Exploring the Therapeutic Potential of MIR-140-3p in Osteoarthritis: Targeting CILP and Ferroptosis for Novel Treatment Strategies

Abstract

Osteoarthritis (OA) is a prevalent and debilitating joint disorder that affects millions of individuals worldwide, severely impairing mobility, independence, and quality of life. Emerging evidence suggests that ferroptosis is a critical factor in OA pathogenesis. However, its precise involvement and underlying mechanisms remain poorly understood. In this study, we first identified that cartilage intermediate layer protein (CILP) mediates the regulation of ferroptosis-related genes in OA through hdWGCNA analysis combined with single-cell RNA sequencing. Further investigation revealed a significant upregulation of CILP protein expression in C28/I2 cells under LPS induction. Mechanistically, bioinformatics analysis identified differentially expressed miRNAs; qRT-PCR combined with a dual-luciferase experiment revealed that miR-140-3p was downregulated and directly targets CILP. Experimental data further demonstrated that miR-140-3p regulates ferroptosis, inflammation, and oxidative stress by targeting CILP. These findings offer valuable insights into the molecular mechanisms of the miR-140-3p/CILP axis in regulating ferroptosis, inflammation, and oxidative stress, thus providing a foundation for developing therapeutic strategies for OA.

References

[1] P. Duarte, R. Elisabete, and B. Jaime, “Osteoarthritis,” Acta Médica Portuguesa 28, no. 1 (2015): 99–106.

[2] B. Abramoff and F. Caldera, “Osteoarthritis: Pathology, Diagnosis, and Treatment Options,” Medical Clinics of North America 104, no. 2 (2020): 293–311.

[3] M. Rahmati, G. Nalesso, A. Mobasheri, and M. Mozafari, “Aging and Osteoarthritis: Central Role of the Extracellular Matrix,” Ageing Research Reviews 40 (2017): 20–30.

[4] L. Lu, J. Wang, A. Fan, et al., “Synovial Mesenchymal Stem Cell-Derived Extracellular Vesicles Containing microRN555A-26a-5p Ameliorate Cartilage Damage of Osteoarthritis,” Journal of Gene Medicine 23, no. 11 (2021): e3379.

[5] Q. He, J. Yang, Z. Pan, et al., “Biochanin A Protects Against Iron Overload Associated Knee Osteoarthritis via Regulating Iron Levels and NRF2/System xc-/GPX4 Axis,” Biomedicine & Pharmacotherapy 157 (2023): 113915.

[6] H. Nicholson, N. Sakhrani, J. Rogot, et al., “Effect of Blood on Synovial Joint Tissues: Potential Role of Ferroptosis,” Applied Sciences (Basel, Switzerland) 14, no. 14 (2024): 6292.

[7] M. He, Y. Wang, J. Xie, et al., “MG Modification of FTH1 and Pri-miR-26a Regulates Ferroptosis and Chemotherapy Resistance in Osteosarcoma,” Oncogene 43, no. 5 (2024): 341–353, https://doi.org/10.1038/s41388-023-02882-5.

[8] H. Kong, M. Sun, X. Zhang, and X. Wang, “Crosstalk Among circRNA/lncRNA, miRNA, and mRNA in Osteoarthritis,” Frontiers in Cell and Developmental Biology 9 (2021): 774370.

[9] C. Diener, A. Keller, and E. Meese, “Emerging Concepts of miRNA Therapeutics: From Cells to Clinic,” Trends in Genetics: TIG 38, no. 6 (2022): 613–626.

[10] Q. Yu, Y. Xiao, M. Guan, et al., “Regulation of Ferroptosis in Osteoarthritis and Osteoarthritic Chondrocytes by Typical MicroRNAs in Chondrocytes,” Frontiers in Medicine 11 (2024): 1478153.

[11] M. Zhou, C. Zhai, K. Shen, et al., “miR-1 Inhibits the Ferroptosis of Chondrocyte by Targeting CX43 and Alleviates Osteoarthritis Progression,” Journal of Immunology Research 2023 (2023): 2061071.

