过表达ESCRT系统成员提升细胞应对U18666A诱发胆固醇积聚的能力
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摘要
U18666A是一种细胞通透性的两亲性氨基类固醇,能够抑制胆固醇的转运与多泡体的生成,导致细胞内胆固醇的积聚,造成细胞凋亡。该研究发现, U18666A处理MLE12细胞后转运必需的内体分选复合体(endosomal sorting complexes required for transport, ESCRT)系统相关蛋白表达降低。该实验构建了过表达ESCRT-III系统成员Vps24以及Chmp1b的转基因细胞系,发现与转空载体的细胞相比,转基因细胞对抗U18666A引发的胆固醇积聚以及后续生理效应的能力有所增强。以上结果提示U18666A可以干扰ESCRT系统的一部分功能诱发细胞内胆固醇积聚,为人类与胆固醇积聚相关疾病的防治提供了新的思路。
U18666 A is a cell-permeable amphiphilic aminosteroid that inhibits the transport of cholesterol and the formation of multivesicular bodies, promotes the accumulation of intracellular cholesterol, and causes apoptosis. We found that endosomal sorting complexes required for transport(ESCRT) system-related protein expression decreased in MLE12 cells treated by U18666 A. Transgenic cell lines that overexpressed ESCRT-III system members Vps24 or Chmp1 b were constructed. Compared with cells which transfected empty vector, Vps24 or Chmp1 b over expression transgenic cells have an increased ability to counteract the accumulation of cholesterol and subsequent physiological effects induced by U18666 A. These results suggest that U18666 A may interfere with some functions of ESCRT system and provide a new approach to the prevention and treatment of cholesterol accumulation related diseases.
U18666 A is a cell-permeable amphiphilic aminosteroid that inhibits the transport of cholesterol and the formation of multivesicular bodies, promotes the accumulation of intracellular cholesterol, and causes apoptosis. We found that endosomal sorting complexes required for transport(ESCRT) system-related protein expression decreased in MLE12 cells treated by U18666 A. Transgenic cell lines that overexpressed ESCRT-III system members Vps24 or Chmp1 b were constructed. Compared with cells which transfected empty vector, Vps24 or Chmp1 b over expression transgenic cells have an increased ability to counteract the accumulation of cholesterol and subsequent physiological effects induced by U18666 A. These results suggest that U18666 A may interfere with some functions of ESCRT system and provide a new approach to the prevention and treatment of cholesterol accumulation related diseases.
引文
1 Hurley JH.ESCRTs are everywhere.EMBO J 2015;34(19):2398-407.
2 Frankel EB,Audhya A.ESCRT-dependent cargo sorting at multivesicular endosomes.Semin Cell Dev Biol 2018;74:4-10.
3 Wollert T,Hurley JH.Molecular mechanism of multivesicular body biogenesis by ESCRT complexes.Nature 2010;464(7290):864-9.
4 Sadler JBA,Wenzel DM,Williams LK,Guindo-Martinez M,Alam SL,Mercader JM,et al.A cancer-associated polymorphism in ESCRT-III disrupts the abscission checkpoint and promotes genome instability.Proc Natl Acad Sci USA 2018;115(38):E8900-8.
5 Lee YS,Pressman S,Andress AP,Kim K,White JL,Cassidy JJ,et al.Silencing by small RNAs is linked to endosomal trafficking.Nat Cell Biol 2009;11(9):1150-6.
6 Wegner CS,Rodahl LM,Stenmark H.ESCRT proteins and cell signalling.Traffic 2011;12(10):1291-7.
7 Dukes JD,Fish L,Richardson JD,Blaikley E,Burns S,Caunt CJ,et al.Functional ESCRT machinery is required for constitutive recycling of claudin-1 and maintenance of polarity in vertebrate epithelial cells.Mol Biol Cell 2011;22(17):3192-205.
8 Jimenez AJ,Maiuri P,Lafaurie-Janvore J,Divoux S,Piel M,Perez F.ESCRT machinery is required for plasma membrane repair.Science 2014;343(6174):1247136.
