Neutrophilic inflammation is a hallmark of many monogenic autoinflammatory diseases; pathomechanisms that regulate extravasation of damaging immune cells into surrounding tissues are poorly understood. Here we identified three unrelated boys with perinatal-onset of neutrophilic cutaneous small vessel vasculitis and systemic inflammation. Two patients developed liver fibrosis in their first year of life. Next-generation sequencing identified two de novo truncating variants in the Src-family tyrosine kinase, LYN, p.Y508*, p.Q507* and a de novo missense variant, p.Y508F, that result in constitutive activation of Lyn kinase. Functional studies revealed increased expression of ICAM-1 on induced patient-derived endothelial cells (iECs) and of β2-integrins on patient neutrophils that increase neutrophil adhesion and vascular transendothelial migration (TEM). Treatment with TNF inhibition improved systemic inflammation; and liver fibrosis resolved on treatment with the Src kinase inhibitor dasatinib. Our findings reveal a critical role for Lyn kinase in modulating inflammatory signals, regulating microvascular permeability and neutrophil recruitment, and in promoting hepatic fibrosis.

2nd Faculty of Medicine Charles University University Hospital Motol Prague Czech Republic

AstraZeneca Research Based Biopharmaceutical Company Waltham MA USA

Clinical Center National Institutes of Health Bethesda MD USA

Collaborative Clinical Research Branch Neutrophil Monitoring Laboratory National Institute of Allergy and Infectious Diseases National Institutes of Health Frederick MD USA

Departments of Pediatrics and Medical Microbiology and Immunology University of Wisconsin Madison Madison WI USA

Great Ormond Street Hospital for Children NHS Foundation Trust London UK

Laboratory of Immunoregulation National Institute of Allergy and Infectious Diseases National Institutes of Health Bethesda MD USA

Laboratory of Leukocyte Signaling Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

Medical University of South Carolina Charleston SC USA

National Cancer Institute National Institutes of Health Bethesda MD USA

National Heart Lung and Blood Institute National Institutes of Health Bethesda MD USA

National Institute of Arthritis and Musculoskeletal and Skin Diseases National Institutes of Health Bethesda MD USA

Raigmore Hospital Inverness Scotland

Royal Hospital for Children Glasgow Scotland

Sanofi Oncology Vitry France

Translational Autoinflammatory Diseases Section National Institute of Allergy and Infectious Diseases National Institutes of Health Bethesda MD USA

Yale University New Haven CT USA

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de Jesus AA, Canna SW, Liu Y, Goldbach-Mansky R. Molecular mechanisms in genetically defined autoinflammatory diseases: disorders of amplified danger signaling. Annu. Rev. Immunol. 2015;33:823–874. doi: 10.1146/annurev-immunol-032414-112227. PubMed DOI PMC

Lin B, Goldbach-Mansky R. Pathogenic insights from genetic causes of autoinflammatory inflammasomopathies and interferonopathies. J. Allergy Clin. Immunol. 2022;149:819–832. doi: 10.1016/j.jaci.2021.10.027. PubMed DOI PMC

Scapini P, Pereira S, Zhang H, Lowell CA. Multiple roles of Lyn kinase in myeloid cell signaling and function. Immunol. Rev. 2009;228:23–40. doi: 10.1111/j.1600-065X.2008.00758.x. PubMed DOI PMC

Lowell CA. Src-family and Syk kinases in activating and inhibitory pathways in innate immune cells: signaling cross talk. Cold Spring Harb. Perspect. Biol. 2011;3:a002352. doi: 10.1101/cshperspect.a002352. PubMed DOI PMC

Ingley E. Functions of the Lyn tyrosine kinase in health and disease. Cell Commun. Signal. 2012;10:21. doi: 10.1186/1478-811X-10-21. PubMed DOI PMC

Stehelin D, Fujita DJ, Padgett T, Varmus HE, Bishop JM. Detection and enumeration of transformation-defective strains of avian sarcoma virus with molecular hybridization. Virology. 1977;76:675–684. doi: 10.1016/0042-6822(77)90250-1. PubMed DOI

Irby RB, et al. Activating SRC mutation in a subset of advanced human colon cancers. Nat. Genet. 1999;21:187–190. doi: 10.1038/5971. PubMed DOI

Turro E, et al. A dominant gain-of-function mutation in universal tyrosine kinase SRC causes thrombocytopenia, myelofibrosis, bleeding, and bone pathologies. Sci. Transl. Med. 2016;8:328ra30. doi: 10.1126/scitranslmed.aad7666. PubMed DOI PMC

