00739nas a2200241 4500008004100000245009400041210006900135260001600204300000900220490000900229653002100238653001200259653001900271653002200290653001200312653002000324100003200344700002000376700002200396700002100418700002300439856003500462 2016 eng d00aFactors regulating the substrate specificity of cytosolic phospholipase A2-alpha in vitro0 aFactors regulating the substrate specificity of cytosolic phosph c2016/07/01/ a15970 v186110aArachidonic acid10aBilayer10aCatalytic site10aMass spectrometry10aMicelle10aPhospholipase A1 aBatchu, Krishna, Chaithanya1 aHänninen, Satu1 aJha, Sawan, Kumar1 aJeltsch, Michael1 aSomerharju, Pentti uhttps://jeltsch.org/Batchu201602911nas a2200109 4500008004100000245008700041210006900128260003000197520249000227100002102717856006302738 2016 eng d00aFrom Molecular Genetics and Biology to Effective Treatments of Lymphatic Disorders0 aFrom Molecular Genetics and Biology to Effective Treatments of L aMulhouse, Francec05/20163 aIn 1971, Judah Folkman proposed the concept of anti-angiogenic tumor therapy. 12 years later, Harold Dvorak isolated the responsible growth factor VEGF. Nine years later, Napoleone Ferrara re­ported the generation of neutralizing monoclonal antibodies against VEGF. Another five years later, Phase I trials started with the humanized version of one of the monoclonals: bevacizumab. Since 2004, when it received FDA approval, it has been marketed under the brand name Avastin. The translation of basic biomedical research into tangible benefits for patients appears sometimes ago­nizingly slow. The public has been promised much by hyped scientific breakthroughs [4]. Scien­tific journals and scientists have played along in over-hyping scientific breakthroughs in the hope of impact factors and citations in order to secure and justify funding and fame. Not surprisingly, practitioners ask when the discoveries from basic research will finally improve the standard of care for their patients. Lymphatic research is no exception. Practitioners are largely still limited to symptomatic treatment and there seems to be still an invisible, but perceptible divide between those who do the molecular biol­ogy research and those who treat patients. The Avastin story is a plea for basic research: it might be compli­cated and it might take time, but it eventually does pay off. How is the lymphatic research community doing concerning the translation of research results into treatment options? Examples of lymphatic re­search in or shortly before the clinical trial stage include: - Growth factor enhanced lymph node transplantation to treat secondary lymphedema - Utilizing the Schlemm channel's lymphatic character in glaucoma treatment - Anti-angiogenic tumor treatment with anti-lymphangiogenic agents Treatment of primary lymphedema with VEGF-C has been proposed. How­ever, our understand­ing of the physiological process of lymph vessel development is far from com­plete, despite sig­nificant re­cent progress in our understanding of devel­opmental lymphangiogenesis and first attempts at tis­sue-engineering lymphatic vessels. If the results from high throughput cancer profilings are predictive of lymphatic conditions, then many patients will feature very individual, multifactorial disease profiles. Even more challenging than the identification of such causes will be the development of treatment regimens that rapidly can be tailored to such individual needs.1 aJeltsch, Michael uhttp://www.eurolymphology.org/JOURNAL/VOL28-N74-2016/#p=1402952nas a2200289 4500008004100000245008800041210006900129260001200198300001200210490000800222520202000230653002802250653002202278653002402300653001702324653004802341100002102389700001302410700002102423700002302444700002102467700002302488700003002511700001802541700002102559856008202580 2016 eng d00aFunctional Importance of a Proteoglycan Co-Receptor in Pathologic Lymphangiogenesis0 aFunctional Importance of a Proteoglycan CoReceptor in Pathologic c05/2016 a210-2210 v1193 aRationale: Lymphatic vessel growth is mediated by major pro-lymphangiogenic factors such as VEGF-C and -D, among other endothelial effectors. Heparan sulfate is a linear polysaccharide expressed on proteoglycan core proteins on cell-membranes and matrix, playing roles in angiogenesis, although little is known regarding any function(s) in lymphatic remodeling in vivo. Objective: To explore the genetic basis and mechanisms whereby heparan sulfate proteoglycans mediate pathologic lymphatic remodeling. Methods and Results: Lymphatic endothelial deficiency in the major heparan sulfate biosynthetic enzyme N-deacetylase/N-sulfotransferase-1 (Ndst1; involved in glycan-chain sulfation) was associated with reduced lymphangiogenesis in pathologic models, including spontaneous neoplasia. Mouse mutants demonstrated tumor-associated lymphatic vessels with apoptotic nuclei. Mutant lymphatic endothelia demonstrated impaired mitogen (Erk) and survival (Akt) pathway signaling as well as reduced VEGF-C mediated protection from starvation-induced apoptosis. Lymphatic endothelial specific Ndst1 deficiency (in Ndst1f/fProx1+/CreERT2 mice) was sufficient to inhibit VEGF-C dependent lymphangiogenesis. Lymphatic heparan sulfate deficiency reduced phosphorylation of the major lymphatic growth receptor VEGFR-3 in response to multiple VEGF-C species. Syndecan-4 was the dominantly expressed