Implementación del estudio farmacogenético de hipersensibilidad al abacavir HLA-B*5701 en Argentina

Introducción: El abacavir (ABC) es un antirretroviral inhibidor de la transcriptasa reversa del virus HIV-1 que está fuertemente asociado al desarrollo de reacciones de hipersensibilidad en individuos portadores del alelo HLA-B*5701. Objetivos: determinar la prevalencia del alelo HLA-B*5701 en pacientes HIV-1 positivos y en una población control de Argentina. Materiales y métodos: desde enero de 2012 hasta octubre de 2013 se estudiaron 869 pacientes HIV-1 positivos y 63 individuos no infectados con HIV-1. La detección del alelo HLA-B*5701 se realizó mediante un ensayo in house basado en la técnica de PCR en tiempo real, diseñado en nuestro laboratorio y validado según guías internacionales. Resultados: el primero de enero de 2012 se implementó el estudio farmacogenético para la detección de hipersensibilidad al ABC en los pacientes incluidos en el Programa VIH/sida de la Dirección de SIDA y ETS, y en los niños infectados con HIV-1 del Hospital Garrahan. Para ello se adoptó un protocolo de envío, recepción y procesado de las muestras, con un informe detallado de los resultados. El alelo HLA-B*5701 se detectó en 42 individuos infectados con HIV-1 y en 3 individuos no infectados. Conclusiones: La prevalencia del alelo HLA-B*5701 en la población de pacientes infectados con HIV-1 y la población control fue similar (4,8 %), lo cual sugiere que la presencia de este alelo no influye en la infección por HIV-1. Esta prevalencia fue similar a la reportada para otras poblaciones de origen caucásico.Fil: Moragas, Matías. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gurevich Messina, Juan Manuel. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Aulicino, Paula. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mecikovsky, Debora. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; ArgentinaFil: Bologna, Rosa. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; ArgentinaFil: Bissio, Emiliano. Ministerio de Salud de la Nación; ArgentinaFil: Falistoco, Carlos. Ministerio de Salud de la Nación; ArgentinaFil: Sen, Luisa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; ArgentinaFil: Mangano, Andrea María Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría ; Argentin

HIV type-1 genotypic resistance profiles in vertically infected patients from Argentina reveal an association between K103N+L100I and L74V mutations.

Background: Patterns and pathways of HIV type-1 (HIV-1) antiretroviral (ARV) drug resistance-associated mutations in clinical isolates are conditioned by ARV history and factors such as viral subtype and fitness. Our aim was to analyse the frequency and association of ARV drug resistance mutations in a group of long-term vertically infected patients from Argentina. Methods: Plasma samples from 71 patients (38 children and 33 adolescents) were collected for genotypic HIV-1 ARV resistance testing during the period between February 2006 and October 2008. Statistically significant pairwise associations between ARV resistance mutations in pol, as well as associations between mutations and drug exposure, were identified using Fisher's exact tests with Bonferroni and false discovery rate corrections. Phylogenetic analyses were performed for subtype assignment. Results: In protease (PR), resistance-associated mutations M46I/L, I54M/L/V/A/S and V82A/F/T/S/M/I were associated with each other and with minor mutations at codons 10, 24 and 71. Mutations V82A/F/T/S/M/I were primarily selected by the administration of ritonavir (RTV) in an historical ARV regimen. In reverse transcriptase, thymidine analogue mutation (TAM)1 profile was more common than TAM2. The non-nucleoside K103N+L100I mutations were observed at high frequency (15.5%) and were significantly associated with the nucleoside mutation L74V in BF recombinants. Conclusions: Associations of mutations at PR sites reflect the frequent use of RTV at an early time in this group of patients and convergent resistance mechanisms driven by the high exposure to protease inhibitors, as well as local HIV-1 diversity. The results provide clinical evidence of a molecular interaction between K103N+L100I and L74V mutations at the reverse transcriptase gene in vivo, limiting the future use of second-generation non-nucleoside reverse transcriptase inhibitors such as etravirine.Fil: Aulicino, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Laboratorio de Biología Celular y Retrovirus; ArgentinaFil: Rocco, Carlos Alberto. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Laboratorio de Biología Celular y Retrovirus; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mecikovsky, Debora. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Laboratorio de Biología Celular y Retrovirus; ArgentinaFil: Bologna, Rosa. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Laboratorio de Biología Celular y Retrovirus; ArgentinaFil: Mangano, Andrea María Mercedes. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Laboratorio de Biología Celular y Retrovirus; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sen, Luisa. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Laboratorio de Biología Celular y Retrovirus; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Impact of the time to achieve viral control on the dynamics of circulating HIV-1 reservoir in vertically infected children with long-term sustained virological suppression: A longitudinal study.

OBJECTIVE:Determine the decay rate of HIV-1 DNA reservoir in vertically infected children during sustained viral suppression (VS) and how it is affected by the age at VS. METHODS:This study included 37 HIV-1 vertically infected children on suppressive antiretroviral therapy for at least 4 years. Children were grouped according to the age of antiretroviral therapy initiation (≤0.5 or >0.5 yrs) and to the age at VS (≤1.5, between >1.5 and 4, and >4 years). Decay of cell-associated HIV-1 DNA (CA-HIV-DNA) level and 2-long terminal repeats (2-LTR) circles frequency were analyzed over 4 years of viral suppression using piecewise linear mixed-effects model with two splines and logistic regression, respectively. RESULTS:CA-HIV-DNA in peripheral blood mononuclear cells had a significant decay during the first two years of VS [-0.26 (95% CI: -0.43, -0.09) log10 copies per one million cells (cpm)/year], and subsequently reached a plateau [-0.06 (95% CI: -0.15, 0.55) log10 cpm/year]. The initial decay was higher in children who achieved VS by 1.5 years of age compared to those who achieved VS between >1.5 and 4 years and those after 4 years of age: -0.51 (95% CI:-0.94, -0.07), -0.35 (95% CI:-0.83, 0.14), and -0.21 (95% CI:-0.39, -0.02) log10cpm PBMC/year, respectively. The 2-LTR circles frequency decayed significantly, from 82.9% at pre-VS to 37.5% and 28.1% at 2 and 4 years of VS, respectively (P = .0009). CONCLUSIONS:These data highlight that achieving VS during the first 18 months of life limit the establishment of HIV-1 reservoirs, reinforcing the clinical benefit of very early effective therapy in children

Prediction of lipids levels under linear mixed models (LMM).

