Screening and validation of Hub genes for iron overload in osteoarthritis based on bioinformatics

doi:10.12307/2025.243

Abstract

Abstract: BACKGROUND: Iron overload refers to excessive accumulation of iron in the body, which can cause pathological changes in various tissues. At present, the molecular mechanism and potential gene targets related to iron overload in osteoarthritis still need to be further studied and explored.
OBJECTIVE: To analyze the key genes of iron overload in osteoarthritis by means of bioinformatics and verify them in animal experiments, so as to provide a new idea for preventing and treating osteoarthritis from the perspective of iron overload.
METHODS: GEO database and GeneCards database were used to screen out genes associated with osteoarthritis and genes associated with iron overload. Then, the intersection of the two was taken to obtain a collection of genes commonly associated with osteoarthritis and iron overload. GO and KEGG enrichment analyses were used to screen the functions and pathways of these genes. To further investigate the interactions between these genes, a protein-protein interaction network was constructed and five computational methods of Cytoscape software were utilized to identify the Hub genes for iron overload in osteoarthritis. Finally, 12 male Sprague-Dawley rats were divided into osteoarthritis group and normal group, with 6 rats in each group. A knee osteoarthritis model was established by the modified Hulth method in the osteoarthritis group. The expression of Hub genes in the knee joint of rats in each group was detected by PCR.

RESULTS AND CONCLUSION: (1) A total of 51 genes associated with iron overload were identified in osteoarthritis. GO enrichment analysis showed that these genes were mainly involved in cytokine receptor binding, chemokine receptor binding, cytokine activity, growth factor receptor binding and oligosaccharide binding. (2) KEGG enrichment analysis showed that genes associated with iron overload in osteoarthritis was mainly involved in tumor necrosis factor signaling pathway and lipid and atherosclerosis signaling pathway. (3) The protein-protein interaction network was constructed, and five Hub genes of iron overload, intercellular cell adhesion molecule-1, tumor necrosis factor superfamily member 11, myelocytomatosis oncogene, janus kinase 2, and interleukin 6, were obtained by further analysis. Animal experiments verified that there were significant differences in the expression of the above Hub genes in the rat knee joint between the control group and the experimental group (P < 0.05). (4) All these findings show that intercellular cell adhesion molecule-1, tumor necrosis factor superfamily member 11, myelocytomatosis oncogene, janus kinase 2, and interleukin 6 can be used as the Hub genes of iron overload in osteoarthritis, which are expected to become new targets for the prevention and treatment of osteoarthritis.

中国组织工程研究杂志出版内容重点：人工关节；骨植入物；脊柱；骨折；内固定；数字化骨科；组织工程

Key words: osteoarthritis, iron overload, Hub gene, differential gene, bioinformatics, animal experiment, experimental verification, biomarker

CLC Number:

R459.9|R363|R364

Deng Keqi, Li Guangdi, Goswami Ashutosh, Liu Xingyu, He Xiaoyong. Screening and validation of Hub genes for iron overload in osteoarthritis based on bioinformatics[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(9): 1972-1980.

