Activin A forms a non-signaling complex with ACVR1 and type II Activin/BMP receptors via its finger 2 tip loop
Senem Aykul, Richard A Corpina, Erich J Goebel, Camille J Cunanan, Alexandra Dimitriou, Hyon Jong Kim, Qian Zhang, Ashique Rafique, Raymond Leidich, Xin Wang, Joyce McClain, Johanna Jimenez, Kalyan C Nannuru, Nyanza J Rothman, John B Lees-Shepard, Erik Martinez-Hackert, Andrew J Murphy, Thomas B Thompson, Aris N Economides, Vincent Idone, Senem Aykul, Richard A Corpina, Erich J Goebel, Camille J Cunanan, Alexandra Dimitriou, Hyon Jong Kim, Qian Zhang, Ashique Rafique, Raymond Leidich, Xin Wang, Joyce McClain, Johanna Jimenez, Kalyan C Nannuru, Nyanza J Rothman, John B Lees-Shepard, Erik Martinez-Hackert, Andrew J Murphy, Thomas B Thompson, Aris N Economides, Vincent Idone
Abstract
Activin A functions in BMP signaling in two ways: it either engages ACVR1B to activate Smad2/3 signaling or binds ACVR1 to form a non-signaling complex (NSC). Although the former property has been studied extensively, the roles of the NSC remain unexplored. The genetic disorder fibrodysplasia ossificans progressiva (FOP) provides a unique window into ACVR1/Activin A signaling because in that disease Activin can either signal through FOP-mutant ACVR1 or form NSCs with wild-type ACVR1. To explore the role of the NSC, we generated 'agonist-only' Activin A muteins that activate ACVR1B but cannot form the NSC with ACVR1. Using one of these muteins, we demonstrate that failure to form the NSC in FOP results in more severe disease pathology. These results provide the first evidence for a biological role for the NSC in vivo and pave the way for further exploration of the NSC's physiological role in corresponding knock-in mice.
Trial registration: ClinicalTrials.gov NCT03188666.
Keywords: ACVR1; ACVR1B; activin A; biochemistry; bone morphogenetic proteins; cell biology; chemical biology; fibrodysplasia ossificans progressiva; mouse; signal transduction.
Conflict of interest statement
SA, RC, CC, AD, HK, QZ, AR, RL, XW, JM, JJ, KN, NR, JL, AM, AE, VI The author is an employee of Regeneron Pharmaceuticals, Inc. Regeneron is currently developing a monoclonal antibody that neutralizes Activin A (REGN2477) as a potential therapy in fibrodysplasia ossificans progressiva (see https://ichgcp.net/clinical-trials-registry/NCT03188666). EG, EM, TT No competing interests declared
© 2020, Aykul et al.
Figures
References
- Allendorph GP, Vale WW, Choe S. Structure of the ternary signaling complex of a TGF-beta superfamily member. PNAS. 2006;103:7643–7648. doi: 10.1073/pnas.0602558103.
- Allendorph GP, Isaacs MJ, Kawakami Y, Izpisua Belmonte JC, Choe S. BMP-3 and BMP-6 structures illuminate the nature of binding specificity with receptors. Biochemistry. 2007;46:12238–12247. doi: 10.1021/bi700907k.
- Andersson O, Reissmann E, Ibáñez CF. Growth differentiation factor 11 signals through the transforming growth factor-beta receptor ALK5 to regionalize the anterior-posterior Axis. EMBO Reports. 2006;7:831–837. doi: 10.1038/sj.embor.7400752.
- Archambeault DR, Yao HH. Activin A, a product of fetal leydig cells, is a unique paracrine regulator of sertoli cell proliferation and fetal testis cord expansion. PNAS. 2010;107:10526–10531. doi: 10.1073/pnas.1000318107.
- Attisano L, Cárcamo J, Ventura F, Weis FM, Massagué J, Wrana JL. Identification of human activin and TGF beta type I receptors that form heteromeric kinase complexes with type II receptors. Cell. 1993;75:671–680. doi: 10.1016/0092-8674(93)90488-C.
- Aykul S, Parenti A, Chu KY, Reske J, Floer M, Ralston A, Martinez-Hackert E. Biochemical and cellular analysis reveals ligand binding specificities, a molecular basis for ligand recognition, and membrane Association-dependent activities of Cripto-1 and cryptic. Journal of Biological Chemistry. 2017;292:4138–4151. doi: 10.1074/jbc.M116.747501.
- Bonecchi R, Garlanda C, Mantovani A, Riva F. Cytokine decoy and scavenger receptors as key regulators of immunity and inflammation. Cytokine. 2016;87:37–45. doi: 10.1016/j.cyto.2016.06.023.
