Bruno Peault – pericytes and MSCs • MSCs, MAPCs, MDSCs, ADSCs – all claimed to be multipotent for mesodermal cells • Superficial similarities with pericytes – also in many tissues • CD146/NG2/PDGFRB • Pericytes express MSC markers in situ and in culture • Pericytes (from muscle AND elsewhere) can regenerate muscle • Single cell-derived cultures are multipotent • MSCs and pericytes are both immunosuppressive in myocardium i.e. inhibit MF infilt. • Pericytes promote hematopoiesis in culture •Ecs inhibit pericyte -> fat differentiation • There are also different MSC-like cells in adventitium – different markers Helmut Augustin – EC quiescence • Postnatal retina Tie2 phalanx>stalk>tip Ang2 tip>stalk>phalanx • Ang1-Tie2 axis N for quiescence (Q) • Ang2 Tg phenocopies Ang1 KO • Loss of Q -> incr. Ang2 expr. In EC • Ang2 rises during melanoma progression • Ang2KO Normal devel., v.low response in Miles and ThG assays Facilitator of TEM • Ang2 KO NG2 and desmin coverage up, aSMA coverage down • Phalanx vessels pericytes NG2 hi, desmin hi, aSMA lo • Ang2 lof -> quiescence • Ang2 gof -> activation Helmut Augustin – Part II - CCM • CCM – loss of quiescence (Q) • oxpr CCM1 -> reduced sprouting & migration reduced apoptosis & proliferation, PAkt up • CCM1 KD -> increased sprouting & migration increased apoptosis & proliferation • CCM1 oxpr – Dll4, Hey1/2 up ie N signaling up • CCM1KD – Dll4, Hey1/2 down • DAPT blunts CCM1 effects CCM1 N Ang Erk Akt proliferation • Can reduce xs angiogenesis with sorafenib (broad tyrK inhibitor) Amin Ghabrial • Junctions between seamless and seamed vessels – HOW? • Heart-of-glass – CCM1/2/3 • D.mel has CCM3 – it binds GC kinaseIII – KD of either -> junction defects Suk-Won Jin - VEGF or BMP? • • • • • • VEGF MO blocks arterial sprouts but not venous BMP2a/2b expr. in V; BMP2b expr. around V Oxpr Noggin3 interferes selectively with CV plexus Oxpr BMP2b -> incr. V sprouting but no response in arteries BMP2a MO -> plexus defects flk1-DN BMPR1 inhibits V connections (no effect in DA) i.e. R is N in ECs • BMP effects are not VEGF-dependent • Mouse EBs – noggin inhibits ang. while BMP2 enhances Christer Betsholtz • • • • Wallgard et al ATVB 2008 – 58 EC-specific genes a couple are Adh. GPCRs (Eldt1, GPR116) – both KOs viable Eldt1 KO – nothing obvious GPR116 KO – capillary dilation, lung enlargement, MF infilt. • SphR1 (Edg1) – KO shows xs sprouting and glomeruli around aorta Nick Gale – organ-specific vasculature • TEM5/GPR124 – KO shows forebrain hemorrhage sim. to avKO but not quite the same (also less severe in SC) • pericytes are around tufts but are aSMA+ • GPR124 expr in ECs and pericytes. • LCM shows tufts expr. Hypoxia, TGFB, hemorrhage, EC, peri genes Mark Kahn • • • • • • • • • • • • ITAMs -> Syk-> SLP76 -> PLCg SLP76 KO – vessel-lymphatic connections. No SLP76 in LECs BMT recapitulates defect – can rescue with GATA1-GFP-SLP Use Vav-Cre to turn on YFP and KO SLP76 – no contrib. of EC to lymphatics Podoplanin +/- enhances Vav-Cre SLP76 cKO PDPN-Fc binds platelets PF4Cre SLP76 cKO – reproduces phenotype – SO PLATELETS CLEC2 is podoplanin receptor – restricted to MKs CLEC2 KO mice reproduce phenotype Can activate WT but not CLEC-KO plts with PDPN-Fc See small plt aggs on neoLEC in WT – plts bind LEC, not BEC No obvious effects of plts on LEC Aniket Gore - Wnt • down the tubes mutant – no ISVs – R-spondin – enhances Wnt FU-FU-TSP - mutant is S>L in TSP • MO phenocopies mutant • Interacts with Kremen (KR-WSC-CUB-TM) • Sign. Looks autocrine • VEGFC is down in mutant • Can restore by inh. GSK3 or by VEGFC mRNA R-spondin Dkk Kremen LRP5/6 VEGF-C Wnt Frz Kareen Kreutziger • • • • hESC-derived cardiomyocyte patches tri-cell constructs – CMyo + HUVEC + MEF/HDF/hMSC two MSC lines differ in their effectiveness difference is in N/DLL4 and some other N/delta Ying Wang - moesin • • • • • MoesinP during EC migration and tube formation In Zf ISV moesin enriched at lumenal membrane and vesicles Colocation of moesin and VEcad is not complete Vecad MO -> no lumens – still lots of vacuoles and moesin Moesin also N for ISV formation and Vecad not org to CCJs or vacuoles