title: Apolipoprotein B100 is required for hepatitis C infectivity and Mipomersen inhibits hepatitis C.
abstract: OBJECTIVE
  To characterize the role of apolipoprotein B100 (apoB100) in hepatitis C viral (HCV) infection.
  
  
  METHODS
  In this study, we utilize a gene editing tool, transcription activator-like effector nucleases (TALENs), to generate human hepatoma cells with a stable genetic deletion ofAPOBto assess of apoB in HCV. Using infectious cell culture-competent HCV, viral pseudoparticles, replicon models, and lipidomic analysis we determined the contribution of apoB to each step of the viral lifecycle. We further studied the effect of mipomersen, an FDA-approved antisense inhibitor of apoB100, on HCV usingin vitrocell-culture competent HCV and determined its impact on viral infectivity with the TCID50 method.
  
  
  RESULTS
  We found that apoB100 is indispensable for HCV infection. Using the JFH-1 fully infectious cell-culture competent virus in Huh 7 hepatoma cells with TALEN-mediated gene deletion of apoB (APOB KO), we found a significant reduction in HCV RNA and protein levels following infection. Pseudoparticle and replicon models demonstrated that apoB did not play a role in HCV entry or replication. However, the virus produced byAPOB KOcells had significantly diminished infectivity as measured by the TCID-50 method compared to wild-type virus. Lipidomic analysis demonstrated that these virions have a fundamentally altered lipidome, with complete depletion of cholesterol esters. We further demonstrate that inhibition of apoB using mipomersen, an FDA-approved anti-sense oligonucleotide, results in a potent anti-HCV effect and significantly reduces the infectivity of the virus.
  
  
  CONCLUSIONS
  ApoB is required for the generation of fully infectious HCV virions, and inhibition of apoB with mipomersen blocks HCV. Targeting lipid metabolic pathways to impair viral infectivity represents a novel host targeted strategy to inhibit HCV.
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Esperance A K
authors.initials: EA
authors.lastname: Schaefer
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: James
authors.initials: J
authors.lastname: Meixiong
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Christina
authors.initials: C
authors.lastname: Mark
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Amy
authors.initials: A
authors.lastname: Deik
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Motola
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Dahlene
authors.initials: D
authors.lastname: Fusco
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Andrew
authors.initials: A
authors.lastname: Yang
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Brisac
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Shadi
authors.initials: S
authors.lastname: Salloum
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Wenyu
authors.initials: W
authors.lastname: Lin
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Clary B
authors.initials: CB
authors.lastname: Clish
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Lee F
authors.initials: LF
authors.lastname: Peng
authors.affiliation: Esperance AK Schaefer, James Meixiong, Christina Mark, Daniel L Motola, Dahlene Fusco, Andrew Yang, Cynthia Brisac, Shadi Salloum, Wenyu Lin, Lee F Peng, Raymond T Chung, Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, United States.
authors.forename: Raymond T
authors.initials: RT
authors.lastname: Chung
mesh_terms: D053299:Apolipoprotein B-100 / Q000037:antagonists & inhibitors / Q000235:genetics*; D002460:Cell Line; D055786:Gene Knockout Techniques; D016174:Hepacivirus; D006526:Hepatitis C / Q000235:genetics* / Q000821:virology; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism* / Q000821:virology; D006801:Humans; D066298:In Vitro Techniques; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology; D009841:Oligonucleotides / Q000494:pharmacology; D012367:RNA, Viral / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: D053299:Apolipoprotein B-100; D020319:Oligodeoxyribonucleotides, Antisense; D009841:Oligonucleotides; D012367:RNA, Viral; D014764:Viral Proteins; C524142:mipomersen
keywords: Apolipoprotein; Gene silencing; Hepatitis C virus; Lipid; Viral replication