Częstość wybranych polimorfizmów pojedynczych nukleotydów związanych z osteoporozą u Polaków zakażonych i niezakażonych HIV

Introduction: Osteoporosis poses significant risk for HIV infected subjects in the era of the long-term antiretroviral treatment. For this study frequency of the selected 11 single nucleotide polymorphisms (SNP single nucleotide polymorphism) previously associated with osteoporosis risk in HIV infected and uninfected cohorts was analysed. Association with the SNP variation and the risk of osteoporosis in the entire study and in HIV-infected cases was investigated. Material and methods: The study included 568 patients (226 women and 342 men): 315 HIV-infected patients and 253 anti-HIV negative cases. Osteoporosis was confirmed using dual energy absorptiometry ionizing radiation (DXA) in eight HIV infected patients and three controls [odds ratio (OR): 7.66, 95% CI: 1.98–29.6; p = 0.001; relative risk (RR): 7.04, 95% CI: 1.98–25.97, p = 0.0019]. SNP assays performed for collagen type 1 (COL1A1) rs1800012, parathyroid hormone (PTH) rs9630182, estrogen receptor gene (ER1) rs2077647, rs3020314 and rs1884051, Vitamin D receptor (VDR) rs1544410 and rs731236, Osteoprotegerin rs4355801, LDL receptor protein (LRP5) rs3736228, RANK rs3018362 and CYP19A1 (aromatase) rs700518 using TaqMan SNP Genotyping Assay (Applied Biosystems) according to the manufacturer’s protocol. For statistics Statistica 12 software was used. Results: Majority of allele frequencies for the studied polymorphisms were consistent with the Hardy-Weinberg equilibrium. CC homozygotes for ER1 rs2077647 were notably more common in HIV (+) cases compared to controls [OR: 2.29, 95%CI: 1.25–4.19, p = 0.003; RR: 2.11, 1.22–3.68, p = 0.0072]. Also GG homozygotes for ER1 rs1884051 were more common both in HIV(+) [OR: 2.57, 1.33–4.94, p = 0.0016; RR: 2.37, 1.29–4.36, p = 0.002] and in all patients with osteoporosis [OR 5.04, 1.24–20.4, p = 0.025; RR 3.94, 1.38–11.24, p = 0.043] . Additionally, in HIV (+) patients parathyroid hormone rs9630182 T allele was notably more common [OR: 1.4, 1.0–1.97, p = 0.024; RR: 1.15, 0.99–1.33, p = 0.029]. Conclusions: Genetic variability for the osteoporosis-associated SNPs was similar in HIV-infected patients and uninfected persons. ER1rs1884051 variants may be associated with the increased osteoporosis risk, but increased incidence of osteoporosis in HIV-infected compared to uninfected people seems to be weakly associated with investigated single nucleotide polymorphisms. Variation in the gene for the estrogen receptor ER1rs1884051 was significantly more frequent in patients with osteoporosis and more common in HIV infection.Wstęp: Osteoporoza staje się istotnym zagrożeniem dla pacjentów zakażonych HIV, szczególnie w erze długotrwałego leczenia antyretrowirusowego. W badaniu oceniono częstość wybranych 11 polimorfizmów pojedynczych nukleotydów (single nucleotide polymorphism, SNP) związanych z osteoporozą u pacjentów zakażonych HIV i bez tego zakażenia. Zbadano, czy zmienność genetyczna wybranych SNP wpływa na ryzyko osteoporozy w całej badanej populacji i u zakażonych HIV. Materiał i metody: Do badania włączono 568 osób (226 kobiet i 342 mężczyzn): 315 pacjentów zakażonych HIV i 253 osoby anty-HIV ujemne. Osteoporozę potwierdzono w badaniu za pomocą absorpcjometrii podwójnej energii promieniowania jonizującego (Dual-energy X-ray absorptiometry, DXA) u 8 pacjentów zakażonych HIV i 3 z grupy kontrolnej [iloraz szans (odds ratio, OR): 7,66; 95% Cl: 1,98–29,6; p = 0,0010; ryzyko względne (relative risk, RR): 7,04; 95% Cl: 1,98–25,97; p = 0,0019]. Zbadano SNP dla genów: kolagenu typu I (COLIA1) rs1800012, parathormonu (PTH) rs9630182, receptora estrogenów typu 1 (ER1) rs2077647 rs3020314 i rs1884051, receptora witaminy D (VDR) rs1544410 i rs731236, osteoprotegeryny (OPG) rs4355801, białka receptorowego dla LDL (LRP5) rs3736228, RANK rs3018362 i aromatazy (CYP19A1) rs700518 przy użyciu zestawów TaqMan SNP Genotyping Assay (AppliedBiosystems) zgodnie z protokołem producenta. Wyniki uzyskanych badań opracowano statystycznie w programie Statistica 12. Wyniki: Częstości alleli większości badanych polimorfizmów były zgodne z prawem Hardy-Weinberga. Analizując występowanie poszczególnych genotypów, stwierdzono istotnie częstsze wstępowanie u zakażonych HIV w porównaniu z grupą kontrolną homozygoty CC ER1 rs2077647 (OR: 2,29; 1,25–4,19; p = 0,003; RR: 2,11; 1,22–3,68; p = 0,0072). Także częściej występowała homozygota GG ER1 rs1884051 u zakażonych HIV (OR: 2,57; 1,33–4,94; p = 0,0016; RR: 2,37; 1,29–4,36; p = 0,002) oraz u pacjentów z osteoporozą (OR: 5,04; 1,24–20,4; p = 0,025; RR: 3,94; 1,38–11,24; p = 0,043). Dodatkowo u pacjentów zakażonych HIV istotnie częściej (p = 0,047) występował allel T genu parathormonu rs9630182 (OR: 1,4; 1,0–1,97; p = 0,024; RR: 1,15; 0,99–1,33; p = 0,029). Wnioski: Zmienność genetyczna pojedynczych nukleotydów związanych z występowaniem osteoporozy była podobna u pacjentów zakażonych HIV i osób niezakażonych. Warianty ER1 rs1884051 mogą mieć związek ze zwiększonym ryzykiem osteoporozy, ale wyższe ryzyko osteoporozy u pacjentów zakażonych HIV w porównaniu z osobami niezakażonymi wydaje się mieć niewielki związek z badanymi polimorfizmami pojedynczych nukleotydów. Zmienność w genie dla receptora estrogenów ER1rs1884051 istotnie częściej wystąpiła u pacjentów z osteoporozą i u zakażonych HIV

