PMID- 26276683 OWN - NLM STAT- MEDLINE DCOM- 20170801 LR - 20211204 IS - 1468-3288 (Electronic) IS - 0017-5749 (Print) IS - 0017-5749 (Linking) VI - 65 IP - 12 DP - 2016 Dec TI - Host cell mTORC1 is required for HCV RNA replication. PG - 2017-2028 LID - 10.1136/gutjnl-2014-308971 [doi] AB - OBJECTIVE: Chronically HCV-infected orthotopic liver transplantation (OLT) recipients appear to have improved outcomes when their immunosuppressive regimen includes a mammalian target of rapamycin (mTOR) inhibitor. The mechanism underlying this observation is unknown. DESIGN: We used virological assays to investigate mTOR signalling on the HCV replication cycle. Furthermore, we analysed HCV RNA levels of 42 HCV-positive transplanted patients treated with an mTOR inhibitor as part of their immunosuppressive regimen. RESULTS: The mTOR inhibitor rapamycin was found to be a potent inhibitor for HCV RNA replication in Huh-7.5 cells as well as primary human hepatocytes. Half-maximal inhibition was observed at 0.01 microg/mL, a concentration that is in the range of serum levels seen in transplant recipients and does not affect cell proliferation. Early replication cycle steps such as cell entry and RNA translation were not affected. Knockdown of raptor, an essential component of mTORC1, but not rictor, an essential component of mTORC2, inhibited viral RNA replication. In addition, overexpression of raptor led to higher viral RNA replication, demonstrating that mTORC1, but not mTORC2, is required for HCV RNA replication. In 42 HCV-infected liver-transplanted or kidney-transplanted patients who were switched to an mTOR inhibitor, we could verify that mTOR inhibition decreased HCV RNA levels in vivo. CONCLUSIONS: Our data identify mTORC1 as a novel HCV replication factor. These findings suggest an underlying mechanism for the observed benefits of mTOR inhibition in HCV-positive OLT recipients and potentiate further investigation of mTOR-containing regimens in HCV-positive recipients of solid organ transplants. CI - Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/. FAU - Stohr, Stefanie AU - Stohr S AD - Department of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule Hannover, Hannover, Germany. AD - German Center for Infection Research (DZIF), Hannover, Germany. FAU - Costa, Rui AU - Costa R AD - Department of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule Hannover, Hannover, Germany. FAU - Sandmann, Lisa AU - Sandmann L AD - Department of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule Hannover, Hannover, Germany. AD - German Center for Infection Research (DZIF), Hannover, Germany. FAU - Westhaus, Sandra AU - Westhaus S AD - Department of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule Hannover, Hannover, Germany. AD - German Center for Infection Research (DZIF), Hannover, Germany. AD - Institute for Molecular Biology, Medizinische Hochschule Hannover, Hannover, Germany. FAU - Pfaender, Stephanie AU - Pfaender S AD - Division of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Hannover, Germany. FAU - Anggakusuma AU - Anggakusuma AD - Division of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Hannover, Germany. FAU - Dazert, Eva AU - Dazert E AD - Cell Growth and Development Biozentrum, Universitat Basel, Basel, Switzerland. FAU - Meuleman, Philip AU - Meuleman P AD - Center for Vaccinology, Ghent University, Ghent University Hospital, Ghent, Belgium. FAU - Vondran, Florian W R AU - Vondran FW AD - German Center for Infection Research (DZIF), Hannover, Germany. AD - Department of General, Visceral and Transplantation Surgery, Medizinische Hochschule Hannover, Hannover, Germany. FAU - Manns, Michael P AU - Manns MP AD - Department of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule Hannover, Hannover, Germany. AD - German Center for Infection Research (DZIF), Hannover, Germany. FAU - Steinmann, Eike AU - Steinmann E AD - Division of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), Hannover, Germany. FAU - von Hahn, Thomas AU - von Hahn T AD - Department of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule Hannover, Hannover, Germany. AD - German Center for Infection Research (DZIF), Hannover, Germany. AD - Institute for Molecular Biology, Medizinische Hochschule Hannover, Hannover, Germany. FAU - Ciesek, Sandra AU - Ciesek S AD - Department of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule Hannover, Hannover, Germany. AD - German Center for Infection Research (DZIF), Hannover, Germany. AD - Integrated Research and Treatment Centre-Transplantation (IFB-Tx), Hannover, Germany. LA - eng PT - Journal Article DEP - 20150814 PL - England TA - Gut JT - Gut JID - 2985108R RN - 0 (Multiprotein Complexes) RN - 0 (RNA, Viral) RN - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 1) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) SB - IM MH - Gene Silencing/drug effects MH - Hepacivirus/*drug effects MH - Hepatitis C, Chronic/therapy MH - Hepatocytes/*drug effects MH - Humans MH - Liver Transplantation MH - Mechanistic Target of Rapamycin Complex 1 MH - Multiprotein Complexes/*pharmacology MH - RNA, Viral/*drug effects MH - TOR Serine-Threonine Kinases/*pharmacology MH - Treatment Outcome MH - Virus Replication/*drug effects PMC - PMC5136730 OTO - NOTNLM OT - HEPATITIS C OT - LIVER TRANSPLANTATION COIS- Conflicts of Interest: None declared. EDAT- 2015/08/16 06:00 MHDA- 2017/08/02 06:00 PMCR- 2016/12/05 CRDT- 2015/08/16 06:00 PHST- 2014/12/08 00:00 [received] PHST- 2015/07/10 00:00 [accepted] PHST- 2015/08/16 06:00 [pubmed] PHST- 2017/08/02 06:00 [medline] PHST- 2015/08/16 06:00 [entrez] PHST- 2016/12/05 00:00 [pmc-release] AID - gutjnl-2014-308971 [pii] AID - 10.1136/gutjnl-2014-308971 [doi] PST - ppublish SO - Gut. 2016 Dec;65(12):2017-2028. doi: 10.1136/gutjnl-2014-308971. Epub 2015 Aug 14.