[12] D. Wang, Y. Fang, L. Lin, et al., “Upregulating miR-181b Promotes Ferroptosis in Osteoarthritic Chondrocytes by Inhibiting SLC7A11,” BMC Musculoskeletal Disorders 24, no. 1 (2023): 862.

[13] S. Li, J. Han, J. Cao, et al., “ADORA2B, Transcriptionally Suppressing by MYC, Promotes Ferroptosis of Chondrocytes via Inhibition of the PI3K/Akt Pathway in Mice With Osteoarthritis,” Environmental Toxicology 39, no. 5 (2024): 2487–2501, https://doi.org/10.1002/tox.24131.

[14] L. Wang, S. Ye, J. Qin, M. Tang, M. Dong, and J. Fang, “Ferroptosis-Related Genes LPCAT3 and PGD Are Potential Diagnostic Biomarkers for Osteoarthritis,” Journal of Orthopaedic Surgery and Research 18, no. 1 (2023): 699.

[15] L. Liu, J. He, C. Liu, et al., “Cartilage Intermediate Layer Protein Affects the Progression of Intervertebral Disc Degeneration by Regulating the Extracellular Microenvironment (Review),” International Journal of Molecular Medicine 47, no. 2 (2021): 475–484, https://doi.org/10.3892/ijmm.2020.4832.

[16] D. Dou, X. Ren, M. Han, et al., “Circ_0008039 Supports Breast Cancer Cell Proliferation, Migration, Invasion, and Glycolysis by Regulating the miR-140-3p/SKA2 Axis,” Molecular Oncology 15, no. 2 (2021): 697–709.

[17] S. Cheng, Z. Nie, J. Cao, and H. Peng, “Circ_0136474 Promotes the Progression of Osteoarthritis by Sponging Mir-140-3p and Upregulating MECP2,” Journal of Molecular Histology 54, no. 1 (2023): 1–12.

[18] Y. Hu, H. Liu, D. Xu, X. Xue, and X. Xu, “The Anti-Inflammatory Effect of miR-140-3p in BMSCs-Exosomes on Osteoarthritis,” Acta Chirurgiae Orthopaedicae et Traumatologiae Cechoslovaca 90, no. 4 (2023): 267–276.

[19] J. Chen, X. Li, Y. Yang, S. Xu, Q. Chen, and J. Xu, “Competing Endogenous RNA Network Analysis of the Molecular Mechanisms of Ischemic Stroke,” BMC Genomics 24, no. 1 (2023): 67.

[20] J. Hao, A. Ma, L. Wang, et al., “General Requirements for Stem Cells,” Cell Proliferation 53, no. 12 (December 2020): e12926.

[21] S. Niu, C. Xia, D. Huang, et al., “Requirements for Human Natural Killer Cells,” Cell Proliferation 57, no. 5 (May 2024): e13588.

[22] L. Wang, S. Liu, K. Li, et al., “General Requirements for the Production of Extracellular Vesicles Derived From Human Stem Cells,” Cell Proliferation 57, no. 3 (March 2024): e13554.

[23] X. Nan, B. Zhang, J. Hao, et al., “Requirements for Human Haematopoietic Stem/Progenitor Cells,” Cell Proliferation 55, no. 4 (April 2022): e13152.

[24] Z. Peng, J. Wu, S. Hu, et al., “Requirments for Primary Human Hepatocyte,” Cell Proliferation 55, no. 4 (April 2022): e13147.

[25] M. Yu, W. Lei, J. Cao, et al., “Requirements for Human Cardiomyocytes,” Cell Proliferation 55, no. 4 (April 2022): e13150.

[26] Y. K. Wang, L. Feng, A. J. Ma, et al., “Human midbrain dopaminergic progenitors,” Cell Proliferation 57, no. 4 (April 2024): e13563.

[27] Y. K. Wang, J. Yu, T. T. Zhang, et al., “Human neural stem cells,” Cell Proliferation 57, no. 4 (April 2024): e13564.

[28] X. Chen, J. Huang, J. Wu, et al., “Human Mesenchymal Stem Cells,” Cell Proliferation 55, no. 4 (April 2022): e13141.

