June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Mitochondrial DNA sensing by STING pathway mediates complement, inflammation and epigenetic genes
Author Affiliations & Notes
  • Cristina M Kenney
    Ophthalmology, Gavin Herbert Eye Inst, UC Irvine, Irvine, California, United States
  • Kevin Schneider
    Ophthalmology, Gavin Herbert Eye Inst, UC Irvine, Irvine, California, United States
  • Shari R. Atilano
    Ophthalmology, Gavin Herbert Eye Inst, UC Irvine, Irvine, California, United States
  • Marilyn Chwa
    Ophthalmology, Gavin Herbert Eye Inst, UC Irvine, Irvine, California, United States
  • Sonali R Nashine
    Ophthalmology, Gavin Herbert Eye Inst, UC Irvine, Irvine, California, United States
  • Stephanie Yiwei Lu
    Ophthalmology, Gavin Herbert Eye Inst, UC Irvine, Irvine, California, United States
    Ophthalmology, Long Beach VA Medical Center, Long Beach, California, United States
  • Anthony B Nesburn
    Ophthalmology, Gavin Herbert Eye Inst, UC Irvine, Irvine, California, United States
  • Baruch D Kuppermann
    Ophthalmology, Gavin Herbert Eye Inst, UC Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Cristina Kenney, None; Kevin Schneider, None; Shari R. Atilano, None; Marilyn Chwa, None; Sonali Nashine, None; Stephanie Lu, None; Anthony Nesburn, None; Baruch Kuppermann, Alcon, Allergan, Apellis, Genentech, GSK, Ophthotech, Regeneron (F), Alcon, Allergan, Catalyst, Genentech, Novartis, Ophthotech, Regeneron (C), Allergan, Genentech, Novartis, Regeneron (R)
  • Footnotes
    Support  Funding Supported by Discovery Eye Foundation, Polly and Michael Smith, Iris and B. Gerald Cantor Foundation, Max Factor Family Foundation. Supported by an RPB Unrestricted Grant
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 5251. doi:
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      Cristina M Kenney, Kevin Schneider, Shari R. Atilano, Marilyn Chwa, Sonali R Nashine, Stephanie Yiwei Lu, Anthony B Nesburn, Baruch D Kuppermann; Mitochondrial DNA sensing by STING pathway mediates complement, inflammation and epigenetic genes. Invest. Ophthalmol. Vis. Sci. 2017;58(8):5251.

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

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Abstract

Purpose : Mitochondrial (mt) DNA haplogroups represent populations of diverse geographic origins. H is the most common European haplogroup, while K haplogroup is highly associated with the Ashkenazi Jewish population. Previously we showed that although the RPE cybrids (cytoplasmic hybrids) have identical nuclei, cybrids with K haplogroup mtDNA have: (1) significantly increased expression of ApoE, a critical lipid transporter molecule; (2) increased expression of complement inhibitors and inflammation-related genes; and (3) differential RNA expression of epigenetic genes compared to H cybrids (Thaker et al, 2016 Neurobio Dis 93:64-77). Our study was designed to determine if the STING (Stimulator of Interferon Genes) sensor system (TMEM173 gene) is involved in retrograde signaling from mitochondria to the nuclei with respect to lipid transport, complement and inflammation pathways.

Methods : Cybrids were generated by fusing platelets from either H or K haplogroups subjects with Rho0 (lacking mtDNA) human ARPE-19 cells. STING was knockdown (KD) in H cybrids (n=5) and K cybrids (n=5) by transfecting the cells with 30pmol siRNA or Silencer negative control (Lipofectamine RNAiMAX , Invitrogen, Life Technologies) according to the manufacturer’s protocol. After 48 hrs, RNA was isolated (PureLink RNA Isolation Kit, Life Technology). Expression levels of genes were measured by qRT-PCR. Statistical analyses were performed using unpaired t-test.

Results : STING expression knockdown (KD) was 87.2%, p=0.0028 in H cybrids and 90.5%, p<0.0001 in K cybrids. After STING-KD, K cybrids showed decreases in CFI (36%, p=0.005) and CFH (21%, p=0.036) but increased transcription levels in IL6 (187%, p=0.02) and IFNb (148%, p=0.46) compared to K Control cybrids (100%). DNMT1 expression levels were increased significantly in both cybrids (H: 175%, p<0.0001; K: 210%, p=0.0009). There were decreased levels in the STING-KD cybrids for DNMT3A (H: 34%, p=0.05; K: 36%, p=0.03) and HDAC1 (H; 32%, p=0.016; K: 32%, p=0.0009) vs Control cybrids (100%). STING-KD did not affect expression levels for APOE, APOC1, CD55, CD59, DNMT3B, TRDMT1, HDAC2, HDAC4, HDAC6, HDAC11 or HAT1.

Conclusions : Within cells, the intracellular STING DNA sensor system plays a role in regulation of 3 epigenetic genes (DNMT1, DNMT3A and HDAC1), 2 complement inhibitors (CFI and CFH) and 2 pro-inflammatory genes (IL6 and IFNb).

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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