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Apr 01, 2025;

Targeting HPK1 inhibits neutrophil responses to mitigate post-stroke lung and cerebral injuries.

Tingting Zhang, Ying Sun, Jing Xia, Hongye Fan, Dingfang Shi, Qian Wu, Ming Huang, Xiao-Yu Hou
Author Information
  1. Tingting Zhang: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, 211198, China. ORCID
  2. Ying Sun: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, 211198, China.
  3. Jing Xia: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, 211198, China.
  4. Hongye Fan: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, 211198, China.
  5. Dingfang Shi: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, 211198, China.
  6. Qian Wu: The Department of Neurology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China. ORCID
  7. Ming Huang: Research Center for Biochemistry and Molecular Biology, Xuzhou Medical University, Xuzhou, Jiangsu, 221004, China. huangming88@xzhmu.edu.cn. ORCID
  8. Xiao-Yu Hou: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, 211198, China. xyhou@cpu.edu.cn. ORCID
PMID: 40169896 DOI: 10.1038/s44321-025-00220-8

Abstract

Circulating neutrophils are responsible for poor neurological outcomes and have been implicated in respiratory morbidity after acute ischemic stroke (AIS). However, the molecular mechanisms regulating neutrophil responses and their pathological relevance in post-stroke complications remain unclear. In this study, we investigated the involvement of hematopoietic progenitor kinase 1 (HPK1) in neutrophil responses and mobilization, as well as subsequent lung and cerebral injuries following AIS. We found that lipopolysaccharide treatment triggered neutrophil activation in an HPK1-dependent manner. HPK1 enhanced intrinsic NF-��B/STAT3/p38-MAPK pathways and gasdermin D cleavage, leading to neutrophil hyperactivation. Following AIS, HPK1 promoted the mobilization of CXCR2 bone marrow neutrophils. HPK1 loss inhibited peripheral neutrophil hyperactivation, neutrophil infiltration, and aggregation of neutrophil extracellular traps, progressively alleviating systemic inflammation and impairments in mouse pulmonary and neurological functions. Furthermore, HPK1 pharmacological inhibition attenuated post-stroke pulmonary and neurological impairments in mice. Our findings revealed that HPK1 upregulates neutrophil mobilization and various responses, promoting post-stroke systemic inflammation and tissue injury. This study highlights HPK1 as a therapeutic target for improving pulmonary and neurological functions after AIS.

Keywords

Acute Ischemic Stroke Gasdermin D Neutrophil Extracellular Traps Neutrophil Mobilization Systemic Inflammation

References

  1. Bai W, Li W, Ning YL, Li P, Zhao Y, Yang N, Jiang YL, Liang ZP, Jiang DP, Wang Y et al (2018) Blood glutamate levels are closely related to acute lung injury and prognosis after stroke. Front Neurol 8:755 [PMID: 29403427]
  2. Bi Y, Shen J, Chen SC, Chen JX, Xia YP (2021) Prognostic value of neutrophil to lymphocyte ratio in acute ischemic stroke after reperfusion therapy. Sci Rep 11:6177 [PMID: 33731740]
  3. Borregaard N (2010) Neutrophils, from marrow to microbes. Immunity 33:657���670 [PMID: 21094463]
  4. Broz P, Pelegr��n P, Shao F (2020) The gasdermins, a protein family executing cell death and inflammation. Nat Rev Immunol 20:143���157 [PMID: 31690840]
  5. Chuang HC, Wang X, Tan TH (2016) MAP4K family kinases in immunity and inflammation. Adv Immunol 129:277���314 [PMID: 26791862]
  6. Denorme F, Portier I, Rustad JL, Cody MJ, de Araujo CV, Hoki C, Alexander MD, Grandhi R, Dyer MR, Neal MD et al (2022) Neutrophil extracellular traps regulate ischemic stroke brain injury. J Clin Invest 132:e154225 [PMID: 35358095]
  7. Deva S, Zhou C, Bishnoi SK, Lau PKH, Tran B, Ba Y, Galsky MD, Wang Y, Zhang Y, Luo S et al (2024) A first-in-human phase 1a dose-escalation study of BGB-15025 (HPK1 inhibitor) as monotherapy and in combination with tislelizumab (TIS; anti-PD-1 antibody) in patients (pts) with advanced solid tumors. J Clin Oncol 42:2585���2585 [DOI: 10.1200/JCO.2024.42.16_suppl.2585]
  8. Dong R, Huang R, Wang J, Liu H, Xu Z (2021) Effects of microglial activation and polarization on brain injury after stroke. Front Neurol 12:620948 [PMID: 34276530]
  9. Eash KJ, Greenbaum AM, Gopalan PK, Link DC (2010) CXCR2 and CXCR4 antagonistically regulate neutrophil trafficking from murine bone marrow. J Clin Invest 120:2423���2431 [PMID: 20516641]
  10. Elkind MSV, Veltkamp R, Montaner J, Johnston SC, Singhal AB, Becker K, Lansberg MG, Tang W, Kasliwal R, Elkins J (2020) Natalizumab in acute ischemic stroke (ACTION II): a randomized, placebo-controlled trial. Neurology 95:e1091���e1104 [PMID: 32591475]
  11. Endres M, Moro MA, Nolte CH, Dames C, Buckwalter MS, Meisel A (2022) Immune pathways in etiology, acute phase, and chronic sequelae of ischemic stroke. Circ Res 130:1167���1186 [PMID: 35420915]
  12. Fielding CA, McLoughlin RM, McLeod L, Colmont CS, Najdovska M, Grail D, Ernst M, Jones SA, Topley N, Jenkins BJ (2008) IL-6 regulates neutrophil trafficking during acute inflammation via STAT3. J Immunol 181:2189���2195 [PMID: 18641358]
  13. Futosi K, Fodor S, M��csai A (2013) Neutrophil cell surface receptors and their intracellular signal transduction pathways. Int Immunopharmacol 17:638���650 [PMID: 23994464]
  14. He WT, Wan H, Hu L, Chen P, Wang X, Huang Z, Yang ZH, Zhong CQ, Han J (2015) Gasdermin D is an executor of pyroptosis and required for interleukin-1�� secretion. Cell Res 25:1285���1298 [PMID: 26611636]
  15. Heilig R, Dick MS, Sborgi L, Meunier E, Hiller S, Broz P (2018) The Gasdermin-D pore acts as a conduit for IL-1�� secretion in mice. Eur J Immunol 48:584���592 [PMID: 29274245]
  16. Hernandez S, Qing J, Thibodeau RH, Du X, Park S, Lee H-M, Xu M, Oh S, Navarro A, Roose-Girma M et al (2018) The kinase activity of hematopoietic progenitor kinase 1 is essential for the regulation of T cell function. Cell Rep 25:80���94 [PMID: 30282040]
  17. Hu MC, Qiu WR, Wang X, Meyer CF, Tan TH (1996) Human HPK1, a novel human hematopoietic progenitor kinase that activates the JNK/SAPK kinase cascade. Genes Dev 10:2251���2264 [PMID: 8824585]