<p>The contribution of each factor was evaluated with Wald test on 127 individuals with full haplotype characterization; p-values are depicted. Significant p-values (p<0.003125, after Bonferroni correction) are indicated in bold numbers. Correlation sign is depicted between parentheses for p-values below 0.05. NA: variable excluded by stepwise backward elimination. ¹Alternative 1 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039678#pone.0039678.s001" target="_blank">Figure S1</a>). ²Alternative 2 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039678#pone.0039678.s001" target="_blank">Figure S1</a>). ³Alternative 3 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039678#pone.0039678.s001" target="_blank">Figure S1</a>).</p

Hypercholesterolemia Is Associated with the Apolipoprotein C-III (<em>APOC3</em>) Genotype in Children Receiving HAART: An Eight-Year Retrospective Study

<div><p>Polymorphisms in apolipoprotein genes have shown to be predictors of plasma lipid levels in adult cohorts receiving highly active antiretroviral therapy (HAART). Our objective was to confirm the association between the <em>APOC3</em> genotype and plasma lipid levels in an HIV-1-infected pediatric cohort exposed to HAART. A total of 130 HIV-1-infected children/adolescents that attended a reference center in Argentina were selected for an 8-year longitudinal study with retrospective data collection. Longitudinal measurements of plasma triglycerides, total cholesterol, HDL-C and LDL-C were analyzed under linear or generalized linear mixed models. The contribution of the <em>APOC3</em> genotype at sites −482, −455 and 3238 to plasma lipid levels prediction was tested after adjusting for potential confounders. Four major <em>APOC3</em> haplotypes were observed for sites −482/−455/3238, with estimated frequencies of 0.60 (C/T/C), 0.14 (T/C/C), 0.11 (C/C/C), and 0.11 (T/C/G). The <em>APOC3</em> genotype showed a significant effect only for the prediction of total cholesterol levels (p<0.0001). However, the magnitude of the differences observed was dependent on the drug combination (p = 0.0007) and the drug exposure duration at the time of the plasma lipid measurement (p = 0.0002). A lower risk of hypercholesterolemia was predicted for double and triple heterozygous individuals, mainly at the first few months after the initiation of Ritonavir-boosted protease inhibitor-based regimens. We report for the first time a significant contribution of the genotype to total cholesterol levels in a pediatric cohort under HAART. The genetic determination of <em>APOC3</em> might have an impact on a large portion of HIV-1-infected children at the time of choosing the treatment regimens or on the counter-measures against the adverse effects of drugs.</p> </div

Predicted risk of dyslipidemia according to different treatment options after one year of exposure.

<p>Odds ratio (OR) estimates for individuals with no <i>APOC3</i> variants (“000” haplotype) were adjusted by the variables listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039678#pone.0039678.s004" target="_blank">Table S1</a> under Generalized Liner Mixed-effects Model (GLMM), and after backward elimination algorithm. Dots depict punctual estimations for the mean effect of the exposure to each drug while lines depict the 95% confidence intervals.</p

Frequency of haplotype pairs (loci−482, 455 and 3238 on the −<i>APOC3</i> gene, respectively) as observed in 127 HIV-1-infected pediatric patients.

<p>Inset on top right shows haplotype frequency estimation by an Expectation-Maximization (EM) algorithm.</p

Generalized Linear Mixed-effects Model (GLMM) projections of hypercholesterolemia risk for patients carrying the indicated haplotype pairs.

<p>GLMM projections for a ARV-experienced male (treatment regimen starting after 62 months on HAART, the observed mean time on HAART at new regimen initiation) under a RTV-boosted PI regimen without D4T. Upper and lower panels depict contrasts to WT before exposure or at the same exposure time. Haplotype notation indicates the gene dose at each locus. Dots represent punctual contrast projections. Dots with labels indicate haplotype pairs statistically significantly different from WT at the same time (only lower panel). Odds ratios for common haplotype pairs “020”, “120” and “121” could not be estimated due to power issues. The contribution of all the factors included in GLMM were included in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039678#pone.0039678.s007" target="_blank">Table S4</a>.</p

Hierarchical test for differences in absolute plasma levels (LMM)¹.

1<p>Bonferroni corrected significance level was α* = 0.003125.</p>2<p>Test for the contribution of <i>APOC3</i> genotypes taking into account interactions with specific treatment scheme (inclusion of PIs boosted with RTV and/or D4T) and time of exposure (alternative 3 vs. null, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039678#pone.0039678.s001" target="_blank">Figure S1</a>).</p>3<p>Test for the contribution of the interaction between <i>APOC3</i> genotypes and time of exposure (alternative 3 vs. alternative 2).</p>4<p>Test for the contribution of the interaction between <i>APOC3</i> genotypes and treatment scheme (alternative 2 vs. alternative 1).</p>5<p>Test for the contribution of <i>APOC3</i> genotypes without interaction (alternative 1 vs. null).</p