyocytes and increased death of the transgenic mice, apparently because of mitochondrial lipotoxicity in the heart. These results suggest that VEGF-B regulates lipid metabolism, an unexpected function for an angiogenic growth factor.1 aKarpanen, Terhi1 aBry, Maija1 aOllila, Hanna, M1 aSeppänen-Laakso, Tuulikki1 aLiimatta, Erkki1 aLeskinen, Hanna1 aKivelä, Riikka1 aHelkamaa, Teemu1 aMerentie, Mari1 aJeltsch, Michael1 aPaavonen, Karri1 aAndersson, Leif, C1 aMervaala, Eero1 aHassinen, Ilmo, E1 aYlä-Herttuala, Seppo1 aOresic, Matej1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1875782702098nas a2200385   4500008004100000245011800041210006900159260001400228300001400242490000700256520095200263100001301215700001501228700001901243700001901262700001601281700002001297700002101317700002001338700002401358700002901382700002001411700002901431700001601460700002501476700002501501700002301526700002101549700001901570700001801589700001801607700002101625700001701646856004901663       2008                        eng d00aReevaluation of the role of VEGF-B suggests a restricted role in the revascularization of the ischemic myocardium0 aReevaluation of the role of VEGFB suggests a restricted role in   c2008/Sep/  a1614 - 200 v283 aOBJECTIVE: The endogenous role of the VEGF family member vascular endothelial growth factor-B (VEGF-B) in pathological angiogenesis remains unclear. METHODS AND RESULTS: We studied the role of VEGF-B in various models of pathological angiogenesis using mice lacking VEGF-B (VEGF-B(-/-)) or overexpressing VEGF-B(167). After occlusion of the left coronary artery, VEGF-B deficiency impaired vessel growth in the ischemic myocardium whereas, in wild-type mice, VEGF-B(167) overexpression enhanced revascularization of the infarct and ischemic border zone. By contrast, VEGF-B deficiency did not affect vessel growth in the wounded skin, hypoxic lung, ischemic retina, or ischemic limb. Moreover, VEGF-B(167) overexpression failed to enhance vascular growth in the skin or ischemic limb. CONCLUSIONS: VEGF-B appears to have a relatively restricted angiogenic activity in the ischemic heart. These insights might offer novel therapeutic opportunities.1 aLi, Xuri1 aTjwa, Marc1 aVan Hove, Inge1 aEnholm, Berndt1 aNeven, Elke1 aPaavonen, Karri1 aJeltsch, Michael1 aJuan, Toni Diez1 aSievers, Richard, E1 aChorianopoulos, Emmanuel1 aWada, Hiromichi1 aVanwildemeersch, Maarten1 aNoel, Agnes1 aFoidart, Jean-Michel1 aSpringer, Matthew, L1 aDegenfeld, Georges1 aDewerchin, Mieke1 aBlau, Helen, M1 aAlitalo, Kari1 aEriksson, Ulf1 aCarmeliet, Peter1 aMoons, Lieve  uhttp://view.ncbi.nlm.nih.gov/pubmed/1851169902705nas a2200229   4500008004100000245014300041210006900184260001400253300001400267490000700281520193200288100002502220700002202245700002102267700002102288700002102309700002602330700002202356700003002378700001802408856004902426       2008                        eng d00aThe tyrosine kinase inhibitor cediranib blocks ligand-induced vascular endothelial growth factor receptor-3 activity and lymphangiogenesis0 atyrosine kinase inhibitor cediranib blocks ligandinduced vascula  c2008/Jun/  a4754 - 620 v683 aSolid tumors express a range of factors required to sustain their growth and promote their dissemination. Among these are vascular endothelial growth factor-A (VEGF-A), the key angiogenic stimulant, and VEGF-C, a primary mediator of lymphangiogenesis. Small molecule tyrosine kinase inhibitors offer the potential to inhibit more than one kinase and impede tumor growth by multiple mechanisms. However, their potency toward individual targets can vary. Cediranib (RECENTIN; AZD2171) is an inhibitor of VEGF signaling that has been shown in experimental models to prevent VEGF-A-induced angiogenesis and primary tumor growth, yet the effects of cediranib on VEGF receptor (VEGFR)-3-mediated endothelial cell function and lymphangiogenesis are unknown. To better understand the activity of cediranib against VEGFR-3 and its associated signaling events compared with its activity against VEGFR-2, we used the receptor-specific ligands VEGF-E and VEGF-C156S. In human endothelial cells, cediranib inhibited VEGF-E-induced phosphorylation of VEGFR-2 and VEGF-C156S-induced phosphorylation of VEGFR-3 at concentrations of </=1nmol/L and inhibited activation of downstream signaling molecules. Additionally, cediranib blocked VEGF-C156S-induced and VEGF-E-induced proliferation, survival, and migration of lymphatic and blood vascular endothelial cells. In vivo, cediranib (6 mg/kg/d) prevented angiogenesis and lymphangiogenesis induced by VEGF-E-expressing and VEGF-C156S-expressing adenoviruses, respectively. Cediranib (6 mg/kg/day) also blocked angiogenesis and lymphangiogenesis induced by adenoviruses expressing VEGF-A or VEGF-C and compromised the blood and lymphatic vasculatures of VEGF-C-expressing tumors. Cediranib may, therefore, be an effective means of preventing tumor progression, not only by inhibiting VEGFR-2 activity and angiogenesis, but also by concomitantly inhibiting VEGFR-3 activity and lymphangiogenesis.1 aHeckman, Caroline, A1 aHolopainen, Tanja1 aWirzenius, Maria1 aKeskitalo, Salla1 aJeltsch, Michael1 aYlä-Herttuala, Seppo1 aWedge, Stephen, R1 aJürgensmeier, Juliane, M1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1855952202160nas a2200217   4500008004100000245017800041210006900219260001400288300001400302490000800316520140200324100002001726700001501746700002401761700002101785700002301806700002001829700002601849700001801875856004901893       