- Boyce BF, Xing L. Biology of RANK, RANKL, and osteoprotegerin. Arthritis Research & Therapy. 2007;9:ar2165. doi: 10.1186/ar2165.
- Cash JN, Rejon CA, McPherron AC, Bernard DJ, Thompson TB. The structure of myostatin:follistatin 288: insights into receptor utilization and heparin binding. The EMBO Journal. 2009;28:2662–2676. doi: 10.1038/emboj.2009.205.
- Cash JN, Angerman EB, Kattamuri C, Nolan K, Zhao H, Sidis Y, Keutmann HT, Thompson TB. Structure of myostatin·follistatin-like 3: n-terminal domains of follistatin-type molecules exhibit alternate modes of binding. The Journal of Biological Chemistry. 2012;287:1043–1053. doi: 10.1074/jbc.M111.270801.
- Dey D, Bagarova J, Hatsell SJ, Armstrong KA, Huang L, Ermann J, Vonner AJ, Shen Y, Mohedas AH, Lee A, Eekhoff EM, van Schie A, Demay MB, Keller C, Wagers AJ, Economides AN, Yu PB. Two tissue-resident progenitor lineages drive distinct phenotypes of heterotopic ossification. Science Translational Medicine. 2016;8:366ra163. doi: 10.1126/scitranslmed.aaf1090.
- Gibson DG, Glass JI, Lartigue C, Noskov VN, Chuang RY, Algire MA, Benders GA, Montague MG, Ma L, Moodie MM, Merryman C, Vashee S, Krishnakumar R, Assad-Garcia N, Andrews-Pfannkoch C, Denisova EA, Young L, Qi ZQ, Segall-Shapiro TH, Calvey CH, Parmar PP, Hutchison CA, Smith HO, Venter JC. Creation of a bacterial cell controlled by a chemically synthesized genome. Science. 2010;329:52–56. doi: 10.1126/science.1190719.
- Gille C, Fähling M, Weyand B, Wieland T, Gille A. Alignment-Annotator web server: rendering and annotating sequence alignments. Nucleic Acids Research. 2014;42:W3–W6. doi: 10.1093/nar/gku400.
- Goebel EJ, Corpina RA, Hinck CS, Czepnik M, Castonguay R, Grenha R, Boisvert A, Miklossy G, Fullerton PT, Matzuk MM, Idone VJ, Economides AN, Kumar R, Hinck AP, Thompson TB. Structural characterization of an activin class ternary receptor complex reveals a third paradigm for receptor specificity. PNAS. 2019;116:15505–15513. doi: 10.1073/pnas.1906253116.
- Greenwald J, Vega ME, Allendorph GP, Fischer WH, Vale W, Choe S. A flexible activin explains the membrane-dependent cooperative assembly of TGF-beta family receptors. Molecular Cell. 2004;15:485–489. doi: 10.1016/j.molcel.2004.07.011.
- Harrington AE, Morris-Triggs SA, Ruotolo BT, Robinson CV, Ohnuma S, Hyvönen M. Structural basis for the inhibition of activin signalling by follistatin. The EMBO Journal. 2006;25:1035–1045. doi: 10.1038/sj.emboj.7601000.
- Harrison CA, Gray PC, Fischer WH, Donaldson C, Choe S, Vale W. An activin mutant with disrupted ALK4 binding blocks signaling via type II receptors. Journal of Biological Chemistry. 2004;279:28036–28044. doi: 10.1074/jbc.M402782200.
- Hatsell SJ, Idone V, Wolken DM, Huang L, Kim HJ, Wang L, Economides AN. ACVR1R206H receptor mutation causes fibrodysplasia ossificans progressiva by imparting responsiveness to activin A. Science Translational Medicine. 2015;303:ra137. doi: 10.1126/scitranslmed.aac4358.
- Hinck AP, Mueller TD, Springer TA. Structural biology and evolution of the TGF-beta family. Cold Spring Harbor Perspectives in Biology. 2016;12:a022103. doi: 10.1101/cshperspect.a022103.
- Hino K, Ikeya M, Horigome K, Matsumoto Y, Ebise H, Nishio M, Sekiguchi K, Shibata M, Nagata S, Matsuda S, Toguchida J. Neofunction of ACVR1 in fibrodysplasia ossificans progressiva. PNAS. 2015;112:15438–15443. doi: 10.1073/pnas.1510540112.
- Hüning I, Gillessen-Kaesbach G. Fibrodysplasia ossificans progressiva: clinical course, genetic mutations and genotype-phenotype correlation. Molecular Syndromology. 2014;5:201–211. doi: 10.1159/000365770.
