Histone demethylase JARID1C/KDM5C regulates Th17 cells by increasing IL-6 expression in diabetic plasmacytoid dendritic cells
Christopher O. Audu, Sonya J. Wolf, Amrita D. Joshi, Jadie Y. Moon, William J. Melvin, Sriganesh B. Sharma, Frank M. Davis, Andrea T. Obi, Rachel Wasikowski, Lam C. Tsoi, Emily C. Barrett, Kevin D. Mangum, Tyler M. Bauer, Steven L. Kunkel, Beth B. Moore, Katherine A. Gallagher
Christopher O. Audu, Sonya J. Wolf, Amrita D. Joshi, Jadie Y. Moon, William J. Melvin, Sriganesh B. Sharma, Frank M. Davis, Andrea T. Obi, Rachel Wasikowski, Lam C. Tsoi, Emily C. Barrett, Kevin D. Mangum, Tyler M. Bauer, Steven L. Kunkel, Beth B. Moore, Katherine A. Gallagher
View: Text | PDF
Research Article Immunology Inflammation

Histone demethylase JARID1C/KDM5C regulates Th17 cells by increasing IL-6 expression in diabetic plasmacytoid dendritic cells

Abstract

Plasmacytoid dendritic cells (pDCs) are first responders to tissue injury, where they prime naive T cells. The role of pDCs in physiologic wound repair has been examined, but little is known about pDCs in diabetic wound tissue and their interactions with naive CD4+ T cells. Diabetic wounds are characterized by increased levels of inflammatory IL-17A cytokine, partly due to increased Th17 CD4+ cells. This increased IL-17A cytokine, in excess, impairs tissue repair. Here, using human tissue and murine wound healing models, we found that diabetic wound pDCs produced excess IL-6 and TGF-β and that these cytokines skewed naive CD4+ T cells toward a Th17 inflammatory phenotype following cutaneous injury. Further, we identified that increased IL-6 cytokine production by diabetic wound pDCs is regulated by a histone demethylase, Jumonji AT-rich interactive domain 1C histone demethylase (JARID1C). Decreased JARID1C increased IL-6 transcription in diabetic pDCs, and this process was regulated upstream by an IFN-I/TYK2/JAK1,3 signaling pathway. When inhibited in nondiabetic wound pDCs, JARID1C skewed naive CD4+ T cells toward a Th17 phenotype and increased IL-17A production. Together, this suggests that diabetic wound pDCs are epigenetically altered to increase IL-6 expression that then affects T cell phenotype. These findings identify a therapeutically manipulable pathway in diabetic wounds.

Authors

Christopher O. Audu, Sonya J. Wolf, Amrita D. Joshi, Jadie Y. Moon, William J. Melvin, Sriganesh B. Sharma, Frank M. Davis, Andrea T. Obi, Rachel Wasikowski, Lam C. Tsoi, Emily C. Barrett, Kevin D. Mangum, Tyler M. Bauer, Steven L. Kunkel, Beth B. Moore, Katherine A. Gallagher

×

Figure 4

Diabetic wound pDCs promote T cell differentiation toward Th17 phenotype.

Options: View larger image (or click on image) Download as PowerPoint
Diabetic wound pDCs promote T cell differentiation toward Th17 phenotype...
(A) Flow cytometry analysis of RORγt expression in naive CD4+ T cells cocultured for 48–72 hours with DIO and ND wound pDCs. (B) Flow cytometry analysis of IL-17A in CD4+ T cells cocultured for 48–72 hours with DIO and ND wound pDCs. (C) RORγt expression in CD4+ T cells after 48- to 72-hour coculture with DIO wound pDCs under several conditions — naive CD4+ T cells (control), naive CD4+ T cells with IL-6 preinhibition (LMT-28, 200 nM, 1 hour), TGF-β receptor–deficient naive CD4+ T cells (Alk5fl/flCD4Cre+ T cells), and Alk5fl/flCD4Cre+ CD4+ T cells with IL-6 receptor preinhibition (LMT-28, 200 nM, 1 hour). (D) IL-17A expression in CD4+ T cells after 48- to 72-hour coculture with DIO wound pDCs under various conditions, as detailed in C. For these coculture experiments, each mouse received 3–4 wounds, and N = 3–5 mice/group, pooled and repeated in triplicate. (E) Cluster uniform manifold approximation and projection (UMAP) of scRNA-Seq from human T2D and non-T2D wounds showed 10 unique cell clusters (representative). (F) scRNA-Seq of human wound T-cell population demonstrating RORγt expression in T2D versus non-T2D controls (N = 42). Dot size corresponds to proportion of cells within the group expressing RORγt, while dot color corresponds to expression level. (G) Intracellular flow cytometry quantifying intracellular RORγt in ND and DIO wound CD4+ T cells. (N = 3–5 mice/group, pooled and repeated in triplicate.) (H) Protein expression by ELISA of IL-17A in ND versus DIO wound CD4+ T cells (N = 5/group, pooled and repeated in triplicate; day 5 wounds). (I) Wound healing curve in global knockout, diabetic IL17A–/– mice compared with age-matched, littermate controls (N = 3/group, pooled and repeated in triplicate). (J) Representative wound healing images and histology. Data are presented as the mean ± SEM. For C and D, data were analyzed using 1-way ANOVA with Holm-Šídák multiple-comparison test. Data for I were analyzed using 2-way repeated measures ANOVA. For all other panels, data were analyzed using 2-tailed Student’s t test once normality was assessed.