Editas Medicine, Inc. · NASDAQ:EDIT
Editas Medicine — TIDES USA 2021
25 slides from Editas Medicine's conference presentation, Sep 23, 2021
Slide 1: An Engineered AsCas12a Nuclease Facilitates the Rapid Generation of Therapeutic Cell Medicines
An Engineered AsCas12a Nuclease Facilitates the Rapid Generation of Therapeutic Cell Medicines Dr. John Zuris Associate Director, Editing Technologies Editas Medicine editasmedicine.com © 2021 Editas Medicine 1
Slide 2: Disclosure
Disclosure § I am an employee and shareholder of Editas Medicine © 2021 Editas Medicine 2
Slide 3: Overview of Presentation
Overview of Presentation § Collaboration with IDT to try and engineer a highly active AsCas12a variant GENERATION OF A HIGHLY § Assessment of AsCas12a editing across targets and cell types ACTIVE ASCAS12A VARIANT § Assessment of how AsCas12a Ultra specificity compared to WT nuclease CALLED ASCAS12A
Slide 4: This Work was Part of a Collaboration with Integrated DNA Technologies
This Work was Part of a Collaboration with Integrated DNA Technologies DIRECTED EVOLUTION OF ENGINEERED OPTIMIZATION OF CLINICALLY RELEVANT KNOCK ASCAS12A AND TESTING IN CANCER CELL LINES OUT AND KNOCK-IN EDITS IN PRIMARY CELLS § Liyang Zhang § Ramya Viswanathan § Chris Vakulskas § Jasmine Edelstein
Slide 5: Multiplexed Gene Editing is Key for the Next Generation of Cell Medicines
Multiplexed Gene Editing is Key for the Next Generation of Cell Medicines Proposed Allogeneic T cell Edits Proposed iNK Edits MHC TCR Class I Host CIITA Host CD8 Epithelial B2M Response T Cell Cell + Engineered MHC T Cell Class II Host CD4 T Cell Edit MHC Class I Host CD8 TCR T Cell Host CIITA Epith
Slide 6: There are Several CRISPR Nuclease Options for the Potential Generation of Engineered Cell Medicines
There are Several CRISPR Nuclease Options for the Potential Generation of Engineered Cell Medicines Editas general suite of CRISPR nucleases Variant PAM Frequency (bp) Cas9 SpCas9 NGG 1 in 8 SaCas9 NNGRRT 1 in 32 SaCas9 KKH NNNRRT 1 in 8 AsCas12a TTTV 1 in 43 Cas12a AsCas12a RR TYCV/CCCC 1 in 18 AsC
Slide 7: The AsCas12a Nuclease has Higher Intrinsic Specificity and Higher Sequence Fidelity
The AsCas12a Nuclease has Higher Intrinsic Specificity and Higher Sequence Fidelity in the Shorter Chemically Synthesized Guide RNA Specificity: Guide RNA synthesis: Matched Target Site (20-Ns): Chemical synthesis of gRNAs occurs in the 3’ à 5’ direction and purity TTTVNNNNNNNNNNNNNNNNNNNNNGG and yi
Slide 8: Given the Clear Specificity Advantages of AsCas12a Why has it Not Been Widely Adopted
Given the Clear Specificity Advantages of AsCas12a Why has it Not Been Widely Adopted in Ex Vivo Applications in Research and the Clinic? Assessment of WT AsCas12a editing and HDR efficiency Cancer cell lines Primary cells HDR in cancer cell lines WT AsCas12a is generally inefficient for both gene K
Slide 9: Bacterial Selection Experiment Revealed a Mutant AsCas12a Protein with Potentially Higher Activity
Bacterial Selection Experiment Performed by IDT Team Revealed a Mutant AsCas12a Protein with Potentially Higher Activity M537R and F870L point mutations were combined and taken forward to make AsCas12a “Ultra” Zhang et al. Nature Communications 2021 © 2021 Editas Medicine 9
Slide 10: AsCas12a Ultra is 40-fold More Potent Compared to WT AsCas12a
AsCas12a Ultra is 40-fold More Potent On Average Compared to WT AsCas12a Across All Potential Targets at the B2M Locus in T cells 40-fold increase in EC50 © 2021 Editas Medicine 10
Slide 11: AsCas12a Ultra Shows Highly Efficient Editing Across All Guides Tested
AsCas12a Ultra Shows Highly Efficient Editing Across All Guides Tested Across Different Loci and Across All Tested Cell Types Assessment of WT AsCas12a editing and HDR efficiency Cancer cell lines Primary cells HDR in cancer cell lines WT Ultra © 2021 Editas Medicine 11
Slide 12: AsCas12a Ultra Enables the Generation of Engineered Cell Medicines
AsCas12a Ultra Enables the Generation of Engineered Cell Medicines Zhang et al. Nature Communications 2021 © 2021 Editas Medicine 12
Slide 13: Specificity is Retained with AsCas12a Ultra
Specificity is Retained with AsCas12a Ultra On-target Editing in GUIDE-Seq Experiment No Off-targets Detected by GUIDE-Seq We continually see this high specificity across current or potential future clinical targets Zhang et al. Nature Communications 2021 © 2021 Editas Medicine 13
