Slide 1: Stereopure AIMers for the treatment of Alpha-1 antitrypsin deficiency (AATD)
Stereopure AIMers for the treatment of Alpha-1 antitrypsin deficiency (AATD) Paloma H. Giangrande, PhD VP Platform Discovery Sciences, Biology TIDES US: May 9-12, 2022
Slide 2: Forward looking statements
Forward looking statements This document contains forward-looking statements. All statements other than statements of historical facts contained in this document, including statements regarding possible or assumed future results of operations, preclinical and clinical studies, business strategies, r
Slide 3: Unlocking RNA editing with PRISM to develop GalNAc-AIMers
Unlocking RNA editing with PRISMTM to develop GalNAc-AIMers: A-to-I editing oligonucleotides Optimize AIMer design for endogenous transcripts and GalNAc conjugation GalNAc-mediated uptake ADAR enzymes PRISM-driven gains • First publication (1995) using oligonucleotide to Primary human edit RNA with
Slide 4: Proof-of-concept RNA editing in NHP liver is robust, durable, and specific
Proof-of-concept RNA editing in NHP liver is robust, durable, and specific AIMers detected in liver of NHP Durable editing in NHP liver in ADAR editing with ACTB AIMer is at day 50 vivo highly specific pre-dose RNA editing within full transcriptome (primary human hepatocytes) GalNAc AIMers Day 50 Co
Slide 5: Proof-of-concept preclinical RNA editing data published in Nature Biotechnology (March 2022)
Proof-of-concept preclinical RNA editing data published in Nature Biotechnology (March 2022) • Foundational AIMer SAR • GalNAc conjugation • In vitro-in vivo translation (NHPs) • Specificity in vitro & in vivo (NHPs) Monian et al., 2022 published online Mar 7, 2022; doi: 10.1038.s41587-022-01225-1 S
Slide 6: AIMers: Realizing the potential of therapeutic RNA editing by harnessing endogenous ADAR
AIMers: Realizing the potential of therapeutic RNA editing by harnessing endogenous ADAR Solved for key therapeutic attributes for potential best-in-class RNA editing therapeutics Delivery and Efficient ADAR Efficient ADAR recruitment Stability intracellular Beyond liver recruitment trafficking • A
Slide 7: SERPINA1 Z mutation: The most common cause of Alpha-1 antitrypsin deficiency (AATD)
SERPINA1 Z mutation: The most common cause of Alpha-1 antitrypsin deficiency (AATD) Wild-type M-AAT protein AAT is an acute phase protein Serum AAT protects lungs predominately produced against neutrophil elastase by hepatocytes Strnad et al., 2020 N Engl J Med 382:1443-55; Stoller et al., 1993 Alph
Slide 8: SERPINA1 Z mutation: The most common cause of Alpha-1 antitrypsin deficiency (AATD)
SERPINA1 Z mutation: The most common cause of Alpha-1 antitrypsin deficiency (AATD) Wild-type M-AAT protein AAT is an acute phase protein Serum AAT protects lungs predominately produced against neutrophil elastase by hepatocytes Gain-of-function and loss-of-function disease Liver Disease Lung Diseas
Slide 9: RNA editing is uniquely suited to address the therapeutic goals of AATD
RNA editing is uniquely suited to address the therapeutic goals of AATD Wave ADAR editing approach addresses all treatment goals: 1) Restore circulating, 2) Reduce Z-AAT protein 3) Retain M-AAT physiological functional wild-type M-AAT aggregation in liver regulation Risk of disease Null Highest (no
Slide 10: Robust SERPINA1 RNA editing in mouse model for AATD restores M-AAT protein in serum
Robust SERPINA1 RNA editing in mouse model for AATD restores M-AAT protein in serum RNA editing in mice day 7 AAT serum protein concentration in mice day 7 (3x5 mg/kg, SC) (3x10 mg/kg, SC) * 60 1000 1000 MZ (50%) % SERPINA1 editing 800 800 Serum AAT protein (ug/ml) 40 (Mean, s.e.m) 600 11 µM 600 11
Slide 11: Bi-weekly AIMer treatment results in serum AAT protein levels in mice above anticipated therapeutic threshold
Bi-weekly AIMer treatment results in serum AAT protein levels in mice above anticipated therapeutic threshold1 GalNAc-AIMer results in serum AAT protein levels >11 uM at week 19 1200 18.5 uM Serum AAT protein (µg/mL) 1000 SA1-5 (Mean, SEM) 800 600 11 uM ~5-fold increase 400 in AAT protein 200 PBS 0
Slide 12: Bi-weekly AIMer treatment supports robust RNA editing and M-AAT protein expression in mice
Bi-weekly AIMer treatment supports robust RNA editing and M-AAT protein expression in mice GalNAc-AIMer results in serum AAT protein levels >11 uM at week 19 1200 18.5 uM ~60% RNA editing & ~70% serum M-AAT protein in Serum AAT protein (µg/mL) 1000 SA1-5 mice (week 19 data) 80 100 % Serum AAT Isofor
Slide 13: Restoration of functional serum M-AAT protein that neutralizes protease activity in mice
Restoration of functional serum M-AAT protein that neutralizes protease activity in mice ~3-fold in increase in neutrophil elastase inhibition at Correction of loss-of-function week 19 phenotypes *** % Relative elastase inhibition 100 PBS 80 SA1-5 (mean, sem) 60 40 Restored M-AAT reaches 20 lungs to
Slide 14: SA1-5 alleviates aggregation of Z-AAT and inflammation in mouse liver
SA1-5 alleviates aggregation of Z-AAT and inflammation in mouse liver Lobular inflammation (19 weeks) Correction of gain-of-function phenotypes PBS SA1-5 Z-AAT aggregation causes progressive liver disease Fibrosis Cirrhosis Hepatocellular Carcinoma PAS-D staining PAS-D-positive area (19 weeks) L
Slide 15: AIMer-directed editing is highly specific in mice
AIMer-directed editing is highly specific in mice No bystander editing observed on SERPINA1 transcript RNA editing only detected at PiZ mutation site in SERPINA1 transcript (mouse liver) RNA editing across transcriptome (mouse liver) C 0% SERPINA1 PBS (PiZ mutation site) T 100% Coverage Coverage C 4
Slide 16: Opportunity for novel and innovative AIMer therapeutics
Opportunity for novel and innovative AIMer therapeutics Correct driver mutations with AIMers Free-uptake of chemically modified oligonucleotides Restore or correct protein function Examples AATD Rett syndrome Recessive or dominant genetically defined diseases Examples Haploinsufficient diseases Loss
Slide 17: Summary
Summary • Our RNA base editing platform capability allows for correction of the most common causative mutation for AATD in preclinical models • We have developed RNA editing oligonucleotides – AIMers – intended to correct homozygous "ZZ" mutations back to "MZ" heterozygous state • Investigational AI
Slide 18: Acknowledgements
Acknowledgements • Colleagues and contributors from Wave Life Sciences 18