- Katagiri T, Tsukamoto S, Kuratani M. Heterotopic bone induction via BMP signaling: potential therapeutic targets for fibrodysplasia ossificans progressiva. Bone. 2018;109:241–250. doi: 10.1016/j.bone.2017.07.024.
- Kokabu S, Gamer L, Cox K, Lowery J, Tsuji K, Raz R, Economides A, Katagiri T, Rosen V. BMP3 suppresses osteoblast differentiation of bone marrow stromal cells via interaction with Acvr2b. Molecular Endocrinology. 2012;26:87–94. doi: 10.1210/me.2011-1168.
- Korupolu RV, Muenster U, Read JD, Vale W, Fischer WH. Activin A/bone morphogenetic protein (BMP) chimeras exhibit BMP-like activity and antagonize activin and myostatin. Journal of Biological Chemistry. 2008;283:3782–3790. doi: 10.1074/jbc.M704530200.
- Krieger E, Joo K, Lee J, Lee J, Raman S, Thompson J, Tyka M, Baker D, Karplus K. Improving physical realism, stereochemistry, and side-chain accuracy in homology modeling: four approaches that performed well in CASP8. Proteins: Structure, Function, and Bioinformatics. 2009;77 Suppl 9:114–122. doi: 10.1002/prot.22570.
- Latres E, Pangilinan J, Miloscio L, Bauerlein R, Na E, Potocky TB, Huang Y, Eckersdorff M, Rafique A, Mastaitis J, Lin C, Murphy AJ, Yancopoulos GD, Gromada J, Stitt T. Myostatin blockade with a fully human monoclonal antibody induces muscle hypertrophy and reverses muscle atrophy in young and aged mice. Skeletal Muscle. 2015;5:34. doi: 10.1186/s13395-015-0060-8.
- Lees-Shepard JB, Nicholas SE, Stoessel SJ, Devarakonda PM, Schneider MJ, Yamamoto M, Goldhamer DJ. Palovarotene reduces heterotopic ossification in juvenile FOP mice but exhibits pronounced skeletal toxicity. eLife. 2018a;7:e40814. doi: 10.7554/eLife.40814.
- Lees-Shepard JB, Yamamoto M, Biswas AA, Stoessel SJ, Nicholas SE, Cogswell CA, Devarakonda PM, Schneider MJ, Cummins SM, Legendre NP, Yamamoto S, Kaartinen V, Hunter JW, Goldhamer DJ. Activin-dependent signaling in fibro/adipogenic progenitors causes fibrodysplasia ossificans progressiva. Nature Communications. 2018b;9:471. doi: 10.1038/s41467-018-02872-2.
- Macdonald LE, Karow M, Stevens S, Auerbach W, Poueymirou WT, Yasenchak J, Frendewey D, Valenzuela DM, Giallourakis CC, Alt FW, Yancopoulos GD, Murphy AJ. Precise and in situ genetic humanization of 6 mb of mouse immunoglobulin genes. PNAS. 2014;111:5147–5152. doi: 10.1073/pnas.1323896111.
- Macías-Silva M, Hoodless PA, Tang SJ, Buchwald M, Wrana JL. Specific activation of Smad1 signaling pathways by the BMP7 type I receptor, ALK2. Journal of Biological Chemistry. 1998;273:25628–25636. doi: 10.1074/jbc.273.40.25628.
- Makanji Y, Zhu J, Mishra R, Holmquist C, Wong WP, Schwartz NB, Mayo KE, Woodruff TK. Inhibin at 90: from discovery to clinical application, a historical review. Endocrine Reviews. 2014;35:747–794. doi: 10.1210/er.2014-1003.
- Matzuk MM, Kumar TR, Vassalli A, Bickenbach JR, Roop DR, Jaenisch R, Bradley A. Functional analysis of activins during mammalian development. Nature. 1995;374:354–356. doi: 10.1038/374354a0.
- Mishra SK, Funair L, Cressley A, Gittes GK, Burns RC. High-affinity Dkk1 receptor Kremen1 is internalized by clathrin-mediated endocytosis. PLOS ONE. 2012;7:e52190. doi: 10.1371/journal.pone.0052190.
- Mukherjee A, Sidis Y, Mahan A, Raher MJ, Xia Y, Rosen ED, Bloch KD, Thomas MK, Schneyer AL. FSTL3 deletion reveals roles for TGF-beta family ligands in glucose and fat homeostasis in adults. PNAS. 2007;104:1348–1353. doi: 10.1073/pnas.0607966104.