HIV-1 Subtype D Infections among Caucasians from Northwestern Poland—Phylogenetic and Clinical Analysis

Background: HIV-1 subtype D infections, which are associated with a faster rate of progression and lymphocyte CD4 decline, cognitive deficit and higher mortality, have rarely been found in native Europeans. In Northwestern Poland, however, infections with this subtype had been identified. This study aimed to analyze the sequence and clinical data for patients with subtype D using molecular phylogeography and identify transmission clusters and ancestry, as well as drug resistance, baseline HIV tropism and antiretroviral treatment efficacy. Methods: Phylogenetic analyses of local HIV-1 subtype D sequences were performed, with time to the most recent common ancestor inferred using Bayesian modeling. Sequence and drug resistance data were linked with the clinical and epidemiological information. Results: Subtype D was found in 24 non-immigrant Caucasian, heterosexually infected patients (75 % of females, median age at diagnosis of 49.5 years; IQR: 29–56 years). Partial pol sequences clustered monophyletically with the clades of Ugandan origin and no evidence of transmission from other European countries was found. Time to the most common recent ancestor was 1989.24 (95 % HPD: 1968.83–1994.46). Baseline drug resistance to nucleoside reverse transcriptase inhibitors was observed in 54.5 % of cases (mutations: M41L, K103N, T215S/D) with evidence of clustering, no baseline integrase or protease resistance and infrequent non-R5 tropism (13.6%). Virologic failure was observed in 60 % of cases an

