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A noncanonical role for the engulfment gene ELMO1 in neutrophils that promotes inflammatory arthritis

Abstract

Rheumatoid arthritis is characterized by progressive joint inflammation and affects ~1% of the human population. We noted single-nucleotide polymorphisms (SNPs) in the apoptotic cell–engulfment genes ELMO1, DOCK2, and RAC1 linked to rheumatoid arthritis. As ELMO1 promotes cytoskeletal reorganization during engulfment, we hypothesized that ELMO1 loss would worsen inflammatory arthritis. Surprisingly, Elmo1-deficient mice showed reduced joint inflammation in acute and chronic arthritis models. Genetic and cell-biology studies revealed that ELMO1 associates with receptors linked to neutrophil function in arthritis and regulates activation and early neutrophil recruitment to the joints, without general inhibition of inflammatory responses. Further, neutrophils from the peripheral blood of human donors that carry the SNP in ELMO1 associated with arthritis display increased migratory capacity, whereas ELMO1 knockdown reduces human neutrophil migration to chemokines linked to arthritis. These data identify ‘noncanonical’ roles for ELMO1 as an important cytoplasmic regulator of specific neutrophil receptors and promoter of arthritis.

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Fig. 1: Engulfment protein ELMO1 contributes to inflammatory arthritis.
Fig. 2: ELMO1 expression in neutrophils regulates disease severity in arthritis.
Fig. 3: The neutrophil-specific ELMO1 protein interactome includes regulators of chemotaxis linked to human arthritis.
Fig. 4: ELMO1 promotes neutrophil migration to inflamed joints.
Fig. 5: Elmo1 function regulates cell type-specific transcriptional programs.
Fig. 6: ELMO1 SNP rs11984075 promotes migration of human neutrophils.

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Data availability

The data that support the findings from this study, including the R code used for bioinformatics analysis and heatmap generation, are available from the corresponding authors upon reasonable request. Sequencing data have been deposited in the GEO database under accession number GSE122412. Reagents used in this study are also listed in the Nature Research Reporting Summary.

References

  1. McInnes, I. B. & Schett, G. The pathogenesis of rheumatoid arthritis. N. Eng. J. Med. 365, 2205–2219 (2011).

    CAS  Google Scholar 

  2. Wright, H. L., Moots, R. J. & Edwards, S. W. The multifactorial role of neutrophils in rheumatoid arthritis. Nat. Rev. Rheumatol. 10, 593–601 (2014).

    CAS  PubMed  Google Scholar 

  3. Moots, R. J. & Naisbett-Groet, B. The efficacy of biologic agents in patients with rheumatoid arthritis and an inadequate response to tumour necrosis factor inhibitors: a systematic review. Rheumatology (Oxford) 51, 2252–2261 (2012).

    CAS  Google Scholar 

  4. Ding, J., Eyre, S. & Worthington, J. Genetics of RA susceptibility, what comes next? RMD Open 1, e000028 (2015).

    PubMed  PubMed Central  Google Scholar 

  5. Arandjelovic, S. & Ravichandran, K. S. Phagocytosis of apoptotic cells in homeostasis. Nat. Immunol. 16, 907–917 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Highton, J., Hessian, P. A., Kean, A. & Chin, M. Cell death by apoptosis is a feature of the rheumatoid nodule. Ann. Rheum. Dis. 62, 77–80 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Pope, R. M. Apoptosis as a therapeutic tool in rheumatoid arthritis. Nat. Rev. Immunol. 2, 527–535 (2002).

    CAS  PubMed  Google Scholar 

  8. Nagata, S., Hanayama, R. & Kawane, K. Autoimmunity and the clearance of dead cells. Cell 140, 619–630 (2010).

    CAS  PubMed  Google Scholar 

  9. Elliott, M. R. & Ravichandran, K. S. The dynamics of apoptotic cell clearance. Dev. Cell. 38, 147–160 (2016).

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Erwig, L. P. & Henson, P. M. Clearance of apoptotic cells by phagocytes. Cell Death Differ. 15, 243–250 (2008).

    CAS  PubMed  Google Scholar 

  11. Gregory, C. D. & Pound, J. D. Microenvironmental influences of apoptosis in vivo and in vitro. Apoptosis 15, 1029–1049 (2010).

    CAS  PubMed  Google Scholar 

  12. Miyanishi, M. et al. Identification of Tim4 as a phosphatidylserine receptor. Nature 450, 435–439 (2007).

    CAS  PubMed  Google Scholar 

  13. Park, D. et al. BAI1 is an engulfment receptor for apoptotic cells upstream of the ELMO/Dock180/Rac module. Nature 450, 430–434 (2007).

