AAV Exploits Subcellular Stress Associated with Inflammation, Endoplasmic Reticulum Expansion, and Misfolded Proteins in Models of Cystic Fibrosis

English version České info

Barriers to infection act at multiple levels to prevent viruses, bacteria, and parasites from commandeering host cells for their own purposes. An intriguing hypothesis is that if a cell experiences stress, such as that elicited by inflammation, endoplasmic reticulum (ER) expansion, or misfolded proteins, then subcellular barriers will be less effective at preventing viral infection. Here we have used models of cystic fibrosis (CF) to test whether subcellular stress increases susceptibility to adeno-associated virus (AAV) infection. In human airway epithelium cultured at an air/liquid interface, physiological conditions of subcellular stress and ER expansion were mimicked using supernatant from mucopurulent material derived from CF lungs. Using this inflammatory stimulus to recapitulate stress found in diseased airways, we demonstrated that AAV infection was significantly enhanced. Since over 90% of CF cases are associated with a misfolded variant of Cystic Fibrosis Transmembrane Conductance Regulator (ΔF508-CFTR), we then explored whether the presence of misfolded proteins could independently increase susceptibility to AAV infection. In these models, AAV was an order of magnitude more efficient at transducing cells expressing ΔF508-CFTR than in cells expressing wild-type CFTR. Rescue of misfolded ΔF508-CFTR under low temperature conditions restored viral transduction efficiency to that demonstrated in controls, suggesting effects related to protein misfolding were responsible for increasing susceptibility to infection. By testing other CFTR mutants, G551D, D572N, and 1410X, we have shown this phenomenon is common to other misfolded proteins and not related to loss of CFTR activity. The presence of misfolded proteins did not affect cell surface attachment of virus or influence expression levels from promoter transgene cassettes in plasmid transfection studies, indicating exploitation occurs at the level of virion trafficking or processing. Thus, we surmised that factors enlisted to process misfolded proteins such as ΔF508-CFTR in the secretory pathway also act to restrict viral infection. In line with this hypothesis, we found that AAV trafficked to the microtubule organizing center and localized near Golgi/ER transport proteins. Moreover, AAV infection efficiency could be modulated with siRNA-mediated knockdown of proteins involved in processing ΔF508-CFTR or sorting retrograde cargo from the Golgi and ER (calnexin, KDEL-R, β-COP, and PSMB3). In summary, our data support a model where AAV exploits a compromised secretory system and, importantly, underscore the gravity with which a stressed subcellular environment, under internal or external insults, can impact infection efficiency.

Vyšlo v časopise: AAV Exploits Subcellular Stress Associated with Inflammation, Endoplasmic Reticulum Expansion, and Misfolded Proteins in Models of Cystic Fibrosis. PLoS Pathog 7(5): e32767. doi:10.1371/journal.ppat.1002053
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002053

Souhrn

Zdroje

1. O'SullivanBPFreedmanSD 2009 Cystic fibrosis. Lancet 373 1891 1904

2. SferraTJCollinsFS 1993 The molecular biology of cystic fibrosis. Annu Rev Med 44 133 144

3. QuBHStricklandEHThomasPJ 1997 Localization and suppression of a kinetic defect in cystic fibrosis transmembrane conductance regulator folding. J Biol Chem 272 15739 15744

4. RiordanJR 2005 Assembly of functional CFTR chloride channels. Annu Rev Physiol 67 701 718

5. GelmanMSKannegaardESKopitoRR 2002 A principal role for the proteasome in endoplasmic reticulum-associated degradation of misfolded intracellular cystic fibrosis transmembrane conductance regulator. J Biol Chem 277 11709 11714

6. JohnstonJAWardCLKopitoRR 1998 Aggresomes: a cellular response to misfolded proteins. J Cell Biol 143 1883 1898

7. FuLSztulE 2009 ER-associated complexes (ERACs) containing aggregated cystic fibrosis transmembrane conductance regulator (CFTR) are degraded by autophagy. Eur J Cell Biol 88 215 226

8. RibeiroCMParadisoAMCarewMAShearsSBBoucherRC 2005 Cystic fibrosis airway epithelial Ca2+ i signaling: the mechanism for the larger agonist-mediated Ca2+ i signals in human cystic fibrosis airway epithelia. J Biol Chem 280 10202 10209

9. RibeiroCMParadisoAMSchwabUPerez-VilarJJonesL 2005 Chronic airway infection/inflammation induces a Ca2+i-dependent hyperinflammatory response in human cystic fibrosis airway epithelia. J Biol Chem 280 17798 17806

10. BartoszewskiRRabAJurkuvenaiteAMazurMWakefieldJ 2008 Activation of the unfolded protein response by deltaF508 CFTR. Am J Respir Cell Mol Biol 39 448 457

