Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.5.4.4 (adenosine deaminase)
 5,136 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Human umbilical cord blood is an abundant source of long term repopulating stem cells and therefore we investigated the utilization of these cells as targets for genetic manipulation directed towards human gene therapy. Using two different retroviral vectors, one which transfers the neomycin resistance gene and the other which transfers therapeutically relevant adenosine deaminase gene, we have demonstrated increased gene transfer efficiency into committed progenitor cells (CPCs) and long term culture-initiating cells (LTC-IC) derived from cord blood versus adult bone marrow. We further identified a chymotryptic fragment of the extracellular matrix molecule fibronectin (FN 30/35), to which primitive hematopoietic cells adhere. Gene transfer efficiency into hematopoietic cells adherent to FN 30/35 is significantly increased when compared to infection on bovine serum albumin-coated control plates. Utilization of this fragment allowed retroviral mediated gene transfer into cord blood derived CPCs and LTC-ICs with high efficiencies, similar to that observed after coculture of hematopoietic cells on virus producer cells. These data imply cord blood may be a promising source for efficient gene delivery to the human hematopoietic system, and the utilization of the FN 30/35 fibronectin molecule may provide a clinically applicable protocol to achieve this aim.
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PMID:Umbilical cord blood stem cells as targets for genetic modification: new therapeutic approaches to somatic gene therapy. 774 18

Human cord blood (CB) contains large numbers of both committed and primitive hematopoietic progenitor cells and has been shown to have the capacity to reconstitute the lympho-hematopoietic system in transplant protocols. To investigate the potential usefulness of CB stem and progenitor cell populations to deliver new genetic material into the blood and immune systems, we have transduced these cells using retroviral technology and compared the efficiency of gene transfer into CB cells with normal adult human bone marrow cells using a variety of infection protocols. Using two retroviral vectors which differ significantly in both recombinant viral titers and vector design, low density CB or adult bone marrow (ABM) cells were infected, and committed progenitor and more primitive hematopoietic cells were analyzed for gene expression by G418 drug resistance (G418r) of neophosphotransferase and protein analysis for murine adenosine deaminase (mADA). Standard methylcellulose progenitor assays were used to quantitate transduction efficiency of committed progenitor cells, and the long term culture-initiating cell (LTC-IC) assay was used to quantitate transduction efficiency of more primitive cells. Our results indicate that CB cells were more efficiently transduced via retroviral-mediated gene transfer as compared with ABM-derived cells. In addition, stable expression of the introduced gene sequences, including the ADA cDNA, was demonstrated in the progeny of infected LTC-ICs after 5 wk in long-term marrow cultures. Expression of the introduced ADA cDNA was higher than the endogenous human ADA gene in the LTC-IC-derived colonies examined. These studies demonstrate that CB progenitor and stem cells can be efficiently infected using retroviral vectors and suggest that CB cells may provide a suitable target population in gene transfer protocols for some genetic diseases.
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PMID:Human cord blood cells as targets for gene transfer: potential use in genetic therapies of severe combined immunodeficiency disease. 834 Jul 57