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. 1997 Sep;71(9):6662–6670. doi: 10.1128/jvi.71.9.6662-6670.1997

Drug resistance during indinavir therapy is caused by mutations in the protease gene and in its Gag substrate cleavage sites.

Y M Zhang 1, H Imamichi 1, T Imamichi 1, H C Lane 1, J Falloon 1, M B Vasudevachari 1, N P Salzman 1
PMCID: PMC191944  PMID: 9261388

Abstract

Two different responses to the therapy were observed in a group of patients receiving the protease inhibitor indinavir. In one, suppression of virus replication occurred and has persisted for 90 weeks (bDNA, < 500 human immunodeficiency virus type 1 [HIV-1] RNA copies/ml). In the second group, a rebound in virus levels in plasma followed the initial sharp decline observed at the start of therapy. This was associated with the emergence of drug-resistant variants. Sequence analysis of the protease gene during the course of therapy revealed that in this second group there was a sequential acquisition of protease mutations at amino acids 46, 82, 54, 71, 89, and 90. In the six patients in this group, there was also an identical mutation in the gag p7/p1 gag protease cleavage site. In three of the patients, this change was seen as early as 6 to 10 weeks after the start of therapy. In one patient, a second mutation occurred at the gag p1/p6 cleavage site, but it appeared 18 weeks after the time of appearance of the p7/p1 mutation. Recombinant HIV-1 variants containing two or three mutations in the protease gene were constructed either with mutations at the p7/p1 cleavage site or with wild-type (WT) gag sequences. When recombinant HIV-1-containing protease mutations at 46 and 82 was grown in MT2 cells, there was a 68% reduction in its rate of replication compared to the WT virus. Introduction of an additional mutation at the gag p7/p1 protease cleavage site compensated for the partially defective protease gene. Similarly, rates of replication of viruses with mutations M46L/I, I54V, and V82A in protease were enhanced both in the presence and in the absence of Indinavir when combined with mutations in the gag p7/p1 and the gag p1/p6 cleavage sites. Optimal rates of virus replication require protease cleavage of precursor polyproteins. A mutation in the cleavage site that enhanced the availability of a protein that was rate limiting for virus maturation would confer on that virus a significant growth advantage and may explain the uniform emergence of viruses with alterations at the p7/p1 cleavage site. This is the first report of the emergence of mutations in the gag p7/p1 protease cleavage sites in patients receiving protease therapy and identifies this change as an important determinant of HIV-1 resistance to protease inhibitors in patient populations.