Abe K, et al. Gain-of-function mutations in a member of the Src family kinases cause autoinflammatory bone disease in mice and humans. Proc. Natl Acad. Sci. USA. 2019;116:11872–11877. doi: 10.1073/pnas.1819825116. PubMed DOI PMC

Kanderova V, et al. Early-onset pulmonary and cutaneous vasculitis driven by constitutively active SRC-family kinase HCK. J. Allergy Clin. Immunol. 2022;149:1464–1472.e3. doi: 10.1016/j.jaci.2021.07.046. PubMed DOI

Zarrin AA, Bao K, Lupardus P, Vucic D. Kinase inhibition in autoimmunity and inflammation. Nat. Rev. Drug Discov. 2021;20:39–63. doi: 10.1038/s41573-020-0082-8. PubMed DOI PMC

Williams NK, Lucet IS, Klinken SP, Ingley E, Rossjohn J. Crystal structures of the Lyn protein tyrosine kinase domain in its Apo- and inhibitor-bound state. J. Biol. Chem. 2009;284:284–291. doi: 10.1074/jbc.M807850200. PubMed DOI

Roskoski R., Jr. Src protein-tyrosine kinase structure and regulation. Biochem. Biophys. Res. Commun. 2004;324:1155–1164. doi: 10.1016/j.bbrc.2004.09.171. PubMed DOI

Draber P, et al. SCIMP, a transmembrane adaptor protein involved in major histocompatibility complex class II signaling. Mol. Cell Biol. 2011;31:4550–4562. doi: 10.1128/MCB.05817-11. PubMed DOI PMC

Kralova J, et al. The transmembrane adaptor protein SCIMP facilitates sustained dectin-1 signaling in dendritic cells. J. Biol. Chem. 2016;291:16530–16540. doi: 10.1074/jbc.M116.717157. PubMed DOI PMC

Luo L, Lucas RM, Liu L, Stow JL. Signalling, sorting and scaffolding adaptors for Toll-like receptors. J. Cell Sci. 2019;133:jcs239194. doi: 10.1242/jcs.239194. PubMed DOI

Boras M, et al. Skap2 is required for beta2 integrin-mediated neutrophil recruitment and functions. J. Exp. Med. 2017;214:851–874. doi: 10.1084/jem.20160647. PubMed DOI PMC

Wynn TA. Common and unique mechanisms regulate fibrosis in various fibroproliferative diseases. J. Clin. Invest. 2007;117:524–529. doi: 10.1172/JCI31487. PubMed DOI PMC

Pandey E, Nour AS, Harris EN. Prominent receptors of liver sinusoidal endothelial cells in liver homeostasis and disease. Front. Physiol. 2020;11:873. doi: 10.3389/fphys.2020.00873. PubMed DOI PMC

Del Campo JA, Gallego P, Grande L. Role of inflammatory response in liver diseases: therapeutic strategies. World J. Hepatol. 2018;10:1–7. doi: 10.4254/wjh.v10.i1.1. PubMed DOI PMC

Benedicto A, et al. Liver sinusoidal endothelial cell ICAM-1 mediated tumor/endothelial crosstalk drives the development of liver metastasis by initiating inflammatory and angiogenic responses. Sci. Rep. 2019;9:13111. doi: 10.1038/s41598-019-49473-7. PubMed DOI PMC

Babbs C, et al. Endothelial cell transformation in primary biliary cirrhosis: a morphological and biochemical study. Hepatology. 1990;11:723–729. doi: 10.1002/hep.1840110503. PubMed DOI

Ramachandran P, et al. Resolving the fibrotic niche of human liver cirrhosis at single-cell level. Nature. 2019;575:512–518. doi: 10.1038/s41586-019-1631-3. PubMed DOI PMC

Su T, et al. Single-cell transcriptomics reveals zone-specific alterations of liver sinusoidal endothelial cells in cirrhosis. Cell Mol. Gastroenterol. Hepatol. 2021;11:1139–1161. doi: 10.1016/j.jcmgh.2020.12.007. PubMed DOI PMC

Li Y, Xiong L, Gong J. Lyn kinase enhanced hepatic fibrosis by modulating the activation of hepatic stellate cells. Am. J. Transl. Res. 2017;9:2865–2877. PubMed PMC

Gavard J, Gutkind JS. VEGF controls endothelial-cell permeability by promoting the beta-arrestin-dependent endocytosis of VE-cadherin. Nat. Cell Biol. 2006;8:1223–1234. doi: 10.1038/ncb1486. PubMed DOI