    CAS  PubMed  Google Scholar 

  14. Lemke, G. & Rothlin, C. V. Immunobiology of the TAM receptors. Nat. Rev. Immunol. 8, 327–336 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Savill, J. Recognition and phagocytosis of cells undergoing apoptosis. Br. Med. Bull. 53, 491–508 (1997).

    CAS  PubMed  Google Scholar 

  16. Penberthy, K. K. & Ravichandran, K. S. Apoptotic cell recognition receptors and scavenger receptors. Immunol. Rev. 269, 44–59 (2016).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Brugnera, E. et al. Unconventional Rac-GEF activity is mediated through the Dock180–ELMO complex. Nat. Cell Biol. 4, 574–582 (2002).

    CAS  PubMed  Google Scholar 

  18. Zhernakova, A. et al. Meta-analysis of genome-wide association studies in celiac disease and rheumatoid arthritis identifies fourteen non-HLA shared loci. PLoS Genet. 7, e1002004 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Whitaker, J. W. et al. Integrative omics analysis of rheumatoid arthritis identifies non-obvious therapeutic targets. PLoS ONE 10, e0124254 (2015).

    PubMed  PubMed Central  Google Scholar 

  20. Grimsley, C. M. et al. Dock180 and ELMO1 proteins cooperate to promote evolutionarily conserved Rac-dependent cell migration. J. Biol. Chem. 279, 6087–6097 (2004).

    CAS  PubMed  Google Scholar 

  21. Gumienny, T. L. et al. CED12/ELMO, a novel member of the CrkII/Dock180/Rac pathway, is required for phagocytosis and cell migration. Cell 107, 27–41 (2001).

    CAS  PubMed  Google Scholar 

  22. Kouskoff, V. et al. Organ-specific disease provoked by systemic autoimmunity. Cell 87, 811–822 (1996).

    CAS  PubMed  Google Scholar 

  23. Korganow, A. S. et al. From systemic T cell self-reactivity to organ-specific autoimmune disease via immunoglobulins. Immunity 10, 451–461 (1999).

    CAS  PubMed  Google Scholar 

  24. Elliott, M. R. et al. Unexpected requirement for ELMO1 in clearance of apoptotic germ cells in vivo. Nature 467, 333–337 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Brand, D. D., Latham, K. A. & Rosloniec, E. F. Collagen-induced arthritis. Nat. Protoc. 2, 1269–1275 (2007).

    CAS  PubMed  Google Scholar 

  26. Clausen, B. E., Burkhardt, C., Reith, W., Renkawitz, R. & Forster, I. Conditional gene targeting in macrophages and granulocytes using LysMcre mice. Transgenic Res. 8, 265–277 (1999).

    CAS  PubMed  Google Scholar 

  27. Passegue, E., Wagner, E. F. & Weissman, I. L. JunB deficiency leads to a myeloproliferative disorder arising from hematopoietic stem cells. Cell 119, 431–443 (2004).

    CAS  PubMed  Google Scholar 

  28. Yona, S. et al. Fate mapping reveals origins and dynamics of monocytes and tissue macrophages under homeostasis. Immunity 38, 79–91 (2013).

    CAS  PubMed  Google Scholar 

  29. Lewis, A. J., Seymour, C. W. & Rosengart, M. R. Current murine models of sepsis. Surg. Infect. (Larchmt) 17, 385–393 (2016).

    Google Scholar 

  30. Xiong, H. et al. Distinct contributions of neutrophils and CCR2+ monocytes to pulmonary clearance of different Klebsiella pneumoniae strains. Infect. Immun. 83, 3418–3427 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Pierson, E. R., Wagner, C. A. & Goverman, J. M. The contribution of neutrophils to CNS autoimmunity. Clin. Immunol. 189, 23–28 (2018).

    CAS  PubMed  Google Scholar 

  32. Szklarczyk, D. et al. The STRING database in 2017: quality-controlled protein–protein association networks, made broadly accessible. Nucleic Acids Res. 45, D362–D368 (2017).

    CAS  PubMed  Google Scholar 

  33. Chou, R. C. et al. Lipid-cytokine-chemokine cascade drives neutrophil recruitment in a murine model of inflammatory arthritis. Immunity 33, 266–278 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Lammermann, T. et al. Neutrophil swarms require LTB4 and integrins at sites of cell death in vivo. Nature 498, 371–375 (2013).

    PubMed  Google Scholar 

  35. Mancardi, D. A. et al. Cutting edge: the murine high-affinity IgG receptor FcγRIV is sufficient for autoantibody-induced arthritis. J. Immunol. 186, 1899–1903 (2011).

