The kinesin KIF4 mediates HBV/HDV entry through the regulation of surface NTCP localization and can be targeted by RXR agonists in vitro

被引：12

作者：

Gad, Sameh A. ^{[1
,2
,3
]}

Sugiyama, Masaya ^{[4
]}

Tsuge, Masataka ^{[5
]}

Wakae, Kosho ^{[1
]}

Fukano, Kento ^{[1
]}

Oshima, Mizuki ^{[1
,6
]}

Sureau, Camille ^{[7
]}

Watanabe, Noriyuki ^{[1
]}

Kato, Takanobu ^{[1
]}

Murayama, Asako ^{[1
]}

Li, Yingfang ^{[1
]}

Shoji, Ikuo ^{[8
]}

Shimotohno, Kunitada ^{[9
]}

Chayama, Kazuaki ^{[10
,11
,12
]}

Muramatsu, Masamichi ^{[1
]}

Wakita, Takaji ^{[1
]}

Nozaki, Tomoyoshi ^{[2
]}

Aly, Hussein H. ^{[1
]}

机构：

[1] Natl Inst Infect Dis, Dept Virol 2, Tokyo, Japan

[2] Univ Tokyo, Grad Sch Med, Dept Biomed Chem, Tokyo, Japan

[3] Minia Univ, Fac Pharm, Dept Microbiol & Immunol, Al Minya, Egypt

[4] Natl Ctr Global Hlth & Med, Genome Med Sci Project, Chiba, Japan

[5] Hiroshima Univ, Nat Sci Ctr Basic Res & Dev, Hiroshima, Japan

[6] Tokyo Univ Sci, Grad Sch Sci & Technol, Noda, Chiba, Japan

[7] Inst Natl Transfus Sanguine, Paris, France

[8] Kobe Univ, Ctr Infect Dis, Grad Sch Med, Kobe, Hyogo, Japan

[9] Natl Ctr Global Hlth & Med, Ctr Hepatitis & Immunol, Chiba, Japan

[10] Hiroshima Univ, Grad Sch Biomed & Hlth Sci, Collaborat Res Lab Med Innovat, Hiroshima, Japan

[11] Hiroshima Univ, Res Ctr Hepatol & Gastroenterol, Grad Sch Biomed & Hlth Sci, Hiroshima, Japan

[12] RIKEN Ctr Integrat Med Sci, Yokohama, Kanagawa, Japan

来源：

PLOS PATHOGENS | 2022年 / 18卷 / 03期

基金：

日本学术振兴会;

关键词：

HEPATITIS-B-VIRUS; MEMBRANE TRANSPORTER; ENVELOPE PROTEIN; LIVER; DNA; HYPERCHOLANEMIA; REPLICATION; INFECTIVITY; HEPATOCYTES; INITIATION;

D O I：

10.1371/journal.ppat.1009983

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Intracellular transport via microtubule-based dynein and kinesin family motors plays a key role in viral reproduction and transmission. We show here that Kinesin Family Member 4 (KIF4) plays an important role in HBV/HDV infection. We intended to explore host factors impacting the HBV life cycle that can be therapeutically addressed using siRNA library transfection and HBV/NLuc (HBV/NL) reporter virus infection in HepG2-hNTCP cells. KIF4 silencing resulted in a 3-fold reduction in luciferase activity following HBV/NL infection. KIF4 knockdown suppressed both HBV and HDV infection. Transient KIF4 depletion reduced surface and raised intracellular NTCP (HBV/HDV entry receptor) levels, according to both cellular fractionation and immunofluorescence analysis (IF). Overexpression of wild-type KIF4 but not ATPase-null KIF4 mutant regained the surface localization of NTCP and significantly restored HBV permissiveness in these cells. IF revealed KIF4 and NTCP colocalization across microtubule filaments, and a co-immunoprecipitation study revealed that KIF4 interacts with NTCP. KIF4 expression is regulated by FOXM1. Interestingly, we discovered that RXR agonists (Bexarotene, and Alitretinoin) down-regulated KIF4 expression via FOXM1-mediated suppression, resulting in a substantial decrease in HBV-Pre-S1 protein attachment to HepG2-hNTCP cell surface and subsequent HBV infection in both HepG2-hNTCP and primary human hepatocyte (PXB) (Bexarotene, IC50 1.89 +/- 0.98 mu M) cultures. Overall, our findings show that human KIF4 is a critical regulator of NTCP surface transport and localization, which is required for NTCP to function as a receptor for HBV/HDV entry. Furthermore, small molecules that suppress or alleviate KIF4 expression would be potential antiviral candidates targeting HBV and HDV entry. Author summaryUnderstanding HBV/HDV entry machinery and the mechanism by which NTCP (HBV/HDV entry receptor) surface expression is regulated is crucial to develop antiviral entry inhibitors. We found that NTCP surface transport is mainly controlled by the motor kinesin KIF4. Surprisingly, KIF4 was negatively regulated by RXR receptors through FOXM1-mediated suppression. This study not only mechanistically correlated the role of RXR receptors in regulating HBV/HDV entry but also suggested a novel approach to develop therapeutic rexinoids for preventing HBV and/or HDV infections in important clinical situations, such as in patients undergoing liver transplantation or those who are at a high risk of HBV infection and unresponsive to HBV vaccination.

引用

页数：26