Distinct cellular roles for PDCD10 define a gut-brain axis in cerebral cavernous malformation
Alan T Tang, Katie R Sullivan, Courtney C Hong, Lauren M Goddard, Aparna Mahadevan, Aileen Ren, Heidy Pardo, Amy Peiper, Erin Griffin, Ceylan Tanes, Lisa M Mattei, Jisheng Yang, Li Li, Patricia Mericko-Ishizuka, Le Shen, Nicholas Hobson, Romuald Girard, Rhonda Lightle, Thomas Moore, Robert Shenkar, Sean P Polster, Claudia J Rödel, Ning Li, Qin Zhu, Kevin J Whitehead, Xiangjian Zheng, Amy Akers, Leslie Morrison, Helen Kim, Kyle Bittinger, Christopher J Lengner, Markus Schwaninger, Anna Velcich, Leonard Augenlicht, Salim Abdelilah-Seyfried, Wang Min, Douglas A Marchuk, Issam A Awad, Mark L Kahn, Alan T Tang, Katie R Sullivan, Courtney C Hong, Lauren M Goddard, Aparna Mahadevan, Aileen Ren, Heidy Pardo, Amy Peiper, Erin Griffin, Ceylan Tanes, Lisa M Mattei, Jisheng Yang, Li Li, Patricia Mericko-Ishizuka, Le Shen, Nicholas Hobson, Romuald Girard, Rhonda Lightle, Thomas Moore, Robert Shenkar, Sean P Polster, Claudia J Rödel, Ning Li, Qin Zhu, Kevin J Whitehead, Xiangjian Zheng, Amy Akers, Leslie Morrison, Helen Kim, Kyle Bittinger, Christopher J Lengner, Markus Schwaninger, Anna Velcich, Leonard Augenlicht, Salim Abdelilah-Seyfried, Wang Min, Douglas A Marchuk, Issam A Awad, Mark L Kahn
Abstract
Cerebral cavernous malformation (CCM) is a genetic, cerebrovascular disease. Familial CCM is caused by genetic mutations in KRIT1, CCM2, or PDCD10 Disease onset is earlier and more severe in individuals with PDCD10 mutations. Recent studies have shown that lesions arise from excess mitogen-activated protein kinase kinase kinase 3 (MEKK3) signaling downstream of Toll-like receptor 4 (TLR4) stimulation by lipopolysaccharide derived from the gut microbiome. These findings suggest a gut-brain CCM disease axis but fail to define it or explain the poor prognosis of patients with PDCD10 mutations. Here, we demonstrate that the gut barrier is a primary determinant of CCM disease course, independent of microbiome configuration, that explains the increased severity of CCM disease associated with PDCD10 deficiency. Chemical disruption of the gut barrier with dextran sulfate sodium augments CCM formation in a mouse model, as does genetic loss of Pdcd10, but not Krit1, in gut epithelial cells. Loss of gut epithelial Pdcd10 results in disruption of the colonic mucosal barrier. Accordingly, loss of Mucin-2 or exposure to dietary emulsifiers that reduce the mucus barrier increases CCM burden analogous to loss of Pdcd10 in the gut epithelium. Last, we show that treatment with dexamethasone potently inhibits CCM formation in mice because of the combined effect of action at both brain endothelial cells and gut epithelial cells. These studies define a gut-brain disease axis in an experimental model of CCM in which a single gene is required for two critical components: gut epithelial function and brain endothelial signaling.
Conflict of interest statement
Competing interests: The authors declare no competing financial interests. IA is Chairman of the Scientific Advisory Board for Angioma Alliance and provides expert opinions related to clinical care of cerebral cavernous malformations.
Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.
Figures
References
- Benakis C, Brea D, Caballero S, Faraco G, Moore J, Murphy M, Sita G, Racchumi G, Ling L, Pamer EG, Iadecola C, Anrather J, Commensal microbiota affects ischemic stroke outcome by regulating intestinal gammadelta T cells. Nat Med 22, 516–523 (2016); published online EpubMay (10.1038/nm.4068).
