Projects / Programmes source: ARIS

Identification of Subpopulations of Haematopoietic Stem Cells and Their Impact on Therapy of Dilated Cardiomyopathy

Research activity

Code Science Field Subfield
3.06.00  Medical sciences  Cardiovascular system   

Code Science Field
B530  Biomedical sciences  Cardiovascular system 

Code Science Field
3.02  Medical and Health Sciences  Clinical medicine 
Cardiomyopathy, Heart failure, Stem cells, Very Small Embryonic-like Stem Cells (VSEL)
Evaluation (rules)
source: COBISS
Researchers (14)
no. Code Name and surname Research area Role Period No. of publicationsNo. of publications
1.  19331  PhD Tina Cirman  Microbiology and immunology  Researcher  2013 - 2016  51 
2.  20611  MSc Marko Cukjati  Microbiology and immunology  Researcher  2013 - 2016  91 
3.  20605  PhD Tadeja Dovč Drnovšek  Biochemistry and molecular biology  Researcher  2013 - 2016  71 
4.  38276  Katerina Jazbec  Microbiology and immunology  Researcher  2016  49 
5.  30968  PhD Mojca Jež  Microbiology and immunology  Researcher  2013 - 2016  101 
6.  21525  PhD Polona Klemenc  Microbiology and immunology  Researcher  2013 - 2016  40 
7.  21227  PhD Metka Krašna  Biotechnology  Researcher  2013 - 2016  49 
8.  21228  PhD Elvira Maličev  Microbiology and immunology  Researcher  2013 - 2016  142 
9.  32517  PhD Renata Okrajšek  Cardiovascular system  Researcher  2013 - 2016  101 
10.  28021  PhD Gregor Poglajen  Cardiovascular system  Researcher  2013 - 2016  326 
11.  28510  PhD Jasmina Živa Rožman  Human reproduction  Researcher  2014 - 2015  44 
12.  12336  PhD Primož Rožman  Microbiology and immunology  Head  2013 - 2016  476 
13.  19345  PhD Miran Šebeštjen  Cardiovascular system  Researcher  2013 - 2016  260 
14.  18826  PhD Bojan Vrtovec  Medical sciences  Researcher  2013 - 2016  481 
Organisations (2)
no. Code Research organisation City Registration number No. of publicationsNo. of publications
1.  0311  Blood Transfusion Centre of Slovenia  Ljubljana  5053960  1,733 
2.  0312  University Medical Centre Ljubljana  Ljubljana  5057272000  74,860 
Identification of Subpopulations of Hematopoietic Stem Cells and Their Impact on Therapy of Dilated Cardiomyopathy   INTRODUCTION: Currently, heart diseases can be treated by transplantation of autologous hematopoietic stem cells (HSCs). The principle of this treatment is based on the hypothesis that HSCs after transplantation into the heart can transform into vascular cells or cardiomyocytes (Orlic 2001). HSCs express markers CD34, CD38, CD90, and CD133 ( Meregalli 2010). In adults, stem cells with embryonic characteristics have been reported (very small embryonic-like stem cells - VSEL) (Kucia 2006). They probably take part in generation of vessels, myogenesis and prevention of scarring (Wojakowski 2011), however the only clinical study performed did not confirm their clinical effectiveness (Tendera 2009). In therapeutic products VSELs are very rare, which is also due to their loss during isolation (Bhartiya 2011). Dilated cardiomyopathy (DCM) is the main cause of heart defects. Currently there is no causative treatment available, except transplantation of an allogeneic heart. One of the newer means is cell therapy by using autologous HSCs from the bone marro (BM) or mobilized peripheral blood (mPB) (Arnous 2011). In patients with DCM only a few pilot studies have been done, reporting a modest improvements of left ventricular ejection fraction (LVEF), i.e. 3% to maximum 11.5% (Seth 2010). In our investigation we intend to clarify how the cellular content of the HSC graft influences the effect of HSCs in the treatment of DCM patients. The composition and stemness of the transplants will be studied and compared to their clinical effectiveness. We expect that the transplants containing higher numbers of pluripotent SCs will exhibit a superior clinical effect. METHODS: During preclinical study, following will be performed: a) collection of graft by apheresis from the G-CSF mobilized patients, b) Determination of SC subpopulations in autologous grafts by pluripotency differentiation markers (CD34, CD133 and CD184 (CXCR4); c) Quantification of pluripotency markers SSEA4, TRA-1-60 and TRA-1-81, CD117 (c-kit receptor), and CRIPTO-1 