Mesenchymal stem cell-derived exosomes alleviate obliterative bronchiolitis after lung transplantation by regulating macrophage pyroptosis_Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Authors：

LIU Jie , ZHAO Tian ,  CHEN Guohan

Department of Thoracic Surgery, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, 200120, P. R. China;

Corresponding author：

CHEN Guohan, Email: chenguohan0160@126.com

Keywords：

Mesenchymal stem cell-derived exosome; macrophage; COL5A1; pyroptosis; lung transplantation

DOI：

10.7507/1007-4848.202504089

Video：

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Objective To investigate the regulatory role of MSC-derived exosomes in obliterative bronchiolitis after lung transplantation. Methods The murine lung transplantation model was established with male C57BL/6 mice, and the mice were divided into a sham group (sham, n=6), a surgery group (OB, n=6), and a treatment group (OB+MSC-exo, n=6). The in vitro model was created by stimulating RAW264.7 with lipopolysaccharide+nigericin (LPS+Nigericin), and comprised a PBS group, a LPS+Nigericin group, and a LPS+Nigericin+MSC-exo group. Immunofluorescence and hematoxylin-eosin (HE) staining were used to analyze gasdermin D (GSDMD) expression, as well as lumen stenosis in lung grafts. Bioinformatics methods were employed to predict and screen target gene collagen type V alpha 1 (COL5A1). Q-PCR was used to measure mRNA expression levels of interleukin (IL)-1β, IL-18, IL-6, tumor necrosis factor-α (TNF-α), and COL5A1 in lung grafts and macrophages. Western blot was performed to detect Cleaved-Caspase 1 protein expression in lung grafts and GSDMD protein expression in macrophages. Results Immunofluorescence and HE staining revealed that in vivo infusion of MSC-exo reduced GSDMD expression in grafts, ameliorated tracheal epithelial cilia loss and lumen stenosis, and decreased Cleaved-Caspase 1 protein as well as IL-1β and IL-18 mRNA expression. MSC-exo treatment or COL5A1 knockdown reduced IL-1β, IL-18, IL-6, and TNF-α mRNA expression in macrophages, with comparable efficacy. MSC-exo infusion also decreased the number of COL5A1⁺ cells and mRNA expression levels in lung grafts. Conclusion MSC-derived exosomes alleviate obliterative bronchiolitis after lung transplantation by inhibiting COL5A1.

