Analysis of Testes and Semen from Rabbits Treated by Intravenous Injection with a Retroviral Vector Encoding the Human Factor VIII Gene: No Evidence of Germ Line Transduction
- 10 December 2000
- journal article
- research article
- Published by Mary Ann Liebert Inc in Human Gene Therapy
- Vol. 11 (18) , 2529-2540
- https://doi.org/10.1089/10430340050208000
Abstract
In a phase 1 clinical trial, we are evaluating a murine leukemia virus (MuLV)-based retroviral vector encoding the human factor VIII gene [hFVIII(V)], administered intravenously, as a therapy for hemophilia A. Preclinical biolocalization studies in adult rabbits revealed vector-specific PCR signals in testis tissue at low levels. In follow-up animal studies we used PCR to (1) estimate the frequency with which a given cell in testis tissue is transduced, and (2) determine whether a positive PCR signal could be detected in semen samples from animals treated with hFVIII(V). Using the 99% confidence bound, results indicate that the probability that a given cell within the testis was transduced is less than 1/709,000 (97 days after treatment). This probability decreased with time after hFVIII(V) administration. Moreover, the rate of provector sequence detection in semen samples collected weekly throughout two cycles of spermatogenesis was 3/4281 reactions (0.07%), which is lower than the rate of false positives (1/800, 0.125%) observed for control animals. Using PCR assays with single-copy sensitivity, we have shown that the small number of transduced cells present in testis tissue does not give rise to detectable transduced cells in semen.Keywords
This publication has 31 references indexed in Scilit:
- Direct Exposure of Mouse Spermatogenic Cells to High Doses of Adenovirus Gene Therapy Vector Does Not Result in Germ Cell TransductionHuman Gene Therapy, 2000
- Regulatory Issues: Department of Health and Human Services. Recombinant DNA Advisory Committee National Institutes of Health. Minutes of Meeting March 11–12, 1999Human Gene Therapy, 2000
- Gene Therapy of Human Severe Combined Immunodeficiency (SCID)-X1 DiseaseScience, 2000
- Long-Term Persistence of Human Bone Marrow Stromal Cells Transduced with Factor VIII-Retroviral Vectors and Transient Production of Therapeutic Levels of Human Factor VIII in Nonmyeloablated Immunodeficient MiceHuman Gene Therapy, 2000
- Distribution of retroviral vectors and vector producer cells using two routes of administration in ratsGene Therapy, 1999
- Efficient Gene Transfer by Hybrid Retroviral Vectors to Murine Spermatogenic CellsHuman Gene Therapy, 1999
- Increased gene transfer into human CD34+ progenitor cells using retroviral vectors produced by a canine packaging cell lineTransplantation and Cellular Therapy, 1998
- Adenovirus-Mediated Hepatic Gene Transfer in Mice: Comparison of Intravascular and Biliary AdministrationHuman Gene Therapy, 1996
- In Vivo Gene Therapy of Hemophilia B: Sustained Partial Correction in Factor IX-Deficient DogsScience, 1993
- Long-Term Improvement of Hypercholesterolemia After Ex Vivo Gene Therapy in LDLR-Deficient RabbitsScience, 1991