- Stanley D, Mason LJ, Mackin KE, Srikhanta YN, Lyras D, Prakash MD, Nurgali K, Venegas A, Hill MD, Moore RJ, Wong CH, Translocation and dissemination of commensal bacteria in post-stroke infection. Nat Med 22, 1277–1284 (2016); published online EpubNov (10.1038/nm.4194).
- Faraco G, Brea D, Garcia-Bonilla L, Wang G, Racchumi G, Chang H, Buendia I, Santisteban MM, Segarra SG, Koizumi K, Sugiyama Y, Murphy M, Voss H, Anrather J, Iadecola C, Dietary salt promotes neurovascular and cognitive dysfunction through a gut-initiated TH17 response. Nature neuroscience 21, 240–249 (2018); published online EpubFeb (10.1038/s41593-017-0059-z).
- Sampson TR, Debelius JW, Thron T, Janssen S, Shastri GG, Ilhan ZE, Challis C, Schretter CE, Rocha S, Gradinaru V, Chesselet MF, Keshavarzian A, Shannon KM, Krajmalnik-Brown R, Wittung-Stafshede P, Knight R, Mazmanian SK, Gut Microbiota Regulate Motor Deficits and Neuroinflammation in a Model of Parkinson’s Disease. Cell 167, 1469–1480 e1412 (2016); published online EpubDec 01 (10.1016/j.cell.2016.11.018).
- Wang PY, Caspi L, Lam CK, Chari M, Li X, Light PE, Gutierrez-Juarez R, Ang M, Schwartz GJ, Lam TK, Upper intestinal lipids trigger a gut-brain-liver axis to regulate glucose production. Nature 452, 1012–1016 (2008); published online EpubApr 24 (10.1038/nature06852).
- Perry RJ, Peng L, Barry NA, Cline GW, Zhang D, Cardone RL, Petersen KF, Kibbey RG, Goodman AL, Shulman GI, Acetate mediates a microbiome-brain-beta-cell axis to promote metabolic syndrome. Nature 534, 213–217 (2016); published online EpubJun 9 (10.1038/nature18309).
- Bonaz BL, Bernstein CN, Brain-gut interactions in inflammatory bowel disease. Gastroenterology 144, 36–49 (2013); published online EpubJan (10.1053/j.gastro.2012.10.003).
- Sharon G, Garg N, Debelius J, Knight R, Dorrestein PC, Mazmanian SK, Specialized metabolites from the microbiome in health and disease. Cell Metab 20, 719–730 (2014); published online EpubNov 4 (10.1016/j.cmet.2014.10.016).
- Fung TC, Olson CA, Hsiao EY, Interactions between the microbiota, immune and nervous systems in health and disease. Nature neuroscience 20, 145–155 (2017); published online EpubFeb (10.1038/nn.4476).
- Spadoni I, Fornasa G, Rescigno M, Organ-specific protection mediated by cooperation between vascular and epithelial barriers. Nat Rev Immunol 17, 761–773 (2017); published online EpubDec (10.1038/nri.2017.100).
- Spiegler S, Rath M, Paperlein C, Felbor U, Cerebral Cavernous Malformations: An Update on Prevalence, Molecular Genetic Analyses, and Genetic Counselling. Molecular syndromology 9, 60–69 (2018); published online EpubFeb (10.1159/000486292).
- Akers A, Al-Shahi Salman R, Dahlem AAI,K, Flemming K, Hart B, Kim H, Jusue-Torres I, Kondziolka D, Lee C, Morrison L, Rigamonti D, Rebeiz T, Tournier-Lasserve E, Waggoner D, Whitehead K, Synopsis of Guidelines for the Clinical Management of Cerebral Cavernous Malformations: Consensus Recommendations Based on Systematic Literature Review by the Angioma Alliance Scientific Advisory Board Clinical Experts Panel. Neurosurgery 80, 665–680 (2017); published online EpubMay 01 (10.1093/neuros/nyx091).
- Zhou Z, Tang AT, Wong WY, Bamezai S, Goddard LM, Shenkar R, Zhou S, Yang J, Wright AC, Foley M, Arthur JS, Whitehead KJ, Awad IA, Li DY, Zheng X, Kahn ML, Cerebral cavernous malformations arise from endothelial gain of MEKK3-KLF2/4 signalling. Nature 532, 122–126 (2016); published online EpubApr 7 (10.1038/nature17178).