Benn A, Bredow C, Casanova I, Vukicevic S, Knaus P. VE-cadherin facilitates BMP-induced endothelial cell permeability and signaling. J. Cell Sci. 2016;129:206–218. PubMed PMC

Sindrilaru A, et al. Site of blood vessel damage and relevance of CD18 in a murine model of immune complex-mediated vasculitis. J. Invest. Dermatol. 2007;127:447–454. doi: 10.1038/sj.jid.5700563. PubMed DOI

Hibbs ML, et al. Sustained activation of Lyn tyrosine kinase in vivo leads to autoimmunity. J. Exp. Med. 2002;196:1593–1604. doi: 10.1084/jem.20020515. PubMed DOI PMC

Williams MR, Azcutia V, Newton G, Alcaide P, Luscinskas FW. Emerging mechanisms of neutrophil recruitment across endothelium. Trends Immunol. 2011;32:461–469. doi: 10.1016/j.it.2011.06.009. PubMed DOI PMC

Phillipson M, et al. Intraluminal crawling of neutrophils to emigration sites: a molecularly distinct process from adhesion in the recruitment cascade. J. Exp. Med. 2006;203:2569–2575. doi: 10.1084/jem.20060925. PubMed DOI PMC

Yang L, et al. ICAM-1 regulates neutrophil adhesion and transcellular migration of TNF-alpha-activated vascular endothelium under flow. Blood. 2005;106:584–592. doi: 10.1182/blood-2004-12-4942. PubMed DOI PMC

Piwnica-Worms H, Saunders KB, Roberts TM, Smith AE, Cheng SH. Tyrosine phosphorylation regulates the biochemical and biological properties of pp60c-src. Cell. 1987;49:75–82. doi: 10.1016/0092-8674(87)90757-4. PubMed DOI

Jin LL, et al. Tyrosine phosphorylation of the Lyn Src homology 2 (SH2) domain modulates its binding affinity and specificity. Mol. Cell Proteomics. 2015;14:695–706. doi: 10.1074/mcp.M114.044404. PubMed DOI PMC

Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis. Adv. Drug Deliv. Rev. 2017;121:27–42. doi: 10.1016/j.addr.2017.05.007. PubMed DOI PMC

Wheeler DL, Iida M, Dunn EF. The role of Src in solid tumors. Oncologist. 2009;14:667–678. doi: 10.1634/theoncologist.2009-0009. PubMed DOI PMC

Cai H, et al. Differential transformation capacity of Src family kinases during the initiation of prostate cancer. Proc. Natl Acad. Sci. USA. 2011;108:6579–6584. doi: 10.1073/pnas.1103904108. PubMed DOI PMC

Kohlhas V, Hallek M, Nguyen PH. Constitutive activation of Lyn kinase enhances BCR responsiveness, but not the development of CLL in Emicro-TCL1 mice. Blood Adv. 2020;4:6106–6116. doi: 10.1182/bloodadvances.2020002584. PubMed DOI PMC

Louvrier, C. et al. De novo gain-of-function variations in LYN lead to an early onset systemic autoinflammatory disorder. Arthritis Rheumatol.10.1002/art.42354 (2022). PubMed

Han J, et al. A critical role for Lyn kinase in strengthening endothelial integrity and barrier function. Blood. 2013;122:4140–4149. doi: 10.1182/blood-2013-03-491423. PubMed DOI PMC

Lee D, et al. Dasatinib inhibits Lyn and Fyn Src-family kinases in mast cells to suppress type I hypersensitivity in mice. Biomol. Ther. 2020;28:456–464. doi: 10.4062/biomolther.2020.013. PubMed DOI PMC

Rivera-Torres J, San Jose E. Src tyrosine kinase inhibitors: new perspectives on their immune, antiviral, and senotherapeutic potential. Front. Pharmacol. 2019;10:1011. doi: 10.3389/fphar.2019.01011. PubMed DOI PMC

Carmona-Rivera C, et al. A role for muscarinic receptors in neutrophil extracellular trap formation and levamisole-induced autoimmunity. JCI Insight. 2017;2:e89780. doi: 10.1172/jci.insight.89780. PubMed DOI PMC

Chu PH, et al. Stem cell-derived endothelial cell model that responds to tobacco smoke like primary endothelial cells. Chem. Res. Toxicol. 2020;33:751–763. doi: 10.1021/acs.chemrestox.9b00363. PubMed DOI PMC