Risk of All-Cause Mortality in HIV Infected Patients Is Associated with Clinical, Immunologic Predictors and the CCR5 Δ32 Deletion

OBJECTIVE: Investigation of the interplay between the CCR5 Δ32/wt genotype and demographic, epidemiological, clinical and immunological factors associated with mortality in the cART era. DESIGN: Longitudinal data from 507 HIV-infected patients following the Δ32 allele detection were analyzed. METHODS: Cumulative 15 years mortality was calculated using Kaplan-Meyer methodology. Hazard ratios were estimated using univariate Cox models. Basing on Akakie information criteria and statistical significance multivariate Cox model was constructed and effect plots presenting adjusted hazard ratio time-dependency were drawn. Analysis of the association of all-cause mortality and CCR5 Δ32/wt genotype prior to the antiretroviral treatment (cART) initiation (n = 507) and on the therapy (n = 422) was also performed. RESULTS: A mortality rate of 2.66 (CI 2.57-3.19) per 100 person-years was observed. Univariate analysis factors modifying the risk of death included the CCR5 genotype, gender, history of cART, AIDS diagnosis and also CD4 lymphocyte nadir, zenith, the latest CD4 count and stable levels >500 cells/µl. For multivariate analysis the following predictors were selected: CCR5 genotype (HR for wt/wt 2.53, CI 1.16-5.53, p = 0.02), gender (HR for males 1.91, 95%CI 1.1-3.36, p = 0.023), introduction of combined antiretroviral treatment (HR 4.85, CI 3.0-7.89, if untreated or treated <1 month, p<0.0001) CD4 count of 500 cells/µl for six months or more (HR 4.16, CI 1.95-8.88 if not achieved, p = 0.028), the latest CD4 count (HR 5.44, CI 3.39-8.74 for <100 cells/µl, p<0.0001) and history of AIDS (HR 1.69, CI 1.03-2.79, p = 0.039). Among untreated individuals the Δ32/wt genotype was associated with notably better survival (p = 0.026), while among cART treated individuals the Δ32 mutation did not correlate significantly with higher survival rates (p = 0.23). CONCLUSIONS: The Δ32 CCR5 allele is associated with a reduction of the risk of all-cause mortality in HIV (+) patients alongside clinical and immunologic predictors such as AIDS, history of cART, lymphocyte CD4 cell count and gender

Relationships between the subtype D sequences inferred using maximum likelihood phylogeny (GenBank deposited <i>pol</i> sequences).

<p>Country-specific sequences are marked with the same color: red – Poland, blue – Uganda, green – Tanzania, yellow – Europe (except Poland), brown – Cameroon, magenta - Senegal, cyan- Sudan, dark green – other African countries, violet – South America, grey – Asia, orange – North America.</p

Frequency of secondary drug resistance mutations in patients failing antiretroviral therapy.

<p>Frequency of secondary drug resistance mutations in patients failing antiretroviral therapy.</p

Drug resistance and treatment efficacy in the group infected with subtype D.

<p>*Percentage expressed adherence based on the number of months of medications dispensed by the number of months of follow-up; clinician assigned adherence based on the patient's statement regarding missed doses and treatment interruptions.</p

Phylogenetic trees of the subtype D sequences from Northwestern Poland.

<p>Figure a - maximum likelihood tree with bootstrap values for 1000 replicates drawn at the branches. Figure b – time scaled Bayesian MCMC tree. On the tree branches estimated time to the most recent common ancestor (tMRCA) and posterior probabilities expressed as percentage are shown. For both figures clustered sequences are marked in red and four identified clusters indicated as blue boxes and numbered are drawn on the right. Drug resistance mutations are marked at the tip nodes after the sequence identifier. *source patient for the transmission of the drug resistance within the cluster.</p

Frequency of baseline drug resistance mutations among treatment-naive patients.

<p>Frequency of baseline drug resistance mutations among treatment-naive patients.</p

Bayesian skyline plot for estimation of the number of subtype D HIV-1 cases in the local population.

<p>95% CI are marked in blue. Y-axis: predicted number of cases (log scale), X-axis: timescale (years).</p