2007                        eng d00aDistinct architecture of lymphatic vessels induced by chimeric vascular endothelial growth factor-C/vascular endothelial growth factor heparin-binding domain fusion proteins0 aDistinct architecture of lymphatic vessels induced by chimeric v  c2007/May/  a1468 - 750 v1003 aVascular endothelial growth factor (VEGF)-C and VEGF-D are composed of the receptor-binding VEGF homology domain and a carboxy-terminal silk homology domain that requires proteolytic cleavage for growth factor activation. Here, we explored whether the C-terminal heparin-binding domain of the VEGF(165) or VEGF(189) isoform also containing neuropilin-binding sequences could substitute for the silk homology domain of VEGF-C. Such VEGF-C/VEGF-heparin-binding domain chimeras were produced and shown to activate VEGF-C receptors, and, when expressed in tissues via adenovirus or adeno-associated virus vectors, stimulated lymphangiogenesis in vivo. However, both chimeras induced a distinctly different pattern of lymphatic vessels when compared with VEGF-C. Whereas VEGF-C-induced vessels were initially a dense network of small diameter vessels, the lymphatic vessels induced by the chimeric growth factors tended to form directly along tissue borders, along basement membranes that are rich in heparan sulfate. For example, in skeletal muscle, the chimeras induced formation of lumenized lymphatic vessels more efficiently than wild-type VEGF-C. We conclude that the matrix-binding domain of VEGF can target VEGF-C activity to heparin-rich basement membrane structures. These properties may prove useful for tissue engineering and attempts to regenerate lymphatic vessels in lymphedema patients.1 aTammela, Tuomas1 aHe, Yulong1 aLyytikkä, Johannes1 aJeltsch, Michael1 aMarkkanen, Johanna1 aPajusola, Katri1 aYlä-Herttuala, Seppo1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1747873301875nas a2200217   4500008004100000245015700041210006900198260001400267300001300281490000800294520113500302100002101437700002001458700002301478700002001501700002101521700002301542700002501565700001801590856004901608       2007                        eng d00aEnhanced capillary formation stimulated by a chimeric vascular endothelial growth factor/vascular endothelial growth factor-C silk domain fusion protein0 aEnhanced capillary formation stimulated by a chimeric vascular e  c2007/May/  a1460 - 70 v1003 aVascular endothelial growth factor (VEGF)-C and VEGF-D require proteolytic cleavage of the carboxy terminal silk-homology domain for activation. To study the functions of the VEGF-C propeptides, we engineered a chimeric growth factor protein, VEGF-CAC, composed of the amino- and carboxy-terminal propeptides of VEGF-C fused to the receptor-activating core domain of VEGF. Like VEGF-C, VEGF-CAC underwent proteolytic cleavage, and like VEGF, it bound to and activated VEGF receptor-1 and VEGF receptor-2, but not the VEGF-C receptor VEGF receptor-3. VEGF-CAC also bound to neuropilins in a heparin-dependent manner. Strikingly, when VEGF-CAC was expressed via an adenovirus vector in the ear skin of immunodeficient mice, it proved to be a more potent inducer of capillary angiogenesis than VEGF. The VEGF-CAC-induced vessels differed greatly from those induced by VEGF, as they formed a very dense and fine network of pericyte and basement membrane-covered capillaries that were functional, as shown by lectin perfusion experiments. VEGF-CAC could prove useful in proangiogenic therapies in patients experiencing tissue ischemia.1 aKeskitalo, Salla1 aTammela, Tuomas1 aLyytikka, Johannes1 aKarpanen, Terhi1 aJeltsch, Michael1 aMarkkanen, Johanna1 aYla-Herttuala, Seppo1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1747873401805nas a2200217   4500008004100000245007400041210006900115260001400184300001400198490000700212520115600219100002201375700002501397700002101422700002101443700001801464700001801482700002001500700001801520856004901538       2006                        eng d00aFunctional interaction of VEGF-C and VEGF-D with neuropilin receptors0 aFunctional interaction of VEGFC and VEGFD with neuropilin recept  c2006/Jul/  a1462 - 720 v203 aLymphatic vascular development is regulated by vascular endothelial growth factor receptor-3 (VEGFR-3), which is activated by its ligands VEGF-C and VEGF-D. Neuropilin-2 (NP2), known to be involved in neuronal development, has also been implicated to play a role in lymphangiogenesis. We aimed to elucidate the mechanism by which NP2 is involved in lymphatic endothelial cell signaling. By in vitro binding studies we found that both VEGF-C and VEGF-D interact with NP2, VEGF-C in a heparin-independent and VEGF-D in a heparin-dependent manner. We also mapped the domains of VEGF-C and NP2 required for their binding. The functional importance of the interaction of NP2 with the lymphangiogenic growth factors was demonstrated by cointernalization of NP2 along with VEGFR-3 in endocytic vesicles of lymphatic endothelial cells upon stimulation with VEGF-C or VEGF-D. NP2 also interacted with VEGFR-3 in coprecipitation studies. Our results show that NP2 is directly involved in an active signaling complex with the key regulators of lymphangiogenesis and thus suggest a mechanism by which NP2 functions in the development of the lymphatic vasculature.1 aKärpänen, Terhi1 aHeckman, Caroline, A1 aKeskitalo, Salla1 aJeltsch, Michael1 aOllila, Hanna1 aNeufeld, Gera1 aTamagnone, Luca1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1681612101519nas a2200193   