Document Type
Article
Publication Date
10-22-2019
Abstract
BACKGROUND: PIE12-trimer is a highly potent D-peptide HIV-1 entry inhibitor that broadly targets group M isolates. It specifically binds the three identical conserved hydrophobic pockets at the base of the gp41 N-trimer with sub-femtomolar affinity. This extremely high affinity for the transiently exposed gp41 trimer provides a reserve of binding energy (resistance capacitor) to prevent the viral resistance pathway of stepwise accumulation of modest affinity-disrupting mutations. Such modest mutations would not affect PIE12-trimer potency and therefore not confer a selective advantage. Viral passaging in the presence of escalating PIE12-trimer concentrations ultimately selected for PIE12-trimer resistant populations, but required an extremely extended timeframe (> 1 year) in comparison to other entry inhibitors. Eventually, HIV developed resistance to PIE12-trimer by mutating Q577 in the gp41 pocket.
RESULTS: Using deep sequence analysis, we identified three mutations at Q577 (R, N and K) in our two PIE12-trimer resistant pools. Each point mutant is capable of conferring the majority of PIE12-trimer resistance seen in the polyclonal pools. Surface plasmon resonance studies demonstrated substantial affinity loss between PIE12-trimer and the Q577R-mutated gp41 pocket. A high-resolution X-ray crystal structure of PIE12 bound to the Q577R pocket revealed the loss of two hydrogen bonds, the repositioning of neighboring residues, and a small decrease in buried surface area. The Q577 mutations in an NL4-3 backbone decreased viral growth rates. Fitness was ultimately rescued in resistant viral pools by a suite of compensatory mutations in gp120 and gp41, of which we identified seven candidates from our sequencing data.
CONCLUSIONS: These data show that PIE12-trimer exhibits a high barrier to resistance, as extended passaging was required to develop resistant virus with normal growth rates. The primary resistance mutation, Q577R/N/K, found in the conserved gp41 pocket, substantially decreases inhibitor affinity but also damages viral fitness, and candidate compensatory mutations in gp160 have been identified.
Recommended Citation
Smith, Amanda R; Weinstock, Matthew T; Siglin, Amanda E; Whitby, Frank G; Francis, J Nicholas; Hill, Christopher P; Eckert, Debra M; Root, Michael J; and Kay, Michael S, "Characterization of resistance to a potent D-peptide HIV entry inhibitor." (2019). Department of Biochemistry and Molecular Biology Faculty Papers. Paper 162.
https://jdc.jefferson.edu/bmpfp/162
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
PubMed ID
31640718
Language
English
Comments
This article has been peer-reviewed. It is the author's final published version in Retrovirology, Volume 16, Issue 1, Oct. 2019, Article number 28.
The published version is available at https://doi.org/10.1186/s12977-019-0489-7. Copyright © Smith et.al.