title: CD133-targeted gene transfer into long-term repopulating hematopoietic stem cells.
abstract: Gene therapy for hematological disorders relies on the genetic modification of CD34(+) cells, a heterogeneous cell population containing about 0.01% long-term repopulating cells. Here, we show that the lentiviral vector CD133-LV, which uses a surface marker on human primitive hematopoietic stem cells (HSCs) as entry receptor, transfers genes preferentially into cells with high engraftment capability. Transduction of unstimulated CD34(+) cells with CD133-LV resulted in gene marking of cells with competitive proliferative advantage in vitro and in immunodeficient mice. The CD133-LV-transduced population contained significantly more cells with repopulating capacity than cells transduced with vesicular stomatitis virus (VSV)-LV, a lentiviral vector pseudotyped with the vesicular stomatitis virus G protein. Upon transfer of a barcode library, CD133-LV-transduced cells sustained gene marking in vivo for a prolonged period of time with a 6.7-fold higher recovery of barcodes compared to transduced control cells. Moreover, CD133-LV-transduced cells were capable of repopulating secondary recipients. Lastly, we show that this targeting strategy can be used for transfer of a therapeutic gene into CD34(+) cells obtained from patients suffering of X-linked chronic granulomatous disease. In conclusion, direct gene transfer into CD133(+) cells allows for sustained long-term engraftment of gene corrected cells.
authors.affiliation: 1] Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt, Germany [2] Current address: Division of Pediatric Hematology/Oncology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Brendel
authors.affiliation: Department of Medicine, Hematology/Oncology, JW-Goethe-University, Frankfurt/M, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Goebel
authors.affiliation: Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt, Germany.
authors.forename: Abriss
authors.initials: A
authors.lastname: Daniela
authors.affiliation: Department of Immunohematology and Blood Transfusion (IHB) Leiden University Medical Center, Leiden, Netherlands.
authors.forename: Martijn
authors.initials: M
authors.lastname: Brugman
authors.affiliation: 1] Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany [2] Current address: Division of Haematology/Transfusion Medicine, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Kneissl
authors.affiliation: Institute for Transfusion Medicine and Immune Hematology, Clinics of the Johann Wolfgang Goethe University, German Red Cross Blood Donor Service Baden-Wuerttemberg-Hessen, Frankfurt am Main, Hessen, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Schwäble
authors.affiliation: Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt, Germany.
authors.forename: Kerstin B
authors.initials: KB
authors.lastname: Kaufmann
authors.affiliation: Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt, Germany.
authors.forename: Uta
authors.initials: U
authors.lastname: Müller-Kuller
authors.affiliation: Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt, Germany.
authors.forename: Hana
authors.initials: H
authors.lastname: Kunkel
authors.affiliation: 1] Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt, Germany [2] Current address: Experimental Cell Therapy and Hematology, Department of Transfusion Medicine, Cell Therapy and Haemostaseology, Ludwig Maximilian University Hospital Munich, Munich, Germany.
authors.forename: Linping
authors.initials: L
authors.lastname: Chen-Wichmann
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Abel
authors.affiliation: Department of Medicine, Hematology/Oncology, JW-Goethe-University, Frankfurt/M, Germany.
authors.forename: Hubert
authors.initials: H
authors.lastname: Serve
authors.affiliation: Department of Cell Biology, University Medical Center Groningen, Groningen, Netherlands.
authors.forename: Leonid
authors.initials: L
authors.lastname: Bystrykh
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Buchholz
authors.affiliation: Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt, Germany.
authors.forename: Manuel
authors.initials: M
authors.lastname: Grez
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000235:genetics* / Q000276:immunology; D018952:Antigens, CD34 / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology; D006105:Granulomatous Disease, Chronic / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000276:immunology* / Q000473:pathology; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010455:Peptides / Q000235:genetics* / Q000276:immunology; D061251:Primary Cell Culture; D014161:Transduction, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; C028391:G protein, vesicular stomatitis virus; D006023:Glycoproteins; D008562:Membrane Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins