Nestin prevents mesenchymal stromal cells from apoptosis in LPS-induced lung injury via inhibition of unfolded protein response sensor IRE1α

Abstract

The clinical applications of MSC therapy have been intensely investigated in acute respiratory distress syndrome. However, clinical studies have fallen short of expectations despite encouraging preclinical results. One of the key problems is that transplanted stem cells can hardly survive in the harsh inflammatory environment. Prolonging the survival of transplanted MSCs might be a promising strategy to enhance the therapeutic efficacy of MSC therapy. Here, we identified Nestin, a class VI intermediate filament, as a positive regulator of MSC survival in the inflammatory microenvironment. We showed that Nestin knockout led to a significant increase of MSC apoptosis, which hampered the therapeutic effects in an LPS-induced lung injury model. Mechanistically, Nestin knockout induced a significant elevation of endoplasmic reticulum (ER) stress level. Further investigations showed that Nestin could bind to IRE1α and inhibit ER stress-induced apoptosis under stress. Furthermore, pretreatment with IRE1α inhibitor 4μ8C improved MSC survival and improved therapeutic effect. Our data suggests that Nestin enhances stem cell survival after transplantation by inhibiting ER stress-induced apoptosis, improving protection, and repair of the lung inflammatory injury.

Keywords

acute respiratory distress syndrome / LPS-induced lung injury / ER stress / IRE1α / Nestin

References

[1] Bellani G, Laffey JG, Pham T, et al.; LUNG SAFE Investigators. Epidemiology, patterns of care, and mortality for patients with acute respiratory distress syndrome in intensive care units in 50 countries. JAMA 2016;315:788–800.

[2] Beitler JR, Thompson BT, Baron RM, et al. Advancing precision medicine for acute respiratory distress syndrome. Lancet Respir Med 2022;10:107–20.

[3] Markov A, Thangavelu L, Aravindhan S, et al. Mesenchymal stem/stromal cells as a valuable source for the treatment of immune-mediated disorders. Stem Cell Res Ther 2021;12:192.

[4] Walter J, Ware LB, Matthay MA. Mesenchymal stem cells: mechanisms of potential therapeutic benefit in ARDS and sepsis. Lancet Respir Med 2014;2:1016–26.

[5] Coulson-Thomas VJ, Coulson-Thomas YM, Gesteira TF, et al. Extrinsic and intrinsic mechanisms by which mesenchymal stem cells suppress the immune system. Ocul Surf 2016;14:121–34.

[6] Fengyun W, LiXin Z, Xinhua Q, et al. Mesenchymal stromal cells attenuate infection-induced acute respiratory distress syndrome in animal experiments: a meta-analysis. Cell Transplant 2020;29:963689720969186.

[7] Dutra Silva J, Su Y, Calfee CS, et al. Mesenchymal stromal cell extracellular vesicles rescue mitochondrial dysfunction and improve barrier integrity in clinically relevant models of ARDS. Eur Respir J 2021;58:2002978.

[8] Laffey JG, Matthay MA. Fifty years of research in ARDS. Cell-based therapy for acute respiratory distress syndrome. Biology and potential therapeutic value. Am J Respir Crit Care Med 2017;196:266–73.

[9] Xu Z, Huang Y, Zhou J, et al. Current status of cell-based therapies for COVID-19: evidence from mesenchymal stromal cells in sepsis and ARDS. Front Immunol 2021;12:738697.

[10] Matthay MA, Calfee CS, Zhuo H, et al. Treatment with allogeneic mesenchymal stromal cells for moderate to severe acute respiratory distress syndrome (START study): a randomised phase 2a safety trial. Lancet Respir Med 2019;7:154–62.

[11] Hetz C, Zhang K, Kaufman RJ. Mechanisms, regulation and functions of the unfolded protein response. Nat Rev Mol Cell Biol 2020;21:421–38.

[12] Higuchi-Sanabria R, Frankino PA, Paul JW 3rd, et al. A futile battle? Protein quality control and the stress of aging. Dev Cell 2018;44:139–63.

[13] Ariyasu D, Yoshida H, Hasegawa Y. Endoplasmic reticulum (ER) stress and endocrine disorders. Int J Mol Sci 2017;18:382.

[14] Kropski JA, Blackwell TS. Endoplasmic reticulum stress in the pathogenesis of fibrotic disease. J Clin Invest 2018;128:64–73.

[15] Hetz C. The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol 2012;13:89–102.

[16] Morita S, Villalta SA, Feldman HC, et al. Targeting ABL-IRE1α signaling spares ER-stressed pancreatic β cells to reverse autoimmune diabetes. Cell Metab 2017;25:883–897.e8.

[17] Lendahl U, Zimmerman LB, McKay RD. CNS stem cells express a new class of intermediate filament protein. Cell 1990;60:585–95.

[18] Méndez-Ferrer S, Michurina TV, Ferraro F, et al. Mesenchymal and haematopoietic stem cells form a unique bone marrow niche. Nature 2010;466:829–34.

[19] Jiang MH, Cai B, Tuo Y, et al. Characterization of Nestin-positive stem Leydig cells as a potential source for the treatment of testicular Leydig cell dysfunction. Cell Res 2014;24:1466–85.

[20] Bornstein SR, Berger I, Steenblock C. Are Nestin-positive cells responsive to stress? Stress 2020;23:662–6.

[21] Steenblock C, Rubin de Celis MF, Delgadillo Silva LF, et al. Isolation and characterization of adrenocortical progenitors involved in the adaptation to stress. Proc Natl Acad Sci USA 2018;115:12997–3002.