4500008004100000245014600041210006900187260001400256300001500270490000800285520086800293100002101161700002001182700002001202700001501222700002101237700001801258856004901276       2006                        eng d00aVascular endothelial growth factor (VEGF)/VEGF-C mosaic molecules reveal specificity determinants and feature novel receptor binding patterns0 aVascular endothelial growth factor VEGFVEGFC mosaic molecules re  c2006/Apr/  a12187 - 950 v2813 aVascular endothelial growth factors (VEGFs) and their receptors play key roles in angiogenesis and lymphangiogenesis. VEGF activates VEGF receptor-1 (VEGFR-1) and VEGFR-2, whereas VEGF-C activates VEGFR-2 and VEGFR-3. We have created a library of VEGF/VEGF-C mosaic molecules that contains factors with novel receptor binding profiles, notably proteins binding to all three VEGF receptors ("super-VEGFs"). The analyzed super-VEGFs show both angiogenic and lymphangiogenic effects in vivo, although weaker than the parental molecules. The composition of the VEGFR-3 binding molecules and scanning mutagenesis revealed determinants of receptor binding and specificity. VEGFR-2 and VEGFR-3 showed striking differences in their requirements for VEGF-C binding; extracellular domain 2 of VEGFR-2 was sufficient, whereas in VEGFR-3, both domains 1 and 2 were necessary.1 aJeltsch, Michael1 aKarpanen, Terhi1 aStrandin, Tomas1 aAho, Kukka1 aLankinen, Hilkka1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1650548902389nas a2200289   4500008004100000245009100041210006900132260001400201300001300215490000800228520151500236100001701751700002001768700001501788700002301803700001901826700002301845700002101868700002401889700002401913700002301937700002601960700002201986700001802008700002402026856004902050       2005                        eng d00aPathogenesis of persistent lymphatic vessel hyperplasia in chronic airway inflammation0 aPathogenesis of persistent lymphatic vessel hyperplasia in chron  c2005/Feb/  a247 - 570 v1153 aEdema occurs in asthma and other inflammatory diseases when the rate of plasma leakage from blood vessels exceeds the drainage through lymphatic vessels and other routes. It is unclear to what extent lymphatic vessels grow to compensate for increased leakage during inflammation and what drives the lymphangiogenesis that does occur. We addressed these issues in mouse models of (a) chronic respiratory tract infection with Mycoplasma pulmonis and (b) adenoviral transduction of airway epithelium with VEGF family growth factors. Blood vessel remodeling and lymphangiogenesis were both robust in infected airways. Inhibition of VEGFR-3 signaling completely prevented the growth of lymphatic vessels but not blood vessels. Lack of lymphatic growth exaggerated mucosal edema and reduced the hypertrophy of draining lymph nodes. Airway dendritic cells, macrophages, neutrophils, and epithelial cells expressed the VEGFR-3 ligands VEGF-C or VEGF-D. Adenoviral delivery of either VEGF-C or VEGF-D evoked lymphangiogenesis without angiogenesis, whereas adenoviral VEGF had the opposite effect. After antibiotic treatment of the infection, inflammation and remodeling of blood vessels quickly subsided, but lymphatic vessels persisted. Together, these findings suggest that when lymphangiogenesis is impaired, airway inflammation may lead to bronchial lymphedema and exaggerated airflow obstruction. Correction of defective lymphangiogenesis may benefit the treatment of asthma and other inflammatory airway diseases.1 aBaluk, Peter1 aTammela, Tuomas1 aAtor, Erin1 aLyubynska, Natalya1 aAchen, Marc, G1 aHicklin, Daniel, J1 aJeltsch, Michael1 aPetrova, Tatiana, V1 aPytowski, Bronislaw1 aStacker, Steven, A1 aYlä-Herttuala, Seppo1 aJackson, David, G1 aAlitalo, Kari1 aMcDonald, Donald, M  uhttp://view.ncbi.nlm.nih.gov/pubmed/1566873402358nas a2200241   4500008004100000245016500041210006900206260001400275300001400289490000700303520155700310100001501867700001901882700002001901700002101921700002201942700002501964700002001989700001702009700002302026700001802049856004902067       2005                        eng d00aVascular endothelial cell growth factor receptor 3-mediated activation of lymphatic endothelium is crucial for tumor cell entry and spread via lymphatic vessels0 aVascular endothelial cell growth factor receptor 3mediated activ  c2005/Jun/  a4739 - 460 v653 aLymphangiogenic growth factors vascular endothelial growth factor (VEGF)-C and VEGF-D have been shown to promote lymphatic metastasis by inducing tumor-associated lymphangiogenesis. In this study, we have investigated how tumor cells gain access into lymphatic vessels and at what stage tumor cells initiate metastasis. We show that VEGF-C produced by tumor cells induced extensive lymphatic sprouting towards the tumor cells as well as dilation of the draining lymphatic vessels, suggesting an active role of lymphatic endothelial cells in lymphatic metastasis. A significant increase in lymphatic vessel growth occurred between 2 and 3 weeks after tumor xenotransplantation, and lymph node metastasis occurred at the same stage. These processes were blocked dose-dependently by inhibition of VEGF receptor 3 (VEGFR-3) signaling by systemic delivery of a soluble VEGFR-3-immunoglobulin (Ig) fusion protein via adenoviral or adeno-associated viral vectors. However, VEGFR-3-Ig did not suppress lymph node metastasis when the treatment was started at a later stage after the tumor cells had already spread out, suggesting that tumor cell entry into lymphatic vessels is a key step during