9 Skowyra ML,Schlesinger PH,Naismith TV,Hanson PI.Triggered recruitment of ESCRT machinery promotes endolysosomal repair.Science 2018;360(6384):eaar5078.
10 Olmos Y,Hodgson L,Mantell J,Verkade P,Carlton JG.ES-CRT-III controls nuclear envelope reformation.Nature 2015;522(7555):236-9.
11 Gong YN,Guy C,Olauson H,Becker JU,Yang M,Fitzgerald P,et al.ESCRT-III acts downstream of MLKL to regulate necroptotic cell death and its consequences.Cell 2017;169(2):286-300e16.
12 Morita E,Sandrin V,McCullough J,Katsuyama A,Baci Hamilton I,Sundquist WI.ESCRT-III protein requirements for HIV-1 budding.Cell Host Microbe 2011;9(3):235-42.
13 Liu Z,Wang L,Tan H,Zhou S,Fu T,Xia Y,et al.Synthesis of 1H-indazoles from N-tosylhydrazones and nitroaromatic compounds.Chem Commun(Camb)2014;50(39):5061-3.
14 Wollert T,Wunder C,Lippincott-Schwartz J,Hurley JH.Membrane scission by the ESCRT-III complex.Nature 2009;458(7235):172-7.
15 Koh CH,Cheung NS.Cellular mechanism of U18666A-mediated apoptosis in cultured murine cortical neurons:bridging Niemann-Pick disease type C and Alzheimer’s disease.Cell Signal2006;18(11):1844-53.
16 Higgins ME,Davies JP,Chen FW,Ioannou YA.Niemann-Pick C1 is a late endosome-resident protein that transiently associates with lysosomes and the trans-Golgi network.Mol Genet Metab1999;68(1):1-13.
17 Roszell BR,Tao JQ,Yu KJ,Huang S,Bates SR.Characterization of the Niemann-Pick C pathway in alveolar type II cells and lamellar bodies of the lung.Am J Physiol Lung Cell Mol Physiol2012;302(9):L919-32.
18 Chapuy-Regaud S,Subra C,Requena M,de Medina P,Amara S,Delton-Vandenbroucke I,et al.Progesterone and a phospholipase inhibitor increase the endosomal bis(monoacylglycero)phosphate content and block HIV viral particle intercellular transmission.Biochimie 2013;95(9):1677-88.
19 Elgner F,Ren H,Medvedev R,Ploen D,Himmelsbach K,Boller K,et al.The intracellular cholesterol transport inhibitor U18666Ainhibits the exosome-dependent release of mature hepatitis C virus.J Virol 2016;90(24):11181-96.
20 de Duve C,de Barsy T,Poole B,Trouet A,Tulkens P,Van Hoof F.Commentary lysosomotropic agents.Biochem Pharmacol 1974;23(18):2495-531.
21 Marceau F,Bawolak MT,Lodge R,Bouthillier J,Gagne-Henley A,Gaudreault RC,et al.Cation trapping by cellular acidic compartments:beyond the concept of lysosomotropic drugs.Toxicol Appl Pharmacol 2012;259(1):1-12.
22 Corless L,Crump CM,Griffin SD,Harris M.Vps4 and the ESCRT-III complex are required for the release of infectious hepatitis C virus particles.J Gen Virol 2010;91(Pt 2):362-72.
23 Barouch-Bentov R,Neveu G,Xiao F,Beer M,Bekerman E,Schor S,et al.Hepatitis C virus proteins interact with the endosomal sorting complex required for transport(ESCRT)machinery via ubiquitination to facilitate viral envelopment.MBio.2016;7(6):e01456-16.
24 Johnson DS,Bleck M,Simon SM.Timing of ESCRT-III protein recruitment and membrane scission during HIV-1 assembly.Elife2018;7:e36221.
25 Jiang B,Himmelsbach K,Ren H,Boller K,Hildt E.Subviral hepatitis B virus filaments,like infectious viral particles,are released via multivesicular bodies.J Virol 2015;90(7):3330-41.
