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Improving Gene Therapy Efficiency through the Enrichment of Human Hematopoietic Stem Cells.
- Source :
-
Molecular therapy : the journal of the American Society of Gene Therapy [Mol Ther] 2017 Sep 06; Vol. 25 (9), pp. 2163-2175. Date of Electronic Publication: 2017 Jun 27. - Publication Year :
- 2017
-
Abstract
- Lentiviral vector (LV)-based hematopoietic stem cell (HSC) gene therapy is becoming a promising clinical strategy for the treatment of genetic blood diseases. However, the current approach of modifying 1 × 10 <superscript>8</superscript> to 1 × 10 <superscript>9</superscript> CD34 <superscript>+</superscript> cells per patient requires large amounts of LV, which is expensive and technically challenging to produce at clinical scale. Modification of bulk CD34 <superscript>+</superscript> cells uses LV inefficiently, because the majority of CD34 <superscript>+</superscript> cells are short-term progenitors with a limited post-transplant lifespan. Here, we utilized a clinically relevant, immunomagnetic bead (IB)-based method to purify CD34 <superscript>+</superscript> CD38 <superscript>-</superscript> cells from human bone marrow (BM) and mobilized peripheral blood (mPB). IB purification of CD34 <superscript>+</superscript> CD38 <superscript>-</superscript> cells enriched severe combined immune deficiency (SCID) repopulating cell (SRC) frequency an additional 12-fold beyond standard CD34 <superscript>+</superscript> purification and did not affect gene marking of long-term HSCs. Transplant of purified CD34 <superscript>+</superscript> CD38 <superscript>-</superscript> cells led to delayed myeloid reconstitution, which could be rescued by the addition of non-transduced CD38 <superscript>+</superscript> cells. Importantly, LV modification and transplantation of IB-purified CD34 <superscript>+</superscript> CD38 <superscript>-</superscript> cells/non-modified CD38 <superscript>+</superscript> cells into immune-deficient mice achieved long-term gene-marked engraftment comparable with modification of bulk CD34 <superscript>+</superscript> cells, while utilizing ∼7-fold less LV. Thus, we demonstrate a translatable method to improve the clinical and commercial viability of gene therapy for genetic blood cell diseases.<br /> (Copyright © 2017 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)
- Subjects :
- ADP-ribosyl Cyclase 1 metabolism
Animals
Antigens, CD34 metabolism
Gene Expression
Gene Transfer Techniques
Genes, Reporter
Genetic Therapy
Genetic Vectors genetics
Graft Survival
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells cytology
Humans
Immunomagnetic Separation
Immunophenotyping
Lentivirus genetics
Mice
Transgenes
Hematopoietic Stem Cells metabolism
Transduction, Genetic
Subjects
Details
- Language :
- English
- ISSN :
- 1525-0024
- Volume :
- 25
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Molecular therapy : the journal of the American Society of Gene Therapy
- Publication Type :
- Academic Journal
- Accession number :
- 28663101
- Full Text :
- https://doi.org/10.1016/j.ymthe.2017.05.023