Enhancement of Myocardial Regeneration Through Genetic Engineering of Cardiac Progenitor Cells Expressing Pim-1 Kinase

Background— Despite numerous studies demonstrating the efficacy of cellular adoptive transfer for therapeutic myocardial regeneration, problems remain for donated cells with regard to survival, persistence, engraftment, and long-term benefits. This study redresses these concerns by enhancing the regenerative potential of adoptively transferred cardiac progenitor cells (CPCs) via genetic engineering to overexpress Pim-1, a cardioprotective kinase that enhances cell survival and proliferation. Methods and Results— Intramyocardial injections of CPCs overexpressing Pim-1 were given to infarcted female mice. Animals were monitored over 4, 12, and 32 weeks to assess cardiac function and engraftment of Pim-1 CPCs with echocardiography, in vivo hemodynamics, and confocal imagery. CPCs overexpressing Pim-1 showed increased proliferation and expression of markers consistent with cardiogenic lineage commitment after dexamethasone exposure in vitro. Animals that received CPCs overexpressing Pim-1 also produced greater levels of cellular engraftment, persistence, and functional improvement relative to control CPCs up to 32 weeks after delivery. Salutary effects include reduction of infarct size, greater number of c-kit+ cells, and increased vasculature in the damaged region. Conclusions— Myocardial repair is significantly enhanced by genetic engineering of CPCs with Pim-1 kinase. Ex vivo gene delivery to enhance cellular survival, proliferation, and regeneration may overcome current limitations of stem cell–based therapeutic approaches.

[1]  H. Haider,et al.  Strategies to promote donor cell survival: combining preconditioning approach with stem cell transplantation. , 2008, Journal of molecular and cellular cardiology.

[2]  S. Houser,et al.  Pim-1 kinase antagonizes aspects of myocardial hypertrophy and compensation to pathological pressure overload , 2008, Proceedings of the National Academy of Sciences.

[3]  Z. Wang,et al.  Pim kinase-dependent inhibition of c-Myc degradation , 2008, Oncogene.

[4]  S. Houser,et al.  Increased Cardiac Myocyte Progenitors in Failing Human Hearts , 2008, Circulation.

[5]  Mark A Sussman,et al.  Myocardial Induction of Nucleostemin in Response to Postnatal Growth and Pathological Challenge , 2008, Circulation research.

[6]  E. Fiumana,et al.  Local Activation or Implantation of Cardiac Progenitor Cells Rescues Scarred Infarcted Myocardium Improving Cardiac Function , 2008, Circulation research.

[7]  T. Tsuruo,et al.  Pim kinases promote cell cycle progression by phosphorylating and down-regulating p27Kip1 at the transcriptional and posttranscriptional levels. , 2008, Cancer research.

[8]  Mark A Sussman,et al.  Evolution of the c‐kit‐Positive Cell Response to Pathological Challenge in the Myocardium , 2008, Stem cells.

[9]  C. Bearzi,et al.  Activation of Cardiac Progenitor Cells Reverses the Failing Heart Senescent Phenotype and Prolongs Lifespan , 2008, Circulation research.

[10]  W. Frishman,et al.  Myocardial regeneration and stem cell repair. , 2008, Current problems in cardiology.

[11]  Richard T. Lee,et al.  Stem-cell therapy for cardiac disease , 2008, Nature.

[12]  C. Bearzi,et al.  Formation of large coronary arteries by cardiac progenitor cells , 2008, Proceedings of the National Academy of Sciences.

[13]  Marielle Afanassieff,et al.  Self‐Renewal of Murine Embryonic Stem Cells Is Supported by the Serine/Threonine Kinases Pim‐1 and Pim‐3 , 2007, Stem cells.

[14]  N. Magnuson,et al.  Pim-1 Kinase-Dependent Phosphorylation of p21Cip1/WAF1 Regulates Its Stability and Cellular Localization in H1299 Cells , 2007, Molecular Cancer Research.

[15]  M. Ashraf,et al.  HIF-1alpha induced-VEGF overexpression in bone marrow stem cells protects cardiomyocytes against ischemia. , 2007, Journal of molecular and cellular cardiology.

[16]  J. Gearhart,et al.  Stem cells and their potential in cell-based cardiac therapies. , 2007, Progress in cardiovascular diseases.

[17]  Arjun Deb,et al.  Secreted frizzled related protein 2 (Sfrp2) is the key Akt-mesenchymal stem cell-released paracrine factor mediating myocardial survival and repair , 2007, Proceedings of the National Academy of Sciences.

[18]  J. L. Hansen,et al.  Cardiac regeneration by resident stem and progenitor cells in the adult heart , 2007, Basic Research in Cardiology.

