Changwan Ryu, Anjali Walia, Vivian Ortiz, Carrighan Perry, Sam Woo, Benjamin C Reeves, Huanxing Sun, Julia Winkler, Jean E Kanyo, Weiwei Wang, Milica Vukmirovic, Nicholas Ristic, Eric A Stratton, Sita Ram Meena, Maksym Minasyan, Daniel Kurbanov, Xinran Liu, TuKiet T Lam, Giuseppina Farina, Jose L Gomez, Mridu Gulati, Erica L Herzog
Arthritis & rheumatology (Hoboken, N.J.) 2020 NovSystemic sclerosis-associated interstitial lung disease (SSc-ILD) is characterized by variable clinical outcomes, activation of innate immune pattern-recognition receptors (PRRs), and accumulation of α-smooth muscle actin (α-SMA)-expressing myofibroblasts. The aim of this study was to identify an association between these entities and mitochondrial DNA (mtDNA), an endogenous ligand for the intracellular DNA-sensing PRRs Toll-like receptor 9 (TLR-9) and cyclic GMP-AMP synthase/stimulator of interferon genes (cGAS/STING), which has yet to be determined. Human lung fibroblasts (HLFs) from normal donors and SSc-ILD explants were treated with synthetic CpG DNA and assayed for α-SMA expression and extracellular mtDNA using quantitative polymerase chain reaction for the human MT-ATP6 gene. Plasma MT-ATP6 concentrations were evaluated in 2 independent SSc-ILD cohorts and demographically matched controls. The ability of SSc-ILD and control plasma to induce TLR-9 and cGAS/STING activation was evaluated with commercially available HEK 293 reporter cells. Plasma concentrations of type I interferons (IFNs), interleukin-6 (IL-6), and oxidized DNA were measured using electrochemiluminescence and enzyme-linked immunosorbent assay-based methods. Extracellular vesicles (EVs) precipitated from plasma were evaluated for MT-ATP6 concentrations and proteomics via liquid chromatography mass spectrometry. Normal HLFs and SSc-ILD fibroblasts developed increased α-SMA expression and MT-ATP6 release following CpG stimulation. Plasma mtDNA concentrations were increased in the 2 SSc-ILD cohorts, reflective of ventilatory decline, and were positively associated with both TLR-9 and cGAS/STING activation as well as type I IFN and IL-6 expression. Plasma mtDNA was not oxidized and was conveyed by EVs displaying a proteomics profile consistent with a multicellular origin. These findings demonstrate a previously unrecognized connection between EV-encapsulated mtDNA, clinical outcomes, and intracellular DNA-sensing PRR activation in SSc-ILD. Further study of these interactions could catalyze novel mechanistic and therapeutic insights into SSc-ILD and related disorders. © 2020, American College of Rheumatology.
Changwan Ryu, Anjali Walia, Vivian Ortiz, Carrighan Perry, Sam Woo, Benjamin C Reeves, Huanxing Sun, Julia Winkler, Jean E Kanyo, Weiwei Wang, Milica Vukmirovic, Nicholas Ristic, Eric A Stratton, Sita Ram Meena, Maksym Minasyan, Daniel Kurbanov, Xinran Liu, TuKiet T Lam, Giuseppina Farina, Jose L Gomez, Mridu Gulati, Erica L Herzog. Bioactive Plasma Mitochondrial DNA Is Associated With Disease Progression in Scleroderma-Associated Interstitial Lung Disease. Arthritis & rheumatology (Hoboken, N.J.). 2020 Nov;72(11):1905-1915
PMID: 32602227
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