PMID- 28318036 OWN - NLM STAT- MEDLINE DCOM- 20170913 LR - 20211006 IS - 1527-3350 (Electronic) IS - 0270-9139 (Print) IS - 0270-9139 (Linking) VI - 66 IP - 1 DP - 2017 Jul TI - Autophagy determines efficiency of liver-directed gene therapy with adeno-associated viral vectors. PG - 252-265 LID - 10.1002/hep.29176 [doi] AB - Use of adeno-associated viral (AAV) vectors for liver-directed gene therapy has shown considerable success, particularly in patients with severe hemophilia B. However, the high vector doses required to reach therapeutic levels of transgene expression caused liver inflammation in some patients that selectively destroyed transduced hepatocytes. We hypothesized that such detrimental immune responses can be avoided by enhancing the efficacy of AAV vectors in hepatocytes. Because autophagy is a key liver response to environmental stresses, we characterized the impact of hepatic autophagy on AAV infection. We found that AAV induced mammalian target of rapamycin (mTOR)-dependent autophagy in human hepatocytes. This cell response was critically required for efficient transduction because under conditions of impaired autophagy (pharmacological inhibition, small interfering RNA knockdown of autophagic proteins, or suppression by food intake), recombinant AAV-mediated transgene expression was markedly reduced, both in vitro and in vivo. Taking advantage of this dependence, we employed pharmacological inducers of autophagy to increase the level of autophagy. This resulted in greatly improved transduction efficiency of AAV vectors in human and mouse hepatocytes independent of the transgene, driving promoter, or AAV serotype and was subsequently confirmed in vivo. Specifically, short-term treatment with a single dose of torin 1 significantly increased vector-mediated hepatic expression of erythropoietin in C57BL/6 mice. Similarly, coadministration of rapamycin with AAV vectors resulted in markedly enhanced expression of human acid-alpha-glucosidase in nonhuman primates. CONCLUSION: We identified autophagy as a pivotal cell response determining the efficiency of AAVs intracellular processing in hepatocytes and thus the outcome of liver-directed gene therapy using AAV vectors and showed in a proof-of-principle study how this virus-host interaction can be employed to enhance efficacy of this vector system. (Hepatology 2017;66:252-265). CI - (c) 2017 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of the American Association for the Study of Liver Diseases. FAU - Hosel, Marianna AU - Hosel M AD - Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany. AD - German Center for Infection Research (DZIF), Partner sites Bonn-Cologne and Hannover-Braunschweig, Germany. FAU - Huber, Anke AU - Huber A AD - Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany. AD - German Center for Infection Research (DZIF), Partner sites Bonn-Cologne and Hannover-Braunschweig, Germany. FAU - Bohlen, Susanne AU - Bohlen S AD - Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany. FAU - Lucifora, Julie AU - Lucifora J AD - INSERM, U1052, Cancer Research Center of Lyon (CRCL), University of Lyon, Lyon, France. FAU - Ronzitti, Giuseppe AU - Ronzitti G AD - Genethon and INSERM U951, Evry, France. FAU - Puzzo, Francesco AU - Puzzo F AD - Genethon and INSERM U951, Evry, France. FAU - Boisgerault, Florence AU - Boisgerault F AD - Genethon and INSERM U951, Evry, France. FAU - Hacker, Ulrich T AU - Hacker UT AD - University Medicine Leipzig, University Cancer Center Leipzig (UCCL), Leipzig, Germany. FAU - Kwanten, Wilhelmus J AU - Kwanten WJ AUID- ORCID: 0000-0001-9474-6695 AD - Laboratory of Experimental Medicine and Pediatrics (LEMP), University of Antwerp, Antwerp, Belgium. FAU - Kloting, Nora AU - Kloting N AD - IFB Adiposity Diseases, University of Leipzig, Leipzig, Germany. FAU - Bluher, Matthias AU - Bluher M AD - Department of Medicine, University of Leipzig, Leipzig, Germany. FAU - Gluschko, Alexander AU - Gluschko A AD - Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany. AD - Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany. FAU - Schramm, Michael AU - Schramm M AD - Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany. AD - Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany. FAU - Utermohlen, Olaf AU - Utermohlen O AD - Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany. AD - Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany. FAU - Bloch, Wilhelm AU - Bloch W AD - Department of Molecular and Cellular Sport Medicine, German Sport University Cologne, Cologne, Germany. FAU - Mingozzi, Federico AU - Mingozzi F AD - Genethon and INSERM U951, Evry, France. AD - University Pierre and Marie Curie, Paris, France. FAU - Krut, Oleg AU - Krut O AD - Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany. FAU - Buning, Hildegard AU - Buning H AD - Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany. AD - German Center for Infection Research (DZIF), Partner sites Bonn-Cologne and Hannover-Braunschweig, Germany. AD - Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany. LA - eng GR - 617432/ERC_/European Research Council/International PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20170529 PL - United States TA - Hepatology JT - Hepatology (Baltimore, Md.) JID - 8302946 SB - IM MH - Animals MH - Autophagy/*genetics MH - Cells, Cultured MH - Dependovirus/*genetics MH - Disease Models, Animal MH - Female MH - Gene Transfer Techniques MH - Genetic Therapy/*methods MH - Genetic Vectors MH - Hepatocytes/*cytology MH - Humans MH - Mice MH - Mice, Inbred C57BL MH - Random Allocation MH - Transduction, Genetic PMC - PMC5518300 EDAT- 2017/03/21 06:00 MHDA- 2017/09/14 06:00 PMCR- 2017/07/20 CRDT- 2017/03/21 06:00 PHST- 2016/11/08 00:00 [received] PHST- 2017/02/21 00:00 [revised] PHST- 2017/03/16 00:00 [accepted] PHST- 2017/03/21 06:00 [pubmed] PHST- 2017/09/14 06:00 [medline] PHST- 2017/03/21 06:00 [entrez] PHST- 2017/07/20 00:00 [pmc-release] AID - HEP29176 [pii] AID - 10.1002/hep.29176 [doi] PST - ppublish SO - Hepatology. 2017 Jul;66(1):252-265. doi: 10.1002/hep.29176. Epub 2017 May 29.