Selection and Characterization of Drug-Resistant Variants of Human Immunodeficiency Virus (AIDS).
Final rept. 22 Jul 94-30 Sep 95,
DEPARTMENT OF VETERANS AFFAIRS PITTSBURGH PA
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The original aims of this 5 year project were i to complete the molecular characterization of variants HW-1 that are resistant to the pyrophosphate analog foscarnet, ii to identify by in vitro selection variants of HIV-1 that are resistant to new reverse transcriptase RT and protease inhibitors that are in preclinical or early clinical stages of evaluation, and iii to construct a panel of drug-resistant infectious proviral clones for use as standards in HW-1 drug susceptibility assays. During the 14 month period of funding, major progress has been made on each of these aims. First, we have completed a comprehensive analysis of the genetic basis for HW-1 resistance to foscarnet. Six novel mutations in HIV-1 RT have been identified in foscarnet resistant laboratory and clinical isolates. In addition, important interactions between foscarnet and azidothymidine resistance have been identified. Second, we have isolated a variant of HIV-1 that is resistant to the novel RT inhibitor dioxolane guanosine and encodes a K65R mutation in the IKKK motif of HIV-1 RT. Third, we have engineered an HlV-1 proviral clone to encode unique silent restriction sites that allow rapid cloning of mutant protease or RT genes into the provirus. Using this novel vector we have produced stocks of recombinant mutant viruses that are highly resistant to foscarnet, nonnucleoside RT inhibitors, azidothymidine, or oxathiolane cytosine nucleosides e.g. 3TC. In summary, new discoveries and several products of importance to HW research have arisen from this project during the short period of funding.
- Medicine and Medical Research