26 Tang Y,Leao IC,Coleman EM,Broughton RS,Hildreth JE.Deficiency of niemann-pick type C-1 protein impairs release of human immunodeficiency virus type 1 and results in Gag accumulation in late endosomal/lysosomal compartments.J Virol2009;83(16):7982-95.
27 Lloyd-Evans E,Morgan AJ,He X,Smith DA,Elliot-Smith E,Sillence DJ,et al.Niemann-Pick disease type C1 is a sphingosine storage disease that causes deregulation of lysosomal calcium.Nat Med 2008;14(11):1247-55.
28 Scheffer LL,Sreetama SC,Sharma N,Medikayala S,Brown KJ,Defour A,et al.Mechanism of Ca2+-triggered ESCRT assembly and regulation of cell membrane repair.Nat Commun 2014;5:5646.
29 Du X,Kazim AS,Dawes IW,Brown AJ,Yang H.The AAAATPase VPS4/SKD1 regulates endosomal cholesterol trafficking independently of ESCRT-III.Traffic 2013;14(1):107-19.
30 Du X,Kazim AS,Brown AJ,Yang H.An essential role of Hrs/Vps27 in endosomal cholesterol trafficking.Cell Rep 2012;1(1):29-35.
2 Frankel EB,Audhya A.ESCRT-dependent cargo sorting at multivesicular endosomes.Semin Cell Dev Biol 2018;74:4-10.
3 Wollert T,Hurley JH.Molecular mechanism of multivesicular body biogenesis by ESCRT complexes.Nature 2010;464(7290):864-9.
4 Sadler JBA,Wenzel DM,Williams LK,Guindo-Martinez M,Alam SL,Mercader JM,et al.A cancer-associated polymorphism in ESCRT-III disrupts the abscission checkpoint and promotes genome instability.Proc Natl Acad Sci USA 2018;115(38):E8900-8.
5 Lee YS,Pressman S,Andress AP,Kim K,White JL,Cassidy JJ,et al.Silencing by small RNAs is linked to endosomal trafficking.Nat Cell Biol 2009;11(9):1150-6.
6 Wegner CS,Rodahl LM,Stenmark H.ESCRT proteins and cell signalling.Traffic 2011;12(10):1291-7.
7 Dukes JD,Fish L,Richardson JD,Blaikley E,Burns S,Caunt CJ,et al.Functional ESCRT machinery is required for constitutive recycling of claudin-1 and maintenance of polarity in vertebrate epithelial cells.Mol Biol Cell 2011;22(17):3192-205.
8 Jimenez AJ,Maiuri P,Lafaurie-Janvore J,Divoux S,Piel M,Perez F.ESCRT machinery is required for plasma membrane repair.Science 2014;343(6174):1247136.
9 Skowyra ML,Schlesinger PH,Naismith TV,Hanson PI.Triggered recruitment of ESCRT machinery promotes endolysosomal repair.Science 2018;360(6384):eaar5078.
10 Olmos Y,Hodgson L,Mantell J,Verkade P,Carlton JG.ES-CRT-III controls nuclear envelope reformation.Nature 2015;522(7555):236-9.
11 Gong YN,Guy C,Olauson H,Becker JU,Yang M,Fitzgerald P,et al.ESCRT-III acts downstream of MLKL to regulate necroptotic cell death and its consequences.Cell 2017;169(2):286-300e16.
12 Morita E,Sandrin V,McCullough J,Katsuyama A,Baci Hamilton I,Sundquist WI.ESCRT-III protein requirements for HIV-1 budding.Cell Host Microbe 2011;9(3):235-42.
13 Liu Z,Wang L,Tan H,Zhou S,Fu T,Xia Y,et al.Synthesis of 1H-indazoles from N-tosylhydrazones and nitroaromatic compounds.Chem Commun(Camb)2014;50(39):5061-3.
14 Wollert T,Wunder C,Lippincott-Schwartz J,Hurley JH.Membrane scission by the ESCRT-III complex.Nature 2009;458(7235):172-7.