tumor dissemination via the lymphatics. Whereas lymphangiogenesis and lymph node metastasis were significantly inhibited by VEGFR-3-Ig, some tumor cells were still detected in the lymph nodes in some of the treated mice. This indicates that complete blockade of lymphatic metastasis may require the targeting of both tumor lymphangiogenesis and tumor cell invasion.1 aHe, Yulong1 aRajantie, Iiro1 aPajusola, Katri1 aJeltsch, Michael1 aHolopainen, Tanja1 aYla-Herttuala, Seppo1 aHarding, Thomas1 aJooss, Karin1 aTakahashi, Takashi1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1593029201674nas a2200265   4500008004100000245011900041210006900160260001400229300001200243490000600255520085100261100002601112700001701138700001801155700001901173700001901192700002401211700002101235700002201256700001901278700002001297700002401317700001801341856004901359       2004                        eng d00aVascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins0 aVascular endothelial growth factor C is required for sprouting o  c2004/Jan/  a74 - 800 v53 aLymphatic vessels are essential for immune surveillance, tissue fluid homeostasis and fat absorption. Defects in lymphatic vessel formation or function cause lymphedema. Here we show that the vascular endothelial growth factor C (VEGF-C) is required for the initial steps in lymphatic development. In Vegfc-/- mice, endothelial cells commit to the lymphatic lineage but do not sprout to form lymph vessels. Sprouting was rescued by VEGF-C and VEGF-D but not by VEGF, indicating VEGF receptor 3 specificity. The lack of lymphatic vessels resulted in prenatal death due to fluid accumulation in tissues, and Vegfc+/- mice developed cutaneous lymphatic hypoplasia and lymphedema. Our results indicate that VEGF-C is the paracrine factor essential for lymphangiogenesis, and show that both Vegfc alleles are required for normal lymphatic development.1 aKarkkainen, Marika, J1 aHaiko, Paula1 aSainio, Kirsi1 aPartanen, Juha1 aTaipale, Jussi1 aPetrova, Tatiana, V1 aJeltsch, Michael1 aJackson, David, G1 aTalikka, Marja1 aRauvala, Heikki1 aBetsholtz, Christer1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1463464601736nas a2200169   4500008004100000245005000041210005000091260001400141300001200155490000800167520126400175100002101439700002001460700001801480700001901498856004901517       2003                        eng d00aGenesis and pathogenesis of lymphatic vessels0 aGenesis and pathogenesis of lymphatic vessels  c2003/Oct/  a69 - 840 v3143 aThe lymphatic system is generally regarded as supplementary to the blood vascular system, in that it transports interstitial fluid, macromolecules, and immune cells back into the blood. However, in insects, the open hemolymphatic (or lymphohematic) system ensures the circulation of immune cells and interstitial fluid through the body. The Drosophila homolog of the mammalian vascular endothelial growth factor receptor (VEGFR) gene family is expressed in hemocytes, suggesting a close relationship to the endothelium that develops later in phylogeny. Lymph hearts are typical organs for the propulsion of lymph in lower vertebrates and are still transiently present in birds. The lymphatic endothelial marker VEGFR-3 is transiently expressed in embryonic blood vessels and is crucial for their development. We therefore regard the question of whether the blood vascular system or the lymphatic system is primary or secondary as open. Future molecular comparisons should be performed without any bias based on the current prevalence of the blood vascular system over the lymphatic system. Here, we give an overview of the structure, function, and development of the lymphatics, with special emphasis on the recently discovered lymphangiogenic growth factors.1 aJeltsch, Michael1 aTammela, Tuomas1 aAlitalo, Kari1 aWilting, Jörg  uhttp://view.ncbi.nlm.nih.gov/pubmed/1294236202307nas a2200241   4500008004100000245010400041210006900145260001400214300001400228490000700242520156400249100002001813700001801833700002301851700002001874700001801894700001901912700002101931700002101952700002501973700001801998856004902016       2003                        eng d00aIntrinsic versus microenvironmental regulation of lymphatic endothelial cell phenotype and function0 aIntrinsic versus microenvironmental regulation of lymphatic endo  c2003/Nov/  a2006 - 130 v173 aVascular endothelial cells are characterized by a high degree of functional and phenotypic plasticity, which is controlled both by their pericellular microenvironment and their intracellular gene expression programs. To gain further insight into the mechanisms regulating the endothelial cell phenotype, we have compared the responses of lymphatic endothelial cells (LECs) and blood vascular endothelial cells (BECs) to vascular endothelial growth factors (VEGFs). VEGFR-3-specific signals are sufficient for LEC but not BEC proliferation, as shown by the ability of the specific ligand VEGF-C156S to stimulate cell cycle entry only in LECs. On the other hand, we found that VEGFR-3 stimulation did not induce LEC cell shape changes typical of VEGFR-2-stimulated LECs, indicating receptor-specific differences in the cytoskeletal responses. Genes induced via VEGFR-2 also differed between BECs and LECs: angiopoietin-2 (Ang-2) was induced via VEGFR-2 in BECs and LECs, but the smooth muscle cell (SMC) chemoattractant BMP-2 was induced only in BECs. Both BECs and LECs were able to promote SMC chemotaxis, but contact with SMCs led to down-regulation of VEGFR-3 