Ontstekingscellen the good and the bad guy

Endogline heterozygoten herstellen slecht na schade MNC isolation Myocardial infarction We recently showed that MNCs derived from HHT-1 patients are impaired in their capacity to home to an ischemic area and stimulate vessel formation. Moreover, injection of healthy MNCs in Eng+/- mice was sufficient to restore vessel formation and to improve heart function defects associated with reduced levels of endoglin but MNCs from HHT1 patients were not, demonstrating that the etiology of HHT1 is possibly associated with a defect in the ability of MNCs to repair local vessel damage. In our study, we demonstrated that MNC-derived from HHT1 patients have a reduced ability to accumulate to the infarct zone in vivo and to stimulate vessel formation in mice that had undergone experimental myocardial infarction Mouse-cardio-MRI Laake LW van et al, Circulation 2006 Nov 21;114(21):2288-97

Wat gebeurd er na een hartaanval Tuesday, April 04, 2017

Een ontsteking en herstel fase Tuesday, April 04, 2017

Wat voor ontstekingscellen zijn er ? Tuesday, April 04, 2017

HHT-1 MNCs: minder vaten en minder cellen Control HHT-1 Control HHT-1 PBS # Human-UEA+ cells We recently showed that MNCs derived from HHT-1 patients are impaired in their capacity to home to an ischemic area and stimulate vessel formation. Moreover, injection of healthy MNCs in Eng+/- mice was sufficient to restore vessel formation and to improve heart function defects associated with reduced levels of endoglin but MNCs from HHT1 patients were not, demonstrating that the etiology of HHT1 is possibly associated with a defect in the ability of MNCs to repair local vessel damage. In our study, we demonstrated that MNC-derived from HHT1 patients have a reduced ability to accumulate to the infarct zone in vivo and to stimulate vessel formation in mice that had undergone experimental myocardial infarction Laake LW van et al, Circulation 2006 Nov 21;114(21):2288-97

Waarom zijn er minder cellen in het hart Circulating MNCs attracted to injury Chemokine receptors Post, CVR, 2010 Tuesday, April 04, 2017

Aantal monocyten in het bloed is gelijk Post, CVR, 2010 Tuesday, April 04, 2017

Niveau van CD26/DPP-IV is hoog Post, CVR, 2010 Tuesday, April 04, 2017

CD26/DPP-IV remt homing van cellen naar schade 1 day post-MI ↑SDF-1α MNC Homing + CXCR4+ MNC - Homing CXCR4+ CD26+ SDF-1 Increased after MI CXCR4: receptor for SDF-1 CD26= Dipeptidyl peptidase:cleaves SDF-1 and internalize CXCR4 Post, CVR, 2010 Tuesday, April 04, 2017

CD26/DPP-IV remt migratie in vitro Decrease in migration capacity of HHT1-MNC Expression of CD26 on MNC of HHT patients was inversely correlated with migration capacity Post, CVR, 2010 Tuesday, April 04, 2017

CD26 remming stimuleert migratie in vitro Post, CVR, 2010 Tuesday, April 04, 2017

CD26/DPP-IV inhibitie stimuleert migratie van gezonde en HHT cellen Tuesday, April 04, 2017

Systemische CD26/DPP-IV remming stimuleert homing van circulerende monocyten Sitagliptin MONOCYTEN Tuesday, April 04, 2017

Macrofagen en weefsel herstel Na schade zijn er 2 fase van macrofaag influx Phase I: vroege invasie van inflammatie macrofagen (M1)  Degradatie van beschadigd weefsel Phase II: Niet inflammatoire macrofagen (M2)  Regeneratieve capacity 100 80 60 40 20 0 0 2 4 6 8 Days after injury % maximum Phase I Phase II M1 M2 Inflammatory Regenerative Tuesday, April 04, 2017

Wat doen M2 macrofagen Tuesday, April 04, 2017

Maturatie van macrofagen +/- GM-CSF (Macrophage differentiation) Inflammatie status Based on expression of Ly6C on macrophages M1: Inflammatory mph M2: Anti-inflammatory mph Tuesday, April 04, 2017

Maturatie van macrofagen in vivo Spleen Muscle Eng+/+ Muscle Eng+/- Monocytes M2 macrophages CD11b / DAPI (MR = Mannose receptor or CD206) MR / DAPI Tuesday, April 04, 2017

Hoe beinvloed TGFb macrofaag maturatie? M1 Inflammatory M2 Regenerative ? Low CD206 expression High CD206 expression - GM-CSF - GM-CSF + TGFβ 24h - GM-CSF + TGFβ 4 days Tuesday, April 04, 2017

TGFb speelt met de balans tussen M1 en M2 Regenerative status Based on expression of Mannose Receptor (CD206) on macrophages Tuesday, April 04, 2017

Dus…. Eng+/- Tuesday, April 04, 2017

Vinden we dit nu alleen in muis? Human Monocyte/macrophage markers: 1. CD14 ++ / CD16-  Immature monocytes 2. CD14+/ CD16+  Pro-inflammatory macrophages 3. CD14++ / CD16+  Regenerative macrophages 1. Immature 2. Pro-inflammatory 3. Regenerative Tuesday, April 04, 2017

Conclusie Ontstekingscellen voor herstel TGFb/Endoglin is crucial voor bloedvatvorming TGFb/Endoglin is homen van circulerende cellen en vorming van herstellende macrofagen. Tuesday, April 04, 2017

Toekomst Tuesday, April 04, 2017

Thanks to: UMCU, Experimental Cardiology LUMC Dept. Mol. Cell Biol., Simone Post Sandra van den Broek Imo Hoefer LUMC Dept. Mol. Cell Biol., Anke Smits Wineke Bakker Peter ten Dijke Dept. Anatomy & Embryology, Christine Mummery Hubrecht laboratory Linda van Laake Franck Lebrin St. Antonius Hospital, Nieuwegein Hans Mager Repke Snijder Cees Westermann All patients en healthy controls for their blood donations