[22] Park D, Xiang AP, Mao FF, et al. Nestin is required for the proper self-renewal of neural stem cells. Stem Cells 2010;28:2162–71.

[23] Wang J, Lu Q, Cai J, et al. Nestin regulates cellular redox homeostasis in lung cancer through the Keap1-Nrf2 feedback loop. Nat Commun 2019;10:5043.

[24] Sahlgren CM, Pallari HM, He T, et al. A nestin scaffold links Cdk5/p35 signaling to oxidant-induced cell death. EMBO J 2006;25:4808–19.

[25] Zhang Y, Wang J, Huang W, et al. Nuclear Nestin deficiency drives tumor senescence via lamin A/C-dependent nuclear deformation. Nat Commun 2018;9:3613.

[26] Chen H, Cai J, Wang J, et al. Targeting Nestin(+) hepatic stellate cells ameliorates liver fibrosis by facilitating TbetaRI degradation. J Hepatol 2021;74:1176–87.

[27] Wang J, Cai J, Huang Y, et al. Nestin regulates proliferation and invasion of gastrointestinal stromal tumor cells by altering mitochondrial dynamics. Oncogene 2016;35:3139–50.

[28] Wang H, Li D, Zhai Z, et al. Characterization and therapeutic application of mesenchymal stem cells with neuromesodermal origin from human pluripotent stem cells. Theranostics 2019;9:1683–97.

[29] Han J, Li Y, Li Y. Strategies to enhance mesenchymal stem cell-based therapies for acute respiratory distress syndrome. Stem Cells Int 2019;2019:5432134.

[30] Borrello MT, Martin MB, Pin CL. The unfolded protein response: an emerging therapeutic target for pancreatitis and pancreatic ductal adenocarcinoma. Pancreatology 2022;22:148–59.

[31] Li C, Chai Y, Wang L, et al. Programmed cell senescence in skeleton during late puberty. Nat Commun 2017;8:1312.

[32] Sejersen T, Lendahl U. Transient expression of the intermediate filament nestin during skeletal muscle development. J Cell Sci 1993;106:1291–300.

[33] Zhang X, Wei X, Deng Y, et al. Mesenchymal stromal cells alleviate acute respiratory distress syndrome through the cholinergic anti-inflammatory pathway. Signal Transduct Target Ther 2022;7:307.

[34] Jackson MV, Morrison TJ, Doherty DF, et al. Mitochondrial transfer via tunneling nanotubes is an important mechanism by which mesenchymal stem cells enhance macrophage phagocytosis in the in vitro and in vivo models of ARDS. Stem Cells 2016;34:2210–23.

[35] Zheng D, Bhuvan T, Payne NL, et al. Secondary lymphoid organs in mesenchymal stromal cell therapy: more than just a filter. Front Immunol 2022;13:892443.

[36] Pang SHM, D’Rozario J, Mendonca S, et al. Mesenchymal stromal cell apoptosis is required for their therapeutic function. Nat Commun 2021;12:6495.

[37] Ko JH, Lee HJ, Jeong HJ, et al. Mesenchymal stem/stromal cells precondition lung monocytes/macrophages to produce tolerance against allo- and autoimmunity in the eye. Proc Natl Acad Sci USA 2016;113:158–63.

[38] Masterson C, Gonzalez H, Laffey JG. Understanding the impact of the lung microenvironment to enhance the therapeutic potential of mesenchymal stromal cells for acute respiratory distress syndrome. Eur Respir J 2021;58:2100986.

[39] Masterson CH, Tabuchi A, Hogan G, et al. Intra-vital imaging of mesenchymal stromal cell kinetics in the pulmonary vasculature during infection. Sci Rep 2021;11:5265.

[40] Rolandsson Enes S, Hampton TH, Barua J, et al. Healthy versus inflamed lung environments differentially affect mesenchymal stromal cells. Eur Respir J 2021;58:2004149.

[41] Chistiakov DA. Endogenous and exogenous stem cells: a role in lung repair and use in airway tissue engineering and transplantation. J Biomed Sci 2010;17:92.

[42] Li M, Jiang Y, Hou Q, et al. Potential pre-activation strategies for improving therapeutic efficacy of mesenchymal stem cells: current status and future prospects. Stem Cell Res Ther 2022;13:146.

[43] Yang JX, Zhang N, Wang HW, et al. CXCR4 receptor overexpression in mesenchymal stem cells facilitates treatment of acute lung injury in rats. J Biol Chem 2015;290:1994–2006.

[44] Chen X, Wu S, Tang L, et al. Mesenchymal stem cells overexpressing heme oxygenase-1 ameliorate lipopolysaccharide-induced acute lung injury in rats. J Cell Physiol 2019;234:7301–19.

[45] Tsai CC, Chen YJ, Yew TL, et al. Hypoxia inhibits senescence and maintains mesenchymal stem cell properties through down-regulation of E2A-p21 by HIF-TWIST. Blood 2011;117:459–69.

[46] Antebi B, Rodriguez LA 2nd, Walker KP 3rd, et al. Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells. Stem Cell Res Ther 2018;9:265.

[47] Lan YW, Choo KB, Chen CM, et al. Hypoxia-preconditioned mesenchymal stem cells attenuate bleomycin-induced pulmonary fibrosis. Stem Cell Res Ther 2015;6:97.

[48] Ke Q, Li L, Cai B, et al. Connexin 43 is involved in the generation of human-induced pluripotent stem cells. Hum Mol Genet 2013;22:2221–33.

RIGHTS & PERMISSIONS

2022 The Author(s) 2022. Published by Oxford University Press on behalf of Higher Education Press.