    CAS  PubMed  Google Scholar 

  36. Hornum, L. et al. C5a and C5aR are elevated in joints of rheumatoid and psoriatic arthritis patients, and C5aR blockade attenuates leukocyte migration to synovial fluid. PLoS ONE 12, e0189017 (2017).

    PubMed  PubMed Central  Google Scholar 

  37. Park-Min, K. H. et al. Negative regulation of osteoclast precursor differentiation by CD11b and β2 integrin-B-cell lymphoma 6 signaling. J. Bone Miner. Res. 28, 135–149 (2013).

    CAS  PubMed  Google Scholar 

  38. Ji, H. et al. Arthritis critically dependent on innate immune system players. Immunity 16, 157–168 (2002).

    CAS  PubMed  Google Scholar 

  39. Monach, P. A. et al. Neutrophils in a mouse model of autoantibody-mediated arthritis: critical producers of Fc receptor γ, the receptor for C5a, and lymphocyte function-associated antigen 1. Arthritis Rheum. 62, 753–764 (2010).

  40. Wipke, B. T. & Allen, P. M. Essential role of neutrophils in the initiation and progression of a murine model of rheumatoid arthritis. J. Immunol. 167, 1601–1608 (2001).

    CAS  PubMed  Google Scholar 

  41. Morgan, B. P., Griffiths, M., Khanom, H., Taylor, S. M. & Neal, J. W. Blockade of the C5a receptor fails to protect against experimental autoimmune encephalomyelitis in rats. Clin. Exp. Immunol. 138, 430–438 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Rittirsch, D. et al. Functional roles for C5a receptors in sepsis. Nat. Med. 14, 551–557 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Miyabe, Y. et al. Complement C5a receptor is the key initiator of neutrophil adhesion igniting immune complex-induced arthritis. Sci. Immunol. 2, eaaj2195 (2017).

    PubMed  PubMed Central  Google Scholar 

  44. Corr, M. & Crain, B. The role of FcγR signaling in the K/B x N serum transfer model of arthritis. J. Immunol. 169, 6604–6609 (2002).

    CAS  PubMed  Google Scholar 

  45. Kiefer, F. et al. The Syk protein tyrosine kinase is essential for Fcγ receptor signaling in macrophages and neutrophils. Mol. Cell. Biol. 18, 4209–4220 (1998).

  46. Ruzankina, Y. et al. Deletion of the developmentally essential gene ATR in adult mice leads to age-related phenotypes and stem cell loss. Cell Stem Cell 1, 113–126 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  47. Matmati, M. et al. A20 (TNFAIP3) deficiency in myeloid cells triggers erosive polyarthritis resembling rheumatoid arthritis. Nat. Genet. 43, 908–912 (2011).

    CAS  PubMed  Google Scholar 

  48. Hathaway, C. K. et al. High Elmo1 expression aggravates and low Elmo1 expression prevents diabetic nephropathy. Proc. Natl Acad. Sci. USA 113, 2218–2222 (2016).

    CAS  PubMed  PubMed Central  Google Scholar 

  49. Cooper, D. N. Functional intronic polymorphisms: buried treasure awaiting discovery within our genes. Hum. Genomics 4, 284–288 (2010).

    PubMed  PubMed Central  Google Scholar 

  50. Szanto, S., Gal, I., Gonda, A., Glant, T. T. & Mikecz, K. Expression of l-selectin, but not CD44, is required for early neutrophil extravasation in antigen-induced arthritis. J. Immunol. 172, 6723–6734 (2004).

    CAS  PubMed  Google Scholar 

  51. Shrum, B. et al. A robust scoring system to evaluate sepsis severity in an animal model. BMC Res. Notes 7, 233 (2014).

    PubMed  PubMed Central  Google Scholar 

  52. Boyum, A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand. J. Clin. Lab. Invest. Suppl. 97, 77–89 (1968).

    CAS  PubMed  Google Scholar 

  53. Stohl, E. A., Criss, A. K. & Seifert, H. S. The transcriptome response of Neisseria gonorrhoeae to hydrogen peroxide reveals genes with previously uncharacterized roles in oxidative damage protection. Mol. Microbiol. 58, 520–532 (2005).

    CAS  PubMed  PubMed Central  Google Scholar 

  54. Collins, S. J., Ruscetti, F. W., Gallagher, R. E. & Gallo, R. C. Terminal differentiation of human promyelocytic leukemia cells induced by dimethyl sulfoxide and other polar compounds. Proc. Natl Acad. Sci. USA 75, 2458–2462 (1978).