- Murphy AJ, Macdonald LE, Stevens S, Karow M, Dore AT, Pobursky K, Huang TT, Poueymirou WT, Esau L, Meola M, Mikulka W, Krueger P, Fairhurst J, Valenzuela DM, Papadopoulos N, Yancopoulos GD. Mice with megabase humanization of their immunoglobulin genes generate antibodies as efficiently as normal mice. PNAS. 2014;111:5153–5158. doi: 10.1073/pnas.1324022111.
- Namwanje M, Brown CW. Activins and inhibins: roles in development, physiology, and disease. Cold Spring Harbor Perspectives in Biology. 2016;8:a021881. doi: 10.1101/cshperspect.a021881.
- Olsen OE, Wader KF, Hella H, Mylin AK, Turesson I, Nesthus I, Waage A, Sundan A, Holien T. Activin A inhibits BMP-signaling by binding ACVR2A and ACVR2B. Cell Communication and Signaling. 2015;13:27. doi: 10.1186/s12964-015-0104-z.
- Onichtchouk D, Chen YG, Dosch R, Gawantka V, Delius H, Massagué J, Niehrs C. Silencing of TGF-beta signalling by the pseudoreceptor BAMBI. Nature. 1999;401:480–485. doi: 10.1038/46794.
- Palomo J, Dietrich D, Martin P, Palmer G, Gabay C. The interleukin (IL)-1 cytokine family--balance between agonists and antagonists in inflammatory diseases. Cytokine. 2015;76:25–37. doi: 10.1016/j.cyto.2015.06.017.
- Pangas SA, Jorgez CJ, Tran M, Agno J, Li X, Brown CW, Kumar TR, Matzuk MM. Intraovarian activins are required for female fertility. Molecular Endocrinology. 2007;21:2458–2471. doi: 10.1210/me.2007-0146.
- Piek E, Afrakhte M, Sampath K, van Zoelen EJ, Heldin CH, ten Dijke P. Functional antagonism between activin and osteogenic protein-1 in human embryonal carcinoma cells. Journal of Cellular Physiology. 1999;180:141–149. doi: 10.1002/(SICI)1097-4652(199908)180:2<141::AID-JCP1>;2-I.
- Rebbapragada A, Benchabane H, Wrana JL, Celeste AJ, Attisano L. Myostatin signals through a transforming growth factor beta-like signaling pathway to block adipogenesis. Molecular and Cellular Biology. 2003;23:7230–7242. doi: 10.1128/MCB.23.20.7230-7242.2003.
- Sheikh MS, Fornace AJ. Death and decoy receptors and p53-mediated apoptosis. Leukemia. 2000;14:1509–1513. doi: 10.1038/sj.leu.2401865.
- Stamler R, Keutmann HT, Sidis Y, Kattamuri C, Schneyer A, Thompson TB. The structure of FSTL3.activin A complex. differential binding of N-terminal domains influences follistatin-type antagonist specificity. The Journal of Biological Chemistry. 2008;283:32831–32838. doi: 10.1074/jbc.M801266200.
- Thompson TB, Lerch TF, Cook RW, Woodruff TK, Jardetzky TS. The structure of the follistatin:activin complex reveals antagonism of both type I and type II receptor binding. Developmental Cell. 2005;9:535–543. doi: 10.1016/j.devcel.2005.09.008.
- Tsuchida K, Mathews LS, Vale WW. Cloning and characterization of a transmembrane serine kinase that acts as an activin type I receptor. PNAS. 1993;90:11242–11246. doi: 10.1073/pnas.90.23.11242.
- Upadhyay J, Xie L, Huang L, Das N, Stewart RC, Lyon MC, Palmer K, Rajamani S, Graul C, Lobo M, Wellman TJ, Soares EJ, Silva MD, Hesterman J, Wang L, Wen X, Qian X, Nannuru K, Idone V, Murphy AJ, Economides AN, Hatsell SJ. The expansion of heterotopic bone in fibrodysplasia ossificans progressiva is activin A-Dependent. Journal of Bone and Mineral Research. 2017;32:2489–2499. doi: 10.1002/jbmr.3235.
- Waterhouse A, Bertoni M, Bienert S, Studer G, Tauriello G, Gumienny R, Heer FT, de Beer TAP, Rempfer C, Bordoli L, Lepore R, Schwede T. SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Research. 2018;46:W296–W303. doi: 10.1093/nar/gky427.
- Yadin D, Knaus P, Mueller TD. Structural insights into BMP receptors: specificity, activation and inhibition. Cytokine & Growth Factor Reviews. 2016;27:13–34. doi: 10.1016/j.cytogfr.2015.11.005.
Source: PubMed