expression in BECs in a 3-dimensional coculture system. This was consistent with the finding that VEGFR-3 is down-regulated in vivo at sites of endothelial cell-pericyte/smooth muscle cell contacts. Collectively, these data show intrinsic cell-specific differences of BEC and LEC responses to VEGFs and identify a pericellular regulatory mechanism for VEGFR-3 down-regulation in endothelial cells.1 aVeikkola, Tanja1 aLohela, Marja1 aIkenberg, Kristian1 aMäkinen, Taija1 aKorff, Thomas1 aSaaristo, Anne1 aPetrova, Tatania1 aJeltsch, Michael1 aAugustin, Hellmut, G1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1459767001908nas a2200253   4500008004100000245007800041210006900119260001400188300001400202490000800216520113900224100002101363700002101384700002201405700002501427700002201452700002501474700002101499700002601520700001801546700001701564700002401581856004901605       2003                        eng d00aVEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia0 aVEGF guides angiogenic sprouting utilizing endothelial tip cell   c2003/Jun/  a1163 - 770 v1613 aVascular endothelial growth factor (VEGF-A) is a major regulator of blood vessel formation and function. It controls several processes in endothelial cells, such as proliferation, survival, and migration, but it is not known how these are coordinately regulated to result in more complex morphogenetic events, such as tubular sprouting, fusion, and network formation. We show here that VEGF-A controls angiogenic sprouting in the early postnatal retina by guiding filopodial extension from specialized endothelial cells situated at the tips of the vascular sprouts. The tip cells respond to VEGF-A only by guided migration; the proliferative response to VEGF-A occurs in the sprout stalks. These two cellular responses are both mediated by agonistic activity of VEGF-A on VEGF receptor 2. Whereas tip cell migration depends on a gradient of VEGF-A, proliferation is regulated by its concentration. Thus, vessel patterning during retinal angiogenesis depends on the balance between two different qualities of the extracellular VEGF-A distribution, which regulate distinct cellular responses in defined populations of endothelial cells.1 aGerhardt, Holger1 aGolding, Matthew1 aFruttiger, Marcus1 aRuhrberg, Christiana1 aLundkvist, Andrea1 aAbramsson, Alexandra1 aJeltsch, Michael1 aMitchell, Christopher1 aAlitalo, Kari1 aShima, David1 aBetsholtz, Christer  uhttp://view.ncbi.nlm.nih.gov/pubmed/1281070002304nas a2200277   4500008004100000245015900041210006900200260001400269300001300283490000700296520139900303100001901702700002001721700001901741700002001760700001901780700002001799700002001819700002101839700002601860700002501886700002201911700002601933700001801959856004901977       2002                        eng d00aAdenoviral VEGF-C overexpression induces blood vessel enlargement, tortuosity, and leakiness but no sprouting angiogenesis in the skin or mucous membranes0 aAdenoviral VEGFC overexpression induces blood vessel enlargement  c2002/Jul/  a1041 - 90 v163 aVascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are important regulators of blood and lymphatic vessel growth and vascular permeability. The VEGF-C/VEGFR-3 signaling pathway is crucial for lymphangiogenesis, and heterozygous inactivating missense mutations of the VEGFR-3 gene are associated with hereditary lymphedema. However, VEGF-C can have potent effects on blood vessels because its receptor VEGFR-3 is expressed in certain blood vessels and because the fully processed form of VEGF-C also binds to the VEGFR-2 of blood vessels. To characterize the in vivo effects of VEGF-C on blood and lymphatic vessels, we have overexpressed VEGF-C via adenovirus- and adeno-associated virus-mediated transfection in the skin and respiratory tract of athymic nude mice. This resulted in dose-dependent enlargement and tortuosity of veins, which, along with the collecting lymphatic vessels were found to express VEGFR-2. Expression of angiopoietin 1 blocked the increased leakiness of the blood vessels induced by VEGF-C whereas vessel enlargement and lymphangiogenesis were not affected. However, angiogenic sprouting of new blood vessels was not observed in response to AdVEGF-C or AAV-VEGF-C. These results show that virally produced VEGF-C induces blood vessel changes, including vascular leak, but its angiogenic potency is much reduced compared with VEGF in normal skin.1 aSaaristo, Anne1 aVeikkola, Tanja1 aEnholm, Berndt1 aHytönen, Maija1 aArola, Johanna1 aPajusola, Katri1 aTurunen, Païvi1 aJeltsch, Michael1 aKarkkainen, Marika, J1 aKerjaschki, Dontscho1 aBueler, Hansruedi1 aYlä-Herttuala, Seppo1 aAlitalo, Kari  uhttp://view.ncbi.nlm.nih.gov/pubmed/1208706502637nas a2200109   4500008004500000245004200045210004100087490001500128520230100143100002102444856006202465       2002                        Engldsh  00aVEGFR-3 Ligands and Lymphangiogenesis0 aVEGFR3 Ligands and Lymphangiogenesis0 vPhD Thesis3 aMost of us have seen their own blood at one or the other time. The occasion might have been
a small accident or in unfortunate cases a severe blood loss caused by a major injury. We also
can feel our heart beating and the resulting pressure wave, the pulse. The existence of the car-
diovascular system is obvious to us. Unlike the cardiovascular system, the lymphatic system
has, until recently, escaped notable attention not only by the laymen, but also by the scientific
community. It is unclear why the lymphatic system originally developed in higher vertebrates.