    CAS  PubMed  PubMed Central  Google Scholar 

  55. Armaka, M., Gkretsi, V., Kontoyiannis, D. & Kollias, G. A standardized protocol for the isolation and culture of normal and arthritogenic murine synovial fibroblasts. Nat. Protoc. https://doi.org/10.1038/nprot.2009.102 (2009).

  56. Alon, R. & Shulman, Z. Chemokine triggered integrin activation and actin remodeling events guiding lymphocyte migration across vascular barriers. Exp. Cell Res. 317, 632–641 (2011).

    CAS  PubMed  Google Scholar 

  57. Bridges, D. & Moorhead, G. B. 14-3-3 proteins: a number of functions for a numbered protein. Sci. STKE 2005, re10 (2005).

    PubMed  Google Scholar 

  58. Delclaux, C. et al. Role of gelatinase B and elastase in human polymorphonuclear neutrophil migration across basement membrane. Am. J. Respir. Cell Mol. Biol. 14, 288–295 (1996).

    CAS  PubMed  Google Scholar 

  59. Diaz-Alvarez, L. & Ortega, E. The many roles of galectin-3, a multifaceted molecule, in innate immune responses against pathogens. Mediators Inflamm. 2017, 9247574 (2017).

    PubMed  PubMed Central  Google Scholar 

  60. Germena, G., Volmering, S., Sohlbach, C. & Zarbock, A. Mutation in the CD45 inhibitory wedge modulates integrin activation and leukocyte recruitment during inflammation. J. Immunol. 194, 728–738 (2015).

    CAS  PubMed  Google Scholar 

  61. Gittens, B. R., Bodkin, J. V., Nourshargh, S., Perretti, M. & Cooper, D. Galectin-3: a positive regulator of leukocyte recruitment in the inflamed microcirculation. J. Immunol. 198, 4458–4469 (2017).

    CAS  PubMed  Google Scholar 

  62. Glennon-Alty, L., Hackett, A. P., Chapman, E. A. & Wright, H. L. Neutrophils and redox stress in the pathogenesis of autoimmune disease. Free Radic. Biol. Med. 125, 25–35 (2018).

    CAS  PubMed  Google Scholar 

  63. Gough, R. E. & Goult, B. T. The tale of two talins—two isoforms to fine-tune integrin signalling. FEBS Lett. 592, 2108–2125 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  64. Jin, J. et al. Proteomic, functional, and domain-based analysis of in vivo 14-3-3 binding proteins involved in cytoskeletal regulation and cellular organization. Curr. Biol. 14, 1436–1450 (2004).

    CAS  PubMed  Google Scholar 

  65. Kletzien, R. F., Harris, P. K. & Foellmi, L. A. Glucose-6-phosphate dehydrogenase: a “housekeeping” enzyme subject to tissue-specific regulation by hormones, nutrients, and oxidant stress. FASEB J. 8, 174–181 (1994).

    CAS  PubMed  Google Scholar 

  66. Oikonomou, K. G., Zachou, K. & Dalekos, G. N. α-actinin: a multidisciplinary protein with important role in B-cell driven autoimmunity. Autoimmun. Rev. 10, 389–396 (2011).

  67. Sadik, C. D., Miyabe, Y., Sezin, T. & Luster, A. D. The critical role of C5a as an initiator of neutrophil-mediated autoimmune inflammation of the joint and skin. Semin. Immunol. 37, 21–29 (2018).

    CAS  PubMed  Google Scholar 

  68. Sano, H. et al. Human galectin-3 is a novel chemoattractant for monocytes and macrophages. J. Immunol. 165, 2156–2164 (2000).

    CAS  PubMed  Google Scholar 

  69. Springer, T. A. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 76, 301–314 (1994).

    CAS  PubMed  Google Scholar 

  70. Thomas, D. C. The phagocyte respiratory burst: historical perspectives and recent advances. Immunol. Lett. 192, 88–96 (2017).

    CAS  PubMed  Google Scholar 

  71. Wyatt, E. et al. Regulation and cytoprotective role of hexokinase III. PLoS ONE 5, e13823 (2010).

    PubMed  PubMed Central  Google Scholar 

  72. Yang, X. et al. Structural basis for protein–protein interactions in the 14-3-3 protein family. Proc. Natl Acad. Sci. USA 103, 17237–17242 (2006).

  73. Balakrishnan, L. et al. Differential proteomic analysis of synovial fluid from rheumatoid arthritis and osteoarthritis patients. Clin. Proteomics 11, 1 (2014).

    PubMed  PubMed Central  Google Scholar 

  74. Cui, J. et al. Rheumatoid arthritis risk allele PTPRC is also associated with response to anti-tumor necrosis factor alpha therapy. Arthritis Rheum. 62, 1849–1861 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  75. de Rooy, D. P. et al. A genetic variant in the region of MMP-9 is associated with serum levels and progression of joint damage in rheumatoid arthritis. Ann. Rheum. Dis. 73, 1163–1169 (2014).