1.	Kulkarni HS, Cherikh WS, Chambers DC, et al. Bronchiolitis obliterans syndrome-free survival after lung transplantation: an International Society for Heart and Lung Transplantation Thoracic Transplant Registry analysis. J Heart Lung Transplant, 2019, 38(1): 5-16.
2.	Kohmoto J, Nakao A, Kaizu T, et al. Low-dose carbon monoxide inhalation prevents ischemia/reperfusion injury of transplanted rat lung grafts. Surgery, 2006, 140(2): 179-185.
3.	Ladak SS, Ward C, Ali S. The potential role of microRNAs in lung allograft rejection. J Heart Lung Transplant, 2016, 35(5): 550-559.
4.	Nayak DK, Zhou F, Xu M, et al. Zbtb7a induction in alveolar macrophages is implicated in anti-HLA-mediated lung allograft rejection. Sci Transl Med, 2017, 9(398): eaal1243.
5.	Yamada Y, Jang JH, De Meester I, et al. CD26 costimulatory blockade improves lung allograft rejection and is associated with enhanced interleukin-10 expression. J Heart Lung Transplant, 2016, 35(4): 508-517.
6.	Mosser DM, Edwards JP. Exploring the full spectrum of macrophage activation. Nat Rev Immunol, 2008, 8(12): 958-969.
7.	Liu J, Zhou X, Zhan Z, et al. IL-25 regulates the polarization of macrophages and attenuates obliterative bronchiolitis in murine trachea transplantation models. Int Immunopharmacol, 2015, 25(2): 383-392.
8.	Meng Q, Liu J, Lin F, et al. IL-17 contributes to the pathogenesis of obliterative bronchiolitis via regulation of M1 macrophages polarization in murine heterotopic trachea transplantation models. Int Immunopharmacol, 2017, 52: 51-60.
9.	Strowig T, Henao-Mejia J, Elinav E, et al. Inflammasomes in health and disease. Nature, 2012, 481(7381): 278-286.
10.	He X, Qian Y, Li Z, et al. TLR4-upregulated IL-1β and IL-1RI promote alveolar macrophage pyroptosis and lung inflammation through an autocrine mechanism. Sci Rep, 2016, 6: 31663.
11.	Gao J, Peng S, Shan X, et al. Inhibition of AIM2 inflammasome-mediated pyroptosis by Andrographolide contributes to amelioration of radiation-induced lung inflammation and fibrosis. Cell Death Dis, 2019, 10(12): 957.
12.	Guo Z, Zhou X, Li J, et al. Mesenchymal stem cells reprogram host macrophages to attenuate obliterative bronchiolitis in murine orthotopic tracheal transplantation. Int Immunopharmacol, 2013, 15(4): 726-734.
13.	Chan E, Liu XX, Guo DJ, et al. Extract of Scutellaria baicalensis Georgi root exerts protection against myocardial ischemia-reperfusion injury in rats. Am J Chin Med, 2011, 39(4): 693-704.
14.	Willis GR, Fernandez-Gonzalez A, Anastas J, et al. Mesenchymal stromal cell exosomes ameliorate experimental bronchopulmonary dysplasia and restore lung function through macrophage immunomodulation. Am J Respir Crit Care Med, 2018, 197(1): 104-116.
15.	Lee C, Mitsialis SA, Aslam M, et al. Exosomes mediate the cytoprotective action of mesenchymal stromal cells on hypoxia-induced pulmonary hypertension. Circulation, 2012, 126(22): 2601-2611.
