St. Jude Children's Research Hospital

The United States of America as Represented by the Department of Health

Genetic Therapy, Inc.

Elio F Vanin

Arthur W Nienhuis

Yanping Yang

Patrick F Kelly

Gerard C Grosveld

The present invention pertains to a method for efficiently introducing exogenous genes into stem cells, particularly human stem cells. The method optionally includes the steps of inducing the proliferation of target cells by pre-stimulation with cytokines and/or growth factors, followed by incubating these cells with RD114-pseudotyped vector particles. In a specific embodiment, the vector particles are retronectin-immobilized or ultracentrifugation-concentrated retroviral vector particles pseudotyped with the feline endogenous retrovirus (RD114) envelope protein. The present invention further discloses a method for somatic gene therapy, which can be used for various therapeutic applications and involves introducing a gene of interest contained within the retroviral genome into human repopulating stem cells followed by introducing these cells into a human host. Finally, the present invention discloses a method for monitoring the efficiency of the stem cell-mediated gene transfer based on detecting the presence of the genes (or the expression products) of the retroviral vector in various stem cell-derived lineages of the host.

Highly efficient gene transfer into human repopulating stem cells by RD114 pseudotyped retroviral vector particles

The field of the invention is recombinant packaging and producer cell lines for producing infectious retroviral vectors. The invention more specifically relates to the generation of pseudotyped retroviral vectors with a broad host range which can be produced at high titers in specially constructed packaging cell lines. Most specifically, the invention relates to the generation of pseudotyped retroviral vectors having vesicular stomatitis virus-G protein (VSV-G) as the membrane-associated viral envelope protein.

Stable virus packaging cell lines

A retroviral vector which includes a nucleic acid sequence encoding a retroviral envelope. The nucleic acid sequence encoding a retroviral envelope includes a first nucleic acid sequence encoding a first envelope portion which is a portion of MCF viral gp 70 protein, a nucleic acid sequence which encodes xenotropic envelope, a nucleic acid sequence which encodes an amphotropic envelope portion, and a nucleic acid sequence which encodes p15E protein. Such retroviral envelopes encoded by such nucleic acid sequence may be included in infectious viral particles. The infectious viral particles also may include gene(s) encoding therapeutic agents, and thus may be used in gene therapy.

Growing community of inventors

Memphis, TN, United States of America

Elio F Vanin