@article {893, title = {Biomass-derived Lignin Nanoparticles for the Sustained Delivery of Vascular Endothelial Growth Factor-C}, journal = {Molecular Pharmaceutics}, year = {Submitted}, chapter = {submitted/preprint}, doi = {10.1101/2025.04.23.649697}, url = {https://doi.org/10.1101/2025.04.23.649697}, author = {Mavali-Zadeh, Ahmad and Gatto, Emanuela and Lettieri, Raffaella and Bokharaie, Bokharaie and Caravella, Alice and D{\textquoteright}Ottavi, Cadia and di Bartolomeo, Elisabetta and Domenici, Fabio and Sima, Setareh and Correia, Alexandra and Rauniyar, Khushbu and Klose, Anna and Gounani, Zahra and Laaksonen, Timo and K{\"u}nnapuu, Jaana and Jeltsch, Michael} } @article {892, title = {Fetal-like reversion in the regenerating intestine is regulated by mesenchymal asporin}, journal = {Cell Stem Cell}, volume = {32}, year = {2025}, month = {Jan-04-2025}, pages = {613 - 626.e8}, abstract = {Mesenchymal cells and the extracellular matrix (ECM) support epithelium during homeostasis and regeneration. However, the role of the mesenchyme in epithelial conversion into a fetal-like regenerative state after damage is not known. We modeled epithelial regeneration by culturing intestinal epithelium on decellularized small intestinal scaffolds (iECM) and identify asporin (Aspn), an ECM-bound proteoglycan, as a critical mediator of epithelial fetal-like reprogramming. After damage, transient increase in Aspn expression by the pericryptal fibroblasts induces epithelial transforming growth factor b (TGF-b)-signaling via CD44 and promotes timely epithelial reprogramming. Temporal control of Aspn is lost in old mice, and after damage, the persistently high level of Aspn stagnates epithelium in the regenerative state. Increase in Wnt signaling can resolve the stagnated regenerative program of the old epithelium, promoting restoration of tissue function. In summary, we establish a platform for modeling epithelial injury responses ex vivo and show that the mesenchymal Aspn-producing niche modulates tissue repair by regulating epithelial fetal-like reprogramming.}, issn = {19345909}, doi = {10.1016/j.stem.2025.02.009}, url = {https://www.sciencedirect.com/science/article/pii/S1934590925000487}, author = {Iqbal, Sharif and Andersson, Simon and Nesta, Ernesta and Pentinmikko, Nalle and Kumar, Ashish and Jha, Sawan K. and Borshagovski, Daniel and Webb, Anna and Gebert, Nadja and Viitala, Emma W. and Ritchie, Alexandra and Scharaw, Sandra and Kuuluvainen, Emilia and Larsen, Hjalte L. and Saarinen, Tuure and Juuti, Anne and Ristim{\"a}ki, Ari and Jeltsch, Michael and Ori, Alessandro and Varjosalo, Markku and Pietil{\"a}inen, Kirsi H. and Ollila, Saara and Jensen, Kim B. and Oudhoff, Menno J. and Katajisto, Pekka} } @article {903, title = {Targeting Bacterial and Human Levodopa Decarboxylases for Improved Drug Treatment of Parkinson{\textquoteright}s Disease: Discovery and Characterization of New Inhibitors}, journal = {European Journal of Pharmaceutical Sciences}, year = {2025}, month = {Jan-05-2025}, pages = {107133}, issn = {09280987}, doi = {10.1016/j.ejps.2025.107133}, url = {https://authors.elsevier.com/sd/article/S0928-0987(25)00132-0}, author = {Reunanen, Saku and Ghemtio, Leo and Patel, Jayendrakumar Z. and Patel, Darshitkumar R. and Airavaara, Kerttu and Yli-Kauhaluoma, Jari and Jeltsch, Michael and Xhaard, Henri and Piepponen, Petteri and Tammela, P{\"a}ivi} } @article {897, title = {Gliflozins, sucrose and flavonoids are allosteric activators of lecithin-cholesterol acyltransferase}, journal = {Scientific Reports}, volume = {14}, year = {2024}, month = {2024/10/30/}, pages = {26085}, abstract = {Abstract Lecithin-cholesterol acyltransferase (LCAT) serves as a pivotal enzyme in preserving cholesterol homeostasis via reverse cholesterol transport, a process closely associated with the onset of atherosclerosis. Impaired LCAT function can lead to severe LCAT deficiency disorders for which no pharmacological treatment exists. LCAT-based