heparan sulfate proteoglycan in mouse lymphatic endothelia, and pathologic lymphangiogenesis was impaired in Sdc4(-/-) mice. On the lymphatic cell surface, VEGF-C induced robust association between syndecan-4 and VEGFR-3 which was sensitive to glycan disruption. Moreover, VEGFR-3 mitogen and survival signaling was reduced in the setting of Ndst1 or Sdc4 deficiency. Conclusions: These findings demonstrate the genetic importance of heparan sulfate and the major lymphatic proteoglycan syndecan-4 in pathologic lymphatic remodeling. This may introduce novel future strategies to alter pathologic lymphatic-vascular remodeling.10aendothelial cell growth10aglycosaminoglycan10alymphatic capillary10aProteoglycan10avascular endothelial growth factor receptor1 aJohns, Scott, C.1 aYin, Xin1 aJeltsch, Michael1 aBishop, Joseph, R.1 aSchuksz, Manuela1 aGhazal, Roland, El1 aWilcox-Adelman, Sarah, A.1 aAlitalo, Kari1 aFuster, Mark, M. uhttp://circres.ahajournals.org/content/early/2016/05/25/CIRCRESAHA.116.30850403585nas a2200589 4500008004100000022001400041245010200055210006900157260001500226300001400241490000800255520182400263653001202087653001802099653002902117653001302146653003102159653002202190653002802212653004602240653002902286653002702315653001302342653002002355653001702375653001102392653003302403653002202436653001502458653000902473653002102482653001302503653001402516653002002530653004302550653002402593653001702617653003002634653004102664653001402705653002302719100001702742700002602759700002002785700002102805700002602826700001802852700001802870700002702888700002002915856006002935 2015 eng d a1524-457100aFunctional Dissection of the CCBE1 Protein: A Crucial Requirement for the Collagen Repeat Domain.0 aFunctional Dissection of the CCBE1 Protein A Crucial Requirement c2015 May 8 a1660-16690 v1163 a

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.

10aAnimals10aBinding Sites10aCalcium-Binding Proteins10aCollagen10aCraniofacial Abnormalities10aEndothelial Cells10aEpidermal Growth Factor10aGene Expression Regulation, Developmental10aGene Knock-In Techniques10aGenital Diseases, Male10aGenotype10aGestational Age10aHEK293 Cells10aHumans10aLymphangiectasis, Intestinal10aLymphatic Vessels10alymphedema10aMice10aMice, Transgenic10aMutation10aPhenotype10aProtein Binding10aProtein Interaction Domains and Motifs10aSignal Transduction10aTransfection10aTumor Suppressor Proteins10aVascular Endothelial Growth Factor C10aZebrafish10aZebrafish Proteins1 aRoukens, Guy1 aPeterson-Maduro, Josi1 aPadberg, Yvonne1 aJeltsch, Michael1 aLeppänen, Veli-Matti1 aBos, Frank, L1 aAlitalo, Kari1 aSchulte-Merker, Stefan1 aSchulte, Dörte uhttp://circres.ahajournals.org/content/116/10/1660.long00713nas a2200181 4500008004100000245007900041210006900120260007600189520012600265653001200391653001000403653002200413653001500435653001100450653001200461100001500473856004300488 2014 eng d00aFrom molecular biology to a causal treatment of lymphatic system disorders0 aFrom molecular biology to a causal treatment of lymphatic system aHalle (Saale), GermanybDeutsche Gesellschaft für Lymphologiec10/20143 aOriginal Conference Abstracts (in German)10aADAMTS310aCCBE110aLymphangiogenesis10alymphedema10aVEGF-C10aVEGFR-31 aJeltsch, M uhttps://jeltsch.org/abstract-Halle201401805nas 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/1681612102100nas a2200145 4500008004500000245004500045210004300090260003700133490002000170520168300190653001101873653001101884100002101895856003801916 1997 Engldsh 00aFunctional Analysis of VEGF-B and VEGF-C0 aFunctional Analysis of VEGFB and VEGFC aHelsinkibUniveristy of Helsinki0 vMaster's Thesis3 aVascular endothelial growth factor (VEGF) is an important regulator of endothelial cell proliferation and migration dur- ing embryonic vasculogenesis and angiogenesis as well as in pathological angiogenesis. The recently cloned new factors structurally homologous to VEGF were designated as VEGF-B/VRF and VEGF-C/VRP. The receptor for VEGF-B is unknown. VEGF-C is the ligand for FLT4, a receptor tyrosine kinase whose expression becomes restricted largely to lymphatic endothelia during development and that is related to VEGF receptors FLT1 and KDR. In this study keratin 14-promoter-directed VEGF-C overexpression in the basal epidermis of transgenic mice was capa- ble of promoting an abundant growth of extensive lymphatic-like vessel structures in the dermis, including large vessel lacunae resembling in their histopathology the human condition known as lymphangioma. Thus, VEGF-C appears to induce selective angiogenesis of the lymphatic vessels in vivo. In contrast, preliminary data on mice, which overexpress VEGF-B under the same promoter, does not yet allow us draw any conclusions about its possible biological function. Recombinant biologically active human VEGF-C was produced using the baculovirus system. Unpurified and purified VEGF-C were used to confirm the interaction of VEGF-C with KDR, a fact recently missed by others. The recombinant protein is going to be used in a large number of future experiments. The production of VEGF-B seems to be intrinsically difficult in non-mammalian cells. Although quantitatively satisfying results could not be obtained yet, the purified growth factor will be used in experiments to identify its receptor.10aVEGF-B10aVEGF-C1 aJeltsch, Michael uhttp://urn.fi/URN:NBN:fi-fe977347