Now, its main function seems to be to collect fluid that has leaked from the blood vessels and
to return it into the cardiovascular system. Much of our knowledge about the development and
structure of the lymphatic system is of considerable age, and it has been said that there has not
been any progress in our understanding since the fine structure of the lymphatics was described
with the introduction of the electron microscope.
Vascular endothelial growth factor (VEGF) is the principal direct inducer of blood vessel
growth, but it does not promote the growth of lymphatic vessels. This study demonstrates for
the first time specific lymphangiogenesis as a response to the VEGF homologue VEGF-C.
Overexpression of full length VEGF-C under the keratin-14 promoter in the skin of transgenic
mice caused a proliferation of the lymphatic endothelium and lymphatic vessel enlargement. In
the chorioallantoic membrane assay, the mature form of VEGF-C was also largely specific for
lymphatic endothelial cells. A newly discovered close homologue of VEGF-C, VEGF-D was
then shown to have the same receptor-binding pattern as VEGF-C.
Contrary to the interaction of VEGF with its receptors, VEGF-C interaction with VEGFR-3 has
not been analyzed at the molecular level. The structural determinants of VEGFR-3 binding were
characterized in relation to VEGF using a non-random family shuffling approach with VEGF
and VEGF-C as parent molecules. This approach led to the identification of VEGF/VEGF-C
mosaic molecules that showed novel receptor binding profiles and a panel of these molecules
was used to delineate the requirements of specific receptors in the induction of angiogenesis
versus lymphangiogenesis.1 aJeltsch, Michael  uhttp://ethesis.helsinki.fi/julkaisut/mat/bioti/vk/jeltsch01909nas a2200229   4500008004100000245010400041210006900145260001400214300001200228490000700240520124400247100001401491700001601505700001501521700001201536700001601548700001301564700001701577700002101594700001501615856004901630       2001                        eng d00aAdenoviral expression of vascular endothelial growth factor-C induces lymphangiogenesis in the skin0 aAdenoviral expression of vascular endothelial growth factorC ind  c2001/Mar/  a623 - 90 v883 aThe 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.1 aEnholm, B1 aKarpanen, T1 aJeltsch, M1 aKubo, H1 aStenback, F1 aPrevo, R1 aJackson, D G1 aYla-Herttuala, S1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/1128289701998nas a2200265   4500008004100000245012000041210006900161260001400230300001400244490000700258520123100265100001601496700001501512700001501527700001601542700001501558700001701573700001201590700001601602700001801618700001501636700001701651700001501668856004901683       2001                        eng d00aSignalling via vascular endothelial growth factor receptor-3 is sufficient for lymphangiogenesis in transgenic mice0 aSignalling via vascular endothelial growth factor receptor3 is s  c2001/Mar/  a1223 - 310 v203 aVascular 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.1 aVeikkola, T1 aJussila, L1 aMakinen, T1 aKarpanen, T1 aJeltsch, M1 aPetrova, T V1 aKubo, H1 aThurston, G1 aMcDonald, D M1 aAchen, M G1 aStacker, S A1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/1125088900590nas a2200181   4500008004100000245008300041210006900124260003800193100001600231700001700247700001600264700001700280700001700297700001400314700001600328700001600344856004800360       2001                        eng d00aSignalling via VEGFR-3 is sufficient for lymphangiogenesis in transgenic mice.0 aSignalling via VEGFR3 is sufficient for lymphangiogenesis in tra  aMiami Beach, Floridac2001/10/29/1 aJussila, L.1 aVeikkola, T.1 aJeltsch, M.1 aThurston, G.1 aMcDonald, D.1 aAchen, M.1 aStacker, S.1 aAlitalo, K.  uhttps://jeltsch.org/Jussila2001?language=hu02021nas a2200289   4500008004100000245009500041210006900136260001400205300001300219490000700232520122500239100001901464700001501483700001501498700001601513700001501529700001301544700001501557700001401572700001701586700001701603700001201620700001501632700001901647700001601666856004901682       2001                        eng d00aVascular endothelial growth factor-C-mediated lymphangiogenesis promotes tumour metastasis0 aVascular endothelial growth factorCmediated lymphangiogenesis pr  c2001/Feb/  a672 - 820 v203 aMetastasis 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.1 aMandriota, S J1 aJussila, L1 aJeltsch, M1 aCompagni, A1 aBaetens, D1 aPrevo, R1 aBanerji, S1 aHuarte, J1 aMontesano, R1 aJackson, D G1 aOrci, L1 aAlitalo, K1 aChristofori, G1 aPepper, M S  uhttp://view.ncbi.nlm.nih.gov/pubmed/1117921202694nas a2200313   4500008004100000245011900041210006900160260001400229300001300243490000800256520179900264100001802063700001602081700001702097700001502114700001902129700001802148700001702166700001302183700001502196700001902211700001502230700001402245700001802259700001702277700001502294700002202309856004902331       2000                        eng d00aIntravascular adenovirus-mediated VEGF-C gene transfer reduces