M2 macrophages-derived exosomes for osteonecrosis of femoral head treatment: modulating neutrophil extracellular traps formation and endothelial phenotype transition

Abstract

Exosomes have shown good potential in ischemic injury disease treatments. However, evidence about their effect and molecular mechanisms in osteonecrosis of femoral head (ONFH) treatment is still limited. Here, we revealed the cell biology characters of ONFH osteonecrosis area bone tissue in single cell scale and thus identified a novel ONFH treatment approach based on M2 macrophages-derived exosomes (M2-Exos). We further show that M2-Exos are highly effective in the treatment of ONFH by modulating the phenotypes communication between neutrophil and endothelium including neutrophil extracellular traps formation and endothelial phenotype transition. Additionally, we identified that M2-Exos’ therapeutic effect is attributed to the high content of miR-93-5p and constructed miR-93-5p overexpression model in vitro and in vivo based on lentivirus and adeno-associated virus respectively. Then we found miR-93-5p can not only reduce neutrophil extracellular traps formation but also improve angiogenic ability of endothelial cells. These results provided a new theoretical basis for the clinical application of ONFH therapeutic exosomes.

Keywords

Biological Sciences / Biochemistry and Cell Biology

References

[1] ChenCY, et al. . Glucocorticoid-induced loss of beneficial gut bacterial extracellular vesicles is associated with the pathogenesis of osteonecrosis. Sci. Adv., 2022, 8: eabg8335.

[2] LiuG, et al. . Osteogenesis-related long noncoding RNA GAS5 as a novel biomarker for osteonecrosis of femoral head. Front. Cell Dev. Biol., 2022, 10. 857612

[3] Tan, H. et al. Extracorporeal shock wave therapy with imaging examination for early osteonecrosis of the femoral head: A systematic review. Int. J. Surg.111, 1144–1153 (2024).

[4] YoonBH, et al. . The 2019 revised version of association research circulation osseous staging system of osteonecrosis of the femoral head. J. Arthroplast., 2020, 35: 933-940.

[5] NakamuraJ, et al. . Time elapsed from definitive diagnosis to surgery for osteonecrosis of the femoral head: A nationwide observational study in Japan. BMJ Open, 2024, 14. e082342

[6] YuanJ, et al. . Single-cell transcriptomic analysis of the immune microenvironment in pediatric acute leukemia. Cancer Lett., 2024, 596. 217018

[7] BandyopadhyayS, et al. . Mapping the cellular biogeography of human bone marrow niches using single-cell transcriptomics and proteomic imaging. Cell, 2024, 187: 3120-3140 e3129.

[8] BixelMG, et al. . Angiogenesis is uncoupled from osteogenesis during calvarial bone regeneration. Nat. Commun., 2024, 15. 4575

[9] GamblinAL, et al. . Bone tissue formation with human mesenchymal stem cells and biphasic calcium phosphate ceramics: the local implication of osteoclasts and macrophages. Biomaterials, 2014, 35: 9660-9667.

[10] ChenR, et al. . Mesenchymal stem cell-immune cell interaction and related modulations for bone tissue engineering. Stem Cells Int, 2022, 2022. 7153584

[11] ChenZ, et al. . Tuning chemistry and topography of nanoengineered surfaces to manipulate immune response for bone regeneration applications. ACS Nano, 2017, 11: 4494-4506.

[12] ZhaoH, et al. . Prevention and treatment of osteoporosis using chinese medicinal plants: special emphasis on mechanisms of immune modulation. J. Immunol. Res., 2018, 2018. 6345857

[13] CaiB, et al. . N2-polarized neutrophils guide bone mesenchymal stem cell recruitment and initiate bone regeneration: A missing piece of the bone regeneration puzzle. Adv. Sci. (Weinh.), 2021, 8e2100584

[14] MohantyT, et al. . A novel mechanism for NETosis provides antimicrobial defense at the oral mucosa. Blood, 2015, 126: 2128-2137.

[15] CristinzianoL, et al. . Neutrophil extracellular traps in cancer. Semin Cancer Biol., 2022, 79: 91-104.

[16] MartinodK, WagnerDD. Thrombosis: tangled up in NETs. Blood, 2014, 123: 2768-2776.

