Dual stem cell comprising administration of G-CSF and Sitagliptin improves stem cell homing, cardiac function and survival after MI in mice
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1 Dual stem cell comprising administration of G-CSF and Sitagliptin improves stem cell homing, cardiac function and survival after MI in mice Hans D Theiss, Lisa Krieg, Marcus Vallaster, Josef Mueller- Hoecker and Wolfgang-M Franz Medical Department I University of Munich Grosshadern Germany
2 Disclosure This work was funded by Deutsche Forschungsgemeinschaft FöFoLe-Programm of the Ludwig-Maximilians University The Ludwig Maximilians University is the holder of the patents Use of G-CSF for Treating Ischemia (EP and US 6/514,474) and Remedies for Ischemia (EP27/3272 and US 6/792,943).
3 Hypothesis of Dual stem cell therapy G-CSF Genetic DPP-IV inhibition Mobilization of stem cells from bone marrow into peripheral blood KP SDF => Stabilization of SDF-1 Improved cardiac homing of CXCR-4 positive stem cells towards SDF-1 gradient => enhanced cardiac function and survival *Zaruba M, *Theiss H et al & Franz WM. Cell Stem Cell 29
4 Hypothesis of Dual stem cell therapy G-CSF Sitagliptin Mobilization of stem cells from bone marrow into peripheral blood KP SDF Clinically admitted inhibitor of DPP- IV => stabilized SDF-1 Improved cardiac homing of CXCR-4 positive stem cells towards SDF-1 gradient Effects of dual stem cell therapy using Sitaglitpin after LAD-ligation in mice
5 Relative units Oral application of Sitagliptin leads to sufficient levels in peripheral blood after MI using mass spectrometry Saline Sitagliptin 5 mg/kg KG MI+Sitagliptin 5 mg/kg KG Sitagliptin 5 mg/kg KG MI + Sitagliptin 5 mg/kg KG 5
6 Serum: DPP-IV activity in nmol/ml*min Sitagliptin reduces activity of DPP-IV in a dose dependant manner Sitagliptin 5 mg/kg KG Sitagliptin 5 mg/kg KG Sitagliptin 5 mg/kg KG Saline
7 Cells in % Cells in % Cells in % Cells in % Sitagliptin +/- G-CSF stimulate cardiac homing of mobilized stem cells CD 45+/CD 34+/c-kit+ (heart) p=,1 p=,6 p=,2 MI+G-CSF MI+Sitagliptin+G-CSF MI+Saline MI+Sitagliptin CD 45+/CD 34+/CD 31+ (heart) p=,7 p=,2 p=,3 1 1 n=9 n=7 n=9 n=1 n=9 n=7 n=8 n=11 p=,35 CD 45+/ CD34+/ CXCR4+ (heart) p=,6 Sca-1 + / c-kit + / lin - (heart) 8 p=,4 p=,7 p=,3,8 p=,9 p=,9 p=,5 7,7 6, MI+G-CSF MI+Sitagliptin+G-CSF MI+Saline MI+Sitagliptin,5,4,3,2 MI+G-CSF MI+Sitagliptin MI+Saline MI+Sitagliptin 1,1 n=6 n=7 n=7 n=8 p=,3 n=11 n=7 n=8 n=6 p=,15
8 Cells in % Sitagliptin +/- G-CSF stimulate resident cardiac stem cells CD 45 - / CD 34 - / Sca-1 + (heart) p=,9 p=, MI+G-CSF MI+Sitagliptin+G-CSF MI+Saline MI+Sitagliptin n=6 n=7 n=6 n=8 p=,6
9 Cells in % Cells in % Sitagliptin stimulates stem cell homing in a dose dependant manner CD 45+/CD 34+/CD 31+ (heart) CD 45+/CD 34+/Sca-1+ (heart) 4 3,5 3 2,5 2 1,5 1,5 p=,3 p=,2 n.s. MI+Sitagliptin 5mg/kg KG MI+Sitagliptin 5mg/kg KG MI+Sitagliptin 5mg/kg KG MI+Saline p=,1 p=,1 n.s. n=11 n=6 n=4 n=8 n=11 n=6 n=4 n=8