    PubMed  Google Scholar 

  76. Gheita, T. A., Kenawy, S. A., El Sisi, R. W., Gheita, H. A. & Khalil, H. Subclinical reduced G6PD activity in rheumatoid arthritis and Sjogren’s syndrome patients: relation to clinical characteristics, disease activity and metabolic syndrome. Mod. Rheumatol. 24, 612–617 (2014).

    CAS  PubMed  Google Scholar 

  77. Hu, C. Y., Chang, S. K., Wu, C. S., Tsai, W. I. & Hsu, P. N. Galectin-3 gene (LGALS3) +292C allele is a genetic predisposition factor for rheumatoid arthritis in Taiwan. Clin. Rheumatol. 30, 1227–1233 (2011).

    PubMed  Google Scholar 

  78. Huang, R. Y., Huang, Q. C. & Burgering, B. M. Novel insight into the role of alpha-actinin-1 in rheumatoid arthritis. Discov. Med. 17, 75–80 (2014).

    PubMed  Google Scholar 

  79. Maksymowych, W. P. & Marotta, A. 14-3-3eta: a novel biomarker platform for rheumatoid arthritis. Clin. Exp. Rheumatol. 32, S-35–S-39 (2014).

    Google Scholar 

  80. Tsuzaka, K., Itami, Y., Shinozaki, N. & Morishita, T. Plasma talin is a new diagnostic and monitoring marker for rheumatoid arthritis. Arthritis Rheum. 63, S134 (2011).

  81. Warchol, T., Lianeri, M., Lacki, J. K., Olesinska, M. & Jagodzinski, P. P. ITGAM Arg77His is associated with disease susceptibility, arthritis, and renal symptoms in systemic lupus erythematosus patients from a sample of the Polish population. DNA Cell Biol. 30, 33–38 (2011).

    CAS  PubMed  Google Scholar 

  82. Cottier, K. E., Fogle, E. M., Fox, D. A. & Ahmed, S. Noxa in rheumatic diseases: present understanding and future impact. Rheumatology (Oxford) 53, 1539–1546 (2014).

    CAS  Google Scholar 

  83. Cui, J. et al. Genome-wide association study and gene expression analysis identifies CD84 as a predictor of response to etanercept therapy in rheumatoid arthritis. PLoS Genet. 9, e1003394 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  84. Elsby, L. M. et al. Functional evaluation of TNFAIP3 (A20) in rheumatoid arthritis. Clin. Exp. Rheumatol. 28, 708–714 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  85. Howng, S. L. et al. Autoimmunity against hNinein, a human centrosomal protein, in patients with rheumatoid arthritis and systemic lupus erythematosus. Mol. Med. Rep. 4, 825–830 (2011).

    CAS  PubMed  Google Scholar 

  86. Issuree, P. D. et al. iRHOM2 is a critical pathogenic mediator of inflammatory arthritis. J. Clin. Invest. 123, 928–932 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  87. Jin, T., Tarkowski, A., Carmeliet, P. & Bokarewa, M. Urokinase, a constitutive component of the inflamed synovial fluid, induces arthritis. Arthritis. Res. Ther. 5, R9–R17 (2003).

    CAS  PubMed  Google Scholar 

  88. Lopez, M. et al. Tumor necrosis factor and transforming growth factor β regulate clock genes by controlling the expression of the cold inducible RNA-binding protein (CIRBP). J. Biol. Chem. 289, 2736–2744 (2014).

    CAS  PubMed  Google Scholar 

  89. Marotte, H. et al. Blocking of interferon regulatory factor 1 reduces tumor necrosis factor α-induced interleukin-18 bioactivity in rheumatoid arthritis synovial fibroblasts by induction of interleukin-18 binding protein a: role of the nuclear interferon regulatory factor 1–NF-κB–c-jun complex. Arthritis Rheum. 63, 3253–3262 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  90. Ohyama, K. et al. Serum immune complex containing thrombospondin-1: a novel biomarker for early rheumatoid arthritis. Ann. Rheum. Dis. 71, 1916–1917 (2012).

    PubMed  Google Scholar 

  91. Okada, Y. et al. Genetics of rheumatoid arthritis contributes to biology and drug discovery. Nature 506, 376–381 (2014).

    CAS  PubMed  Google Scholar 

  92. Perlman, H. et al. Rheumatoid arthritis synovial macrophages express the Fas-associated death domain-like interleukin-1β-converting enzyme-inhibitory protein and are refractory to Fas-mediated apoptosis. Arthritis Rheum. 44, 21–30 (2001).