[17] ManS, Sanchez DuffhuesG, Ten DijkeP, BakerD. The therapeutic potential of targeting the endothelial-to-mesenchymal transition. Angiogenesis, 2019, 22: 3-13.

[18] GuM, et al. . Patient-specific iPSC-derived endothelial cells uncover pathways that protect against pulmonary hypertension in BMPR2 mutation carriers. Cell Stem Cell, 2017, 20: 490-504 e495.

[19] Pardali, E., Sanchez-Duffhues, G., Gomez-Puerto, M. C. & Ten Dijke, P. TGF-beta-induced endothelial-mesenchymal transition in fibrotic diseases. Int. J. Mol. Sci.18, 2157 (2017).

[20] DingY, et al. . Metformin in cardiovascular diabetology: a focused review of its impact on endothelial function. Theranostics, 2021, 11: 9376-9396.

[21] Yu, S., Liu, J. & Yan, N. Endothelial dysfunction induced by extracellular neutrophil traps plays important role in the occurrence and treatment of extracellular neutrophil traps-related disease. Int. J. Mol. Sci. 23, 5626 (2022).

[22] YangS, et al. . Neutrophil extracellular traps delay diabetic wound healing by inducing endothelial-to-mesenchymal transition via the Hippo pathway. Int. J. Biol. Sci., 2023, 19: 347-361.

[23] DasS, et al. . The extracellular RNA communication consortium: Establishing foundational knowledge and technologies for extracellular RNA research. Cell, 2019, 177: 231-242.

[24] WangD, et al. . Schwann cell-derived EVs facilitate dental pulp regeneration through endogenous stem cell recruitment via SDF-1/CXCR4 axis. Acta Biomater., 2022, 140: 610-624.

[25] VogelpoelLT, et al. . Fc gamma receptor-TLR cross-talk elicits pro-inflammatory cytokine production by human M2 macrophages. Nat. Commun., 2014, 5. 5444

[26] HeJ, PhamTL, KakazuA, BazanHEP. Recovery of corneal sensitivity and increase in nerve density and wound healing in diabetic mice after PEDF plus DHA treatment. Diabetes, 2017, 66: 2511-2520.

[27] YinZ, et al. . Macrophage-derived exosomal microRNA-501-3p promotes progression of pancreatic ductal adenocarcinoma through the TGFBR3-mediated TGF-beta signaling pathway. J. Exp. Clin. Cancer Res., 2019, 38: 310.

[28] WangY, et al. . Macrophage-derived extracellular vesicles: diverse mediators of pathology and therapeutics in multiple diseases. Cell Death Dis., 2020, 11. 924

[29] QianY, et al. . M2 macrophage-derived exosomal miR-26b-5p regulates macrophage polarization and chondrocyte hypertrophy by targeting TLR3 and COL10A1 to alleviate osteoarthritis. J. Nanobiotechnology, 2024, 22. 72

[30] WangZ, SunW, LiR, LiuY. miRNA-93-5p in exosomes derived from M2 macrophages improves lipopolysaccharide-induced podocyte apoptosis by targeting Toll-like receptor 4. Bioengineered, 2022, 13: 7683-7696.

[31] MaT, et al. . Enhanced osteolysis targeted therapy through fusion of exosomes derived from M2 macrophages and bone marrow mesenchymal stem cells: Modulating macrophage polarization. Small, 2024, 20. e2303506

[32] UesugiY, et al. . Quality of life of patients with osteonecrosis of the femoral head: a multicentre study. Int. Orthop., 2018, 42: 1517-1525.

[33] LiZ, et al. . Advances in experimental models of osteonecrosis of the femoral head. J. Orthop. Transl., 2023, 39: 88-99

[34] ChenC, et al. . Imbalanced T-cell subsets may facilitate the occurrence of osteonecrosis of the femoral head. J. Inflamm. Res., 2022, 15: 4159-4169.

[35] TanZ, et al. . The dynamic feature of macrophage M1/M2 imbalance facilitates the progression of non-traumatic osteonecrosis of the femoral head. Front. Bioeng. Biotechnol., 2022, 10. 912133

[36] MaM, et al. . Osteoimmunology and osteonecrosis of the femoral head. Bone Jt. Res, 2022, 11: 26-28.

[37] KongN, et al. . An injectable self-adaptive polymer as a drug carrier for the treatment of nontraumatic early-stage osteonecrosis of the femoral head. Bone Res., 2022, 10: 28.