therapies, such as small molecule positive allosteric modulators (PAMs), against LCAT deficiencies and atherosclerosis hold promise, although their efficacy against atherosclerosis remains challenging. Herein we utilized a quantitative in silico metric to predict the activity of novel PAMs and tested their potencies with in vitro enzymatic assays. As predicted, sodium-glucose cotransporter 2 (SGLT2) inhibitors (gliflozins), sucrose and flavonoids activate LCAT. This has intriguing implications for the mechanism of action of gliflozins, which are commonly used in the treatment of type 2 diabetes, and for the endogenous activation of LCAT. Our results underscore the potential of molecular dynamics simulations in rational drug design.}, isbn = {2045-2322}, doi = {10.1038/s41598-024-77104-3}, url = {https://www.nature.com/articles/s41598-024-77104-3}, author = {Niemel{\"a}, Akseli and Giorgi, Laura and Nouri, Sirine and Yurtta{\c s}, Bet{\"u}l and Rauniyar, Khushbu and Jeltsch, Michael and Koivuniemi, Artturi} } @article {860, title = {Expansion and collapse of VEGF diversity in major clades of the animal kingdom}, journal = {Angiogenesis}, volume = {26}, year = {2023}, month = {Jan-08-2023}, pages = {437 - 461}, issn = {0969-6970}, doi = {10.1007/s10456-023-09874-9}, url = {https://link.springer.com/10.1007/s10456-023-09874-9}, author = {Rauniyar, Khushbu and Bokharaie, Honey and Jeltsch, Michael} } @mastersthesis {863, title = {VEGF-C: The evolutionary origin, activation, and potential as a drug target}, year = {2023}, month = {2023/06/09/}, school = {University of Helsinki}, address = {Helsinki. Finland}, url = {http://hdl.handle.net/10138/357923}, author = {Rauniyar, Khushbu} } @article {861, title = {Bioactive VEGF-C from E. coli}, journal = {Scientific Reports}, volume = {12}, year = {2022}, month = {Jan-12-2022}, doi = {10.1038/s41598-022-22960-0}, url = {https://www.nature.com/articles/s41598-022-22960-0}, author = {Rauniyar, Khushbu and Akhondzadeh, Soheila and G{\k a}ciarz, Anna and K{\"u}nnapuu, Jaana and Jeltsch, Michael} } @article {693, title = {KLK3/PSA and cathepsin D activate VEGF-C and VEGF-D}, journal = {eLife}, volume = {8}, year = {2019}, month = {2019/05/17/}, pages = {e44478}, abstract = {Vascular 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.}, keywords = {cancer biology, Cathepsin D, kallikrein-related peptidases, KLK3/PSA, Lymphangiogenesis, mouse, VEGF-C, VEGF-D}, isbn = {2050-084X}, url = {https://elifesciences.org/articles/44478}, author = {Jha, Sawan K and Rauniyar, Khushbu and Chronowska, Ewa and Mattonet, Kenny and Maina, Eunice W and Koistinen, Hannu and Stenman, Ulf-H{\r a}kan and Alitalo, Kari and Jeltsch, Michael} } @article {611, title = {Biology of Vascular Endothelial Growth Factor C in the Morphogenesis of Lymphatic Vessels}, journal = {Frontiers in Biotechnology and Bioengineering}, volume = {6}, year = {2018}, month = {02/2018}, type = {review}, abstract = {Because virtually all tissues contain blood vessels, the importance of hemevascularization has been long recognized in regenerative medicine and tissue engineering. However, the lymphatic vasculature has only recently become a subject of interest. Central to the task of growing a lymphatic network are lymphatic endothelial cells (LECs), which constitute the innermost layer of all lymphatic vessels. The central molecule that directs proliferation and migration of LECs during embryogenesis is Vascular Endothelial Growth Factor-C (VEGF-C). VEGF-C is, therefore, an important ingredient for LEC culture and attempts to (re)generate lymphatic vessels and networks. During its biosynthesis, VEGF-C undergoes a stepwise proteolytic processing, during which its properties and affinities for its interaction partners change. Many of these fundamental aspects of VEGF-C biosynthesis have only recently been uncovered. So far, most - if not all - applications of VEGF-C do not discriminate between different forms of VEGF-C. However, for lymphatic regeneration and engineering purposes, it appears mandatory to understand these differences, since they relate e.g. to such important aspects as biodistribution and receptor activation potential. In this review, we discuss the molecular biology of VEGF-C as it relates to the growth of LECs and lymphatic vessels. However, the properties of VEGF-C are similarly relevant for the cardiovascular system, since both old and recent data show that VEGF-C can have a profound effect on the blood vasculature.