  18. Jakob SM, Pick R, Brechtefeld D, Nussbaum C, Kiefer F, Sperandio M, Walzog B (2013) Hematopoietic progenitor kinase 1 (HPK1) is required for LFA-1-mediated neutrophil recruitment during the acute inflammatory response. Blood 121:4184���4194 [PMID: 23460610]
  19. Jiang W, Zhao Y, Liu R, Zhang B, Xie Y, Gao B, Shi K, Zou M, Jia D, Ding J et al (2024) Histidine-rich glycoprotein modulates neutrophils and thrombolysis-associated hemorrhagic transformation. EMBO Mol Med 16:2146���2169 [PMID: 39148004]
  20. Jickling GC, Liu D, Ander BP, Stamova B, Zhan X, Sharp FR (2015) Targeting neutrophils in ischemic stroke: translational insights from experimental studies. J Cereb Blood Flow Metab 35:888���901 [PMID: 25806703]
  21. Karmakar M, Minns M, Greenberg EN, Diaz-Aponte J, Pestonjamasp K, Johnson JL, Rathkey JK, Abbott DW, Wang K, Shao F et al (2020) N-GSDMD trafficking to neutrophil organelles facilitates IL-1�� release independently of plasma membrane pores and pyroptosis. Nat Commun 11:2212 [PMID: 32371889]
  22. Katan M, Luft A (2018) Global burden of stroke. Semin Neurol 38:208���211 [PMID: 29791947]
  23. Keshari RS, Verma A, Barthwal MK, Dikshit M (2013) Reactive oxygen species-induced activation of ERK and p38 MAPK mediates PMA-induced NETs release from human neutrophils. J Cell Biochem 114:532���540 [PMID: 22961925]
  24. Kiefer F, Tibbles LA, Anafi M, Janssen A, Zanke BW, Lassam N, Pawson T, Woodgett JR, Iscove NN (1996) HPK1, a hematopoietic protein kinase activating the SAPK/JNK pathway. EMBO J 15:7013���7025 [PMID: 9003777]
  25. Kim D, Haynes CL (2013) The role of p38 MAPK in neutrophil functions: single cell chemotaxis and surface marker expression. Analyst 138:6826���6833 [PMID: 24061073]
  26. Kolaczkowska E, Kubes P (2013) Neutrophil recruitment and function in health and inflammation. Nat Rev Immunol 13:159���175 [PMID: 23435331]
  27. Kulkarni HS, Lee JS, Bastarache JA, Kuebler WM, Downey GP, Albaiceta GM, Altemeier WA, Artigas A, Bates JHT, Calfee CS et al (2022) Update on the features and measurements of experimental acute lung injury in animals: an official American thoracic society workshop report. Am J Respir Cell Mol Biol 66:e1���e14 [PMID: 35103557]
  28. Laridan E, Denorme F, Desender L, Fran��ois O, Andersson T, Deckmyn H, Vanhoorelbeke K, De Meyer SF (2017) Neutrophil extracellular traps in ischemic stroke thrombi. Ann Neurol 82:223���232 [PMID: 28696508]
  29. Li H, Pan P, Su X, Liu S, Zhang L, Wu D, Li H, Dai M, Li Y, Hu C et al (2017) Neutrophil extracellular traps are pathogenic in ventilator-induced lung injury and partially dependent on TLR4. Biomed Res Int 2017:8272504 [PMID: 29387725]
  30. Li J, Luo H, Chen Y, Wu B, Han M, Jia W, Wu Y, Cheng R, Wang X, Ke J et al (2023) Comparison of the predictive value of inflammatory biomarkers for the risk of stroke-associated pneumonia in patients with acute ischemic stroke. Clin Interv Aging 18:1477���1490 [PMID: 37720840]
  31. Liew PX, Kubes P (2019) The neutrophil���s role during health and disease. Physiol Rev 99:1223���1248 [PMID: 30758246]
  32. Linney ID, Kaila N (2021) Inhibitors of immuno-oncology target HPK1���a patent review (2016 to 2020). Expert Opin Ther Pat 31:893���910 [PMID: 33956554]
  33. Liu T, Zhang L, Joo D, Sun SC (2017) NF-��B signaling in inflammation. Signal Transduct Target Ther 2:17023 [PMID: 29158945]
  34. Nam KW, Kim TJ, Lee JS, Kwon HM, Lee YS, Ko SB, Yoon BW (2018a) High neutrophil-to-lymphocyte ratio predicts stroke-associated pneumonia. Stroke 49:1886���1892 [PMID: 29967014]
