Development of Cell Culture Infectious Hepatitis C Virus Genotype 1b Clones and 1b-Induced Angiogenesis and Cholesterol Disorders

Globally, hepatitis C virus (HCV) genotype 1b is most prevalent, and its infection has been found to associate with a higher risk of hepatocellular carcinoma (HCC) than other genotype viruses. However, an efficient infectious HCV genotype 1b culture system is unavailable, which has largely hampered the study of this important genotype virus. In this study, by using a systematic approach combining adaptive mutations and infectious 1a TNcc sequences, we succeeded in culture adaption of two full-length 1b clones for the reference strain Con1 and a clinical isolate A6, and designated as Con1cc and A6cc, respectively. Con1cc and A6cc replicated efficiently in hepatoma Huh7.5.1 cells, released HCV infectivity titers of 104.1 and 103.72 focus forming units per milliliter, respectively, and maintained the engineered mutations after passages. Both viruses responded to sofosbuvir and velpatasvir in a dose-dependent manner. With culture infectious 1b clones, we characterized the transcriptomes of 1b Con1cc-infected cells. Con1cc infection activated angiogenesis and disturbed cholesterol homeostasis compared with uninfected cells. In conclusion, we have developed infectious clones for genotype 1b and shown a novel strategy for culture adaptation of HCV isolates by using a genetically close backbone sequence. Besides, this study provides transcriptional landscape of HCV 1b-infected hepatoma cells facilitating the study of genotype 1b infection. Funding Statement: This work was supported by the National Natural Science Foundation of China (No. 81971938 for Y.-P.L.; 81772923 for Y.Z.), The Innovation Research Team for Basic and Clinical Studies on Chronic Liver Diseases of 2018 High-Level Health Teams of Zhuhai (for Y.-P.L. and F.X.), National Key Basic Research Program of China (No. 2015CB554301 for Y.-P.L.), Entrepreneurial Talent Team Award of Guangdong Province (No. 2016ZT06S252 for Y.-P.L.). Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: The use of patient serum was approved by the Medical Ethics Committee at the Zhongshan School of Medicine, Sun Yat-sen University (No. 2014-072).