- Fisher OS, Deng H, Liu D, Zhang Y, Wei R, Deng Y, Zhang F, Louvi A, Turk BE, Boggon TJ, Su B, Structure and vascular function of MEKK3-cerebral cavernous malformations 2 complex. Nat Commun 6, 7937 (2015)10.1038/ncomms8937).
- Wang X, Hou Y, Deng K, Zhang Y, Wang DC, Ding J, Structural Insights into the Molecular Recognition between Cerebral Cavernous Malformation 2 and Mitogen-Activated Protein Kinase Kinase Kinase 3. Structure, (2015); published online EpubApr 29 (10.1016/j.str.2015.04.003).
- Cullere X, Plovie E, Bennett PM, MacRae CA, Mayadas TN, The cerebral cavernous malformation proteins CCM2L and CCM2 prevent the activation of the MAP kinase MEKK3. Proc Natl Acad Sci U S A, (2015); published online EpubNov 4 (10.1073/pnas.1510495112).
- Zhou Z, Rawnsley DR, Goddard LM, Pan W, Cao XJ, Jakus Z, Zheng H, Yang J, Arthur JS, Whitehead KJ, Li D, Zhou B, Garcia BA, Zheng X, Kahn ML, The cerebral cavernous malformation pathway controls cardiac development via regulation of endocardial MEKK3 signaling and KLF expression. Dev Cell 32, 168–180 (2015); published online EpubJan 26 (10.1016/j.devcel.2014.12.009).
- Denier C, Labauge P, Bergametti F, Marchelli F, Riant F, Arnoult M, Maciazek J, Vicaut E, Brunereau L, Tournier-Lasserve E, Genotype-phenotype correlations in cerebral cavernous malformations patients. Ann Neurol 60, 550–556 (2006); published online EpubNov (
- Shenkar R, Shi C, Rebeiz T, Stockton RA, McDonald DA, Mikati AG, Zhang L, Austin C, Akers AL, Gallione CJ, Rorrer A, Gunel M, Min W, Marcondes de Souza J, Lee C, Marchuk DA, Awad IA, Exceptional aggressiveness of cerebral cavernous malformation disease associated with PDCD10 mutations. Genet Med 17, 188–196 (2015); published online EpubMar (10.1038/gim.2014.97).
- Tang AT, Choi JP, Kotzin JJ, Yang Y, Hong CC, Hobson N, Girard R, Zeineddine HA, Lightle R, Moore T, Cao Y, Shenkar R, Chen M, Mericko P, Yang J, Li L, Tanes C, Kobuley D, Vosa U, Whitehead KJ, Li DY, Franke L, Hart B, Schwaninger M, Henao-Mejia J, Morrison L, Kim H, Awad IA, Zheng X, Kahn ML, Endothelial TLR4 and the microbiome drive cerebral cavernous malformations. Nature 545, 305–310 (2017); published online EpubMay 18 (10.1038/nature22075).
- Akers AL, Johnson E, Steinberg GK, Zabramski JM, Marchuk DA, Biallelic somatic and germline mutations in cerebral cavernous malformations (CCMs): evidence for a two-hit mechanism of CCM pathogenesis. Hum Mol Genet 18, 919–930 (2009); published online EpubMar 1 (10.1093/hmg/ddn430).
- Zawistowski JS, Stalheim L, Uhlik MT, Abell AN, Ancrile BB, Johnson GL, Marchuk DA, CCM1 and CCM2 protein interactions in cell signaling: implications for cerebral cavernous malformations pathogenesis. Hum Mol Genet 14, 2521–2531 (2005); published online EpubSep 1 (10.1093/hmg/ddi256).
- Zhou Z, Tang AT, Wong WY, Bamezai S, Goddard LM, Shenkar R, Zhou S, Yang J, Wright AC, Foley M, Arthur JS, Whitehead KJ, Awad IA, Li DY, Zheng X, Kahn ML, Cerebral cavernous malformations arise from endothelial gain of MEKK3-KLF2/4 signalling. Nature, (2016); published online EpubMar 30 (10.1038/nature17178).