    CAS  PubMed  Google Scholar 

  93. Rico, M. C. et al. Amelioration of inflammation, angiogenesis and CTGF expression in an arthritis model by a TSP1-derived peptide treatment. J. Cell. Physiol. 211, 504–512 (2007).

    CAS  PubMed  Google Scholar 

  94. Yoo, I. S. et al. Serum and synovial fluid concentrations of cold-inducible RNA-binding protein in patients with rheumatoid arthritis. Int. J. Rheum. Dis. 21, 148–154 (2018).

    CAS  PubMed  Google Scholar 

  95. Li, M. J. et al. GWASdb: a database for human genetic variants identified by genome-wide association studies. Nucleic Acids Res. 40, D1047–D1054 (2012).

    CAS  PubMed  Google Scholar 

  96. Pletscher-Frankild, S., Palleja, A., Tsafou, K., Binder, J. X. & Jensen, L. J. DISEASES: text mining and data integration of disease-gene associations. Methods 74, 83–89 (2015).

    CAS  PubMed  Google Scholar 

  97. Davis, A. P. et al. The comparative toxicogenomics database’s 10th year anniversary: update 2015. Nucleic Acids Res. 43, D914–D920 (2015).

    CAS  PubMed  Google Scholar 

  98. Davis, A. P. et al. Comparative toxicogenomics database: a knowledgebase and discovery tool for chemical-gene-disease networks. Nucleic Acids Res. 37, D786–D792 (2009).

    CAS  PubMed  Google Scholar 

  99. Eppig, J. T., Blake, J. A., Bult, C. J., Kadin, J. A. & Richardson, J. E. The Mouse Genome Database (MGD): facilitating mouse as a model for human biology and disease. Nucleic Acids Res. 43, D726–D736 (2015).

    CAS  PubMed  Google Scholar 

  100. Blake, J. A., Richardson, J. E., Bult, C. J., Kadin, J. A. & Eppig, J. T. MGD: the mouse genome database. Nucleic Acids Res. 31, 193–195 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors thank members of the Ravichandran laboratory for discussions and critical reading of the manuscript, A. Criss and A. Smirnov for human neutrophils, S. T. Fleury for assistance with neutrophil purification, and K. Koster for assistance with bone marrow preparations. This work is supported by grants to K.S.R. from NIGMS R35GM122542, NIMH (MH096484), NHLBI (P01HL120840), NICHD (HD07498), and the Center for Cell Clearance/University of Virginia School of Medicine, and the Odysseus Award from the FWO, Belgium, to M.R.E. from NIAID (AI114554), to M.K. from NIAID (P01 AI102851), and to A.G. (NS083542 and NS101281). Additional support was provided by the Philip S. Magaram, Esq. Research Award from the Arthritis Foundation to S.A.. J.S.A.P. is supported by a Mark Foundation award from the Cancer Research Institute and a postdoctoral enrichment award from Burroughs Wellcome and was previously supported by a postdoctoral fellowship through a T32 Cancer Training Grant. C.D.L. is supported by an award from The Wellcome Trust (206566/Z/17/Z). L.S.S. is supported by the Roaring Fork Valley Postdoctoral Research Fellowship (PF-17-098-01-CSM) from the American Cancer Society, and K.K.P. is supported by an NHLBI F30 award (F30 HL126385) and was previously supported by an NIH T32 Immunology Training Grant (T32 AI007496).

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Authors

Contributions

Conceptualization, S.A. and K.S.R.; methodology, S.A. and K.S.R.; software, J.S.A.P.; investigation, S.A., J.S.A.P., C.D.L., K.K.P., T.-H.K., M.Z., D.A R., T.-Y.C., A.M.B., L.S.S., A.H.C., and A.G.; data curation, J.S.A.P.; writing, S.A. and K.S.R.; resources, A.G., T.P.C., M.K., M.R.E. and K.S.R.; funding acquisition, K.S.R.

Corresponding authors

Correspondence to Sanja Arandjelovic or Kodi S. Ravichandran.

