1. 15. ročník kongresu s medzinárodnou účasťou
Slovenská spoločnosť pre laboratórnu medicínu Slovenská zdravotnícka univerzita Slovenská lekárska spoločnosť Slovenská komora iných zdravotníckych pracovníkov
LABORATÓRNA MEDICÍNA 2014 
15. ročník kongresu s medzinárodnou účasťou
Kongres konaný pod záštitou EUMS – European Union of Medical Specialists
Možnosti bunkovej terapie u pacientov s ischemickou chorobou srdca
Ján Kyselovič, Peter Musil
Katedra farmakológie a toxikológie, Farmaceutická fakulta
Ústav farmakológie a klinickej farmakológie, Lekárska fakulta
Univerzita Komenského v Bratislave
Martin 29. novembra 2014

2. Background
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 Medicine
Materials Science
Stem cells are the seeds of tissue repair and regeneration and a promising source for novel therapies.

3. Introduction
Elena Matsa & Chris Denning, J. of Cardiovasc. Trans. Res. (2012) 5:581–592

4. Intramyocardial CD133+ Cell Therapy

5. Introduction

6. Introduction
CABG
CABG +CD133+

7. Stem Cell CD133+ preparation and transplamtation

8. Stem Cell CD133+ preparation and transplamtation

9. Meta/analysis of Cell Therapy Clincial Trails

10. Ten years of Cardiac Regeneration
Perception of the General Cardiology Community

11. Mechanisms of cardiac regeneration after cell therapy?

12. Ten years of Cardiac Regeneration
Current status
Ten years of Cardiac Regeneration
Perception of the General Cardiology Community
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

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

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

15. Regulation of differentiation
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.

16. Regulation of differentiation endogenic factors

17. Open questions of cell therapy
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.

18. Molekulárna farmakológia Ca antagonistov
SHRSP NaCl +
lacidipine
NaCl
0, mg/kg/day
140
160
180
200
220
240
260
BP mmHg ns
SHR-SP + Lacidipine
H2O *
Tlak krvi 7 8 9 10 11 12
SHRSP NaCl 25 50 75
100
% prežívanie potkanov
týždne liečby
Hmotnosť srdca
NaCl
0, mg/kg/day
3,0
3,2
3,4
3,6
3,8
4,0
4,2
Heart/body *
SHR-SP + Lacidipine
H2O

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

20. Isoproterenol-induced heart failure – Growth factors
IL-1b
Cardiotropin -1
TNFa
HGF
LIF
VEGF
SDF-1a

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

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

23. Results – Human Explanted Heart
Hierarchical clustering:
Spearman’s correlation matrix
of all data (before selection)

24. Regulation of differentiation – small molecules
Qian et al,. Neth Heart J, 2008; 16:

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

26. Summary of Pharmacological intervention
Stem cells:
the modification of the 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

27. Co authors
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 Hrabovská, PharmD, PhD
Marek Matus, PharmD, PhD
Assistant of professors
Peter Musil, PhD Adrea Gazova PharmD, PhD
Ondrej Sprušansky , PhD Adela Čorejová, Pharm, PhD.
Dana Kučerová, PhD Miriam Verčinská, PharmD. PhD
PhD students
Lenka Pivackova Hanka Cernecka Lucia Mesarosova
Lea Dugovicova Katarina Tucekova Kristina Stahlova
Bea Gajdacova Jana Mlynarova Matej Kucera
Gabriel Doka Lucia Mensikova
DPT
FaF UK Bratislava
MUDr. Michal Hulman
doc. MUDr.. Eva Goncalvesova

28. Ďakujem za pozornosť