Human umbilical cord mesenchymal stem cell-derived nanovesicles ameliorate acute lung injury by hsa-let-7g-5p inhibition of NF-κB/NLRP3 pathway

Abstract

Aim: Acute lung injury (ALI), marked by vigorous inflammatory reactions and elevated incidence, is a grave clinical issue for which efficacious pharmaceutical interventions are still lacking. This research delves into the anti-inflammatory actions and the associated pathways of nanovesicles originating from human umbilical cord mesenchymal stem cells (UCMSC-NVs) within a model of ALI induced by lipopolysaccharide (LPS).

Methods: UCMSC-NVs were prepared via serial extrusion and characterized using transmission electron microscopy and dynamic light scattering. Their effects on pulmonary inflammation and injury were evaluated in an LPS-induced ALI mouse model. Anti-inflammatory effects were analyzed using Western blot, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence, focusing on the role of hsa-let-7g-5p in regulating the nuclear factor κB (NF-κB)/NOD-like receptor protein 3 (NLRP3) pathway.

Results: The UCMSC-NVs acquired through serial extrusion displayed bilayer vesicle structures, with a mean diameter of around 100 nm. Moreover, these vesicles exhibited elevated expression levels of CD9, CD63, and CD81. Administration of UCMSC-NVs significantly alleviated lung damage, accompanied by a reduction in alveolar leakage and neutrophil infiltration. Furthermore, it significantly downregulated the expression of pro-inflammatory cytokines, such as interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α). UCMSC-NVs reduced macrophage infiltration in the lungs of ALI mice by inhibiting the NF-κB/NLRP3 signaling, leading to a shift in macrophage polarization, with decreased M1 and increased M2 polarization. Our findings demonstrated a novel therapeutic mechanism wherein hsa-let-7g-5p encapsulated within UCMSC-NVs alleviates inflammation by inhibiting NF-κB/NLRP3 expression, thereby mitigating ALI.

Conclusion: These results provide a foundation for the development of novel cell-free therapies with clinical potential for treating inflammatory lung diseases such as ALI and acute respiratory distress syndrome (ARDS).

Keywords

Umbilical cord mesenchymal stem cell-derived nanovesicles / acute lung injury / inflammation / microRNA / NF-κB/NLRP3 inflammasome pathway

References

[1] Rajasekaran S,Rajaguru P,Thimmulappa RK.MicroRNA regulation of acute lung injury and acute respiratory distress syndrome.J Cell Physiol2016;231:2097-106

[2] Ragaller M.Acute lung injury and acute respiratory distress syndrome.J Emerg Trauma Shock2010;3:43-51 PMCID:PMC2823143

[3] Bhatia M.Role of inflammatory mediators in the pathophysiology of acute respiratory distress syndrome.J Pathol2004;202:145-56

[4] Kaku S,Htet NN.Acute respiratory distress syndrome: etiology, pathogenesis, and summary on management.J Intensive Care Med2020;35:723-37

[5] Huppert LA,Ware LB.Pathogenesis of acute respiratory distress syndrome.Semin Respir Crit Care Med2019;40:31-9 PMCID:PMC7060969

[6] Console L,Indiveri C.Exosomes in inflammation and role as biomarkers.Clin Chim Acta2019;488:165-71

[7] Kolhe R,Liu S.Gender-specific differential expression of exosomal miRNA in synovial fluid of patients with osteoarthritis.Sci Rep2017;7:2029 PMCID:PMC5435729

[8] Abdelgawad M,Farghali AA,Lotfy A.Mesenchymal stem cell-based therapy and exosomes in COVID-19: current trends and prospects.Stem Cell Res Ther2021;12:469 PMCID:PMC8379570

[9] Askenase PW.COVID-19 therapy with mesenchymal stromal cells (MSC) and convalescent plasma must consider exosome involvement: do the exosomes in convalescent plasma antagonize the weak immune antibodies?.J Extracell Vesicles2020;10:e12004 PMCID:PMC7710130

[10] Ma YY,Chen JY.Umbilical cord mesenchymal‐stem‐cell‐derived nanovesicles as a novel strategy to promote wound healing in diabetes.Nano Select2023;4:170-80

[11] Park KS,Shelke GV.Mesenchymal stromal cell-derived nanovesicles ameliorate bacterial outer membrane vesicle-induced sepsis via IL-10.Stem Cell Res Ther2019;10:231 PMCID:PMC6676541

[12] Zhang J,Li L.Exosome and exosomal microRNA: trafficking, sorting, and function.Genomics Proteomics Bioinformatics2015;13:17-24 PMCID:PMC4411500

[13] Deng L,Stevens H.MicroRNA-143 activation regulates smooth muscle and endothelial cell crosstalk in pulmonary arterial hypertension.Circ Res2015;117:870-83 PMCID:PMC4620852

[14] Wang M,Xie L.Mesenchymal stem cell-based immunomodulation: properties and clinical application.Stem Cells Int2018;2018:3057624 PMCID:PMC6022321

