title: Transient detection of beta-galactosidase activity in hematopoietic cells, following reinjection of retrovirally marked autologous blood progenitors in patients with breast or ovarian cancer receiving high-dose chemotherapy.
abstract: OBJECTIVE
  The aim of this report is to demonstrate the feasibility and safety of genetically modifying autologous human blood CD34(+) cells in vitro, with a retroviral vector that encodes a marker gene. The fate of genetically modified cells and their progeny was followed in vivo, after reinfusion in patients treated with high-dose chemotherapy for poor-prognosis breast or ovarian carcinomas.
  
  
  METHODS
  Six patients received genetically modified autologous peripheral blood progenitors, together with unmanipulated aphereses, following high-dose chemotherapy. CD34(+) cells were immunoselected from aphereses, and retrovirally transduced by coculture with the retroviral vector producing cell line, to express a nuclear localized version of E. coli beta-galactosidase, encoded by a defective Moloney-murine leukemia virus-derived retroviral vector. Cells were reinfused to the patients after myeloablation, without prior ex vivo selection.
  
  
  RESULTS
  Five out of six patients showed the transient presence of low numbers of beta-galactosidase(+) cells, as detected with an immunocytochemical assay, in the peripheral blood, during the first month following infusion. One patient had beta-galactosidase(+) clonogenic progenitors in her marrow at two months after transplantation, including HPP-CFC; intriguingly, this patient had the lowest percentage of X-gal(+) cells in her graft. Patients experienced side effects that are often observed after high-dose chemotherapy.
  
  
  CONCLUSIONS
  Feasibility and safety of genetic modification of human hematopoietic stem and progenitor cells are demonstrated by this study. Ex vivo or in vivo selection is not mandatory, even in clinical situations where transduced cells have no survival advantage over wild-type cells; however, significant improvements in gene transfer technology are needed to achieve potentially useful levels of expression in such clinical situations.
authors.affiliation: Institut Paoli-Calmettes, Centre Régional de Lutte Contre le Cancer Provence-Alpes-Côte d'Azur, Marseille, France.
authors.forename: Claude
authors.initials: C
authors.lastname: Bagnis
authors.forename: Christian
authors.initials: C
authors.lastname: Chabannon
authors.forename: Gwenaelle
authors.initials: G
authors.lastname: Gravis
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Imbert
authors.forename: Christine
authors.initials: C
authors.lastname: Maroc
authors.forename: Florence
authors.initials: F
authors.lastname: Bardin
authors.forename: Patrick
authors.initials: P
authors.lastname: Ladaique
authors.forename: Frédéric
authors.initials: F
authors.lastname: Viret
authors.forename: Dominique
authors.initials: D
authors.lastname: Genre
authors.forename: Catherine
authors.initials: C
authors.lastname: Faucher
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Stoppa
authors.forename: Norbert
authors.initials: N
authors.lastname: Vey
authors.forename: Didier
authors.initials: D
authors.lastname: Blaise
authors.forename: Dominique
authors.initials: D
authors.lastname: Maraninchi
authors.forename: Patrice
authors.initials: P
authors.lastname: Viens
authors.forename: Patrice
authors.initials: P
authors.lastname: Mannoni
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use*; D001943:Breast Neoplasms / Q000473:pathology / Q000628:therapy*; D003131:Combined Modality Therapy; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D008875:Middle Aged; D008979:Moloney murine leukemia virus / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000473:pathology / Q000628:therapy*; D014161:Transduction, Genetic; D014182:Transplantation, Autologous; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D001616:beta-Galactosidase