}, keywords = {ADAMTS3, CCBE1, growth factor signaling, growth factors, Lymphatic Vessels, lymphedema, proteolytic processing, Tissue Engineering, VEGF receptors, VEGF-C}, issn = {2296-4185}, doi = {10.3389/fbioe.2018.00007}, url = {https://www.frontiersin.org/articles/10.3389/fbioe.2018.00007/full}, author = {Khushbu Rauniyar and Sawan K Jha and Michael Jeltsch} } @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}, author = {Jha, SK and Rauniyar, K and Jeltsch, M} } @article {589, title = {Efficient activation of the lymphangiogenic growth factor VEGF-C requires the C-terminal domain of VEGF-C and the N-terminal domain of CCBE1}, journal = {Scientific Reports}, volume = {7}, year = {2017}, month = {2017/07/07/}, pages = {4916}, doi = {10.1038/s41598-017-04982-1}, url = {https://www.nature.com/articles/s41598-017-04982-1}, author = {Jha, SK and Rauniyar, Khushbu and K{\"a}rp{\"a}nen, Terhi and Lepp{\"a}nen, Veli-Matti and Brouillard, Pascal and Vikkula, Miikka and Alitalo, Kari and Jeltsch, Michael} } @article {527, title = {Lymphatic Vessels in Regenerative Medicine and Tissue Engineering}, journal = {Tissue Engineering Part B}, volume = {22}, year = {2016}, month = {2016}, pages = {1-13}, type = {Review}, abstract = {Once a DOI is available for this article, the final publication will be available from Mary Ann Liebert, Inc., publishers at http://dx.doi.org/10.1089/TEN.TEB.2016.0034. The postprint manuscript is available from here and for the next 30 days also from the publisher via this bit.ly shortcut: http://bit.ly/1VKjjMk. }, doi = {10.1089/ten.TEB.2016.0034}, url = {http://online.liebertpub.com/doi/10.1089/ten.TEB.2016.0034}, author = {Schaupper, Mira V. and Jeltsch, Michael and Rohringer, Sabrina and Redl, Heinz and Holnthoner, Wolfgang} } @article {502, title = {Functional Dissection of the CCBE1 Protein: A Crucial Requirement for the Collagen Repeat Domain.}, journal = {Circ Res}, volume = {116}, year = {2015}, month = {2015 May 8}, pages = {1660-1669}, abstract = {

RATIONALE: Collagen- and calcium-binding EGF domain-containing protein 1 (CCBE1) is essential for lymphangiogenesis in vertebrates and has been associated with Hennekam syndrome. Recently, CCBE1 has emerged as a crucial regulator of vascular endothelial growth factor-C (VEGFC) signaling.

OBJECTIVE: CCBE1 is a secreted protein characterized by 2 EGF domains and 2 collagen repeats. The functional role of the different CCBE1 protein domains is completely unknown. Here, we analyzed the functional role of the different CCBE1 domains in vivo and in vitro.

METHODS AND RESULTS: We analyzed the functionality of several CCBE1 deletion mutants by generating knock-in mice expressing these mutants, by analyzing their ability to enhance Vegfc signaling in vivo in zebrafish, and by testing their ability to induce VEGFC processing in vitro. We found that deleting the collagen domains of CCBE1 has a much stronger effect on CCBE1 activity than deleting the EGF domains. First, although CCBE1ΔCollagen mice fully phenocopy CCBE1 knock-out mice, CCBE1ΔEGF knock-in embryos still form rudimentary lymphatics. Second, Ccbe1ΔEGF, but not Ccbe1ΔCollagen, could partially substitute for Ccbe1 to enhance Vegfc signaling in zebrafish. Third, CCBE1ΔEGF, similarly to CCBE1, but not CCBE1ΔCollagen could activate VEGFC processing in vitro. Furthermore, a Hennekam syndrome mutation within the collagen domain has a stronger effect than a Hennekam syndrome mutation within the EGF domain.