neointima formation in balloon-denuded rabbit aorta0 aIntravascular adenovirusmediated VEGFC gene transfer reduces neo  c2000/Oct/  a2262 - 80 v1023 aBACKGROUND: 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.1 aHiltunen, M O1 aLaitinen, M1 aTurunen, M P1 aJeltsch, M1 aHartikainen, J1 aRissanen, T T1 aLaukkanen, J1 aNiemi, M1 aKossila, M1 aHäkkinen, T P1 aKivelä, A1 aEnholm, B1 aMansukoski, H1 aTurunen, A M1 aAlitalo, K1 aYlä-Herttuala, S  uhttp://view.ncbi.nlm.nih.gov/pubmed/1105610300881nas a2200169   4500008004100000245004100041210003900082260001400121300001300135490000700148520044500155100001600600700001500616700001600631700001500647856004900662       1999                        eng d00aCurrent biology of VEGF-B and VEGF-C0 aCurrent biology of VEGFB and VEGFC  c1999/Dec/  a528 - 350 v103 aEndothelial 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.1 aOlofsson, B1 aJeltsch, M1 aEriksson, U1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/1060068901922nas a2200253   4500008004100000245014100041210006900182260001400251300001500265490000700280520116200287100001601449700001901465700001601484700001901500700001201519700001301531700001301544700001701557700001501574700001501589700001601604856004801620       1998                        eng d00aVascular endothelial growth factor B (VEGF-B) binds to VEGF receptor-1 and regulates plasminogen activator activity in endothelial cells0 aVascular endothelial growth factor B VEGFB binds to VEGF recepto  c1998/Sep/  a11709 - 140 v953 aThe vascular endothelial growth factor (VEGF) family has recently expanded by the identification and cloning of three additional members, namely VEGF-B, VEGF-C, and VEGF-D. In this study we demonstrate that VEGF-B binds selectively to VEGF receptor-1/Flt-1. This binding can be blocked by excess VEGF, indicating that the interaction sites on the receptor are at least partially overlapping. Mutating the putative VEGF receptor-1/Flt-1 binding determinants Asp63, Asp64, and Glu67 to alanine residues in VEGF-B reduced the affinity to VEGF receptor-1 but did not abolish binding. Mutational analysis of conserved cysteines contributing to VEGF-B dimer formation suggest a structural conservation with VEGF and platelet-derived growth factor. Proteolytic processing of the 60-kDa VEGF-B186 dimer results in a 34-kDa dimer containing the receptor-binding epitopes. The binding of VEGF-B to its receptor on endothelial cells leads to increased expression and activity of urokinase type plasminogen activator and plasminogen activator inhibitor 1, suggesting a role for VEGF-B in the regulation of extracellular matrix degradation, cell adhesion, and migration.1 aOlofsson, B1 aKorpelainen, E1 aPepper, M S1 aMandriota, S J1 aAase, K1 aKumar, V1 aGunji, Y1 aJeltsch, M M1 aShibuya, M1 aAlitalo, K1 aEriksson, U  uhttp://view.ncbi.nlm.nih.gov/pubmed/975173001686nas a2200217   4500008004100000245013700041210006900178260001400247300001300261490000700274520102000281100001501301700001501316700001201331700001601343700001401359700001501373700001501388700001701403856004801420       1998                        eng d00aVascular endothelial growth factor D (VEGF-D) is a ligand for the tyrosine kinases VEGF receptor 2 (Flk1) and VEGF receptor 3 (Flt4)0 aVascular endothelial growth factor D VEGFD is a ligand for the t  c1998/Jan/  a548 - 530 v953 aWe 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.1 aAchen, M G1 aJeltsch, M1 aKukk, E1 aMäkinen, T1 aVitali, A1 aWilks, A F1 aAlitalo, K1 aStacker, S A  uhttp://view.ncbi.nlm.nih.gov/pubmed/943522902306nas a2200181   4500008004100000245020900041210006900250260001400319300001300333490000800346520164400354100001601998700001902014700001502033700001302048700001502061856004802076       1998                        eng d00aVascular 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 activity0 aVascular endothelial growth factor VEGFC synergizes with basic f  c1998/Dec/  a439 - 520 v1773 aVascular 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.1 aPepper, M S1 aMandriota, S J1 aJeltsch, M1 aKumar, V1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/980815201905nas a2200217   4500008004100000245009100041210006900132260001400201300001500215490000800230520128700238100001401525700001201539700001301551700001501564700001601579700001501595700001401610700001501624856004801639       1997                        eng d00aGenomic organization of human and mouse genes for vascular endothelial growth factor C0 aGenomic organization of human and mouse genes for vascular endot  c1997/Oct/  a25176 - 830 v2723 aWe 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.1 aChilov, D1 aKukk, E1 aTaira, S1 aJeltsch, M1 aKaukonen, J1 aPalotie, A1 aJoukov, V1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/931213001376nas a2200241   4500008004100000245006300041210006200104260001400166300001300180490000800193520074000201100001500941700001700956700001400973700001200987700001300999700001501012700001401027700001601041700001401057700001501071856004801086       1997                        eng d00aHyperplasia