title: Semi-automated closed system manufacturing of lentivirus gene-modified haematopoietic stem cells for gene therapy.
abstract: Haematopoietic stem cell (HSC) gene therapy has demonstrated potential to treat many diseases. However, current state of the art requires sophisticated ex vivo gene transfer in a dedicated Good Manufacturing Practices facility, limiting availability. An automated process would improve the availability and standardized manufacture of HSC gene therapy. Here, we develop a novel program for semi-automated cell isolation and culture equipment to permit complete benchtop generation of gene-modified CD34+blood cell products for transplantation. These cell products meet current manufacturing quality standards for both mobilized leukapheresis and bone marrow, and reconstitute human haematopoiesis in immunocompromised mice. Importantly, nonhuman primate autologous gene-modified CD34+cell products are capable of stable, polyclonal multilineage reconstitution with follow-up of more than 1 year. These data demonstrate proof of concept for point-of-care delivery of HSC gene therapy. Given the many target diseases for gene therapy, there is enormous potential for this approach to treat patients on a global scale.
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, Washington 98109, USA.|Departments of Medical Oncology and Pathology, University of Washington, 1410 Campus Parkway, Seattle, Washington 98195, USA.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Adair
authors.affiliation: Miltenyi Biotec Inc., 2303 Lindbergh St, Auburn, California 95602, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Waters
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, Washington 98109, USA.
authors.forename: Kevin G
authors.initials: KG
authors.lastname: Haworth
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, Washington 98109, USA.
authors.forename: Sara P
authors.initials: SP
authors.lastname: Kubek
authors.affiliation: Department of Pharmaceutical Sciences, Washington State University Spokane, PO Box 1495, Spokane, Washington 99210, USA.
authors.forename: Grant D
authors.initials: GD
authors.lastname: Trobridge
authors.affiliation: Department of Pharmaceutical Sciences, Washington State University Spokane, PO Box 1495, Spokane, Washington 99210, USA.
authors.forename: Jonah D
authors.initials: JD
authors.lastname: Hocum
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, Washington 98109, USA.
authors.forename: Shelly
authors.initials: S
authors.lastname: Heimfeld
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, Washington 98109, USA.|Departments of Medical Oncology and Pathology, University of Washington, 1410 Campus Parkway, Seattle, Washington 98195, USA.
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Kiem
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism; D057205:Automation, Laboratory / Q000295:instrumentation* / Q000592:standards; D015415:Biomarkers / Q000378:metabolism; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D057809:HEK293 Cells; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D008254:Macaca nemestrina; D051379:Mice; D013194:Staining and Labeling / Q000379:methods; D014161:Transduction, Genetic / Q000379:methods* / Q000592:standards
chemical_list: D018952:Antigens, CD34; D015415:Biomarkers