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Selected References

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  1. Adachi A., Gendelman H. E., Koenig S., Folks T., Willey R., Rabson A., Martin M. A. Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone. J Virol. 1986 Aug;59(2):284–291. doi: 10.1128/jvi.59.2.284-291.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Coffin J. M. HIV population dynamics in vivo: implications for genetic variation, pathogenesis, and therapy. Science. 1995 Jan 27;267(5197):483–489. doi: 10.1126/science.7824947. [DOI] [PubMed] [Google Scholar]
  3. Collier A. C., Coombs R. W., Schoenfeld D. A., Bassett R. L., Timpone J., Baruch A., Jones M., Facey K., Whitacre C., McAuliffe V. J. Treatment of human immunodeficiency virus infection with saquinavir, zidovudine, and zalcitabine. AIDS Clinical Trials Group. N Engl J Med. 1996 Apr 18;334(16):1011–1017. doi: 10.1056/NEJM199604183341602. [DOI] [PubMed] [Google Scholar]
  4. Condra J. H., Holder D. J., Schleif W. A., Blahy O. M., Danovich R. M., Gabryelski L. J., Graham D. J., Laird D., Quintero J. C., Rhodes A. Genetic correlates of in vivo viral resistance to indinavir, a human immunodeficiency virus type 1 protease inhibitor. J Virol. 1996 Dec;70(12):8270–8276. doi: 10.1128/jvi.70.12.8270-8276.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Condra J. H., Schleif W. A., Blahy O. M., Gabryelski L. J., Graham D. J., Quintero J. C., Rhodes A., Robbins H. L., Roth E., Shivaprakash M. In vivo emergence of HIV-1 variants resistant to multiple protease inhibitors. Nature. 1995 Apr 6;374(6522):569–571. doi: 10.1038/374569a0. [DOI] [PubMed] [Google Scholar]
  6. Croteau G., Doyon L., Thibeault D., McKercher G., Pilote L., Lamarre D. Impaired fitness of human immunodeficiency virus type 1 variants with high-level resistance to protease inhibitors. J Virol. 1997 Feb;71(2):1089–1096. doi: 10.1128/jvi.71.2.1089-1096.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Darke P. L., Huff J. R. HIV protease as an inhibitor target for the treatment of AIDS. Adv Pharmacol. 1994;25:399–454. doi: 10.1016/s1054-3589(08)60438-x. [DOI] [PubMed] [Google Scholar]
  8. Dewar R. L., Highbarger H. C., Sarmiento M. D., Todd J. A., Vasudevachari M. B., Davey R. T., Jr, Kovacs J. A., Salzman N. P., Lane H. C., Urdea M. S. Application of branched DNA signal amplification to monitor human immunodeficiency virus type 1 burden in human plasma. J Infect Dis. 1994 Nov;170(5):1172–1179. doi: 10.1093/infdis/170.5.1172. [DOI] [PubMed] [Google Scholar]
  9. Doyon L., Croteau G., Thibeault D., Poulin F., Pilote L., Lamarre D. Second locus involved in human immunodeficiency virus type 1 resistance to protease inhibitors. J Virol. 1996 Jun;70(6):3763–3769. doi: 10.1128/jvi.70.6.3763-3769.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Embretson J., Zupancic M., Ribas J. L., Burke A., Racz P., Tenner-Racz K., Haase A. T. Massive covert infection of helper T lymphocytes and macrophages by HIV during the incubation period of AIDS. Nature. 1993 Mar 25;362(6418):359–362. doi: 10.1038/362359a0. [DOI] [PubMed] [Google Scholar]
  11. Faulkner D. V., Jurka J. Multiple aligned sequence editor (MASE). Trends Biochem Sci. 1988 Aug;13(8):321–322. doi: 10.1016/0968-0004(88)90129-6. [DOI] [PubMed] [Google Scholar]
  12. Greer J., Erickson J. W., Baldwin J. J., Varney M. D. Application of the three-dimensional structures of protein target molecules in structure-based drug design. J Med Chem. 1994 Apr 15;37(8):1035–1054. doi: 10.1021/jm00034a001. [DOI] [PubMed] [Google Scholar]
  13. Haertle T., Carrera C. J., Wasson D. B., Sowers L. C., Richman D. D., Carson D. A. Metabolism and anti-human immunodeficiency virus-1 activity of 2-halo-2',3'-dideoxyadenosine derivatives. J Biol Chem. 1988 Apr 25;263(12):5870–5875. [PubMed] [Google Scholar]
  14. Harada S., Koyanagi Y., Yamamoto N. Infection of HTLV-III/LAV in HTLV-I-carrying cells MT-2 and MT-4 and application in a plaque assay. Science. 1985 Aug 9;229(4713):563–566. doi: 10.1126/science.2992081. [DOI] [PubMed] [Google Scholar]