of lymphatic vessels in VEGF-C transgenic mice0 aHyperplasia of lymphatic vessels in VEGFC transgenic mice  c1997/May/  a1423 - 50 v2763 aNo 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.1 aJeltsch, M1 aKaipainen, A1 aJoukov, V1 aMeng, X1 aLakso, M1 aRauvala, H1 aSwartz, M1 aFukumura, D1 aJain, R K1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/916201101810nas a2200229   4500008004100000245008100041210006900122260001400191300001500205490000700220520117000227100001401397700001301411700001301424700001501437700002201452700001101474700001501485700001701500700001501517856004801532       1997                        eng d00aProteolytic processing regulates receptor specificity and activity of VEGF-C0 aProteolytic processing regulates receptor specificity and activi  c1997/Jul/  a3898 - 9110 v163 aThe 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 'mature' 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.1 aJoukov, V1 aSorsa, T1 aKumar, V1 aJeltsch, M1 aClaesson-Welsh, L1 aCao, Y1 aSaksela, O1 aKalkkinen, N1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/923380000565nas a2200205   4500008004100000245005800041210005600099260001400155300001200169490000800181100001400189700001700203700001500220700001600235700001600251700001300267700001600280700001500296856004800311       1997                        eng d00aVascular endothelial growth factors VEGF-B and VEGF-C0 aVascular endothelial growth factors VEGFB and VEGFC  c1997/Nov/  a211 - 50 v1731 aJoukov, V1 aKaipainen, A1 aJeltsch, M1 aPajusola, K1 aOlofsson, B1 aKumar, V1 aEriksson, U1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/936552402815nas a2200217   4500008004100000245013100041210006900172260001400241300001300255490000800268520214700276100001202423700001702435700002002452700001802472700001502490700001402505700001502519700001502534856004802549       1997                        eng d00aVEGF and VEGF-C: specific induction of angiogenesis and lymphangiogenesis in the differentiated avian chorioallantoic membrane0 aVEGF and VEGFC specific induction of angiogenesis and lymphangio  c1997/Aug/  a96 - 1090 v1883 aThe lymphangiogenic potency of endothelial growth factors has not been studied to date. This is partially due to the lack of in vivo lymphangiogenesis assays. We have studied the lymphatics of differentiated avian chorioallantoic membrane (CAM) using microinjection of Mercox resin, semi- and ultrathin sectioning, immunohistochemical detection of fibronectin and alpha-smooth muscle actin, and in situ hybridization with VEGFR-2 and VEGFR-3 probes. CAM is drained by lymphatic vessels which are arranged in a regular pattern. Arterioles and arteries are accompanied by a pair of interconnected lymphatics and form a plexus around bigger arteries. Veins are also associated with lymphatics, particularly larger veins, which are surrounded by a lymphatic plexus. The lymphatics are characterized by an extremely thin endothelial lining, pores, and the absence of a basal lamina. Patches of the extracellular matrix can be stained with an antibody against fibronectin. Lymphatic endothelial cells of differentiated CAM show ultrastructural features of this cell type. CAM lymphatics do not possess mediae. In contrast, the lymphatic trunks of the umbilical stalk are invested by a single but discontinuous layer of smooth muscle cells. CAM lymphatics express VEGFR-2 and VEGFR-3. Both the regular pattern and the typical structure of these lymphatics suggest that CAM is a suitable site to study the in vivo effects of potential lymphangiogenic factors. We have studied the effects of VEGF homo- and heterodimers, VEGF/PlGF heterodimers, and PlGF and VEGF-C homodimers on Day 13 CAM. All the growth factors containing at least one VEGF chain are angiogenic but do not induce lymphangiogenesis. PlGF-1 and PlGF-2 are neither angiogenic nor lymphangiogenic. VEGF-C is the first lymphangiogenic factor and seems to be highly chemoattractive for lymphatic endothelial cells. It induces proliferation of lymphatic endothelial cells and development of new lymphatic sinuses which are directed immediately beneath the chorionic epithelium. Our studies show that VEGF and VEGF-C are specific angiogenic and lymphangiogenic growth factors, respectively.1 aOh, S J1 aJeltsch, M M1 aBirkenhäger, R1 aMcCarthy, J E1 aWeich, H A1 aChrist, B1 aAlitalo, K1 aWilting, J  uhttp://view.ncbi.nlm.nih.gov/pubmed/924551502083nas a2200205   4500008004100000245011700041210006900158260001400227300001400241490000800255520146100263100001201724700001901736700001301755700001701768700001501785700001401800700001501814856004801829       1996                        eng d00aVEGF-C receptor binding and pattern of expression with VEGFR-3 suggests a role in lymphatic vascular development0 aVEGFC receptor binding and pattern of expression with VEGFR3 sug  c1996/Dec/  a3829 - 370 v1223 aThe 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.1 aKukk, E1 aLymboussaki, A1 aTaira, S1 aKaipainen, A1 aJeltsch, M1 aJoukov, V1 aAlitalo, K  uhttp://view.ncbi.nlm.nih.gov/pubmed/9012504