10 Infarct size in % Infarct size in % Sitagliptin +/- G-CSF reduces infarct size 3 days after MI Size of infarction (day 6) Size of infarction (day 3) 5 45 n.s. n.s. n.s. 4 p=,27 p=,7 p=, n=7 n=7 n=6 n=5 MI+G-CSF MI+Sitagliptin+G-CSF MI+Saline MI+Sitagliptin n=6 n=7 n=5 n=11
11 CD31+ capillaries per HPF Sitagliptin +/- G-CSF increases neovascularization in the border zone (immunohistochemistry of CD31+ capillaries) 8 p=,27 p=,2 p=, G-CSF Sitagliptin + G-CSF Saline Sitagliptin Saline n=3 n=6 n=3 n=4 p=,8 G-CSF+Sitagliptin
12 Apoptotic CMs/HPF Sitagliptin + G-CSF reduce apoptosis (TUNEL assay) Sitagliptin + G-CSF Saline
13 Sitagliptin + G-CSF enhance cardiac function 3 days after MI (assessed by Millar-tip catheter) MI MI + G-CSF MI + G-CSF +Sitagliptin MI + Sitagliptin
14 ml/min mmhg/ml bpm Ejection fraction in % Sitagliptin + G-CSF enhance cardiac function 3 days after MI (assessed by Millar-tip catheter) Heart Rate (day 3) Ejection Fraction (day 3) 6 n.s. n.s. n.s. 7 p=,4 p=,9 p=,3 5 4 MI+G-CSF MI+Sitagliptin+G-CSF MI+G-C MI+Sita 3 2 MI+Saline MI+Sitagliptin 3 2 MI+Sali MI+Sita 1 n=6 n=7 n=6 n=7 Cardiac Output (day 3) 1 n=5 n=6 n=5 n=7 Arterial Elastance (day 3) 6 p=,31 p=,44 p=,14 2 p=,44 p=,46 p=, MI+G-CSF MI+Sitagliptin+G-CSF MI+Saline MI+Sitagliptin MI+G- MI+Sit MI+Sa MI+Sit n=6 n=6 n=5 n=6 n=6 n=6 n=5 n=6
15 Sitagliptin + G-CSF improve survival 3 days after MI
16 Effects specific to the SDF1-CXCR4-axis? => antagonization of CXCR-4 using AMD31 CXCR-4 antagonization => inhibited stem cell homing AMD31
17 Sitagliptin + G-CSF improve survival 3 days after MI => reversed by CXCR4- antagonization using AMD31
18 Conclusion G-CSF Sitagliptin Mobilization of stem cells from bone marrow into peripheral blood KP SDF Improved stem cell homing Inhibition of DPP- IV => stabilized SDF-1 Cardiac homing of CXCR-4 positive stem cells towards SDF-1 gradient Resident cardiac stem cells Cardiac remodeling Neovascularization Cardiac function Survival specific to the SDF1-CXCR4-axis
19 Transfer from bench to bedside: SITAGRAMI-Trial Safety and efficacy of SITAgliptin plus GRanulocyte-colony-stimulating factor in patients suffering from Acute Myocardial Infarction EudraCT-Nummer: G-CSF Sitagliptin Theiss HD et al. & Franz WM Int J Card 21
20 ... Thank you for your attention... Medical Department I University of Munich Grosshadern Prof. Dr. Wolfgang-M. Franz Lisa Krieg Markus Vallaster Dr. Marc-Michael Zaruba Dr. Christoph Brenner Rebekka Fischer Judith Arcifa, MTA Barbara Markieton, BTA Prof. Dr. Gerhard Steinbeck Institute of Pathology (University of Munich ) Prof. Dr. Josef Müller-Höcker Dr. Gerald Assmann
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