CONCLUSIONS: We propose that the collagen domains of CCBE1 are crucial for the activation of VEGFC in vitro and in vivo. The EGF domains of CCBE1 are dispensable for regulation of VEGFC processing in vitro, however, they are necessary for full lymphangiogenic activity of CCBE1 in vivo.

}, keywords = {Animals, Binding Sites, Calcium-Binding Proteins, Collagen, Craniofacial Abnormalities, Endothelial Cells, Epidermal Growth Factor, Gene Expression Regulation, Developmental, Gene Knock-In Techniques, Genital Diseases, Male, Genotype, Gestational Age, HEK293 Cells, Humans, Lymphangiectasis, Intestinal, Lymphatic Vessels, lymphedema, Mice, Mice, Transgenic, Mutation, Phenotype, Protein Binding, Protein Interaction Domains and Motifs, Signal Transduction, Transfection, Tumor Suppressor Proteins, Vascular Endothelial Growth Factor C, Zebrafish, Zebrafish Proteins}, issn = {1524-4571}, doi = {10.1161/CIRCRESAHA.116.304949}, url = {http://circres.ahajournals.org/content/116/10/1660.long}, author = {Roukens, M Guy and Peterson-Maduro, Josi and Padberg, Yvonne and Jeltsch, Michael and Lepp{\"a}nen, Veli-Matti and Bos, Frank L and Alitalo, Kari and Schulte-Merker, Stefan and Schulte, D{\"o}rte} } @article {505, title = {Ischemia-Reperfusion Injury Enhances Lymphatic Endothelial VEGFR3 and Rejection in Cardiac Allografts}, journal = {American Journal of Transplantation}, volume = {16}, year = {2015}, month = {12/2015}, pages = {1160-1172}, abstract = {Organ damage and innate immunity during heart transplantation may evoke adaptive immunity with serious consequences. Because lymphatic vessels bridge innate and adaptive immunity, they are critical in immune surveillance; however, their role in ischemia{\textendash}reperfusion injury (IRI) in allotransplantation remains unknown. We investigated whether the lymphangiogenic VEGF-C/VEGFR3 pathway during cardiac allograft IRI regulates organ damage and subsequent interplay between innate and adaptive immunity. We found that cardiac allograft IRI, within hours, increased graft VEGF-C expression and lymphatic vessel activation in the form of increased lymphatic VEGFR3 and adhesion protein expression. Pharmacological VEGF-C/VEGFR3 stimulation resulted in early lymphatic activation and later increase in allograft inflammation. In contrast, pharmacological VEGF-C/VEGFR3 inhibition during cardiac allograft IRI decreased early lymphatic vessel activation with subsequent dampening of acute and chronic rejection. Genetic deletion of VEGFR3 specifically in the lymphatics of the transplanted heart recapitulated the survival effect achieved by pharmacological VEGF-C/VEGFR3 inhibition. Our results suggest that tissue damage rapidly changes lymphatic vessel phenotype, which, in turn, may shape the interplay of innate and adaptive immunity. Importantly, VEGF-C/VEGFR3 inhibition during solid organ transplant IRI could be used as lymphatic-targeted immunomodulatory therapy to prevent acute and chronic rejection.}, doi = {10.1111/ajt.13564}, url = {http://onlinelibrary.wiley.com/doi/10.1111/ajt.13564/abstract}, author = {Dashkevich, A. and Raissadati, A. and Syrj{\"a}l{\"a}, S. O. and Zarkada, G. and Ker{\"a}nen, M. A. I. and Tuuminen, R. and Krebs, R. and Anisimov, A. and Jeltsch, M. and Lepp{\"a}nen, V.-M. and Alitalo, K. and Nyk{\"a}nen, A. I. and Lemstr{\"o}m, K. B.} } @article {427, title = {Critical role of VEGF-C/VEGFR-3 signaling in innate and adaptive immune responses in experimental obliterative bronchiolitis.}, journal = {Am J Pathol}, volume = {181}, year = {2012}, month = {2012 Nov}, pages = {1607-20}, abstract = {

