June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Self-DNA induces inflammation in myoepithelial cells of the lacrimal gland by AIM2 inflammasome activation
Author Affiliations & Notes
  • Menglu Yang
    Ophthalmology, Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
  • Darlene A Dartt
    Ophthalmology, Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Menglu Yang None; Darlene Dartt None
  • Footnotes
    Support  RO1 EY019470, RO1 EY026202
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 2000 – A0330. doi:
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    • Get Citation

      Menglu Yang, Darlene A Dartt; Self-DNA induces inflammation in myoepithelial cells of the lacrimal gland by AIM2 inflammasome activation. Invest. Ophthalmol. Vis. Sci. 2022;63(7):2000 – A0330.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Primary Sjögren's Syndrome (pSS) is an autoimmune disease that mainly affects the lacrimal glands and salivary glands. Antibodies against nuclear materials have been found in patients but the mechanism is unclear. When foreign DNA enters the cells, several mechanisms will be activated to defend against viral infection or local tissue damage. In this study, we focus on one of the mechanisms, namely Absent in Melanoma (AIM)2 inflammasome, in myoepithelial cells of the lacrimal gland. AIM2 non-selectively binds the double-stranded DNA in the cytoplasm, causing the formation of the AIM2 inflammasome to activate caspase-1, which cleaves immature pro-IL-1ß to mature IL-1ß that contribute to inflammation. We hypothesize that in pSS patients, self-DNA from the bloodstream of SS patients triggers the activation of AIM2 inflammasome which in turn causes inflammation to damage the healthy lacrimal gland.

Methods : Primary MECs culture was acquired from female wildtype C57Bl/6 mice aged 4-6 weeks. Genomic DNA (gDNA) extracted from the same culture was used to resemble the self-DNA. A random double-stranded DNA poly A:T was used as a positive control for AIM2 activation. Immunofluorescent (IF) was used to detect the assembly of inflammasomes; RT-PCR was used to detect the expression of AIM2. The activation of caspase-1 was measured using FAM-FLICA assay.

Results : The MECs were treated with gDNA for 6 hours, with random double-stranded DNA poly A:T as a positive control. Immunofluorescent (IF) staining result shows the accumulation of AIM2 signals and a speck of Apoptosis-associated speck-like protein containing a CARD (ASC), indicating the assembly of AIM2 inflammasome in both gDNA and poly AT treated groups. RT-PCR showed a significantly increased expression of Aim2 in both gDNA and poly AT treated groups compared to the non-treated group. FAM-FLICA assay showed that the activation of caspase-1 is significantly increased in both poly AT and gDNA treated groups compared to the non-treated group. We next applied the inhibitor of AIM2, A151, to the MECs together with gDNA or poly AT, and the ratio of AIM2 or ASC speck positive cells was measured from each group. We found that A151 significantly terminated the presence of AIM2 and ASC speck compared to the gDNA or poly AT group.

Conclusions : Self-DNA induces caspase-1 activation in MECs through AIM2 inflammasome, which may be a key mechanism in pSS.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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