- Ridder DA, Lang MF, Salinin S, Roderer JP, Struss M, Maser-Gluth C, Schwaninger M, TAK1 in brain endothelial cells mediates fever and lethargy. J Exp Med 208, 2615–2623 (2011); published online EpubDec 19 (10.1084/jem.20110398).
- Cuttano R, Rudini N, Bravi L, Corada M, Giampietro C, Papa E, Morini MF, Maddaluno L, Baeyens N, Adams RH, Jain MK, Owens GK, Schwartz M, Lampugnani MG, Dejana E, KLF4 is a key determinant in the development and progression of cerebral cavernous malformations. EMBO Mol Med, (2015); published online EpubNov 26 (10.15252/emmm.201505433).
- Renz M, Otten C, Faurobert E, Rudolph F, Zhu Y, Boulday G, Duchene J, Mickoleit M, Dietrich AC, Ramspacher C, Steed E, Manet-Dupe S, Benz A, Hassel D, Vermot J, Huisken J, Tournier-Lasserve E, Felbor U, Sure U, Albiges-Rizo C, Abdelilah-Seyfried S, Regulation of beta1 integrin-Klf2-mediated angiogenesis by CCM proteins. Dev Cell 32, 181–190 (2015); published online EpubJan 26 (10.1016/j.devcel.2014.12.016).
- Choi JP, Foley M, Zhou Z, Wong WY, Gokoolparsadh N, Arthur JS, Li DY, Zheng X, Micro-CT Imaging Reveals Mekk3 Heterozygosity Prevents Cerebral Cavernous Malformations in Ccm2-Deficient Mice. PLoS One 11, e0160833 (2016)10.1371/journal.pone.0160833).
- Girard R, Zeineddine HA, Orsbon C, Tan H, Moore T, Hobson N, Shenkar R, Lightle R, Shi C, Fam MD, Cao Y, Shen L, Neander AI, Rorrer A, Gallione C, Tang AT, Kahn ML, Marchuk DA, Luo ZX, Awad IA, Micro-computed tomography in murine models of cerebral cavernous malformations as a paradigm for brain disease. J Neurosci Methods 271, 14–24 (2016); published online EpubSep 15 (10.1016/j.jneumeth.2016.06.021).
- Wu GD, Chen J, Hoffmann C, Bittinger K, Chen YY, Keilbaugh SA, Bewtra M, Knights D, Walters WA, Knight R, Sinha R, Gilroy E, Gupta K, Baldassano R, Nessel L, Li H, Bushman FD, Lewis JD, Linking long-term dietary patterns with gut microbial enterotypes. Science 334, 105–108 (2011); published online EpubOct 7 (10.1126/science.1208344).
- Rothschild D, Weissbrod O, Barkan E, Kurilshikov A, Korem T, Zeevi D, Costea PI, Godneva A, Kalka IN, Bar N, Shilo S, Lador D, Vila AV, Zmora N, Pevsner-Fischer M, Israeli D, Kosower N, Malka G, Wolf BC, Avnit-Sagi T, Lotan-Pompan M, Weinberger A, Halpern Z, Carmi S, Fu J, Wijmenga C, Zhernakova A, Elinav E, Segal E, Environment dominates over host genetics in shaping human gut microbiota. Nature, (2018); published online EpubFeb 28 (10.1038/nature25973).
- Zeineddine HA, Girard R, Saadat L, Shen L, Lightle R, Moore T, Cao Y, Hobson N, Shenkar R, Avner K, Chaudager K, Koskimaki J, Polster SP, Fam MD, Shi C, Lopez-Ramirez MA, Tang AT, Gallione C, Kahn ML, Ginsberg M, Marchuk DA, Awad IA, Phenotypic characterization of murine models of cerebral cavernous malformations. Lab Invest, (2018); published online EpubJun 26 (10.1038/s41374-018-0030-y).
- Johansson ME, Phillipson M, Petersson J, Velcich A, Holm L, Hansson GC, The inner of the two Muc2 mucin-dependent mucus layers in colon is devoid of bacteria. Proc Natl Acad Sci U S A 105, 15064–15069 (2008); published online EpubSep 30 (10.1073/pnas.0803124105).
