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Monosti bunkovej terapie u pacientov s ischemickou chorobou srdca Jn Kyselovi, Peter Musil Katedra farmakolgie a toxikolgie, Farmaceutick fakulta stav farmakolgie a klinickej farmakolgie, Lekrska fakulta Univerzita Komenskho v Bratislave Slovensk spolonos pre laboratrnu medicnu Slovensk zdravotncka univerzita Slovensk lekrska spolonos Slovensk komora inch zdravotnckych pracovnkov LABORATRNA MEDICNA 2014 15. ronk kongresu s medzinrodnou asou Kongres konan pod ztitou EUMS European Union of Medical Specialists Martin 29. novembra 2014

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Regenerative Medicine is the process of creating living, functional tissues to repair or replace tissue or organ function lost due to age, disease, damage, or congenital defects. Medicine Bioengineering Chemical Engineering Developmental Biology Molecular Biology Drug Delivery Physiology Biochemistry Genomics Proteomics Nanotechnology Regenerative MedicineMaterials Science Background Stem cells are the seeds of tissue repair and regeneration and a promising source for novel therapies.

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Elena Matsa & Chris Denning, J. of Cardiovasc. Trans. Res. (2012) 5:581592 Introduction

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Intramyocardial CD133+ Cell Therapy

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Introduction

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CABGCABG +CD133+

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Stem Cell CD133+ preparation and transplamtation

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Meta/analysis of Cell Therapy Clincial Trails

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Ten years of Cardiac Regeneration Perception of the General Cardiology Community

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Mechanisms of cardiac regeneration after cell therapy ?

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Clinical trials demonstrate excellent safety profile Yet, Efficacy is modest and inconsistent across the trials with inter-trial heterogeneity Promise remains controversial and does not meet clinical expectations Ten years of Cardiac Regeneration Perception of the General Cardiology Community Current status

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Pluripotency of Hematopoietic and Stromal Adult Stem Cell Trans-differentiation or plasticity of adult stem cells Specific induction

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Jonathan A. Epstein N A T U R E | V O L 4 7 3 | 1 9 M A Y 2 0 1 1 Michael A. Laflamme1 & Charles E. Murry Regulation of cardiac differentiation

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Cells can interact with each other and with their environment This interaction turns specific signaling paths ON or OFF At the present time we do not fully understand the mechanisms by which any mammalian gene is switched on and off during differentiation and development. This involves the interaction of key cis-elements e.g. enhancers, promoters and silencers with trans-acting proteins (e.g. transcription factors, coactivators, adaptors and chromatin modifiers) that give rise to changes in gene expression, It is associated with changes in epigenetic modification - methylation, replication, histone acetylation and nuclear position. The multiple growth factors and chemokines that have been shown to promote endogenous stem cell mobilization, proliferation, and homing to the injured myocardium, the development of growth factor/chemokine cocktails seems an attractive therapeutic approach for enhancing cardiac regeneration. Regulation of differentiation

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Regulation of differentiation endogenic factors

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However, apart from hematopoietic stem cell (HSC) transplantation, essentially all other stem cell treatments remain experimental. Numerous challenges, technical and clinical barriers must be overcome before novel stem cell therapies can achieve meaningful clinical impact. In recent years, unproven therapies have been widely practiced outside the standard clinical trial network, threatening the cause of legitimate clinical investigation. High hopes have inspired numerous clinical trials, but it has been difficult to obtain unequivocal evidence for robust clinical benefit. When clinical investigation proceeds largely empirically, and without a deeper understanding therapeutic mechanisms, it is difficult to reformulate therapeutic strategies after clinical failures. Open questions of cell therapy

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Molekulrna farmakolgia Ca antagonistov SHRSP NaCl + lacidipine 789101112 0 SHRSP NaCl 25 50 75 100 % prevanie potkanov tdne lieby NaCl 0,3 1 3 mg/kg/day 140 160 180 200 220 240 260 BP mmHg ns SHR-SP + Lacidipine H2OH2O *** Tlak krvi NaCl 0,3 1 3 mg/kg/day 3,0 3,2 3,4 3,6 3,8 4,0 4,2 Heart/body * SHR-SP + Lacidipine H2OH2O *** Hmotnos srdca

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Pharmacological intervention to gene expression DFT CU Bratislava Kyselovic et al. J Hypertension, 1998 Krenek, Kyselovic et al., Hyperten 2001

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Isoproterenol-induced heart failure Growth factors Cardiotropin -1 IL-6 LIF IL-1 TNF HGF VEGF SDF-1

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Isoproterenol-induced heart failure Transcription factors c-fos STAT 3 DPT FaF UK Bratislava Krenek, et al., Eur J Heart Failure. 2009, (11), 140

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Pharmacological intervention to gene expression Masart et al. Hypertension. 1999 Kyselovic et al, J Hyperten. 2004 Simko et al. Pharmazie. 1999a, b, 2001 Kyselovic et al. JPEP. 2005 Krenek et al. Hypertension. 2001 Cagalinec et al. JCP. 2006 Kyselovic et al. Br. J. Pharmacol. 2001 Krenek et al.Cnd J Physiol. 2006 Bcl-2Bax 8h control ISO 2days 8 days Bacharova et al. Clin. Exp. Hyperten.2004, 2005 Bacharova et al.Electrocardioly 2008 Mackoviov et a.l- Can.J. Pharmacol. 2011 Krenek et al. EU J Hert failure, 2009 Klimas et al., Basic Clin Pharmacol, 2012Jankyova et. al., Pathol Res Prac. 2012

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Hierarchical clustering : Spearmans correlation matrix of all data (before selection) Results Human Explanted Heart

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Regulation of differentiation small molecules Qian et al,. Neth Heart J, 2008; 16: 337-343

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Open questions of genomics 6th September 2012 - The ENCODE (Encyclopedia of DNA Elements) Numerous challenges and technical barriers must be overcome before novel stem cell therapies can achieve meaningful clinical impact

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Summary of Pharmacological intervention Stem cells: the modification of t he multiple growth factors and chemokines that have been shown to promote endogenous stem cell mobilization, proliferation, and homing to the injured myocardium the interaction with gene expression via enhancers, promoters and silencers, transcription factors, coactivators, adaptors and chromatin modifiers the association with changes in epigenetic modification - methylation, replication, histone acetylation and nuclear position. Myocardium: pharmacological treatments target disease mechanisms at the molecular/cellular and whole-organ level. it is also complex of genetic, neurohumoral, immunological a metabolic changes modify function of cardiomyocytes, fibroblast or interstitium. Patient: Therapeutic, adverse and unwanted effects organs systems and whole body

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Department of Pharmacology and Toxicology, Faculty of Pharmacy Comenius University Bratislava, Slovakia Associate professors: Peter Krenek PharmD, PhD Peter Ochodnicky PharmD, PhD Jan Klimas, PharmD, PhD Adriana Adameova PharmD, PhD Anna Hrabovsk, PharmD, PhD Marek Matus, PharmD, PhD Assistant of professors Peter Musil, PhD Adrea Gazova PharmD, PhD Ondrej Spruansky, PhDAdela orejov, Pharm, PhD. Dana Kuerov, PhDMiriam Verinsk, PharmD. PhD PhD students Lenka PivackovaHanka CerneckaLucia Mesarosova Lea DugovicovaKatarina TucekovaKristina Stahlova Bea GajdacovaJana MlynarovaMatej Kucera Gabriel DokaLucia Mensikova Co authors DPT FaF UK Bratislava MUDr. Michal Hulman doc. MUDr.. Eva Goncalvesova

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akujem za pozornos