June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
RNAi BASED APPROACH FOR MYOC-ASSOCIATED GLAUCOMA
Author Affiliations & Notes
  • Gaurang Patel
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • Brijeshkumar Patel
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • Charleen Hunt
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • Sarthak R Zaveri
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • Sabrina Walley
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • rajeevalochan wudali
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • Sven-Moller Tank
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • James Mcninch
    Alnylam Pharmaceuticals Inc, Cambridge, Massachusetts, United States
  • Mark Schlegel
    Alnylam Pharmaceuticals Inc, Cambridge, Massachusetts, United States
  • Adam Castoreno
    Alnylam Pharmaceuticals Inc, Cambridge, Massachusetts, United States
  • Elena Castellanos-Rizaldos
    Alnylam Pharmaceuticals Inc, Cambridge, Massachusetts, United States
  • Abigail Liebow
    Alnylam Pharmaceuticals Inc, Cambridge, Massachusetts, United States
  • Kavita Praveen
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • Ying Hu
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • Vasant Jadhav
    Alnylam Pharmaceuticals Inc, Cambridge, Massachusetts, United States
  • Carl Romano
    Regeneron Pharmaceuticals Inc, Tarrytown, New York, United States
  • Footnotes
    Commercial Relationships   Gaurang Patel Regeneron, Code E (Employment); Brijeshkumar Patel Regeneron, Code E (Employment); Charleen Hunt Regeneron, Code E (Employment); Sarthak Zaveri Regeneron, Code E (Employment); Sabrina Walley Regeneron, Code E (Employment); rajeevalochan wudali Regeneron, Code E (Employment); Sven-Moller Tank Regeneron, Code E (Employment); James Mcninch Alnylam, Code E (Employment); Mark Schlegel Alnylam, Code E (Employment); Adam Castoreno Alnylam, Code E (Employment); Elena Castellanos-Rizaldos Alnylam, Code E (Employment); Abigail Liebow Alnylam, Code E (Employment); Kavita Praveen Regeneron, Code E (Employment); Ying Hu Regeneron, Code E (Employment); Vasant Jadhav Alnylam, Code E (Employment); Carl Romano Regeneron, Code E (Employment)
  • Footnotes
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Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3092. doi:
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      Gaurang Patel, Brijeshkumar Patel, Charleen Hunt, Sarthak R Zaveri, Sabrina Walley, rajeevalochan wudali, Sven-Moller Tank, James Mcninch, Mark Schlegel, Adam Castoreno, Elena Castellanos-Rizaldos, Abigail Liebow, Kavita Praveen, Ying Hu, Vasant Jadhav, Carl Romano; RNAi BASED APPROACH FOR MYOC-ASSOCIATED GLAUCOMA. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3092.

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

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Abstract

Purpose : Mutations in Myocilin (MYOC) are responsible for the most common genetic cause of glaucoma, accounting for 8-10% of autosomal dominant familial Juvenile Open Angle Glaucoma cases, as well as 2-3% of POAG cases. MYOC toxic gain of function mutant proteins aggregate intracellularly, leading to trabecular meshwork (TM) stress, elevated IOP, and glaucoma. In this study, we tested the hypothesis that silencing expression of disease-causing MYOC variants could lead to decreased TM stress and lowered IOP.

Methods : First, we made humanized MYOC mice to screen different siRNA duplexes in vivo. Next, we developed humanized MYOC.Y437H mice over-expressing human mutant MYOC using a CRISPRa approach (SAM-MYOC mice). We also validated the model using IOP lowering drugs (timolol and Rhopressa). We then tested several lead MYOC siRNA duplexes in the SAM-MYOC model to assess the IOP lowering efficacy. In addition, we also used ex vivo human eye perfusion culture model to screen this lead siRNA duplexes

Results : First, we found the several best siRNAs duplexes to lower both MYOC mRNA and protein. To develop animal model, out of several CRISPR guides delivered via AAV or Lentivirus in mouse eye, one guide led to significant upregulation (15-20-fold) of mutant MYOC and high IOP (5-6 mmHg from baseline). We showed that both timolol and Rhopressa lowered IOP in these mice, further validating the model. Next, 5 weeks after virally inducing high IOP in SAM-MYOC, we injected either control or MYOC siRNA intravitreally. While the control siRNA had no effect on IOP, in the MYOC siRNA group IOP was lowered to baseline within a week and this effect persisted for more than 5.5 months, when IOP elevated again gradually. Second, we used the ex vivo human eye perfusion culture system to explore the efficacy of MYOC siRNA to knockdown MYOC mRNA and protein in the TM and MYOC protein levels in the perfusate. Over 90% knockdown of both RNA and protein was achieved.

Conclusions : These results suggest that siRNA based knockdown of MYOC is a feasible therapeutic approach to treat human MYOC based glaucoma.

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

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