  35. Nam KW, Kim TJ, Yoon BW (2018b) Response by Nam et al to letters regarding article, "High neutrophil-to-lymphocyte ratio predicts stroke-associated pneumonia". Stroke 49:e323���e324 [PMID: 30355227]
  36. Nguyen-Jackson H, Panopoulos AD, Zhang H, Li HS, Watowich SS (2010) STAT3 controls the neutrophil migratory response to CXCR2 ligands by direct activation of G-CSF-induced CXCR2 expression and via modulation of CXCR2 signal transduction. Blood 115:3354���3363 [PMID: 20185584]
  37. Papayannopoulos V (2018) Neutrophil extracellular traps in immunity and disease. Nat Rev Immunol 18:134���147 [PMID: 28990587]
  38. Robba C, Battaglini D, Samary CS, Silva PL, Ball L, Rocco PRM, Pelosi P (2020) Ischaemic stroke-induced distal organ damage: pathophysiology and new therapeutic strategies. Intensive Care Med Exp 8:23 [PMID: 33336314]
  39. Rodr��guez-Ruiz L, Lozano-Gil JM, Naranjo-S��nchez E, Mart��nez-Balsalobre E, Mart��nez-L��pez A, Lachaud C, Blanquer M, Phung TK, Garc��a-Moreno D, Cayuela ML et al (2023) ZAK��/P38 kinase signaling pathway regulates hematopoiesis by activating the NLRP1 inflammasome. EMBO Mol Med 15:e18142 [PMID: 37675820]
  40. Ross AM, Hurn P, Perrin N, Wood L, Carlini W, Potempa K (2007) Evidence of the peripheral inflammatory response in patients with transient ischemic attack. J Stroke Cerebrovasc Dis 16:203���207 [PMID: 17845917]
  41. Samary CS, Ramos AB, Maia LA, Rocha NN, Santos CL, Magalh��es RF, Clevelario AL, Pimentel-Coelho PM, Mendez-Otero R, Cruz FF et al (2018) Focal ischemic stroke leads to lung injury and reduces alveolar macrophage phagocytic capability in rats. Crit Care 22:249 [PMID: 30290827]
  42. Schymeinsky J, Gerstl R, Mannigel I, Niedung K, Frommhold D, Panthel K, Heesemann J, Sixt M, Quast T, Kolanus W et al (2009) A fundamental role of mAbp1 in neutrophils: impact on beta(2) integrin-mediated phagocytosis and adhesion in vivo. Blood 114:4209���4220 [PMID: 19638624]
  43. Senger K, Pham VC, Varfolomeev E, Hackney JA, Corzo CA, Collier J, Lau VWC, Huang Z, Hamidzhadeh K, Caplazi P et al (2017) The kinase TPL2 activates ERK and p38 signaling to promote neutrophilic inflammation. Sci Signal 10:eaah4273 [PMID: 28420753]
  44. Sharma D, Spring KJ, Bhaskar SMM (2021) Neutrophil-lymphocyte ratio in acute ischemic stroke: immunopathology, management, and prognosis. Acta Neurol Scand 144:486���499 [PMID: 34190348]
  45. Shi D, Bai Y, Long R, Xia J, Xu W, Qin D, Yang X, Ding M, Hou XY (2025) Neuronal LAMP2A-mediated reduction of adenylyl cyclases induces acute neurodegenerative responses and neuroinflammation after ischemic stroke. Cell Death Differ 32:337���352 [PMID: 39341961]
  46. Shi Y, Liu T, Nieman DC, Cui Y, Li F, Yang L, Shi H, Chen P (2020) Aerobic exercise attenuates acute lung injury through NET inhibition. Front Immunol 11:409 [PMID: 32265910]
  47. Si J, Shi X, Sun S, Zou B, Li Y, An D, Lin X, Gao Y, Long F, Pang B et al (2020) Hematopoietic progenitor kinase1 (HPK1) mediates T cell dysfunction and is a druggable target for T cell-based immunotherapies. Cancer Cell 38:551���566 [PMID: 32860752]
  48. Simats A, Liesz A (2022) Systemic inflammation after stroke: implications for post-stroke comorbidities. EMBO Mol Med 14:e16269 [PMID: 35971650]
  49. Sollberger G, Choidas A, Burn GL, Habenberger P, Di Lucrezia R, Kordes S, Menninger S, Eickhoff J, Nussbaumer P, Klebl B et al (2018) Gasdermin D plays a vital role in the generation of neutrophil extracellular traps. Sci Immunol 3:eaar6689 [PMID: 30143555]
