Cardiac stem cell genetic engineering using the alphaMHC promoter

Regen Med. 2009 Nov;4(6):823-33. doi: 10.2217/rme.09.51.

Abstract

Aims: Cardiac stem cells (CSCs) show potential as a cellular therapeutic approach to blunt tissue damage and facilitate reparative and regenerative processes after myocardial infarction. Despite multiple published reports of improvement, functional benefits remain modest using normal stem cells delivered by adoptive transfer into damaged myocardium. The goal of this study is to enhance survival and proliferation of CSCs that have undergone lineage commitment in early phases as evidenced by expression of proteins driven by the alpha-myosin heavy chain (alphaMHC) promoter. The early increased expression of survival kinases augments expansion of the cardiogenic CSC pool and subsequent daughter progeny.

Materials & methods: Normal CSCs engineered with fluorescent reporter protein constructs under control of the alphaMHC promoter show transgene protein expression, confirming activity of the promoter in CSCs. Cultured CSCs from both nontransgenic and cardiac-specific transgenic mice expressing survival kinases driven by the alphaMHC promoter were analyzed to characterize transgene expression following treatments to promote differentiation in culture.

Results & conclusion: Therapeutic genes controlled by the alphaMHC promoter can be engineered into and expressed in CSCs and cardiomyocyte progeny with the goal of improving the efficacy of cardiac stem cell therapy.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Differentiation
  • Cell Lineage
  • Cell Proliferation
  • Cells, Cultured
  • Flow Cytometry
  • Genetic Engineering*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Immunoenzyme Techniques
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, Transgenic
  • Myocardium / cytology*
  • Myocytes, Cardiac / cytology*
  • Myosin Heavy Chains / genetics*
  • Promoter Regions, Genetic / genetics*
  • Proto-Oncogene Proteins c-akt / physiology
  • Proto-Oncogene Proteins c-pim-1 / physiology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells / physiology*
  • Transgenes / physiology

Substances

  • Luminescent Proteins
  • RNA, Messenger
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Pim1 protein, mouse
  • Proto-Oncogene Proteins c-akt
  • Proto-Oncogene Proteins c-pim-1
  • Myosin Heavy Chains