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Integrated supplementary information

Supplementary Figure 1 Loss of ELMO1, but not BAI1, reduces disease severity in arthritis.

a) Mice with spontaneous arthritis (K/BxN) or their healthy littermates were used to analyze engulfment machinery expression in isolated paws. b) Expression of apoptotic cell clearance components genes in total paw extracts from K/BxN mice by qRT-PCR. Each data point represents one mouse. c) Incidence of CIA in male Elmo1+/+-DBA (n = 3, white symbols), Elmo1+/– -DBA (n = 12, black symbols) and Elmo1–/–-DBA (n = 6, green symbols). d) Swelling of the paws (left panel) and clinical scores (right panel) were measured in male Elmo1+/+-DBA (n = 3, white symbols), Elmo1+/– -DBA (n = 12, black symbols) and Elmo1–/–-DBA (n = 5, green symbols). e) Incidence of CIA in female Elmo1+/+-DBA (n = 5, white symbols), Elmo1+/– -DBA (n = 8, black symbols) and Elmo1–/–-DBA (n = 7, green symbols). f) Swelling of the paws (left panel) and clinical scores (right panel) were measured in female Elmo1+/+-DBA (n = 5, white symbols), Elmo1+/–-DBA (n = 8, black symbols) and Elmo1–/–-DBA (n = 5, green symbols). g) Elmo1, Elmo2 and Elmo3 expression was analyzed by quantitative RT-PCR in the total paw extracts of Elmo1+/– (black symbols, n = 7) and Elmo1–/– (green symbols, n = 7) mice on day 10 after K/BxN serum injection. Each symbol represents an individual animal. h) Schematic of BAI1 binding to ELMO1-DOCK2, leading to the activation of GTPase RAC1. i) Paw swelling and clinical scores of BAI1+/+ (n = 4, black symbols) and BAI1–/– mice (n = 11, blue symbols) injected with 150 μl of K/BxN serum on day 0.

Supplementary Figure 2 ELMO1 expression in neutrophils regulates disease severity in arthritis.

a) Immunoblot analysis of ELMO1 protein expression in resident peritoneal macrophages from indicated mice. The blot was cropped to show relevant bands. b)Paw swelling and clinical scores of (n = 4, black symbols) and Lyz2-Cre (n = 3, blue symbols) mice injected with 150 μl of K/BxN serum on day 0 and 2. c) Paw swelling and clinical scores of Elmo1fl/fl (Control, n = 7) or Elmo1fl/flLyz2-Cre (n = 5) mice injected with 150 μl of K/BxN serum on day 0 and 2. d) Flow cytometry analysis of paws from of Elmo1fl/fl (n = 3) or Elmo1fl/flLyz2-Cre (n = 3) mice on day 10 after K/BxN serum injection. Cells in the singlet gate are shown. e) Paw swelling and clinical scores of Elmo1fl/fl (Control, n = 5) or Elmo1fl/flCx3cr1-Cre (n = 5) mice injected with 150 μl of K/BxN serum on day 0 and 2. f) Representative hind paw ankle sections stained with hematoxylin and eosin on day 10 after K/BxN serum injection (left panels). Areas of inflammatory cell infiltration are indicated with yellow asterisks. Scale bar = 1 mm. Quantification of inflammation (right panel) in Elmo1fl/fl (Control, n = 5) or Elmo1fl/flCx3cr1-Cre (n = 5). g) Paw swelling and clinical scores of Elmo1fl/fl (Control, n = 6) or Elmo1fl/flMrp8-Cre (n = 3) mice injected with 150 μl of K/BxN serum on day 0 and 2. h) Immunoblot analysis of ELMO1 protein expression in the Ly6G+ cells purified from the bone marrow of indicated mouse strains, as described in the Methods. The blot was cropped to show relevant bands. i) Paw swelling and clinical scores of (n = 8, black symbols) and Mrp8-Cre (n = 3, magenta symbols) mice were injected with 150 μl of K/BxN serum on day 0 and 2. j) Immunoblotting of ELMO1 protein expression in neutrophils and fibroblast like synoviocytes (FLS) from the indicated strains of mice. Each lane represents an individual animal. The blot was cropped to show relevant bands.

Supplementary Figure 3 Bacterial-challenge response and EAE are not changed in Elmo1–/–mice.

a) Mice were challenged with LPS administered intranasally as described in the Methods, and mobilization of neutrophils was analyzed 8 hours later by flow cytometry of cells in the BAL. b) Quantification of 7-AAD-CD11b+Ly6C+Ly6G+ neutrophils in the BAL of Elmo1+/+, Elmo1–/–, Elmo1fl/fl and Elmo1fl/flMrp8-Cre mice. Each symbol represents an animal. c) Elmo1+/+ (n = 18) and Elmo1–/– (n = 19) mice were injected with fecal contents (1.5 mg/g) intraperitoneally to induce fecal-induced peritonitis (FIP) and disease parameters were monitored as described in Methods. d) Number of Ly6G+ neutrophils in the peritoneum 4 hours post FIP induction. Each symbol represents an animal. e) Temperature of mice in c). f) Clinical scores of mice in c) were measured as described in Methods. g) Survival curves of mice in c). All mice alive at 72 hours post FIP induction exhibited complete recovery. h) Purified neutrophils from Elmo1+/+ (n = 4) and Elmo1–/– (n = 4) mice were incubated with Klebsiella pneumoniae at a 1:2 ratio for 1 hour, as described in the Methods, and bacterial killing was analyzed. Each symbol represents neutrophils from an individual animal. i) Elmo1+/– (n = 8, black symbols) and Elmo1–/– (n = 8, green symbols) mice were immunized to induce EAE and disease was scored over the indicated time as described in the Methods. j) Luxol Fast Blue staining of the spinal cords on day 26 after EAE induction (left panel). Demyelination was scored at four different levels of the spinal cord and composite result is shown (right panel). Each symbol represents an individual animal.