on protein level; d) Quantification of molecular markers of pluripotency OCT4, SOX2, NANOG, and CRIPTO-1; e) Determination of alkaline phosphatase, and the activity and expression of the TERT telomerase complex (TERT); f) Determination of the epigenetic metilation profile of target genes; and g) Determination of the miRNA profile of the graft. During the clinical study, a) 80 DCM patients will be included; b) Inclusion criteria will be age of 18-65 years, clinical status of advanced cardiac failure (NYHA III or IV) not reacting to optimal medical treatment during 6 months, and left ventricular ejection fraction (LVEF) 20%-40%; c) Exclusion criteria are defined at the ClinicalTrials.gov; d) Intramyocardial injections of 20x 0.5 mL will be used; e) Effectiveness of the therapy will be monitored in correlation with the cell subpopulation composition; and f) After 1 year, correlation analysis of the effectiveness vs. transplant composition will be made. EXPECTED RESULTS: Four initial hypotheses will be tested, i.e. that the injection of the SCs into the myocardium results in 1) better systolic function of left ventricle, 2) improvement of physical capability, 3) decreased concentration of pro-BNP (pro-brain natriuretic peptide) and inflammation parameters (TNF-α, IL-6), and 4) improved electrophysiology (shortened QTc interval and lowered QTVI in the high-discrimination ECG). The overall goal will be to determine which cell subpopulation in the graft could produce the best therapeutic effects. Positive results could enable treatment with SCs to become an alternative to the heart transplantation.
Significance for science
Subpopulation composition of CD34+ cell grafts and their contribution to the clinical efficacy of transplantation are still unknown. We believe that our findings allowed for a meaningful evaluation of both. The results are original, because so far no one has conducted similar studies in both preclinical and clinical setting. The research results have improved the current cell therapy and thus enable the treatment with hematopoietic stem cells to become a serious alternative to extremely expensive and difficult heart transplantation. The results of this research have been published in many high ranking professional and scientific journals and expose high citation index (See Paragraph 13).
Significance for the country
The understanding of the regenerative capacity of of stem cell subpopulations allows us to optimize cell therapy and leads to further development of regenerative medicine, which replaces the conventional transplantation. Collection of HSC is has been routinely carried out at the Blood Transfusion Centre of Slovenia for more than 20 years, whereas their clinical use is conducted at the Clinical Department of Cardiology, University Medical Centre Ljubljana. Both institutions have all necessary licences and accreditations by national competent authority (agency JAZMP) which allows their further development in this field. The project was successfully completed and represents full affirmation of cell therapies in Slovenia. In strategic terms, the Slovenian medicine entered a major step forward by developing advanced techniques and methods of modern cell therapy, which will constitute the key medical activity in the future. Needless to stress is that such advanced therapy could become a basis for the development of health tourism, etc. Since our activities are based exclusively on high knowledge and high technology, they have no harmful effects on the environment. Finally, we should stressed the main feature of the project - it was not theoretical, but had an immediate translational applicability into the treatment of a number of Slovenian patients, and that the results are evident in epidemiological terms. The effect of the project for the development of Slovenian medicine is extremely tangible in terms of improving health indicators, since cardiovascular disease represent the primary cause of mortality in Slovenia.
Most important scientific results Annual report 2013, 2014, 2015, final report
Most important socioeconomically and culturally relevant results Annual report 2013, 2014, 2015, final report
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