11. YoungerJMFanCYChenLRosserMFPattersonC 2005 Cystic fibrosis transmembrane conductance regulator as a model substrate to study endoplasmic reticulum protein quality control in mammalian cells. Methods Mol Biol 301 293 303

12. Garcia-MataRGaoYSSztulE 2002 Hassles with taking out the garbage: aggravating aggresomes. Traffic 3 388 396

13. LeopoldPLPfisterKK 2006 Viral strategies for intracellular trafficking: motors and microtubules. Traffic 7 516 523

14. WilemanT 2007 Aggresomes and pericentriolar sites of virus assembly: cellular defense or viral design? Annu Rev Microbiol 61 149 167

15. CuschieriLNguyenTVogelJ 2007 Control at the cell center: the role of spindle poles in cytoskeletal organization and cell cycle regulation. Cell Cycle 6 2788 2794

16. BadanoJLTeslovichTMKatsanisN 2005 The centrosome in human genetic disease. Nat Rev Genet 6 194 205

17. FabunmiRPWigleyWCThomasPJDeMartinoGN 2000 Activity and regulation of the centrosome-associated proteasome. J Biol Chem 275 409 413

18. BernalesSMcDonaldKLWalterP 2006 Autophagy counterbalances endoplasmic reticulum expansion during the unfolded protein response. PLoS Biol 4 e423

19. BernsKIGiraudC 1996 Biology of adeno-associated virus. Curr Top Microbiol Immunol 218 1 23

20. SanliogluSMonickMMLuleciGHunninghakeGWEngelhardtJF 2001 Rate limiting steps of AAV transduction and implications for human gene therapy. Curr Gene Ther 1 137 147

21. DingWYanZZakRSaavedraMRodmanDM 2003 Second-strand genome conversion of adeno-associated virus type 2 (AAV-2) and AAV-5 is not rate limiting following apical infection of polarized human airway epithelia. J Virol 77 7361 7366

22. HauckBZhaoWHighKXiaoW 2004 Intracellular viral processing, not single-stranded DNA accumulation, is crucial for recombinant adeno-associated virus transduction. J Virol 78 13678 13686

23. Bantel-SchaalUHubBKartenbeckJ 2002 Endocytosis of adeno-associated virus type 5 leads to accumulation of virus particles in the Golgi compartment. J Virol 76 2340 2349

24. BartlettJSWilcherRSamulskiRJ 2000 Infectious entry pathway of adeno-associated virus and adeno-associated virus vectors. J Virol 74 2777 2785

25. DuanDLiQKaoAWYueYPessinJE 1999 Dynamin is required for recombinant adeno-associated virus type 2 infection. J Virol 73 10371 10376

26. DingWZhangLYanZEngelhardtJF 2005 Intracellular trafficking of adeno-associated viral vectors. Gene Ther 12 873 880

27. KelkarSDeBPGaoGWilsonJMCrystalRG 2006 A common mechanism for cytoplasmic dynein-dependent microtubule binding shared among adeno-associated virus and adenovirus serotypes. J Virol 80 7781 7785

28. DuanDYueYYanZYangJEngelhardtJF 2000 Endosomal processing limits gene transfer to polarized airway epithelia by adeno-associated virus. J Clin Invest 105 1573 1587

29. HansenJQingKKwonHJMahCSrivastavaA 2000 Impaired intracellular trafficking of adeno-associated virus type 2 vectors limits efficient transduction of murine fibroblasts. J Virol 74 992 996

30. MartinoMEOlsenJCFulcherNBWolfgangMCO'NealWK 2009 Airway epithelial inflammation-induced endoplasmic reticulum Ca2+ store expansion is mediated by X-box binding protein-1. J Biol Chem 284 14904 14913

31. LiWZhangLJohnsonJSZhijianWGriegerJC 2009 Generation of Novel AAV Variants by Directed Evolution for Improved CFTR Delivery to Human Ciliated Airway Epithelium. Mol Ther 12 2067 2077

32. DenningGMAndersonMPAmaraJFMarshallJSmithAE 1992 Processing of mutant cystic fibrosis transmembrane conductance regulator is temperature-sensitive. Nature 358 761 764

33. FrenchPJvan DoorninckJHPetersRHVerbeekEAmeen NA etal 1996 A delta F508 mutation in mouse cystic fibrosis transmembrane conductance regulator results in a temperature-sensitive processing defect in vivo. J Clin Invest 98 1304 1312

34. YangYDevorDCEngelhardtJFErnstSAStrongTV 1993 Molecular basis of defective anion transport in L cells expressing recombinant forms of CFTR. Hum Mol Genet 2 1253 1261