15 Koh CH,Cheung NS.Cellular mechanism of U18666A-mediated apoptosis in cultured murine cortical neurons:bridging Niemann-Pick disease type C and Alzheimer’s disease.Cell Signal2006;18(11):1844-53.
16 Higgins ME,Davies JP,Chen FW,Ioannou YA.Niemann-Pick C1 is a late endosome-resident protein that transiently associates with lysosomes and the trans-Golgi network.Mol Genet Metab1999;68(1):1-13.
17 Roszell BR,Tao JQ,Yu KJ,Huang S,Bates SR.Characterization of the Niemann-Pick C pathway in alveolar type II cells and lamellar bodies of the lung.Am J Physiol Lung Cell Mol Physiol2012;302(9):L919-32.
18 Chapuy-Regaud S,Subra C,Requena M,de Medina P,Amara S,Delton-Vandenbroucke I,et al.Progesterone and a phospholipase inhibitor increase the endosomal bis(monoacylglycero)phosphate content and block HIV viral particle intercellular transmission.Biochimie 2013;95(9):1677-88.
19 Elgner F,Ren H,Medvedev R,Ploen D,Himmelsbach K,Boller K,et al.The intracellular cholesterol transport inhibitor U18666Ainhibits the exosome-dependent release of mature hepatitis C virus.J Virol 2016;90(24):11181-96.
20 de Duve C,de Barsy T,Poole B,Trouet A,Tulkens P,Van Hoof F.Commentary lysosomotropic agents.Biochem Pharmacol 1974;23(18):2495-531.
21 Marceau F,Bawolak MT,Lodge R,Bouthillier J,Gagne-Henley A,Gaudreault RC,et al.Cation trapping by cellular acidic compartments:beyond the concept of lysosomotropic drugs.Toxicol Appl Pharmacol 2012;259(1):1-12.
22 Corless L,Crump CM,Griffin SD,Harris M.Vps4 and the ESCRT-III complex are required for the release of infectious hepatitis C virus particles.J Gen Virol 2010;91(Pt 2):362-72.
23 Barouch-Bentov R,Neveu G,Xiao F,Beer M,Bekerman E,Schor S,et al.Hepatitis C virus proteins interact with the endosomal sorting complex required for transport(ESCRT)machinery via ubiquitination to facilitate viral envelopment.MBio.2016;7(6):e01456-16.
24 Johnson DS,Bleck M,Simon SM.Timing of ESCRT-III protein recruitment and membrane scission during HIV-1 assembly.Elife2018;7:e36221.
25 Jiang B,Himmelsbach K,Ren H,Boller K,Hildt E.Subviral hepatitis B virus filaments,like infectious viral particles,are released via multivesicular bodies.J Virol 2015;90(7):3330-41.
26 Tang Y,Leao IC,Coleman EM,Broughton RS,Hildreth JE.Deficiency of niemann-pick type C-1 protein impairs release of human immunodeficiency virus type 1 and results in Gag accumulation in late endosomal/lysosomal compartments.J Virol2009;83(16):7982-95.
27 Lloyd-Evans E,Morgan AJ,He X,Smith DA,Elliot-Smith E,Sillence DJ,et al.Niemann-Pick disease type C1 is a sphingosine storage disease that causes deregulation of lysosomal calcium.Nat Med 2008;14(11):1247-55.
28 Scheffer LL,Sreetama SC,Sharma N,Medikayala S,Brown KJ,Defour A,et al.Mechanism of Ca2+-triggered ESCRT assembly and regulation of cell membrane repair.Nat Commun 2014;5:5646.
29 Du X,Kazim AS,Dawes IW,Brown AJ,Yang H.The AAAATPase VPS4/SKD1 regulates endosomal cholesterol trafficking independently of ESCRT-III.Traffic 2013;14(1):107-19.
30 Du X,Kazim AS,Brown AJ,Yang H.An essential role of Hrs/Vps27 in endosomal cholesterol trafficking.Cell Rep 2012;1(1):29-35.