- Johansson ME, Gustafsson JK, Sjoberg KE, Petersson J, Holm L, Sjovall H, Hansson GC, Bacteria penetrate the inner mucus layer before inflammation in the dextran sulfate colitis model. PLoS One 5, e12238 (2010); published online EpubAug 18 (10.1371/journal.pone.0012238).
- Johansson ME, Larsson JM, Hansson GC, The two mucus layers of colon are organized by the MUC2 mucin, whereas the outer layer is a legislator of host-microbial interactions. Proc Natl Acad Sci U S A 108 Suppl 1, 4659–4665 (2011); published online EpubMar 15 (10.1073/pnas.1006451107).
- Chassaing B, Srinivasan G, Delgado MA, Young AN, Gewirtz AT, Vijay-Kumar M, Fecal lipocalin 2, a sensitive and broadly dynamic non-invasive biomarker for intestinal inflammation. PLoS One 7, e44328 (2012)10.1371/journal.pone.0044328).
- Bergstrom KS, Kissoon-Singh V, Gibson DL, Ma C, Montero M, Sham HP, Ryz N, Huang T, Velcich A, Finlay BB, Chadee K, Vallance BA, Muc2 protects against lethal infectious colitis by disassociating pathogenic and commensal bacteria from the colonic mucosa. PLoS Pathog 6, e1000902 (2010); published online EpubMay 13 (10.1371/journal.ppat.1000902).
- Birchenough GM, Nystrom EE, Johansson ME, Hansson GC, A sentinel goblet cell guards the colonic crypt by triggering Nlrp6-dependent Muc2 secretion. Science 352, 1535–1542 (2016); published online EpubJun 24 (10.1126/science.aaf7419).
- Van der Sluis M, De Koning BA, De Bruijn AC, Velcich A, Meijerink JP, Van Goudoever JB, Buller HA, Dekker J, Van Seuningen I, Renes IB, Einerhand AW, Muc2-deficient mice spontaneously develop colitis, indicating that MUC2 is critical for colonic protection. Gastroenterology 131, 117–129 (2006); published online EpubJul (10.1053/j.gastro.2006.04.020).
- Schneider MR, Dahlhoff M, Horst D, Hirschi B, Trulzsch K, Muller-Hocker J, Vogelmann R, Allgauer M, Gerhard M, Steininger S, Wolf E, Kolligs FT, A key role for E-cadherin in intestinal homeostasis and Paneth cell maturation. PLoS One 5, e14325 (2010); published online EpubDec 14 (10.1371/journal.pone.0014325).
- Draheim KM, Fisher OS, Boggon TJ, Calderwood DA, Cerebral cavernous malformation proteins at a glance. J Cell Sci 127, 701–707 (2014); published online EpubFeb 15 (10.1242/jcs.138388).
- Sonnenburg JL, Xu J, Leip DD, Chen CH, Westover BP, Weatherford J, Buhler JD, Gordon JI, Glycan foraging in vivo by an intestine-adapted bacterial symbiont. Science 307, 1955–1959 (2005); published online EpubMar 25 (10.1126/science.1109051).
- Jakobsson HE, Rodriguez-Pineiro AM, Schutte A, Ermund A, Boysen P, Bemark M, Sommer F, Backhed F, Hansson GC, Johansson ME, The composition of the gut microbiota shapes the colon mucus barrier. EMBO Rep 16, 164–177 (2015); published online EpubFeb (10.15252/embr.201439263).
- Mamantopoulos M, Ronchi F, Van Hauwermeiren F, Vieira-Silva S, Yilmaz B, Martens L, Saeys Y, Drexler SK, Yazdi AS, Raes J, Lamkanfi M, McCoy KD, Wullaert A, Nlrp6- and ASC-Dependent Inflammasomes Do Not Shape the Commensal Gut Microbiota Composition. Immunity 47, 339–348 e334 (2017); published online EpubAug 15 (10.1016/j.immuni.2017.07.011).
- Chassaing B, Koren O, Goodrich JK, Poole AC, Srinivasan S, Ley RE, Gewirtz AT, Dietary emulsifiers impact the mouse gut microbiota promoting colitis and metabolic syndrome. Nature 519, 92–96 (2015); published online EpubMar 5 (10.1038/nature14232).