Supplementary Figure 4 Elmo1 deletion in neutrophils inhibits neutrophil migration.

a) Total bone marrow cells were purified as described in the Methods, and cell migration toward the indicated concentrations of LTB4 or CXCL1 was evaluated after 3 hours at 37 °C by flow cytometry. Each symbol represents an individual animal. b) Transcript levels of the LTB4 receptor Blt1 are analyzed by qPCR. Each symbol represents an individual animal. c) Flow cytometry analysis of cell surface levels of CXCR2 in Elmo1+/+ and Elmo1–/– mice on Ly6G+ neutrophils in the blood (top panels, n = 9, 11) and the bone marrow (bottom panels, n = 3, 3). MFI, mean fluorescence index. FMO, fluorescence minus one. Each symbol represents an individual animal. d) Flow cytometry analysis of cell surface levels of FcγRIII and FcγRIV on CD11b+Ly6G+ neutrophils in the blood (top panels) and the bone marrow (bottom panels) of Elmo1+/+ and Elmo1–/– mice. e) Immunoblotting of FcγRI protein expression in Ly6G+ neutrophils purified from the bone marrow of Elmo1+/+ and Elmo1–/– mice. The blot was cropped to show relevant bands in non-adjacent lanes. f) Immunoblotting of Syk protein expression in Ly6G+ neutrophils purified from the bone marrow of Elmo1+/+ and Elmo1–/– mice. The blot was cropped to show relevant bands in non-adjacent lanes.

Supplementary Figure 5 Inducible deletion of Elmo1 in ongoing arthritis.

(a) Paw swelling of male and female Elmo1fl/flUbc-CreERt2 (n = 9 for males, n = 7 for females, green symbols) and littermate control (n = 12 for males, n = 10 for females, black symbols) mice with K/BxN serum transfer induced arthritis and tamoxifen administration (arrows). b) Paw swelling (left panel) and clinical scores (right panel) of Elmo1fl/fl (control, n = 5) and Ubc-CreERt2/Elmo1fl/fl (n = 3) mice with K/BxN serum transfer induced arthritis without tamoxifen administration. c) Schematic of the K/BxN serum transfer arthritis induction and treatment with tamoxifen to induce deletion of Elmo1 during ongoing arthritis. d) Paw swelling of male Ubc-CreERt2/Elmo1fl/fl (n = 3, green symbols) and littermate control (n = 6, black symbols) mice with K/BxN serum transfer induced arthritis and tamoxifen administration (arrows). e) ELMO1 protein expression in human peripheral blood neutrophils (duplicates are shown) and buffy coat cells from two different donors (1 and 2). ERK protein expression is used as a loading control. The blot was cropped to show relevant bands.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5 and Supplementary Tables 1–3

Reporting Summary

Supplementary Video 1 (Elmo1KO): Reduced migration of Elmo1–/– neutrophils toward the site of laser induced injury.

Representative time-lapse videos showing movement of neutrophils in response to laser-induced injury, as described in the Methods. Damaged area is indicated by the yellow dotted line. Second harmonic generation is shown in gray, neutrophils are shown in green. Time-scale, min:sec, Scale bar = 20µm.

Supplementary Video 2 (Elmo1WT): Reduced migration of Elmo1–/– neutrophils toward the site of laser induced injury.

Representative time-lapse videos showing movement of neutrophils in response to laser-induced injury, as described in the Methods. Damaged area is indicated by the yellow dotted line. Second harmonic generation is shown in gray, neutrophils are shown in green. Time-scale, min:sec, Scale bar = 20µm.

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Arandjelovic, S., Perry, J.S.A., Lucas, C.D. et al. A noncanonical role for the engulfment gene ELMO1 in neutrophils that promotes inflammatory arthritis. Nat Immunol 20, 141–151 (2019). https://doi.org/10.1038/s41590-018-0293-x

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