16.	Kourembanas S. Exosomes: vehicles of intercellular signaling, biomarkers, and vectors of cell therapy. Annu Rev Physiol, 2015, 77: 13-27.
17.	Burrello J, Monticone S, Gai C, et al. Stem cell-derived extracellular vesicles and immune-modulation. Front Cell Dev Biol, 2016, 4: 83.
18.	Akbari A, Rezaie J. Potential therapeutic application of mesenchymal stem cell-derived exosomes in SARS-CoV-2 pneumonia. Stem Cell Res Ther, 2020, 11(1): 356.
19.	Wang W, Liu Z, Su J, et al. Macrophage micro-RNA-155 promotes lipopolysaccharide-induced acute lung injury in mice and rats. Am J Physiol Lung Cell Mol Physiol, 2016, 311(2): L494-L506.
20.	Ferguson SW, Wang J, Lee CJ, et al. The microRNA regulatory landscape of MSC-derived exosomes: a systems view. Sci Rep, 2018, 8(1): 1419.
21.	Parker MW, Rossi D, Peterson M, et al. Fibrotic extracellular matrix activates a profibrotic positive feedback loop. J Clin Invest, 2014, 124(4): 1622-1635.
22.	Montgomery RL, Yu G, Latimer PA, et al. MicroRNA mimicry blocks pulmonary fibrosis. EMBO Mol Med, 2014, 6(10): 1347-1356.
23.	Kim KK, Sisson TH, Horowitz JC. Fibroblast growth factors and pulmonary fibrosis: it's more complex than it sounds. J Pathol, 2017, 241(1): 6-9.
24.	Vittal R, Fan L, Greenspan DS, et al. IL-17 induces type V collagen overexpression and EMT via TGF-β-dependent pathways in obliterative bronchiolitis. Am J Physiol Lung Cell Mol Physiol, 2013, 304(6): L401-L414.
25.	Xu Z, Chen Y, Ma L, et al. Role of exosomal non-coding RNAs from tumor cells and tumor-associated macrophages in the tumor microenvironment. Mol Ther, 2022, 30(10): 3133-3154.
26.	Hua T, Yang M, Song H, et al. Huc-MSCs-derived exosomes attenuate inflammatory pain by regulating microglia pyroptosis and autophagy via the miR-146a-5p/TRAF6 axis. J Nanobiotechnology, 2022, 20(1): 324.
27.	Brugiere O, Verleden SE. Putting the spotlight on macrophage-derived cathepsin in the pathophysiology of obliterative bronchiolitis. Eur Respir J, 2021, 57(5): 2004607.
28.	Cheng P, Li S, Chen H. Macrophages in lung injury, repair, and fibrosis. Cells, 2021, 10(2): 436.
29.	Gao F, Xiong D, Sun Z, et al. ARC@DPBNPs suppress LPS-induced acute lung injury via inhibiting macrophage pyroptosis and M1 polarization by ERK pathway in mice. Int Immunopharmacol, 2024, 131: 111794.
30.	Li B, Zou Z, Meng F, et al. Dust mite-derived Der f 3 activates a pro-inflammatory program in airway epithelial cells via PAR-1 and PAR-2. Mol Immunol, 2019, 109: 1-11.
31.	Liu Q, Lv C, Jiang Y, et al. From hair to liver: emerging application of hair follicle mesenchymal stem cell transplantation reverses liver cirrhosis by blocking the TGF-β/Smad signaling pathway to inhibit pathological HSC activation. PeerJ, 2022, 10: e12872.

