SARS-CoV-2 ORF8 drives osteoclastogenesis in preexisting immune-mediated inflammatory diseases
Ivonne Melano, Tamiris Azamor, Camila C.S. Caetano, Nikki M. Meyer, Chineme Onwubueke, Anabelle Visperas, Débora Familiar-Macedo, Gielenny M. Salem, Brandy-Lee Soos, Cassandra M. Calabrese, Youn Jung Choi, Shuyang Chen, Younho Choi, Xianfang Wu, Zilton Vasconcelos, Suzy A.A. Comhair, Karin Nielsen-Saines, Leonard H. Calabrese, M. Elaine Husni, Jae U. Jung, Nicolas S. Piuzzi, Suan-Sin Foo, Weiqiang Chen
Ivonne Melano, Tamiris Azamor, Camila C.S. Caetano, Nikki M. Meyer, Chineme Onwubueke, Anabelle Visperas, Débora Familiar-Macedo, Gielenny M. Salem, Brandy-Lee Soos, Cassandra M. Calabrese, Youn Jung Choi, Shuyang Chen, Younho Choi, Xianfang Wu, Zilton Vasconcelos, Suzy A.A. Comhair, Karin Nielsen-Saines, Leonard H. Calabrese, M. Elaine Husni, Jae U. Jung, Nicolas S. Piuzzi, Suan-Sin Foo, Weiqiang Chen
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Research Article Inflammation Virology

SARS-CoV-2 ORF8 drives osteoclastogenesis in preexisting immune-mediated inflammatory diseases

Abstract

Patients with immune-mediated inflammatory diseases (IMIDs) like rheumatoid arthritis (RA) are at higher risk for severe COVID-19 and long-term complications in bone health. Emerging clinical evidence demonstrated that SARS-CoV-2 infection reduces bone turnover and promotes bone loss, but the mechanism underlying worsened bone health remains elusive. This study sought to identify specific immune mediators that exacerbated preexisting IMIDs after SARS-CoV-2 exposure. Plasma samples from 4 groups were analyzed: healthy, IMID only, COVID-19 only, and COVID-19 + IMID. Using high-throughput multiplexed proteomics, we profiled 1,500 protein biomarkers and identified 148 unique biomarkers in COVID-19 patients with IMIDs, including elevated inflammatory cytokines (e.g., IL-17F) and bone resorption markers. Long-term circulating SARS-CoV-2 ORF8, a virulence factor for COVID-19, was detected in the COVID + IMID group. RA was one of the most common IMIDs in our study. ORF8 treatment of RA-derived human osteoblasts (RA-hOBs) increased levels of inflammatory (TNF, IL6, CCL2) and bone resorption (RANKL/osteoprotegerin ratio) markers compared with healthy controls. Supernatants from ORF8-treated RA-hOBs drove the differentiation of macrophages into osteoclast-like cells. These findings suggest that SARS-CoV-2 exposure can exacerbate IMIDs through ORF8-driven inflammation and osteoclastogenesis, highlighting potential therapeutic targets for managing COVID-19–induced bone pathologies.

Authors

Ivonne Melano, Tamiris Azamor, Camila C.S. Caetano, Nikki M. Meyer, Chineme Onwubueke, Anabelle Visperas, Débora Familiar-Macedo, Gielenny M. Salem, Brandy-Lee Soos, Cassandra M. Calabrese, Youn Jung Choi, Shuyang Chen, Younho Choi, Xianfang Wu, Zilton Vasconcelos, Suzy A.A. Comhair, Karin Nielsen-Saines, Leonard H. Calabrese, M. Elaine Husni, Jae U. Jung, Nicolas S. Piuzzi, Suan-Sin Foo, Weiqiang Chen

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Figure 5

SARS-CoV-2 ORF8 treatment on primary human osteoblasts derived from patients with RA drives overt inflammation and dysregulates bone resorption markers.

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SARS-CoV-2 ORF8 treatment on primary human osteoblasts derived from pati...
(A) Percentage of disease distribution in IMID and COVID + IMID groups. (B) Schematic representation of in vitro SARS-CoV-2 ORF8 stimulation in hOBs derived from healthy controls (H-hOBs) and patients with RA (RA-hOBs) upon SARS-CoV-2 ORF8 stimulation. Cells and supernatants were harvested at 2 days and 4 days posttreatment. (C) Heatmap representation of transcriptional profile of inflammatory factors IL6, IL17A, IL17F, TNF, and CCL2 and bone-associated factors RANKL, OPG, RANKL/OPG ratio, ALP, PTHR1, COL1A1, and COL17A1 in H-hOBs and RA-hOBs stimulated with 10 ng/mL, 20 ng/mL, or 50 ng/mL of purified SARS-CoV-2 ORF8, relative to mock-treated controls. (D and E) Transcriptional profile of inflammatory factors (D) and bone-associated factors (E) in H-hOBs and RA-hOBs stimulated with 20 ng/mL of purified SARS-CoV-2 ORF8, relative to mock-treated controls n = 4–6 per group. (F) Protein levels of RANK, OPG, and RANKL/OPG in the supernatant of mock controls and SARS-CoV-2 ORF8–stimulated H-hOBs and RA-hOBs. n = 4–6 per group. Data are represented as means ± SEM. Statistical analysis was performed using 2-way ANOVA and Bonferroni’s multiple comparisons test. Data were generated from 3 independent experiments. Significant differences among groups are displayed with asterisks (*P < 0.05, **P < 0.01, and ***P < 0.001).