E138K and M184I mutations in HIV-1 reverse transcriptase coemerge as a result of APOBEC3 editing in the absence of drug exposure.
BACKGROUND
Recent clinical trials with rilpivirine combined with emtricitabine and tenofovir revealed that patients failing treatment, frequently, harbored viruses encoding resistance-associated mutations in the HIV-1 reverse transcriptase at position E138K and M184I. We show here that APOBEC3 proteins play a role in the emergence of these drug resistance mutations.


METHODS
We used a Vif mutant that has suboptimal activity against APOBEC3 to assess the in-vitro frequency of APOBEC3-induced resistance mutations in reverse transcriptase. To assess the degree of in-vivo G-to-A viral hypermutation, a large amount of data of HIV-1 RT proviral sequences from peripheral blood mononuclear cells (PBMCs) recovered from infected patients under HAART was analyzed.


RESULTS
In-vitro replication experiments in cell lines with and without APOBEC3 expression suggest that APOBEC3-driven mutagenesis contributes to the generation of both M184I and E138K within HIV proviral repository in the absence of drug exposure. Additionally, analysis of 601 patients PBMCs sequences revealed that the copresence of mutations E138K and M184I were never detected in nonhypermutated sequences, whereas these mutations were found at a high frequency (24%) in the context of APOBEC3 editing and in the absence of exposure to etravirine-rilpivirine.


CONCLUSIONS
We demonstrate using in-vitro experiments and analyzing patients PBMCs sequences that M184I and E138K resistance-associated mutations may pre-exist in proviral reservoir at a high frequency prior to drug exposure, as a result of APOBEC3 editing. Thus, incomplete neutralization of one or more APOBEC3 proteins may favor viral escape to rilpivirine-emtricitabine.
Université Pierre et Marie Curie-Paris, Groupe Hospitalier Pitié Salpêtrière, Laboratoire de Virologie, Paris, France. slim.fourati@psl.aphp.fr
Slim
S
Fourati
Isabelle
I
Malet
Sidonie
S
Lambert
Cathia
C
Soulie
Marc
M
Wirden
Philippe
P
Flandre
Djeneba B
DB
Fofana
Sophie
S
Sayon
Anne
A
Simon
Christine
C
Katlama
Vincent
V
Calvez
Anne-Geneviève
AG
Marcelin
D000071478:APOBEC Deaminases; D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D019380:Anti-HIV Agents / Q000494:pharmacology*; D003564:Cytidine Deaminase; D043525:Cytosine Deaminase / Q000187:drug effects / Q000235:genetics* / Q000276:immunology; D004252:DNA Mutational Analysis; D024882:Drug Resistance, Viral; D005260:Female; D018698:Glutamic Acid; D054303:HIV Reverse Transcriptase / Q000235:genetics* / Q000276:immunology; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D007532:Isoleucine; D008239:Lysine; D008297:Male; D008715:Methionine; D009154:Mutation; D009570:Nitriles / Q000494:pharmacology; D063065:Organophosphonates / Q000494:pharmacology; D011724:Pyridazines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D000068696:Rilpivirine; D000068698:Tenofovir; D017211:Treatment Failure; D014779:Virus Replication
D019380:Anti-HIV Agents; D009570:Nitriles; D063065:Organophosphonates; D011724:Pyridazines; D011743:Pyrimidines; D007532:Isoleucine; C451734:etravirine; D018698:Glutamic Acid; D000068698:Tenofovir; D008715:Methionine; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; D043525:Cytosine Deaminase; D000071478:APOBEC Deaminases; C502889:APOBEC3 proteins, human; D003564:Cytidine Deaminase; D000068696:Rilpivirine; D000225:Adenine; D008239:Lysine