Slide 14: AsCas12a Ultra is More Potent than enCas12a and HiFi Variant
AsCas12a Ultra is More Potent than enCas12a and HiFi Variant enCas12a and enCas12a HiFi shown to have higher activity and expanded PAM targeting range to WT AsCas12a as reported in Kleinstiver et al. Nat Biotech 2019 Key Takeaway: for maximal editing efficiency AsCas12a Ultra should be nuclease of c
Slide 15: AsCas12a Ultra has Superior Potency Across Key Primary Cell Types
AsCas12a Ultra has Superior Potency Across Key Primary Cell Types Potency across a wide range of targets Case example in T cells Case examples in HSPCs AsCas12a Ultra is 4-fold more active than enCas12a Tested at suboptimal Sequence specificities of enCas12a enCas12a-Ultra concentration of RNP enCas
Slide 16: AsCas12a Ultra Allows for Highly Efficient Multiplexed Editing in T cells
AsCas12a Ultra Allows for Highly Efficient Multiplexed Editing in T cells Zhang et al. Nature Communications 2021 © 2021 Editas Medicine 16
Slide 17: AsCas12a Ultra Allows for Highly Efficient Single and Multigene Knock-in
AsCas12a Ultra Allows for Highly Efficient Single and Multigene Knock-in DKO + B2M – mCherry AAV + TRAC – GFP AAV TRAC – GFP AAV only mCherry+ mCherry+ GFP+ GFP+ B2M - mCherry AAV only B2M + TRAC DKO only mCherry+ MHCI+ GFP+ TCR+ Zhang et al. Nature Communications 2021 © 2021 Editas Medicine 17
Slide 18: AsCas12a Ultra Generates Highly Efficient KO of a Tumor Suppressive Cytokine Receptor
AsCas12a Ultra Generates Highly Efficient KO of a Tumor Suppressive Cytokine Receptor Which Leads to Enhanced NK Cell Killing Function TGFBR2 KO to inhibit suppression of NK cells by TGF-b TGFBR2 editing is highly efficiency with AsCas12a Ultra Functional assay shows decreased SMAD phosphorylation I
Slide 19: AsCas12a Ultra Enables Efficient Knock-in of Many Transgenes Including an EGFR CAR
AsCas12a Ultra Enables Efficient Knock-in of Many Transgenes Including an EGFR CAR Which Leads to Increased NK Cell Killing of Tumors Cells Key takeaway: AsCas12a can be leveraged for AAV6-mediated knock-in of a CAR in NK cells Zhang et al. Nature Communications 2021 © 2021 Editas Medicine 19
Slide 20: AsCas12a Ultra Enables Efficient Knock-in Using AAV6 in HSPCs
AsCas12a Ultra Enables Efficient Knock-in Using AAV6 in HSPCs Optimization of Knock-in in HSCs at B2M locus % MHC– mCherry+ by Flow Cytometry 40 4 µM RNP HSPCs 30 20 10 0 1 10 100 MOI x 104 (vg/cell) MHC+ MHC+ MHC+ mCherry+ mCherry+ mCherry+ Zhang et al. Nature Communications 2021 © 2021 Editas Medi
Slide 21: Mutations Incorporated into an Engineered AsCas12a Used in EDIT-301 Sickle Cell Program
AsCas12a Ultra Mutations Incorporated into an Engineered AsCas12a at Editas which is Being Used in Our Sickle Cell Program: EDIT-301 Editing Specificity Editing Efficiency Indel Profile Optimized >100 100 Conditions 10 SpCas9 SpCas9 Cut-sites (Count) NHEJ Indels (%) 80 AsCas12a 80 8 AsCas12a Indels
Slide 22: New HDR Technology Leverages Engineered AsCas12a
New HDR Technology Leverages Engineered AsCas12a SLEEK = SeLection by Essential-gene Exon Knock-in SLEEK Efficiency in T cells Comparison to TRAC Knock-in Allogeneic Healthy Donor Applications ✱✱✱ 100 % GFP+ by Flow Cytometry 80 CAR/eTCR #1 (SLEEK) TRAC KO 60 CAR/eTCR #2 Additional KOs 40 (SLEEK) 20
Slide 23: Summary
Summary § In collaboration with IDT, we generated and tested a highly active variant of AsCas12a across many different clinically relevant targets and cell types § This highly active AsCas12a variant: – Achieves high editing rates with all cell types tested and is uniformly active across on-target s
Slide 24: Other Editas Presentations at TIDES USA 2021
For More Exciting Information on Editas’s CRISPR Platform Please See Our Other Presentations Taking Place at TIDES USA 2021 § Oral presentations: – IND-enabling Small-Scale Guide RNA Production Under GMP for CRISPR Based Cell Therapies – Keith Jarvis – Characterization of gRNAs and Ribonucleoprotein
Slide 25: We're Hiring
We’re Hiring Come Work on Our Engineered AsCas12a and Our Many Other Amazing Technologies and Bring CRISPR Medicines to Patients https://www.editasmedicine.com/careers/ https://www.linkedin.com/in/john-zuris-5a5b2537/ © 2021 Editas Medicine 25