  50. Song C, Li H, Li Y, Dai M, Zhang L, Liu S, Tan H, Deng P, Liu J, Mao Z et al (2019) NETs promote ALI/ARDS inflammation by regulating alveolar macrophage polarization. Exp Cell Res 382:111486 [PMID: 31255598]
  51. Sun JM, Fan HY, Zhu Y, Pan TT, Wu YP, Zhang DY, Hou XY (2023) Glioblastoma cellular MAP4K1 facilitates tumor growth and disrupts T effector cell infiltration. Life Sci Alliance 6:e202301966 [PMID: 37734869]
  52. Swamydas M, Lionakis MS (2013) Isolation, purification and labeling of mouse bone marrow neutrophils for functional studies and adoptive transfer experiments. J Vis Exp 10:e50586
  53. Tian C, Ji Z, Xiang W, Huang X, Wang S, Wu Y, Pan S, Zhou L, Deng Z (2018) Association of lower leukocyte count before thrombolysis with early neurological improvement in acute ischemic stroke patients. J Clin Neurosci 56:44���49 [PMID: 30126695]
  54. Wang J, Zhang J, Ye Y, Xu Q, Li Y, Feng S, Xiong X, Jian Z, Gu L (2022) Peripheral organ injury after stroke. Front Immunol 13:901209 [PMID: 35720359]
  55. Wanrooy BJ, Wen SW, Wong CHY (2021) Dynamic roles of neutrophils in post-stroke neuroinflammation. Immunol Cell Biol 99:924���935 [PMID: 33894069]
  56. Wu ZR, Zhou TQ, Ai SC (2024) Neutrophil extracellular traps correlate with severity and prognosis in patients with ischemic stroke: a systematic review and meta-analysis. Acta Neurol Belg 124:513���522 [PMID: 37950825]
  57. Xia J, Zhang T, Sun Y, Huang Z, Shi D, Qin D, Yang X, Liu H, Yao G, Wei L et al (2024) Suppression of neuronal CDK9/p53/VDAC signaling provides bioenergetic support and improves post-stroke neuropsychiatric outcomes. Cell Mol Life Sci 81:384 [PMID: 39235466]
  58. Xu Q, Ye Y, Wang Z, Zhu H, Li Y, Wang J, Gao W, Gu L (2022) NLRP3 knockout protects against lung injury induced by cerebral ischemia-reperfusion. Oxid Med Cell Longev 2022:6260102 [PMID: 35432726]
  59. Yahya T, Jilani MH, Khan SU, Mszar R, Hassan SZ, Blaha MJ, Blankstein R, Virani SS, Johansen MC, Vahidy F et al (2020) Stroke in young adults: current trends, opportunities for prevention and pathways forward. Am J Prev Cardiol 3:100085 [PMID: 34327465]
  60. Yildiz C, Palaniyar N, Otulakowski G, Khan MA, Post M, Kuebler WM, Tanswell K, Belcastro R, Masood A, Engelberts D et al (2015) Mechanical ventilation induces neutrophil extracellular trap formation. Anesthesiology 122:864���875 [PMID: 25665049]
  61. Zhang J, Ren Q, Song Y, He M, Zeng Y, Liu Z, Xu J (2017) Prognostic role of neutrophil-lymphocyte ratio in patients with acute ischemic stroke. Medicine (Baltimore) 96:e8624 [PMID: 29137097]
  62. Zhang R, Wu X, Hu W, Zhao L, Zhao S, Zhang J, Chu Z, Xu Y (2019) Neutrophil-to-lymphocyte ratio predicts hemorrhagic transformation in ischemic stroke: a meta-analysis. Brain Behav 9:e01382 [PMID: 31429525]
  63. Zhou L, Wang T, Zhang K, Zhang X, Jiang S (2022) The development of small-molecule inhibitors targeting HPK1. Eur J Med Chem 244:114819 [PMID: 36209628]
  64. Zhou X, Zhang C, Wu X, Hu X, Zhang Y, Wang X, Zheng L, Gao P, Du J, Zheng W et al (2022) Dusp6 deficiency attenuates neutrophil-mediated cardiac damage in the acute inflammatory phase of myocardial infarction. Nat Commun 13:6672 [PMID: 36335128]

Grants

  1. 82173081/MOST | National Natural Science Foundation of China (NSFC)
  2. 81673418/MOST | National Natural Science Foundation of China (NSFC)
  3. 18KJA310007/Major Basic Research Project of the Natural Science Foundation of the Jiangsu Higher Education Institutions (Key University of Science Research Project of Jiangsu Province)
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