Chronic inflammation, a hallmark of obliterative bronchiolitis, is known to induce lymphangiogenesis. We therefore studied the role of lymphangiogenic vascular endothelial growth factor C (VEGF-C), its receptor VEGFR-3, and lymphangiogenesis during development of experimental obliterative bronchiolitis [ie, obliterative airway disease (OAD)] in rat tracheal allografts. The functional importance of VEGF-C was investigated by adenovirus-mediated overexpression of VEGF-C (AdVEGF-C), and by inhibition of VEGF-C activity with VEGFR-3-Ig (AdVEGFR-3-Ig). Analyses included histology, immunohistochemistry, and real-time RT-PCR 10 and 30 days after transplantation. In the course of OAD development, lymphangiogenesis was induced in the airway wall during the alloimmune response, which was reversed by cyclosporine A in a dose-dependent fashion. VEGF-C overexpression in tracheal allografts induced epithelial activation, neutrophil chemotaxis, and a shift toward a Th17 adaptive immune response, followed by enhanced lymphangiogenesis and the development of OAD. In contrast, inhibition of VEGF-C activity with VEGFR-3-Ig inhibited lymphangiogenesis and angiogenesis and reduced infiltration of CD4(+) T cells and the development of OAD. Lymphangiogenesis was linked to T-cell responses during the development of OAD, and VEGF-C/VEGFR-3 signaling modulated innate and adaptive immune responses in the development of OAD in rat tracheal allografts. Our results thus suggest VEGFR-3-signaling as a novel strategy to regulate T-cell responses in the development of obliterative bronchiolitis after lung transplantation.

}, keywords = {Adaptive Immunity, Animals, Bronchiolitis Obliterans, Chemotaxis, Cyclosporine, Dendritic Cells, Dose-Response Relationship, Drug, Down-Regulation, Epithelial Cells, Epithelium, Graft Rejection, Immunity, Innate, Immunoglobulins, Inflammation, Lymphangiogenesis, Macrophages, Neutrophils, Rats, Signal Transduction, Th17 Cells, Trachea, Transplantation, Homologous, Up-Regulation, Vascular Endothelial Growth Factor C, Vascular Endothelial Growth Factor Receptor-3}, issn = {1525-2191}, doi = {10.1016/j.ajpath.2012.07.021}, author = {Krebs, Rainer and Tikkanen, Jussi M and Ropponen, Jussi O and Jeltsch, Michael and Jokinen, Janne J and Yl{\"a}-Herttuala, Seppo and Nyk{\"a}nen, Antti I and Lemstr{\"o}m, Karl B} } @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 {30, title = {Vascular endothelial cell growth factor receptor 3-mediated activation of lymphatic endothelium is crucial for tumor cell entry and spread via lymphatic vessels}, journal = {Cancer Res}, volume = {65}, year = {2005}, month = {2005/Jun/}, pages = {4739 - 46}, abstract = {Lymphangiogenic 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.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/15930292}, author = {He, Yulong and Rajantie, Iiro and Pajusola, Katri and Jeltsch, Michael and Holopainen, Tanja and Yla-Herttuala, Seppo and Harding, Thomas and Jooss, Karin and Takahashi, Takashi and Alitalo, Kari} } @article {28, title = {Vascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins}, journal = {Nat Immunol}, volume = {5}, year = {2004}, month = {2004/Jan/}, pages = {74 - 80}, abstract = {Lymphatic 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.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/14634646}, author = {Karkkainen, Marika J and Haiko, Paula and Sainio, Kirsi and Partanen, Juha and Taipale, Jussi and Petrova, Tatiana V and Jeltsch, Michael and Jackson, David G and Talikka, Marja and Rauvala, Heikki and Betsholtz, Christer and Alitalo, Kari} } @article {26, title = {VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia}, journal = {J Cell Biol}, volume = {161}, year = {2003}, month = {2003/Jun/}, pages = {1163 - 77}, abstract = {Vascular 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.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/12810700}, author = {Gerhardt, Holger and Golding, Matthew and Fruttiger, Marcus and Ruhrberg, Christiana and Lundkvist, Andrea and Abramsson, Alexandra and Jeltsch, Michael and Mitchell, Christopher and Alitalo, Kari and Shima, David and Betsholtz, Christer} } @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 {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} }