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Cytotherapy. 2017 Sep;19(9):1096-1112. doi: 10.1016/j.jcyt.2017.06.002. Epub 2017 Jul 18.

Preclinical studies for a phase 1 clinical trial of autologous hematopoietic stem cell gene therapy for sickle cell disease.

Author information

1
Department of Microbiology, Immunology and Molecular Genetics and the Eli & Edythe Broad Stem Cell Research Center, University of California, Los Angeles, California, USA.
2
Translational Core Laboratory, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
3
Consultant, San Francisco, California, USA.
4
UCLA BM/Stem Cell Transplant Laboratory, University of California, Los Angeles, USA.
5
Translational Trials Development and Support Labs, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
6
Department of General Internal Medicine and Health Services Research, University of California, Los Angeles, USA.
7
Department of Microbiology, Immunology and Molecular Genetics and the Eli & Edythe Broad Stem Cell Research Center, University of California, Los Angeles, California, USA. Electronic address: e-dkohn@mednet.ucla.edu.

Abstract

BACKGROUND AIMS:

Gene therapy by autologous hematopoietic stem cell transplantation (HSCT) represents a new approach to treat sickle cell disease (SCD). Optimization of the manufacture, characterization and testing of the transduced hematopoietic stem cell final cell product (FCP), as well as an in depth in vivo toxicology study, are critical for advancing this approach to clinical trials.

METHODS:

Data are shown to evaluate and establish the feasibility of isolating, transducing with the Lenti/βAS3-FB vector and cryopreserving CD34+ cells from human bone marrow (BM) at clinical scale. In vitro and in vivo characterization of the FCP was performed, showing that all the release criteria were successfully met. In vivo toxicology studies were conducted to evaluate potential toxicity of the Lenti/βAS3-FB LV in the context of a murine BM transplant.

RESULTS:

Primary and secondary transplantation did not reveal any toxicity from the lentiviral vector. Additionally, vector integration site analysis of murine and human BM cells did not show any clonal skewing caused by insertion of the Lenti/βAS3-FB vector in cells from primary and secondary transplanted mice.

CONCLUSIONS:

We present here a complete protocol, thoroughly optimized to manufacture, characterize and establish safety of a FCP for gene therapy of SCD.

KEYWORDS:

gene therapy; hematopoietic stem cells; lentiviral vectors; sickle cell disease

PMID:
28733131
DOI:
10.1016/j.jcyt.2017.06.002
[Indexed for MEDLINE]

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