[15] Wei X,Lv H.Correction: MicroRNA-377-3p released by mesenchymal stem cell exosomes ameliorates lipopolysaccharide-induced acute lung injury by targeting RPTOR to induce autophagy.Cell Death Dis2020;11:746 PMCID:PMC7487066

[16] Kadota T,Araya J.Human bronchial epithelial cell-derived extracellular vesicle therapy for pulmonary fibrosis via inhibition of TGF-β-WNT crosstalk.J Extracell Vesicles2021;10:e12124 PMCID:PMC8329991

[17] Alharbi KS,Fuloria S.Nuclear factor-kappa B and its role in inflammatory lung disease.Chem Biol Interact2021;345:109568

[18] Li W,Chen Y.Classic signaling pathways in alveolar injury and repair involved in sepsis-induced ALI/ARDS: new research progress and prospect.Dis Markers2022;2022:6362344 PMCID:PMC9206211

[19] Wang YM,Chen WW.Paclitaxel alleviated sepsis-induced acute lung injury by activating MUC1 and suppressing TLR-4/NF-κB pathway.Drug Des Devel Ther2019;13:3391-404 PMCID:PMC6766586

[20] Lian J,Du J.Extracellular vesicle-transmitted miR-671-5p alleviates lung inflammation and injury by regulating the AAK1/NF-κB axis.Mol Ther2023;31:1365-82 PMCID:PMC10188640

[21] Cui Y,Guo Y.Bioinspired nanovesicles convert the skeletal endothelium-associated secretory phenotype to treat osteoporosis.ACS Nano2022;16:11076-91

[22] Zhang X,Deng Y.Mesenchymal stromal cells alleviate acute respiratory distress syndrome through the cholinergic anti-inflammatory pathway.Signal Transduct Target Ther2022;7:307 PMCID:PMC9441842

[23] Zhang S,Bonvillain RW.Interleukin 6 mediates the therapeutic effects of adipose-derived stromal/stem cells in lipopolysaccharide-induced acute lung injury.Stem Cells2014;32:1616-28 PMCID:PMC4365913

[24] Philipp D,Wahlers T,Paunel-Görgülü A.Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization.Stem Cell Res Ther2018;9:286 PMCID:PMC6202843

[25] Yang S,Feng C.Umbilical cord-derived mesenchymal stem cells: strategies, challenges, and potential for cutaneous regeneration.Front Med2012;6:41-7

[26] Liang B,Ding JN,Xu JG.Dimethyloxaloylglycine-stimulated human bone marrow mesenchymal stem cell-derived exosomes enhance bone regeneration through angiogenesis by targeting the AKT/mTOR pathway.Stem Cell Res Ther2019;10:335 PMCID:PMC6869275

[27] Shao H,Castro CM,Weissleder R.New technologies for analysis of extracellular vesicles.Chem Rev2018;118:1917-50 PMCID:PMC6029891

[28] Zhao S,Zheng B.Highly-metastatic colorectal cancer cell released miR-181a-5p-rich extracellular vesicles promote liver metastasis by activating hepatic stellate cells and remodelling the tumour microenvironment.J Extracell Vesicles2022;11:e12186 PMCID:PMC8765330

[29] Cao M,Yan W.Cancer-cell-secreted extracellular vesicles suppress insulin secretion through miR-122 to impair systemic glucose homeostasis and contribute to tumour growth.Nat Cell Biol2022;24:954-67 PMCID:PMC9233030

[30] He Y,Wang X.Neutrophil-to-hepatocyte communication via LDLR-dependent miR-223-enriched extracellular vesicle transfer ameliorates nonalcoholic steatohepatitis.J Clin Invest2021;131:e141513 PMCID:PMC7843220

[31] Ge L,Li W.Extracellular vesicles derived from hypoxia-preconditioned olfactory mucosa mesenchymal stem cells enhance angiogenesis via miR-612.J Nanobiotechnology2021;19:380 PMCID:PMC8607643

[32] Ding J,Cai X.Extracellular vesicles derived from M1 macrophages deliver miR-146a-5p and miR-146b-5p to suppress trophoblast migration and invasion by targeting TRAF6 in recurrent spontaneous abortion.Theranostics2021;11:5813-30 PMCID:PMC8058722

[33] Ren W,Yang C.Extracellular vesicles secreted by hypoxia pre-challenged mesenchymal stem cells promote non-small cell lung cancer cell growth and mobility as well as macrophage M2 polarization via miR-21-5p delivery.J Exp Clin Cancer Res2019;38:62 PMCID:PMC6367822

[34] Hu Q,Yang Y.Extracellular vesicles in the pathogenesis and treatment of acute lung injury.Mil Med Res2022;9:61 PMCID:PMC9623953

[35] Lv LL,Wu M.Exosomal miRNA-19b-3p of tubular epithelial cells promotes M1 macrophage activation in kidney injury.Cell Death Differ2020;27:210-26 PMCID:PMC7206053

[36] Espinoza-Culupú A,Farfán-López M.Acylpolyamine mygalin as a TLR4 antagonist based on molecular docking and in vitro analyses.Biomolecules2020;10:1624 PMCID:PMC7761503