[19]  Arjun Deb,et al.  Mesenchymal stem cells overexpressing Akt dramatically repair infarcted myocardium and improve cardiac function despite infrequent cellular fusion or differentiation. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[20]  S. Houser,et al.  Pim-1 regulates cardiomyocyte survival downstream of Akt , 2006, Nature Medicine.

[21]  J. Ingwall,et al.  Evidence supporting paracrine hypothesis for Akt‐modified mesenchymal stem cell‐mediated cardiac protection and functional improvement , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[22]  T. Möröy,et al.  The oncogenic serine/threonine kinase Pim-1 directly phosphorylates and activates the G2/M specific phosphatase Cdc25C. , 2006, The international journal of biochemistry & cell biology.

[23]  J. Arthur,et al.  Pim kinases phosphorylate multiple sites on Bad and promote 14-3-3 binding and dissociation from Bcl-XL , 2006, BMC Cell Biology.

[24]  H. Haider,et al.  Cell-based ex vivo delivery of angiogenic growth factors for cardiac repair. , 2005, Arteriosclerosis, thrombosis, and vascular biology.

[25]  D. Torella,et al.  Stem cells in the dog heart are self-renewing, clonogenic, and multipotent and regenerate infarcted myocardium, improving cardiac function. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Daria Nurzynska,et al.  Myocardial regeneration by activation of multipotent cardiac stem cells in ischemic heart failure. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[27]  P. Hammerman,et al.  Pim and Akt oncogenes are independent regulators of hematopoietic cell growth and survival. , 2005, Blood.

[28]  H. Kaneto,et al.  Role of Pim-1 in Smooth Muscle Cell Proliferation* , 2004, Journal of Biological Chemistry.

[29]  P. Koskinen,et al.  Pim‐1 kinase promotes inactivation of the pro‐apoptotic Bad protein by phosphorylating it on the Ser112 gatekeeper site , 2004, FEBS letters.

[30]  Jiahua Jiang,et al.  Ganoderma lucidum Suppresses Growth of Breast Cancer Cells Through the Inhibition of Akt/NF-κB Signaling , 2004, Nutrition and cancer.

[31]  S. Oliviero,et al.  Identification of Flk-1 target genes in vasculogenesis: Pim-1 is required for endothelial and mural cell differentiation in vitro. , 2004, Blood.

[32]  H. Kiem,et al.  Scaffold attachment region-containing retrovirus vectors improve long-term proviral expression after transplantation of GFP-modified CD34+ baboon repopulating cells. , 2003, Blood.

[33]  D. Torella,et al.  Adult Cardiac Stem Cells Are Multipotent and Support Myocardial Regeneration , 2003, Cell.

[34]  J. Ingwall,et al.  Mesenchymal stem cells modified with Akt prevent remodeling and restore performance of infarcted hearts , 2003, Nature Medicine.

[35]  Wenyi Wei,et al.  Phosphorylation of the cell cycle inhibitor p21Cip1/WAF1 by Pim-1 kinase. , 2002, Biochimica et biophysica acta.

[36]  N. Bhattacharya,et al.  Pim-1 associates with protein complexes necessary for mitosis , 2002, Chromosoma.

[37]  Z. Wang,et al.  Pim-1: a serine/threonine kinase with a role in cell survival, proliferation, differentiation and tumorigenesis. , 2001, Journal of veterinary science.

[38]  Federica Limana,et al.  Mobilized bone marrow cells repair the infarcted heart, improving function and survival , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[39]  David M. Bodine,et al.  Bone marrow cells regenerate infarcted myocardium , 2001, Nature.

[40]  I. Plavec,et al.  Effect of scaffold attachment region on transgene expression in retrovirus vector-transduced primary T cells and macrophages. , 1999, Human gene therapy.

[41]  B. Torbett,et al.  Transduction of human CD34+ cells that mediate long-term engraftment of NOD/SCID mice by HIV vectors. , 1999, Science.

[42]  Fred H. Gage,et al.  Development of a Self-Inactivating Lentivirus Vector , 1998, Journal of Virology.

[43]  I. Plavec,et al.  Scaffold Attachment Region-Mediated Enhancement of Retroviral Vector Expression in Primary T Cells , 1998, Journal of Virology.

[44]  M. Ashraf,et al.  Stable therapeutic effects of mesenchymal stem cell-based multiple gene delivery for cardiac repair. , 2008, Cardiovascular research.

[45]  C. Barbas,et al.  T-cell protection and enrichment through lentiviral CCR5 intrabody gene delivery , 2007, Gene Therapy.

[46]  S. Dimmeler,et al.  Restoration of cardiac function with progenitor cells. , 2006, Novartis Foundation symposium.

[47]  K. Deb,et al.  Journal of Translational Medicine Human Embryonic Stem Cells: Preclinical Perspectives , 2022 .