- Desmet SJ, De Bosscher K, Glucocorticoid receptors: finding the middle ground. J Clin Invest 127, 1136–1145 (2017); published online EpubApr 3 (10.1172/JCI88886).
- Goodwin JE, Feng Y, Velazquez H, Sessa WC, Endothelial glucocorticoid receptor is required for protection against sepsis. Proc Natl Acad Sci U S A 110, 306–311 (2013); published online EpubJan 2 (10.1073/pnas.1210200110).
- Goodwin JE, Feng Y, Velazquez H, Zhou H, Sessa WC, Loss of the endothelial glucocorticoid receptor prevents the therapeutic protection afforded by dexamethasone after LPS. PLoS One 9, e108126 (2014)10.1371/journal.pone.0108126).
- Das I, Png CW, Oancea I, Hasnain SZ, Lourie R, Proctor M, Eri RD, Sheng Y, Crane DI, Florin TH, McGuckin MA, Glucocorticoids alleviate intestinal ER stress by enhancing protein folding and degradation of misfolded proteins. J Exp Med 210, 1201–1216 (2013); published online EpubJun 3 (10.1084/jem.20121268).
- Becker DE, Basic and clinical pharmacology of glucocorticosteroids. Anesth Prog 60, 25–31; quiz 32 (2013); published online EpubSpring (10.2344/0003-3006-60.1.25).
- Goudreault M, D’Ambrosio LM, Kean MJ, Mullin MJ, Larsen BG, Sanchez A, Chaudhry S, Chen GI, Sicheri F, Nesvizhskii AI, Aebersold R, Raught B, Gingras AC, A PP2A phosphatase high density interaction network identifies a novel striatin-interacting phosphatase and kinase complex linked to the cerebral cavernous malformation 3 (CCM3) protein. Mol Cell Proteomics 8, 157–171 (2009); published online EpubJan (10.1074/mcp.M800266-MCP200).
- Zhang Y, Tang W, Zhang H, Niu X, Xu Y, Zhang J, Gao K, Pan W, Boggon TJ, Toomre D, Min W, Wu D, A network of interactions enables CCM3 and STK24 to coordinate UNC13D-driven vesicle exocytosis in neutrophils. Dev Cell 27, 215–226 (2013); published online EpubOct 28 (10.1016/j.devcel.2013.09.021).
- Jenny Zhou H, Qin L, Zhang H, Tang W, Ji W, He Y, Liang X, Wang Z, Yuan Q, Vortmeyer A, Toomre D, Fuh G, Yan M, Kluger MS, Wu D, Min W, Endothelial exocytosis of angiopoietin-2 resulting from CCM3 deficiency contributes to cerebral cavernous malformation. Nat Med 22, 1033–1042 (2016); published online EpubSep (10.1038/nm.4169).
- Pal S, Lant B, Yu B, Tian R, Tong J, Krieger JR, Moran MF, Gingras AC, Derry WB, CCM-3 Promotes C. elegans Germline Development by Regulating Vesicle Trafficking Cytokinesis and Polarity. Curr Biol 27, 868–876 (2017); published online EpubMar 20 (10.1016/j.cub.2017.02.028).
- McDonald DA, Shi C, Shenkar R, Stockton RA, Liu F, Ginsberg MH, Marchuk DA, Awad IA, Fasudil decreases lesion burden in a murine model of cerebral cavernous malformation disease. Stroke 43, 571–574 (2012); published online EpubFeb (10.1161/STROKEAHA.111.625467).
- Shenkar R, Shi C, Austin C, Moore T, Lightle R, Cao Y, Zhang L, Wu M, Zeineddine HA, Girard R, McDonald DA, Rorrer A, Gallione C, Pytel P, Liao JK, Marchuk DA, Awad IA, RhoA Kinase Inhibition With Fasudil Versus Simvastatin in Murine Models of Cerebral Cavernous Malformations. Stroke 48, 187–194 (2017); published online EpubJan (10.1161/STROKEAHA.116.015013).
Source: PubMed