1. Kulkarni HS, Cherikh WS, Chambers DC, et al. Bronchiolitis obliterans syndrome-free survival after lung transplantation: an International Society for Heart and Lung Transplantation Thoracic Transplant Registry analysis. J Heart Lung Transplant, 2019, 38(1): 5-16.
2. Kohmoto J, Nakao A, Kaizu T, et al. Low-dose carbon monoxide inhalation prevents ischemia/reperfusion injury of transplanted rat lung grafts. Surgery, 2006, 140(2): 179-185.
3. Ladak SS, Ward C, Ali S. The potential role of microRNAs in lung allograft rejection. J Heart Lung Transplant, 2016, 35(5): 550-559.
4. Nayak DK, Zhou F, Xu M, et al. Zbtb7a induction in alveolar macrophages is implicated in anti-HLA-mediated lung allograft rejection. Sci Transl Med, 2017, 9(398): eaal1243.
5. Yamada Y, Jang JH, De Meester I, et al. CD26 costimulatory blockade improves lung allograft rejection and is associated with enhanced interleukin-10 expression. J Heart Lung Transplant, 2016, 35(4): 508-517.
6. Mosser DM, Edwards JP. Exploring the full spectrum of macrophage activation. Nat Rev Immunol, 2008, 8(12): 958-969.
7. Liu J, Zhou X, Zhan Z, et al. IL-25 regulates the polarization of macrophages and attenuates obliterative bronchiolitis in murine trachea transplantation models. Int Immunopharmacol, 2015, 25(2): 383-392.
8. Meng Q, Liu J, Lin F, et al. IL-17 contributes to the pathogenesis of obliterative bronchiolitis via regulation of M1 macrophages polarization in murine heterotopic trachea transplantation models. Int Immunopharmacol, 2017, 52: 51-60.
9. Strowig T, Henao-Mejia J, Elinav E, et al. Inflammasomes in health and disease. Nature, 2012, 481(7381): 278-286.
10. He X, Qian Y, Li Z, et al. TLR4-upregulated IL-1β and IL-1RI promote alveolar macrophage pyroptosis and lung inflammation through an autocrine mechanism. Sci Rep, 2016, 6: 31663.
11. Gao J, Peng S, Shan X, et al. Inhibition of AIM2 inflammasome-mediated pyroptosis by Andrographolide contributes to amelioration of radiation-induced lung inflammation and fibrosis. Cell Death Dis, 2019, 10(12): 957.
12. Guo Z, Zhou X, Li J, et al. Mesenchymal stem cells reprogram host macrophages to attenuate obliterative bronchiolitis in murine orthotopic tracheal transplantation. Int Immunopharmacol, 2013, 15(4): 726-734.
13. Chan E, Liu XX, Guo DJ, et al. Extract of Scutellaria baicalensis Georgi root exerts protection against myocardial ischemia-reperfusion injury in rats. Am J Chin Med, 2011, 39(4): 693-704.
14. Willis GR, Fernandez-Gonzalez A, Anastas J, et al. Mesenchymal stromal cell exosomes ameliorate experimental bronchopulmonary dysplasia and restore lung function through macrophage immunomodulation. Am J Respir Crit Care Med, 2018, 197(1): 104-116.
15. Lee C, Mitsialis SA, Aslam M, et al. Exosomes mediate the cytoprotective action of mesenchymal stromal cells on hypoxia-induced pulmonary hypertension. Circulation, 2012, 126(22): 2601-2611.
16. Kourembanas S. Exosomes: vehicles of intercellular signaling, biomarkers, and vectors of cell therapy. Annu Rev Physiol, 2015, 77: 13-27.
17. Burrello J, Monticone S, Gai C, et al. Stem cell-derived extracellular vesicles and immune-modulation. Front Cell Dev Biol, 2016, 4: 83.
18. Akbari A, Rezaie J. Potential therapeutic application of mesenchymal stem cell-derived exosomes in SARS-CoV-2 pneumonia. Stem Cell Res Ther, 2020, 11(1): 356.
19. Wang W, Liu Z, Su J, et al. Macrophage micro-RNA-155 promotes lipopolysaccharide-induced acute lung injury in mice and rats. Am J Physiol Lung Cell Mol Physiol, 2016, 311(2): L494-L506.
20. Ferguson SW, Wang J, Lee CJ, et al. The microRNA regulatory landscape of MSC-derived exosomes: a systems view. Sci Rep, 2018, 8(1): 1419.
21. Parker MW, Rossi D, Peterson M, et al. Fibrotic extracellular matrix activates a profibrotic positive feedback loop. J Clin Invest, 2014, 124(4): 1622-1635.
22. Montgomery RL, Yu G, Latimer PA, et al. MicroRNA mimicry blocks pulmonary fibrosis. EMBO Mol Med, 2014, 6(10): 1347-1356.
23. Kim KK, Sisson TH, Horowitz JC. Fibroblast growth factors and pulmonary fibrosis: it's more complex than it sounds. J Pathol, 2017, 241(1): 6-9.
24. Vittal R, Fan L, Greenspan DS, et al. IL-17 induces type V collagen overexpression and EMT via TGF-β-dependent pathways in obliterative bronchiolitis. Am J Physiol Lung Cell Mol Physiol, 2013, 304(6): L401-L414.
25. Xu Z, Chen Y, Ma L, et al. Role of exosomal non-coding RNAs from tumor cells and tumor-associated macrophages in the tumor microenvironment. Mol Ther, 2022, 30(10): 3133-3154.
26. Hua T, Yang M, Song H, et al. Huc-MSCs-derived exosomes attenuate inflammatory pain by regulating microglia pyroptosis and autophagy via the miR-146a-5p/TRAF6 axis. J Nanobiotechnology, 2022, 20(1): 324.
27. Brugiere O, Verleden SE. Putting the spotlight on macrophage-derived cathepsin in the pathophysiology of obliterative bronchiolitis. Eur Respir J, 2021, 57(5): 2004607.
28. Cheng P, Li S, Chen H. Macrophages in lung injury, repair, and fibrosis. Cells, 2021, 10(2): 436.
29. Gao F, Xiong D, Sun Z, et al. ARC@DPBNPs suppress LPS-induced acute lung injury via inhibiting macrophage pyroptosis and M1 polarization by ERK pathway in mice. Int Immunopharmacol, 2024, 131: 111794.
30. Li B, Zou Z, Meng F, et al. Dust mite-derived Der f 3 activates a pro-inflammatory program in airway epithelial cells via PAR-1 and PAR-2. Mol Immunol, 2019, 109: 1-11.
31. Liu Q, Lv C, Jiang Y, et al. From hair to liver: emerging application of hair follicle mesenchymal stem cell transplantation reverses liver cirrhosis by blocking the TGF-β/Smad signaling pathway to inhibit pathological HSC activation. PeerJ, 2022, 10: e12872.

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