Figures/Tables 5

References

[1] HUNTER DJ, MARCH L, CHEW M. Osteoarthritis in 2020 and beyond: a Lancet Commission. Lancet. 2020;396(10264): 1711-1712.
[2] TAMPIERI A, SANDRI M, IAFISCO M, et al. Nanotechnological approach and bio-inspired materials to face degenerative diseases in aging. Aging Clin Exp Res. 2021; 33(4):805-821.
[3] KATZ JN, ARANT KR, LOESER RF. Diagnosis and treatment of hip and knee osteoarthritis: a review. JAMA. 2021;325(6): 568-578.
[4] CAMASCHELLA C, NAI A, SILVESTRI L. Iron metabolism and iron disorders revisited in the hepcidin era. Haematologica. 2020; 105(2):260-272.
[5] BRISSOT P, TROADEC MB, LORÉAL O, et al. Pathophysiology and classification of iron overload diseases; update 2018. Transfus Clin Biol. 2019;26(1):80-88.
[6] 黎少聪,何琪,潘兆丰,等.Micro-CT评价铁超载影响膝骨关节炎模型小鼠软骨类组织的变化[J].中国组织工程研究, 2023,27(29):4658-4663.
[7] CAI C, HU W, CHU T. Interplay between iron overload and osteoarthritis: clinical significance and cellular mechanisms. Front Cell Dev Biol. 2021;9:817104.
[8] BURTON LH, RADAKOVICH LB, MAROLF AJ, et al. Systemic iron overload exacerbates osteoarthritis in the strain 13 guinea pig. Osteoarthritis Cartilage. 2020;28(9): 1265-1275.
[9] WANG J, CHEN L, JIN S, et al. MiR-98 promotes chondrocyte apoptosis by decreasing Bcl-2 expression in a rat model of osteoarthritis. Acta Biochim Biophys Sin (Shanghai). 2016;48(10):923-929.
[10] PRITZKER KP, GAY S, JIMENEZ SA, et al. Osteoarthritis cartilage histopathology: grading and staging. Osteoarthritis Cartilage. 2006;14(1):13-29.
[11] HU F, HU W, XU H. Schisandrin B alleviates LPS induced mitochondrial damage in C28I2 cells. J Membr Biol. 2024. doi:10.1007/s00232-023-00299-5.
[12] OO WM. Prospects of disease-modifying osteoarthritis drugs. Clin Geriatr Med. 2022;38(2):397-432.
[13] RADUSHKEVITZ-FRISHMAN T, CHARNI-NATAN M, GOLDSTEIN I. Dynamic chromatin accessibility during nutritional iron overload reveals a BMP6-independent induction of cell cycle genes. J Nutr Biochem. 2023; 119:109407.
[14] SIMÃO M, GAVAIA PJ, CAMACHO A, et al. Intracellular iron uptake is favored in Hfe-KO mouse primary chondrocytes mimicking an osteoarthritis-related phenotype. Biofactors. 2019;45(4):583-597.
[15] GUILLEM-LLOBAT P, MARÍN M, ROULEAU M, et al. New insights into the pro-inflammatory and osteoclastogenic profile of circulating monocytes in osteoarthritis patients. Int J Mol Sci. 2024;25(3):1710.
[16] KOURTZELIS I, MITROULIS I, VON RENESSE J, et al. From leukocyte recruitment to resolution of inflammation: the cardinal role of integrins. J Leukoc Biol. 2017;102(3): 677-683.
[17] WANG Z, WANG B, ZHANG J, et al. Chemokine (C-C Motif) ligand 2/chemokine receptor 2 (CCR2) axis blockade to delay chondrocyte hypertrophy as a therapeutic strategy for osteoarthritis. Med Sci Monit. 2021;27:e930053.
[18] WANG R, LI J, XU X, et al. Andrographolide attenuates synovial inflammation of osteoarthritis by interacting with tumor necrosis factor receptor 2 trafficking in a rat model. J Orthop Translat. 2021;29:89-99.
[19] 韩建建,徐鹏,韩礼纲,等.关节镜清理术联合透明质酸钠对膝骨关节炎病人关节液细胞间黏附分子1、白细胞介素6的影响[J].蚌埠医学院学报,2021,46(2): 222-225.
[20] SCALZONE A, CERQUENI G, WANG XN, et al. A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis. Front Bioeng Biotechnol. 2023;11:1167623.
[21] WANG Y, LIU J, HUANG B, et al. Mathematical modeling and application of IL-1β/TNF signaling pathway in regulating chondrocyte apoptosis. Front Cell Dev Biol. 2023;11:1288431.
[22] HEIDARI B, BABAEI M, YOSEFGHAHRI B. Prevention of osteoarthritis progression by statins, targeting metabolic and inflammatory aspects: a review. Mediterr J Rheumatol. 2021;32(3):227-236.
[23] IIJIMA H, GILMER G, WANG K, et al. Meta-analysis integrated with multi-omics data analysis to elucidate pathogenic mechanisms of age-related knee osteoarthritis in mice. J Gerontol A Biol Sci Med Sci. 2022;77(7):1321-1334.
[24] ALVARADO-DÍAZ CP, NÚÑEZ MT, DEVOTO L, et al. Iron overload-modulated nuclear factor kappa-B activation in human endometrial stromal cells as a mechanism postulated in endometriosis pathogenesis. Fertil Steril. 2015;103(2):439-447.
[25] LEE KT, SU CH, LIU SC, et al. Cordycerebroside A inhibits ICAM-1-dependent M1 monocyte adhesion to osteoarthritis synovial fibroblasts. J Food Biochem. 2022;46(7):e14108.
[26] LI M, XIAO YB, WANG XT, et al. Proline-serine-threonine phosphatase-interacting protein 2 alleviates diabetes mellitus-osteoarthritis in rats through attenuating synovial inflammation and cartilage injury. Orthop Surg. 2021;13(4):1398-1407.
[27] CHANKAMNGOEN W, KRUNGCHANUCHAT S, THONGBUNCHOO J, et al. Extracellular Fe(2+) and Fe(3+) modulate osteocytic viability, expression of SOST, RANKL and FGF23, and fluid flow-induced YAP1 nuclear translocation. Sci Rep. 2023;13(1):21173.
[28] PAN L, YANG F, CAO X, et al. Identification of five hub immune genes and characterization of two immune subtypes of osteoarthritis. Front Endocrinol (Lausanne). 2023;14:1144258.
[29] MANDALA A, CHEN WJ, ARMSTRONG A, et al. PPARα agonist fenofibrate attenuates iron-induced liver injury in mice by modulating the Sirt3 and β-catenin signaling. Am J Physiol Gastrointest Liver Physiol. 2021;321(4):G262-G269.
[30] JIANG K, JIANG T, CHEN Y, et al. Mesenchymal stem cell-derived exosomes modulate chondrocyte glutamine metabolism to alleviate osteoarthritis progression. Mediators Inflamm. 2021; 2021:2979124.
[31] WANG C, WANG X, SONG G, et al. A high-fructose diet in rats induces systemic iron deficiency and hepatic iron overload by an inflammation mechanism. J Food Biochem. 2021;45(1):e13578.
[32] CHEN B, NING K, SUN ML, et al. Regulation and therapy, the role of JAK2/STAT3 signaling pathway in OA: a systematic review. Cell Commun Signal. 2023;21(1):67.
[33] DI PAOLA A, TORTORA C, ARGENZIANO M, et al. Emerging roles of the iron chelators in inflammation. Int J Mol Sci. 2022;23(14):7977.
[34] HE J, WANG L, DING Y, et al. lncRNA FER1L4 is dysregulated in osteoarthritis and regulates IL-6 expression in human chondrocyte cells. Sci Rep. 2021;11(1): 13032.