Editas Medicine, Inc. · NASDAQ:EDIT
Editas Medicine — Technical Presentation (May 2024)
13 slides from Editas Medicine's technical presentation, May 10, 2024
Slide 1: LNP-Based Delivery of CRISPR/Cas12a for the Potential Treatment of Myocilin-Associated Glaucoma
LNP-Based Delivery of CRISPR/Cas12a for the Potential Treatment of Myocilin-Associated Glaucoma Heather MacLeod May 10, 2024
Slide 2: Disclosure
Disclosure The speaker and the co-authors are current or former employees and stockholders of Editas Medicine, Inc. 2
Slide 3: Disease Background on Primary Open Angle Glaucoma (POAG)
Disease Background on Primary Open Angle Glaucoma (POAG) • Glaucoma is an optic neuropathy resulting in loss of peripheral vision followed by loss of central vison1–3 o A leading cause of irreversible blindness in the developing world1,2 • Primary open-angle glaucoma (POAG) is a subset of glaucoma d
Slide 4: Rationale for In Vivo Gene Editing via CRISPR/Cas Knockout of Myocilin
Rationale for In Vivo Gene Editing via CRISPR/Cas Knockout of Myocilin • The trabecular meshwork (TM) is responsible for regulating the outflow of aqueous humor and maintaining IOP at safe levels1,3 • Myocilin is a secreted protein of unknown function produced by the TM2,3 • Pathogenic gain-of-funct
Slide 5: Development of a Lipid Nanoparticle for Delivery of CRISPR/Cas Editing Machinery to the Trabecular Meshwork
Development of a Lipid Nanoparticle for Delivery of CRISPR/Cas Editing Machinery to the Trabecular Meshwork Lipid Nanoparticle (LNP) Chaotic Final LNP administered by injection into the Key for mixing via anterior chamber of the eye cellular microfluidic uptake device Cargo Figure adapted from Han X
Slide 6: Selection of an Ionizable Lipid that Robustly and Specifically Targets the TM Tissue
Selection of an Ionizable Lipid that Robustly and Specifically Targets the TM Tissue LNPs encapsulating EGFP mRNA were evaluated in mice in vivo for transfection of the TM tissue PBS Ionizable lipid #1 TM TM Ionizable lipid #2 Ionizable lipid #3 TM TM Trabecular meshwork (TM) area is outlined in red
Slide 7: Development of an Optimized LNP Cargo: Selection of gRNA with Chemical Modifications
Development of an Optimized LNP Cargo: Selection of gRNA with Chemical Modifications to Improve Editing In Vitro Editing in primary TM cells in vitro Human myocilin gene 100 Mod 1 Mod 2 80 Exon 1 Exon 2 Exon 3 Mod 3 Mod 4 Chr 1 60 Mod 5 Mod 6 % indel Mod 7 40 AsCas12a cut site 20 0 10-5 0 .0 0 0 0 1
Slide 8: Editing Myocilin In Vitro Reduces ER Stress Associated with Mutant Myocilin
Editing Myocilin In Vitro Reduces ER Stress Associated with Mutant Myocilin MYOC Editing Myocilin protein expression HEK293T cells stably expressed either MYOC normalized to GAPDH 3 100 (intracellular) wildtype or Y437H mutant myocilin 80 2 WT Y437H 60 Indel %%indel MYOC MYOC UNT 40 1 20 MYOC 0 0 10
Slide 9: Mice Expressing Human Myocilin with the Pathogenic Y437H Mutation Exhibit Elevated IOP
Mice Expressing Human Myocilin with the Pathogenic Y437H Mutation Exhibit Elevated IOP Full length human myocilin with the Y437H mutation replaced the mouse myocilin gene Mouse Chr 1 ATG STOP (+) 1 2 3 1.5 kb of hMYOC promoter Y437H IOP 16 **** **** **** HOM Y437H hMYOC IOP (mmHg) WT 14 12 0 2 3 4 5
Slide 10: LNP Delivery of AsCas12a mRNA and Modified gRNA Reduces Expression of hMYOC mRNA In Vivo
LNP Delivery of AsCas12a mRNA and Modified gRNA Reduces Expression of hMYOC mRNA In Vivo Mouse anterior chamber dissection Reduction of hMYOCY437H mRNA % of hMYOCY437H mRNA (compared to Veh) Veh Mod 5 Mod 7 Mod 3 • Data presented as mean ± SEM and analyzed by one-way ANOVA with Tukey’s multiple comp
Slide 11: Editing the Myocilin Gene In Vivo Significantly Reduces IOP in a Mouse Model of Myocilin-Associated POAG
Editing the Myocilin Gene In Vivo Significantly Reduces IOP in a Mouse Model of Myocilin-Associated POAG IOP in hMYOCY437H mice 18 Non-injected Veh 16 IOP (mmHg) 0.025 mg/mL 14 *** 0.1 mg/mL 0.5 mg/mL 12 *** * **** 95% CI of WT IOP 0 Pre- 2 Wk 4 Wk 6 Wk dosing MYOC gene editing resulted in a reducti
Slide 12: Summary and Conclusions
Summary and Conclusions Successful development and optimization of an LNP for in vivo gene editing in myocilin- associated POAG • Identified an ionizable lipid and lipid ratios that robustly and specifically transfects the TM • Established modifications to gRNA that improve in vivo editing • Develop
Slide 13: Thank you
Thank you 13