Centre for Microbiology Research, Kenya Medical Research Institute, Nairobi, Kenya.
Department of Pharmacology and Pharmacognosy, School of Pharmacy, University of Nairobi, Nairobi, Kenya.
PLoS One. 2022 Mar 2;17(3):e0260872. doi: 10.1371/journal.pone.0260872. eCollection 2022.
The effects of genetic variation of cytochrome P450 2B6 (CYP2B6) and constitutive androstane receptor (CAR) on efavirenz (EFV) plasma concentration was evaluated among 312 HIV patients in Nairobi Kenya. The EFV plasma concentration at steady-state were determined using ultra-high-performance liquid chromatography with a tandem quadruple mass spectrometer (LC-MS/MS). Thirteen CYP2B6 (329G>T, 341T>C, 444 G>T/C, 15582C>T, 516G>T, 548T>G, 637T>C, 785A>G, 18492C>T, 835G>C, 1459C>T and 21563C>T) and one CAR (540C>T) single nucleotide polymorphisms (SNPs) were genotyped using real-time polymerase chain reaction. HIV drug resistance mutations were detected using an in-house genotypic assay. The EFV concentration of patients ranged from 4 ng/mL to 332697 ng/mL (median 2739.5 ng/mL, IQR 1878-4891.5 ng/mL). Overall, 22% patients had EFV concentrations beyond therapeutic range of 1000-4000 ng/mL (4.5%% < 1000 ng/mL and 31.7% > 4000 ng/mL). Five SNPs (15582C>T, 516G>T, 785A>G, 983T>C and 21563C>T) were associated with higher EFV plasma concentration while 18492C>T with lower EFV plasma concentration (p<0.05). Strong linkage disequilibrium (LD) was observed for 15582C>T, 516G>T, 785A>G, 18492C>T, 983T>C, 21563C>T, 1459C>T and CAR 540C>T. Sixteen haplotypes were observed and CTGCTTCC, CTGCTTCT, TTGCTTCT and CGACCCCT were associated with high EFV plasma concentration. In multivariate analysis, factors significantly associated with EFV plasma concentration included; the presence of skin rash (β = 1379, 95% confidence interval (CI) = 3216.9-3416.3; p < 0.039), T allele of CYP2B6 516G>T (β = 1868.9, 95% CI 3216.9-3416.3; p < 0.018), the C allele of CYP2B6 983T>C (β = 2638.3, 95% CI = 1348-3929; p < 0.0001), T allele of CYP2B6 21563C>T (β = 1737, 95% CI = 972.2-2681.9; p < 0.0001) and the presence of 5 to 7 numbers of SNPs per patient (β = 570, 95% CI = 362-778; p < 0.0001) and HIV viral load ≤1000 cells/mL (β = -4199.3, 95% CI = -7914.9 --483.6; p = 0.027). About 36.2% of the patients had EFV plasma concentrations beyond therapeutic window, posing high risk of treatment failure or toxicity. The SNPs of CYP2B6 516G>T, CYP2B6 983T>C, 21563C>T, presence of higher numbers of SNPs per patient and haplotypes CTGCTTCC, CTGCTTCT, TTGCTTCT and CGACCCCT could efficiently serves as genetic markers for EFV plasma concentration and could guide personalization of EFV based ART treatment in Kenya.
在肯尼亚内罗毕的 312 名 HIV 患者中，评估了细胞色素 P450 2B6（CYP2B6）和组成型雄烷受体（CAR）的遗传变异对依非韦伦（EFV）血浆浓度的影响。使用超高效液相色谱-串联四极杆质谱联用（LC-MS/MS）法测定稳态时的 EFV 血浆浓度。使用实时聚合酶链反应（PCR）对 13 个 CYP2B6（329G>T、341T>C、444 G>T/C、15582C>T、516G>T、548T>G、637T>C、785A>G、18492C>T、835G>C、1459C>T 和 21563C>T）和 1 个 CAR（540C>T）单核苷酸多态性（SNP）进行基因分型。使用内部基因分型测定法检测 HIV 耐药突变。患者的 EFV 浓度范围为 4ng/mL 至 332697ng/mL（中位数 2739.5ng/mL，IQR 1878-4891.5ng/mL）。总体而言，22%的患者 EFV 浓度超过 1000-4000ng/mL 的治疗范围（4.5%<1000ng/mL 和 31.7%>4000ng/mL）。5 个 SNP（15582C>T、516G>T、785A>G、983T>C 和 21563C>T）与较高的 EFV 血浆浓度相关，而 18492C>T 与较低的 EFV 血浆浓度相关（p<0.05）。15582C>T、516G>T、785A>G、18492C>T、983T>C、21563C>T、1459C>T 和 CAR 540C>T 之间存在强连锁不平衡（LD）。观察到 16 种单倍型，CTGCTTCC、CTGCTTCT、TTGCTTCT 和 CGACCCCT 与较高的 EFV 血浆浓度相关。多变量分析显示，与 EFV 血浆浓度显著相关的因素包括：皮疹（β=1379，95%置信区间（CI）=3216.9-3416.3；p<0.039）、CYP2B6 516G>T 的 T 等位基因（β=1868.9，95%CI=3216.9-3416.3；p<0.018）、CYP2B6 983T>C 的 C 等位基因（β=2638.3，95%CI=1348-3929；p<0.0001）、CYP2B6 21563C>T 的 T 等位基因（β=1737，95%CI=972.2-2681.9；p<0.0001）和患者每例 SNP 数为 5 至 7 个（β=570，95%CI=362-778；p<0.0001）以及 HIV 病毒载量≤1000 个细胞/ml（β=-4199.3，95%CI=-7914.9--483.6；p=0.027）。约 36.2%的患者 EFV 血浆浓度超过治疗窗，存在治疗失败或毒性的高风险。CYP2B6 516G>T、CYP2B6 983T>C、21563C>T、患者每例 SNP 数较高和 CTGCTTCC、CTGCTTCT、TTGCTTCT 和 CGACCCCT 等单倍型可作为 EFV 血浆浓度的遗传标记，可指导肯尼亚 EFV 基于 ART 的个体化治疗。
