The long term objective of this research proposal is to develop methods for treating human immunodeficiency virus (HIV) infection using infusions of peripheral blood lymphocytes (PBL) which have been transduced with retroviral vectors encoding RNA decoys. RNA decoys are short RNA oligonucleotides which mimic the TAR and RRE sequences of the HIV genomic RNA and inhibit the HIV RNA production by sequestering the critical HIV encoded RNA binding proteins tat and rev. Intracellular expression of TAR and RRE decoys causes inhibition of HIV replication and protects against depletion of immortalized human CD4+ lymphocytes following Infection with HIV. In addition, TAR decoys have been demonstrated to protect primary human PBL from HIV. The specific objectives of this proposal are to perform a series of pre- clinical studies designed to define optimal gene transfer procedures and optimal RNA decoy vectors for treatment of HIV infected individuals. To evaluate efficacy and safety issues in populations of primary CD4+ T-cells which are uniformly transduced, studies will be performed using RNA decoy transduced isolated primary CD4+ T-cell clones. Since two clinical applications are envisioned: 1) infusion of RNA transduced syngeneic PBL from healthy donors and, 2) autologous reinfusion of transduced PBL from previously infected individuals lacking syngeneic donors, studies will be performed using bulk PBL obtained from normal and HIV infected individuals. Maximally efficient gene transfer conditions as well as conditions which minimize activation of HIV in latently infected PBL will be determined. The effectiveness of alternative RNA decoys will be compared against laboratory and natural Isolates of HIV. Finally, bulk transduced CD4+ PBL will be evaluated to determine if these cells are safe for therapeutic use. The principal investigator of this grant proposal is a Clinical Assistant attending on the Hematologic Oncology Service at Memorial Sloan Kettering Cancer Center. Beginning In July, 1993, he will be appointed as an Assistant Professor in the Hematology/Oncology Division at Duke University Medical Center. He has 5 years of laboratory research experience primarily involving retroviral mediated gene transfer in human hematopoietic stem cells, hematopoietic stem cell physiology, and HIV inhibition via gene transfer of TAR decoys into primary human PBL. The proposed research will be performed at Duke University Medical Center and will be sponsored by Eli Gilboa, Ph.D.
