title: Gene transfer into human cord blood-derived CD34(+) cells by adeno-associated viral vectors.
abstract: OBJECTIVE
  Bone marrow-derived CD34(+) cells are currently used in clinical trials in patients with ischemic heart disease. An option to enhance activity of injected progenitors may be offered by genetic engineering of progenitor cells with angiogenic growth factors. Recombinant adeno-associated viral vectors (rAAV) have emerged as a leading gene transfer systems. In contrast to other vector systems in use for genetic engineering of CD34(+) cells, rAAV-mediated gene expression does not depend on vector integration. This is relevant for application in regenerative medicine of ischemic tissues, where transient transgene expression is likely sufficient to achieve therapeutic benefits.
  
  
  METHODS
  We compared three different human AAV serotypes, packaged as pseudotypes by a helper virus-free production method, for their transduction efficiency in human cord blood-derived CD34(+) cells. We further assessed the impact of vector genome conformation, of alpha(v)beta(5) and alpha(5)beta(1) integrin availability and of the transcription-modulating drugs retinoic acid and Trichostatin A on rAAV-mediated human CD34(+) cell transduction.
  
  
  RESULTS
  We provide, for the first time, evidence that hCD34(+) cells can be reproducibly transduced with high efficiency by self-complementary rAAV2 without inducing cytotoxicity or interfering with their differentiation potential. We further show the involvement of alpha(5)beta(1) integrin as a crucial AAV2 internalization receptor and a function for transcription-modulating drugs in enhancing rAAV-mediated transgene expression.
  
  
  CONCLUSIONS
  This study represents a first step toward translation of a combined cellular/rAAV-based therapy of ischemic disease.
authors.affiliation: Department I of Internal Medicine, University of Cologne, Cologne, Germany.
authors.forename: Natascha K
authors.initials: NK
authors.lastname: Schuhmann
authors.forename: Ombretta
authors.initials: O
authors.lastname: Pozzoli
authors.forename: Jessica
authors.initials: J
authors.lastname: Sallach
authors.forename: Anke
authors.initials: A
authors.lastname: Huber
authors.forename: Daniele
authors.initials: D
authors.lastname: Avitabile
authors.forename: Luca
authors.initials: L
authors.lastname: Perabo
authors.forename: Gunter
authors.initials: G
authors.lastname: Rappl
authors.forename: Maurizio C
authors.initials: MC
authors.lastname: Capogrossi
authors.forename: Michael
authors.initials: M
authors.lastname: Hallek
authors.forename: Maurizio
authors.initials: M
authors.lastname: Pesce
authors.forename: Hildegard
authors.initials: H
authors.lastname: Büning
mesh_terms: D018952:Antigens, CD34; D000970:Antineoplastic Agents / Q000494:pharmacology; D002986:Clinical Trials as Topic; D000229:Dependovirus; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism*; D005822:Genetic Vectors; D016679:Genome, Viral; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D039081:Integrin alpha5beta1 / Q000096:biosynthesis* / Q000235:genetics; D017202:Myocardial Ischemia / Q000378:metabolism / Q000628:therapy; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D033581:Stem Cell Transplantation / Q000379:methods; D013234:Stem Cells; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D018952:Antigens, CD34; D000970:Antineoplastic Agents; D006877:Hydroxamic Acids; D039081:Integrin alpha5beta1; D011500:Protein Synthesis Inhibitors; C012589:trichostatin A; D014212:Tretinoin