35. GentzschMChoudhuryAChangXBPaganoRERiordanJR 2007 Misassembled mutant DeltaF508 CFTR in the distal secretory pathway alters cellular lipid trafficking. J Cell Sci 120 447 455

36. BuyseFVandenbrandenMRuysschaertJM 2004 Mistargeted MRPdeltaF728 mutant is rescued by intracellular GSH. FEBS Lett 578 145 151

37. WardCLOmuraSKopitoRR 1995 Degradation of CFTR by the ubiquitin-proteasome pathway. Cell 83 121 127

38. PajusolaKGruchalaMJochHLuscherTFYla-Herttuala S etal 2002 Cell-type-specific characteristics modulate the transduction efficiency of adeno-associated virus type 2 and restrain infection of endothelial cells. J Virol 76 11530 11540

39. KartenbeckJStukenbrokHHeleniusA 1989 Endocytosis of simian virus 40 into the endoplasmic reticulum. J Cell Biol 109 2721 2729

40. PelkmansLKartenbeckJHeleniusA 2001 Caveolar endocytosis of simian virus 40 reveals a new two-step vesicular-transport pathway to the ER. Nat Cell Biol 3 473 483

41. SchelhaasMMalmstromJPelkmansLHaugstetterJEllgaardL 2007 Simian Virus 40 depends on ER protein folding and quality control factors for entry into host cells. Cell 131 516 529

42. JohnsonJSSamulskiRJ 2009 Enhancement of adeno-associated virus infection by mobilizing capsids into and out of the nucleolus. J Virol 83 2632 2644

43. KleizenBBraakmanI 2004 Protein folding and quality control in the endoplasmic reticulum. Curr Opin Cell Biol 16 343 349

44. LewisMJPelhamHR 1992 Ligand-induced redistribution of a human KDEL receptor from the Golgi complex to the endoplasmic reticulum. Cell 68 353 364

45. MunroSPelhamHR 1987 A C-terminal signal prevents secretion of luminal ER proteins. Cell 48 899 907

46. PelhamHR 1988 Evidence that luminal ER proteins are sorted from secreted proteins in a post-ER compartment. Embo J 7 913 918

47. LeeMCMillerEAGoldbergJOrciLSchekmanR 2004 Bi-directional protein transport between the ER and Golgi. Annu Rev Cell Dev Biol 20 87 123

48. JohnsonJSLiCDiprimioNWeinbergMSMcCownTJ 2010 Mutagenesis of Adeno-Associated Virus Type 2 Capsid Protein VP1 Uncovers New Roles for Basic Amino Acids in Trafficking and Cell-Specific Transduction. J Virol 84 8888 8902

49. TangBLLowSHHongW 1995 Differential response of resident proteins and cycling proteins of the Golgi to brefeldin A. Eur J Cell Biol 68 199 205

50. DouarAMPoulardKStockholmDDanosO 2001 Intracellular trafficking of adeno-associated virus vectors: routing to the late endosomal compartment and proteasome degradation. J Virol 75 1824 1833

51. FerrariFKSamulskiTShenkTSamulskiRJ 1996 Second-strand synthesis is a rate-limiting step for efficient transduction by recombinant adeno-associated virus vectors. J Virol 70 3227 3234

52. YalkinogluAOHeilbronnRBurkleASchlehoferJRzur HausenH 1988 DNA amplification of adeno-associated virus as a response to cellular genotoxic stress. Cancer Res 48 3123 3129

53. StyersMLO'ConnorAKGrabskiRCormet-BoyakaESztulE 2008 Depletion of beta-COP reveals a role for COP-I in compartmentalization of secretory compartments and in biosynthetic transport of caveolin-1. Am J Physiol Cell Physiol 294 C1485 1498

54. SchwartzOMarechalVFriguetBArenzana-SeisdedosFHeardJM 1998 Antiviral activity of the proteasome on incoming human immunodeficiency virus type 1. J Virol 72 3845 3850

55. GroschelBBushmanF 2005 Cell cycle arrest in G2/M promotes early steps of infection by human immunodeficiency virus. J Virol 79 5695 5704

56. WatDDoullI 2003 Respiratory virus infections in cystic fibrosis. Paediatr Respir Rev 4 172 177

57. RoseneckerJHarmsKHBerteleRMPohl-KoppeAv MutiusE 1996 Adenovirus infection in cystic fibrosis patients: implications for the use of adenoviral vectors for gene transfer. Infection 24 5 8

58. XuZJensenGYenTS 1997 Activation of hepatitis B virus S promoter by the viral large surface protein via induction of stress in the endoplasmic reticulum. J Virol 71 7387 7392

59. SuHLLiaoCLLinYL 2002 Japanese encephalitis virus infection initiates endoplasmic reticulum stress and an unfolded protein response. J Virol 76 4162 4171

