Slide 1: Antisense oligonucleotide mediated increase of OPA1 expression using TANGO for ADOA
Antisense oligonucleotide mediated increase of OPA1 expression using TANGO technology for the treatment of autosomal dominant optic atrophy Association for Research in Vision & Ophthalmology Meeting 2020 Aditya Venkatesh, Ph.D. Senior Scientist 1 © Copyright 2020 Stoke Therapeutics, Inc. All rights
Slide 2: Authors and disclosures
Authors and disclosures Authors: Aditya Venkatesh, Zhiyu Li, Syed Ali, Anne Christiansen, Kian Huat Lim, Jacob Kach, Robert Hufnagel, Jeffrey Hoger, Isabel Aznarez, Gene Liau 1. Stoke Therapeutics, Bedford, MA, United States. 2. Medical Genetics and Ophthalmic Genomics Unit, National Eye Institute,
Slide 3: Disclaimer
Disclaimer This presentation has been prepared by Stoke Therapeutics, Inc. ("Stoke") for informational purposes only and not for any other purpose. Nothing contained in this presentation is, or should be construed as, a recommendation, promise or representation by the presenter or Stoke or any offic
Slide 4: Autosomal dominant optic atrophy and OPA1
Autosomal dominant optic atrophy and OPA1 • Autosomal dominant optic atrophy (ADOA) is the most commonly diagnosed optic nerve disorder and is characterized by retinal ganglion cell loss • Disease affects approximately 1 in 30,000 people and causes progressive and irreversible vision loss • ADOA typ
Slide 5: Applying TANGO for the treatment of autosomal dominant haploinsufficiency diseases
Applying TANGO for the treatment of autosomal dominant haploinsufficiency diseases • Targeted augmentation of nuclear gene output (TANGO) uses antisense oligonucleotides (ASOs) to modulate splicing to precisely upregulate protein expression • TANGO ASOs reduce or prevent the generation of naturally
Slide 6: Advantages of TANGO for the treatment of ocular diseases
Advantages of TANGO for the treatment of ocular diseases • Intravitreal injection of ASOs permits diffusion throughout the eye and the ability to transduce retinal neurons • Potential for long-term efficacy (up to 1 year in mouse retina) after a single intravitreal injection (Kach et al, ARVO Poster
Slide 7: OPA1 non-productive splicing event identification and validation
OPA1 non-productive splicing event identification and validation • Novel exon inclusion event (Exon X) identified in the OPA1 gene which leads to introduction of a premature termination codon (PTC) • This produces a non-productive mRNA transcript that is degraded by non-sense mediated decay (NMD), p
Slide 8: OPA1 non-productive splicing event conservation
OPA1 non-productive splicing event conservation • OPA1 non-productive splicing event is conserved in non-human primates and rabbits • Abundance of event was detected to be approximately 20% in vivo in primate and rabbit ocular tissue Primate Rabbit %NMD exon inclusion P93 P942 Adult New Zealand (3 m
Slide 9: Selected OPA1-targeting ASOs reduce non-productive mRNA and increase productive mRNA in vitro
Selected OPA1-targeting ASOs reduce non-productive mRNA and increase productive mRNA in vitro RT-PCR for non-productive OPA1 mRNA ASO-10 ASO-11 ASO-12 ASO-13 ASO-14 ASO-15 ASO-15 ASO-17 ASO-18 ASO-1 ASO-2 ASO-3 ASO-4 ASO-5 ASO-6 ASO-7 ASO-8 ASO-9 • Cells: HEK293 cells No ASO • ASO conc.: 80nM • Deli
Slide 10: ASO-14 decreases non-productive OPA1 mRNA and increases OPA1 expression in a dose-dependent manner
ASO-14 decreases non-productive OPA1 mRNA and increases OPA1 expression in a dose-dependent manner in vitro Non-productive OPA1 mRNA Productive OPA1 mRNA OPA1 protein * * * Relative OPA1 mRNA expression 25 1.4 Relative OPA1 protein 1.2 * normalized to -actin 20 1.0 15 0.8 10 * 0.6 0.4 5 * 0.2 0 0.0
Slide 11: ASO-14 increases OPA1 protein expression in an OPA1 haploinsufficient cell line
ASO-14 increases OPA1 protein expression in an OPA1 haploinsufficient (OPA1+/-) cell line ASO-14 increases OPA1 protein levels in OPA1+/- HEK293 cells by ~50%, which translates to 75% of wild-type levels OPA1+/- ASO-14 No ASO 1nM 10nM 20nM 40nM 80nM OPA1 β-tubulin * * * • Cells: OPA1 +/- HEK293 cell
Slide 12: In vivo proof of concept study in wild-type rabbits
In vivo proof of concept study in wild-type rabbits Study Design Day 1 Single intravitreal (IVT) injection of vehicle/ASO (n=3/group) Female NZW rabbits Day 16 Euthanize animals and collect retinal tissue Final conc. Dose Dose volume Matrices collected Group N Treatment expected in (ug/eye) (uL/eye)
Slide 13: ASO increases OPA1 protein expression in vivo in wild-type rabbit retinae
ASO increases OPA1 protein expression in vivo in wild-type rabbit retinae Rabbit surrogate ASO decreases non-productive splicing and increases OPA1 protein expression in wild-type rabbit retinae following intravitreal injection. The test ASO was well tolerated for up to 15 days after injection Non-p
Slide 14: Conclusions
Conclusions Preclinical data support the potential use of Stoke’s TANGO technology in ADOA ASO-mediated specific reduction in non-productive OPA1 mRNA, increase in productive OPA1 mRNA and increase in OPA1 protein in a dose-dependent manner in vitro 50% increase in OPA1 protein levels in OPA1+/-
Slide 15: Questions?
Questions? Aditya Venkatesh, Ph.D. Senior Scientist avenkatesh@stoketherapeutics.com Dawn Kalmar Corporate Communications and Investor Relations Contact dkalmar@stoketherapeutics.com 15 © Copyright 2020 Stoke Therapeutics, Inc. All rights reserved