<italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix

ABSTRACT Protease inhibitors (PIs) are the second- and last-line therapy for the majority of HIV-infected patients worldwide. Only around 20% of individuals who fail PI regimens develop major resistance mutations in protease. We sought to explore the role of mutations in gag-pro genotypic and phenot...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Rawlings Datir, Steven Kemp, Kate El Bouzidi, Petra Mlchocova, Richard Goldstein, Judy Breuer, Greg J. Towers, Clare Jolly, Miguel E. Quiñones-Mateu, Patrick S. Dakum, Nicaise Ndembi, Ravindra K. Gupta
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	HIV resistance protease drug Africa antiretroviral Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7199b87aeca649fba51303efebde939d
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:7199b87aeca649fba51303efebde939d
record_format	dspace
spelling	oai:doaj.org-article:7199b87aeca649fba51303efebde939d2021-11-15T15:55:44Z<italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix10.1128/mBio.02036-202150-7511https://doaj.org/article/7199b87aeca649fba51303efebde939d2020-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02036-20https://doaj.org/toc/2150-7511ABSTRACT Protease inhibitors (PIs) are the second- and last-line therapy for the majority of HIV-infected patients worldwide. Only around 20% of individuals who fail PI regimens develop major resistance mutations in protease. We sought to explore the role of mutations in gag-pro genotypic and phenotypic changes in viruses from six Nigerian patients who failed PI-based regimens without known drug resistance-associated protease mutations in order to identify novel determinants of PI resistance. Target enrichment and next-generation sequencing (NGS) with the Illumina MiSeq system were followed by haplotype reconstruction. Full-length Gag-protease gene regions were amplified from baseline (pre-PI) and virologic failure (VF) samples, sequenced, and used to construct gag-pro-pseudotyped viruses. Phylogenetic analysis was performed using maximum-likelihood methods. Susceptibility to lopinavir (LPV) and darunavir (DRV) was measured using a single-cycle replication assay. Western blotting was used to analyze Gag cleavage. In one of six participants (subtype CRF02_AG), we found 4-fold-lower LPV susceptibility in viral clones during failure of second-line treatment. A combination of four mutations (S126del, H127del, T122A, and G123E) in the p17 matrix of baseline virus generated a similar 4-fold decrease in susceptibility to LPV but not darunavir. These four amino acid changes were also able to confer LPV resistance to a subtype B Gag-protease backbone. Western blotting demonstrated significant Gag cleavage differences between sensitive and resistant isolates in the presence of drug. Resistant viruses had around 2-fold-lower infectivity than sensitive clones in the absence of drug. NGS combined with haplotype reconstruction revealed that resistant, less fit clones emerged from a minority population at baseline and thereafter persisted alongside sensitive fitter viruses. We used a multipronged genotypic and phenotypic approach to document emergence and temporal dynamics of a novel protease inhibitor resistance signature in HIV-1 matrix, revealing the interplay between Gag-associated resistance and fitness.Rawlings DatirSteven KempKate El BouzidiPetra MlchocovaRichard GoldsteinJudy BreuerGreg J. TowersClare JollyMiguel E. Quiñones-MateuPatrick S. DakumNicaise NdembiRavindra K. GuptaAmerican Society for MicrobiologyarticleHIVresistanceproteasedrugAfricaantiretroviralMicrobiologyQR1-502ENmBio, Vol 11, Iss 6 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	HIV resistance protease drug Africa antiretroviral Microbiology QR1-502
spellingShingle	HIV resistance protease drug Africa antiretroviral Microbiology QR1-502 Rawlings Datir Steven Kemp Kate El Bouzidi Petra Mlchocova Richard Goldstein Judy Breuer Greg J. Towers Clare Jolly Miguel E. Quiñones-Mateu Patrick S. Dakum Nicaise Ndembi Ravindra K. Gupta <italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix
description	ABSTRACT Protease inhibitors (PIs) are the second- and last-line therapy for the majority of HIV-infected patients worldwide. Only around 20% of individuals who fail PI regimens develop major resistance mutations in protease. We sought to explore the role of mutations in gag-pro genotypic and phenotypic changes in viruses from six Nigerian patients who failed PI-based regimens without known drug resistance-associated protease mutations in order to identify novel determinants of PI resistance. Target enrichment and next-generation sequencing (NGS) with the Illumina MiSeq system were followed by haplotype reconstruction. Full-length Gag-protease gene regions were amplified from baseline (pre-PI) and virologic failure (VF) samples, sequenced, and used to construct gag-pro-pseudotyped viruses. Phylogenetic analysis was performed using maximum-likelihood methods. Susceptibility to lopinavir (LPV) and darunavir (DRV) was measured using a single-cycle replication assay. Western blotting was used to analyze Gag cleavage. In one of six participants (subtype CRF02_AG), we found 4-fold-lower LPV susceptibility in viral clones during failure of second-line treatment. A combination of four mutations (S126del, H127del, T122A, and G123E) in the p17 matrix of baseline virus generated a similar 4-fold decrease in susceptibility to LPV but not darunavir. These four amino acid changes were also able to confer LPV resistance to a subtype B Gag-protease backbone. Western blotting demonstrated significant Gag cleavage differences between sensitive and resistant isolates in the presence of drug. Resistant viruses had around 2-fold-lower infectivity than sensitive clones in the absence of drug. NGS combined with haplotype reconstruction revealed that resistant, less fit clones emerged from a minority population at baseline and thereafter persisted alongside sensitive fitter viruses. We used a multipronged genotypic and phenotypic approach to document emergence and temporal dynamics of a novel protease inhibitor resistance signature in HIV-1 matrix, revealing the interplay between Gag-associated resistance and fitness.
format	article
author	Rawlings Datir Steven Kemp Kate El Bouzidi Petra Mlchocova Richard Goldstein Judy Breuer Greg J. Towers Clare Jolly Miguel E. Quiñones-Mateu Patrick S. Dakum Nicaise Ndembi Ravindra K. Gupta
author_facet	Rawlings Datir Steven Kemp Kate El Bouzidi Petra Mlchocova Richard Goldstein Judy Breuer Greg J. Towers Clare Jolly Miguel E. Quiñones-Mateu Patrick S. Dakum Nicaise Ndembi Ravindra K. Gupta
author_sort	Rawlings Datir
title	<italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix
title_short	<italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix
title_full	<italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix
title_fullStr	<italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix
title_full_unstemmed	<italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix
title_sort	<italic toggle="yes">in vivo</italic> emergence of a novel protease inhibitor resistance signature in hiv-1 matrix
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/7199b87aeca649fba51303efebde939d
work_keys_str_mv	AT rawlingsdatir italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT stevenkemp italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT kateelbouzidi italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT petramlchocova italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT richardgoldstein italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT judybreuer italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT gregjtowers italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT clarejolly italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT miguelequinonesmateu italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT patricksdakum italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT nicaisendembi italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix AT ravindrakgupta italictoggleyesinvivoitalicemergenceofanovelproteaseinhibitorresistancesignatureinhiv1matrix
_version_	1718427154526502912

<italic toggle="yes">In Vivo</italic> Emergence of a Novel Protease Inhibitor Resistance Signature in HIV-1 Matrix

Ejemplares similares