60. IslerJASkaletAHAlwineJC 2005 Human cytomegalovirus infection activates and regulates the unfolded protein response. J Virol 79 6890 6899

61. MulveyMAriasCMohrI 2007 Maintenance of endoplasmic reticulum (ER) homeostasis in herpes simplex virus type 1-infected cells through the association of a viral glycoprotein with PERK, a cellular ER stress sensor. J Virol 81 3377 3390

62. WigleyWCFabunmiRPLeeMGMarinoCRMuallemS 1999 Dynamic association of proteasomal machinery with the centrosome. J Cell Biol 145 481 490

63. YanZZakRLuxtonGWRitchieTCBantel-SchaalU 2002 Ubiquitination of both adeno-associated virus type 2 and 5 capsid proteins affects the transduction efficiency of recombinant vectors. J Virol 76 2043 2053

64. YanZLei-ButtersDCLiuXZhangYZhangL 2006 Unique biologic properties of recombinant AAV1 transduction in polarized human airway epithelia. J Biol Chem 281 29684 29692

65. WangXVenableJLaPointePHuttDMKoulovAV 2006 Hsp90 cochaperone Aha1 downregulation rescues misfolding of CFTR in cystic fibrosis. Cell 127 803 815

66. ZhongLQingKSiYChenLTanM 2004 Heat-shock treatment-mediated increase in transduction by recombinant adeno-associated virus 2 vectors is independent of the cellular heat-shock protein 90. J Biol Chem 279 12714 12723

67. CoeHBedardKGroenendykJJungJMichalakM 2008 Endoplasmic reticulum stress in the absence of calnexin. Cell Stress Chaperones 13 497 507

68. KawaguchiYKovacsJJMcLaurinAVanceJMItoA 2003 The deacetylase HDAC6 regulates aggresome formation and cell viability in response to misfolded protein stress. Cell 115 727 738

69. JohnstonJAIllingMEKopitoRR 2002 Cytoplasmic dynein/dynactin mediates the assembly of aggresomes. Cell Motil Cytoskeleton 53 26 38

70. AkacheBGrimmDShenXFuessSYantSR 2007 A two-hybrid screen identifies cathepsins B and L as uncoating factors for adeno-associated virus 2 and 8. Mol Ther 15 330 339

71. LinJHWalterPYenTS 2008 Endoplasmic reticulum stress in disease pathogenesis. Annu Rev Pathol 3 399 425

72. BridgesJPXuYNaCLWongHRWeaverTE 2006 Adaptation and increased susceptibility to infection associated with constitutive expression of misfolded SP-C. J Cell Biol 172 395 407

73. GregersenNBrossPVangSChristensenJH 2006 Protein misfolding and human disease. Annu Rev Genomics Hum Genet 7 103 124

74. FulcherMLGabrielSBurnsKAYankaskasJRRandellSH 2005 Well-differentiated human airway epithelial cell cultures. Methods Mol Med 107 183 206

75. XiaoXLiJSamulskiRJ 1998 Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus. J Virol 72 2224 2232

76. KantorBMaHWebster-CyriaqueJMonahanPEKafriT 2009 Epigenetic activation of unintegrated HIV-1 genomes by gut-associated short chain fatty acids and its implications for HIV infection. Proc Natl Acad Sci U S A 106 18786 18791

77. ZhangLPeeplesMEBoucherRCCollinsPLPicklesRJ 2002 Respiratory syncytial virus infection of human airway epithelial cells is polarized, specific to ciliated cells, and without obvious cytopathology. J Virol 76 5654 5666

78. GriegerJCJohnsonJSGurda-WhitakerBAgbandje-McKennaMSamulskiRJ 2007 Surface-exposed adeno-associated virus Vp1-NLS capsid fusion protein rescues infectivity of noninfectious wild-type Vp2/Vp3 and Vp3-only capsids but not that of fivefold pore mutant virions. J Virol 81 7833 7843

79. HaddenMKLubbersDJBlaggBS 2006 Geldanamycin, radicicol, and chimeric inhibitors of the Hsp90 N-terminal ATP binding site. Curr Top Med Chem 6 1173 1182

80. MizushimaNLevineBCuervoAMKlionskyDJ 2008 Autophagy fights disease through cellular self-digestion. Nature 451 1069 1075

81. SeglenPOGordonPB 1982 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. Proc Natl Acad Sci U S A 79 1889 1892

82. ChenYPiperPW 1995 Consequences of the overexpression of ubiquitin in yeast: elevated tolerances of osmostress, ethanol and canavanine, yet reduced tolerances of cadmium, arsenite and paromomycin. Biochim Biophys Acta 1268 59 64