  15. Korber B., Myers G. Signature pattern analysis: a method for assessing viral sequence relatedness. AIDS Res Hum Retroviruses. 1992 Sep;8(9):1549–1560. doi: 10.1089/aid.1992.8.1549. [DOI] [PubMed] [Google Scholar]
  16. Lech W. J., Wang G., Yang Y. L., Chee Y., Dorman K., McCrae D., Lazzeroni L. C., Erickson J. W., Sinsheimer J. S., Kaplan A. H. In vivo sequence diversity of the protease of human immunodeficiency virus type 1: presence of protease inhibitor-resistant variants in untreated subjects. J Virol. 1996 Mar;70(3):2038–2043. doi: 10.1128/jvi.70.3.2038-2043.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Markowitz M., Saag M., Powderly W. G., Hurley A. M., Hsu A., Valdes J. M., Henry D., Sattler F., La Marca A., Leonard J. M. A preliminary study of ritonavir, an inhibitor of HIV-1 protease, to treat HIV-1 infection. N Engl J Med. 1995 Dec 7;333(23):1534–1539. doi: 10.1056/NEJM199512073332204. [DOI] [PubMed] [Google Scholar]
  18. Molla A., Korneyeva M., Gao Q., Vasavanonda S., Schipper P. J., Mo H. M., Markowitz M., Chernyavskiy T., Niu P., Lyons N. Ordered accumulation of mutations in HIV protease confers resistance to ritonavir. Nat Med. 1996 Jul;2(7):760–766. doi: 10.1038/nm0796-760. [DOI] [PubMed] [Google Scholar]
  19. Pantaleo G., Graziosi C., Demarest J. F., Butini L., Montroni M., Fox C. H., Orenstein J. M., Kotler D. P., Fauci A. S. HIV infection is active and progressive in lymphoid tissue during the clinically latent stage of disease. Nature. 1993 Mar 25;362(6418):355–358. doi: 10.1038/362355a0. [DOI] [PubMed] [Google Scholar]
  20. Rose R. E., Gong Y. F., Greytok J. A., Bechtold C. M., Terry B. J., Robinson B. S., Alam M., Colonno R. J., Lin P. F. Human immunodeficiency virus type 1 viral background plays a major role in development of resistance to protease inhibitors. Proc Natl Acad Sci U S A. 1996 Feb 20;93(4):1648–1653. doi: 10.1073/pnas.93.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Saksela K., Muchmore E., Girard M., Fultz P., Baltimore D. High viral load in lymph nodes and latent human immunodeficiency virus (HIV) in peripheral blood cells of HIV-1-infected chimpanzees. J Virol. 1993 Dec;67(12):7423–7427. doi: 10.1128/jvi.67.12.7423-7427.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schmit J. C., Ruiz L., Clotet B., Raventos A., Tor J., Leonard J., Desmyter J., De Clercq E., Vandamme A. M. Resistance-related mutations in the HIV-1 protease gene of patients treated for 1 year with the protease inhibitor ritonavir (ABT-538). AIDS. 1996 Aug;10(9):995–999. doi: 10.1097/00002030-199610090-00010. [DOI] [PubMed] [Google Scholar]
  23. Thompson J. D., Higgins D. G., Gibson T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. doi: 10.1093/nar/22.22.4673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vella S., Galluzzo C., Giannini G., Pirillo M. F., Duncan I., Jacobsen H., Andreoni M., Sarmati L., Ercoli L. Saquinavir/zidovudine combination in patients with advanced HIV infection and no prior antiretroviral therapy: CD4+ lymphocyte/plasma RNA changes, and emergence of HIV strains with reduced phenotypic sensitivity. Antiviral Res. 1996 Jan;29(1):91–93. doi: 10.1016/0166-3542(95)00926-4. [DOI] [PubMed] [Google Scholar]
  25. Winslow D. L., Stack S., King R., Scarnati H., Bincsik A., Otto M. J. Limited sequence diversity of the HIV type 1 protease gene from clinical isolates and in vitro susceptibility to HIV protease inhibitors. AIDS Res Hum Retroviruses. 1995 Jan;11(1):107–113. doi: 10.1089/aid.1995.11.107. [DOI] [PubMed] [Google Scholar]
  26. Wlodawer A., Erickson J. W. Structure-based inhibitors of HIV-1 protease. Annu Rev Biochem. 1993;62:543–585. doi: 10.1146/annurev.bi.62.070193.002551. [DOI] [PubMed] [Google Scholar]
  27. Yamaguchi K., Byrn R. A. Clinical isolates of HIV-1 contain few pre-existing proteinase inhibitor resistance-conferring mutations. Biochim Biophys Acta. 1995 Dec 6;1253(2):136–140. doi: 10.1016/0167-4838(95)00167-1. [DOI] [PubMed] [Google Scholar]

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