[29] J. Hao, J. Cao, L. Wang, et al., “Requirements for Human Embryonic Stem Cells,” Cell Proliferation 53, no. 12 (2020): e12925, https://doi.org/10.1111/cpr.12925 2020 Oct 19. Erratum in: Cell Prolif. 2021 May;54(5):e13010.

[30] X. Zhang, L. Hou, Z. Guo, et al., “Lipid Peroxidation in Osteoarthritis: Focusing on 4-Hydroxynonenal, Malondialdehyde, and Ferroptosis,” Cell Death Discovery 9, no. 1 (2023): 320.

[31] J. Martel-Pelletier, A. Barr, F. Cicuttini, et al., “Osteoarthritis,” Nature Reviews Disease Primers 2 (2016): 16072.

[32] H. Wang, T. Yuan, Y. Wang, et al., “Osteoclasts and Osteoarthritis: Novel Intervention Targets and Therapeutic Potentials During Aging,” Aging Cell 23, no. 4 (2024): e14092.

[33] M. Sun, F. Beier, and M. Pest, “Recent Developments in Emerging Therapeutic Targets of Osteoarthritis,” Current Opinion in Rheumatology 29, no. 1 (2017): 96–102.

[34] J. Alexander, K , J. Stephen, R , and M. Tristan, “Inflammation in Osteoarthritis: The Latest Progress and Ongoing Challenges,” Current Opinion in Rheumatology 35, no. 2 (2023): 128–134.

[35] E. Hadzic and F. Beier, “Emerging Therapeutic Targets for Osteoarthritis,” Expert Opinion on Therapeutic Targets 27, no. 2 (2023): 111–120.

[36] Z. Zhao, X. Dai, Z. Wang, Z. Bao, and J. Guan, “MicroRNA-26a Reduces Synovial Inflammation and Cartilage Injury in Osteoarthritis of Knee Joints Through Impairing the NF-κB Signaling Pathway,” Bioscience Reports 39, no. 4 (2019): BSR20182025.

[37] D. Liyan, H. Shasha, G. Nuoqing, T. Wen, Z. Weizhen, and L. Lianxiang, “Molecular Mechanisms of Ferroptosis and Relevance to Inflammation,” Inflammation Research 72, no. 2 (2022): 281–299.

[38] G. Jérémie, J. G. Gregory, and M. P. R. Cecília, “Lytic Cell Death in Metabolic Liver Disease,” Journal of Hepatology 73, no. 2 (2020): 394–408.

[39] D. Szala, M. Kopańska, J. Trojniak, et al., “The Role of MicroRNAs in the Pathophysiology of Osteoarthritis,” International Journal of Molecular Sciences 25, no. 12 (2024): 6352.

[40] C. J. M, “MicroRNAs and Osteoarthritis,” Cells 7, no. 8 (2018): 92.

[41] M. Shakeri, A. Aminian, K. Mokhtari, et al., “Unraveling the Molecular Landscape of Osteoarthritis: A Comprehensive Review Focused on the Role of Non-coding RNAs,” Pathology, Research and Practice 260 (2024): 155446.

[42] J. Meng, Y. Zou, L. Hou, et al., “MiR-140-3p Ameliorates the Inflammatory Response of Airway Smooth Muscle Cells by Targeting HMGB1 to Regulate the JAK2/STAT3 Signaling Pathway,” Cell Journal 24, no. 11 (2022): 673–680.

[43] X. Zhou, Y. Lu, P. Guo, and C. Zhou, “Upregulation of microRNA-140-3p Mediates Dachshund Family Transcription Factor 1 Expression in Immunoglobulin A Nephropathy Through Cell Cycle-Dependent Mechanisms,” Molecular Medicine Reports 23, no. 2 (2021): 134, https://doi.org/10.3892/mmr.2020.11773.

[44] X. Chen, J. Hu, J. Lai, Z. Zhang, and Z. Tang, “Dexmedetomidine Attenuates LPS-Stimulated Alveolar Type II Cells' Injury Through Upregulation of miR-140-3p and Partial Suppression of PD-L1 Involving Inactivating JNK-Bnip3 Pathway,” Canadian Respiratory Journal 2022 (2022): 8433960.

RIGHTS & PERMISSIONS

2025 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.