[38] QuanH, et al. . Application of biomaterials in treating early osteonecrosis of the femoral head: research progress and future perspectives. Acta Biomater., 2023, 164: 15-73.

[39] Wang, Y. et al. Accumulation of fat not responsible for femoral head necrosis, revealed by single-cell RNA sequencing: A preliminary study. Biomolecules13, 171 (2023).

[40] LiaoZ, et al. . Single-cell transcriptome analysis reveals aberrant stromal cells and heterogeneous endothelial cells in alcohol-induced osteonecrosis of the femoral head. Commun. Biol., 2022, 5: 324.

[41] SekheriM, OthmanA, FilepJG. beta2 integrin regulation of neutrophil functional plasticity and fate in the resolution of inflammation. Front. Immunol., 2021, 12. 660760

[42] BergmannA, et al. . Aldosterone abrogates nuclear factor kappaB-mediated tumor necrosis factor alpha production in human neutrophils via the mineralocorticoid receptor. Hypertension, 2010, 55: 370-379.

[43] JiaE, et al. . Neutrophil extracellular traps induce the bone erosion of gout. BMC Musculoskelet. Disord., 2022, 23. 1128

[44] BraileM, et al. . LPS-mediated neutrophil VEGF-A release is modulated by cannabinoid receptor activation. J. Leukoc. Biol., 2021, 109: 621-631.

[45] JiangJ, et al. . Correlational analysis between neutrophil granulocyte levels and osteonecrosis of the femoral head. BMC Musculoskelet. Disord., 2019, 20. 393

[46] KankiY, et al. . Epigenetically coordinated GATA2 binding is necessary for endothelium-specific endomucin expression. EMBO J., 2011, 30: 2582-2595.

[47] HjortnaesJ, et al. . Valvular interstitial cells suppress calcification of valvular endothelial cells. Atherosclerosis, 2015, 242: 251-260.

[48] DongR, et al. . Rutin alleviates EndMT by restoring autophagy through inhibiting HDAC1 via PI3K/AKT/mTOR pathway in diabetic kidney disease. Phytomedicine, 2023, 112. 154700

[49] XuY, et al. . Stem cell therapy for osteonecrosis of femoral head: Opportunities and challenges. Regen. Ther., 2020, 15: 295-304.

[50] TanF, et al. . Clinical applications of stem cell-derived exosomes. Signal Transduct. Target Ther., 2024, 9: 17.

[51] ChenC, et al. . Engineered exosome-functionalized extracellular matrix-mimicking hydrogel for promoting bone repair in glucocorticoid-induced osteonecrosis of the femoral head. ACS Appl Mater. Interfaces, 2023, 15: 28891-28906.

[52] NikfarjamS, RezaieJ, ZolbaninNM, JafariR. Mesenchymal stem cell derived-exosomes: a modern approach in translational medicine. J. Transl. Med., 2020, 18. 449

[53] GaoZS, et al. . Berberine-loaded M2 macrophage-derived exosomes for spinal cord injury therapy. Acta Biomater., 2021, 126: 211-223.

[54] ZaveriTD, et al. . Macrophage integrins modulate response to ultra-high molecular weight polyethylene particles and direct particle-induced osteolysis. Biomaterials, 2017, 115: 128-140.

[55] GaoC, et al. . Treatment of atherosclerosis by macrophage-biomimetic nanoparticles via targeted pharmacotherapy and sequestration of proinflammatory cytokines. Nat. Commun., 2020, 11. 2622

[56] DaneshvarA, NematiP, AzadiA, AmidR, KadkhodazadehM. M2 macrophage-derived exosomes for bone regeneration: A systematic review. Arch. Oral. Biol., 2024, 166. 106034

[57] JiaoY, et al. . Exosomal PGE2 from M2 macrophages inhibits neutrophil recruitment and NET formation through lipid mediator class switching in sepsis. J. Biomed. Sci., 2023, 30: 62.

[58] ChenM, et al. . DanShen Decoction targets miR-93-5p to provide protection against MI/RI by regulating the TXNIP/NLRP3/Caspase-1 signaling pathway. Phytomedicine, 2024, 135. 156225

Funding

National Natural Science Foundation of China (National Science Foundation of China)(No.82272503)

RIGHTS & PERMISSIONS

The Author(s)