Liver Unit, Liver Diseases - Viral Hepatitis, Vall d'Hebron Institut of Research (VHIR), Hospital Universitari Vall d'Hebron (HUVH), Barcelona, Spain; Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Madrid, Spain.
Liver Unit, Liver Diseases - Viral Hepatitis, Vall d'Hebron Institut of Research (VHIR), Hospital Universitari Vall d'Hebron (HUVH), Barcelona, Spain.
Antiviral Res. 2020 Feb;174:104694. doi: 10.1016/j.antiviral.2019.104694. Epub 2019 Dec 16.
A percentage of hepatitis C virus (HCV)-infected patients fail direct acting antiviral (DAA)-based treatment regimens, often because of drug resistance-associated substitutions (RAS). The aim of this study was to characterize the resistance profile of a large cohort of patients failing DAA-based treatments, and investigate the relationship between HCV subtype and failure, as an aid to optimizing management of these patients. A new, standardized HCV-RAS testing protocol based on deep sequencing was designed and applied to 220 previously subtyped samples from patients failing DAA treatment, collected in 39 Spanish hospitals. The majority had received DAA-based interferon (IFN) α-free regimens; 79% had failed sofosbuvir-containing therapy. Genomic regions encoding the nonstructural protein (NS) 3, NS5A, and NS5B (DAA target regions) were analyzed using subtype-specific primers. Viral subtype distribution was as follows: genotype (G) 1, 62.7%; G3a, 21.4%; G4d, 12.3%; G2, 1.8%; and mixed infections 1.8%. Overall, 88.6% of patients carried at least 1 RAS, and 19% carried RAS at frequencies below 20% in the mutant spectrum. There were no differences in RAS selection between treatments with and without ribavirin. Regardless of the treatment received, each HCV subtype showed specific types of RAS. Of note, no RAS were detected in the target proteins of 18.6% of patients failing treatment, and 30.4% of patients had RAS in proteins that were not targets of the inhibitors they received. HCV patients failing DAA therapy showed a high diversity of RAS. Ribavirin use did not influence the type or number of RAS at failure. The subtype-specific pattern of RAS emergence underscores the importance of accurate HCV subtyping. The frequency of "extra-target" RAS suggests the need for RAS screening in all three DAA target regions.
一部分丙型肝炎病毒 (HCV) 感染患者在接受直接作用抗病毒 (DAA) 药物治疗方案后会出现治疗失败，这通常是由于药物耐药相关突变 (RAS) 引起的。本研究旨在对大量 DAA 治疗失败患者的耐药谱进行特征分析，并探讨 HCV 亚型与治疗失败之间的关系，为这些患者的管理提供帮助。我们设计并应用了一种基于深度测序的新型 HCV-RAS 检测标准化方案，对来自 39 家西班牙医院的 220 例先前经亚型鉴定的 DAA 治疗失败患者的样本进行检测。这些患者大多接受了无干扰素 (IFN)α 的 DAA 治疗方案；79%的患者曾接受索非布韦治疗失败。使用亚型特异性引物对编码非结构蛋白 (NS)3、NS5A 和 NS5B（DAA 靶区）的基因组区域进行分析。病毒亚型分布如下：基因型 (G)1，62.7%；G3a，21.4%；G4d，12.3%；G2，1.8%；混合感染，1.8%。总体而言，88.6%的患者至少携带 1 种 RAS，19%的患者在突变谱中携带频率低于 20%的 RAS。有无利巴韦林对 RAS 选择无影响。无论接受何种治疗，每种 HCV 亚型均表现出特定类型的 RAS。值得注意的是，治疗失败的患者中有 18.6%的患者在目标蛋白中未检测到 RAS，30.4%的患者在其接受的抑制剂非作用靶点蛋白中存在 RAS。DAA 治疗失败的 HCV 患者表现出 RAS 的高度多样性。利巴韦林的使用并未影响治疗失败时 RAS 的类型或数量。RAS 出现的亚型特异性模式强调了准确 HCV 亚型鉴定的重要性。“非靶位”RAS 的出现频率提示需要在三个 DAA 靶区均进行 RAS 筛查。
