Editas Medicine, Inc. · NASDAQ:EDIT
Editas Medicine — Keystone Symposia, Engineering the Genome/Emerging Cellular Therapies 2020
25 slides from Editas Medicine's conference presentation, Feb 10, 2020
Slide 1: Ex vivo applications of Cas12a
Ex vivo applications of Cas12a Richard A Morgan Keystone Symposia, Engineering the Genome/Emerging Cellular Therapies Banff AB, Canada February 10, 2020 editasmedicine.com © 2020 Editas Medicine 1
Slide 2: Disclosures
Disclosures ▪ Rick Morgan is a stockowner of and employee at Editas Medicine, Inc. © 2020 Editas Medicine 2
Slide 3: Cas12a: The Other CRISPR Nuclease
Cas12a: The Other CRISPR Nuclease Editas general suite of nucleases Variant PAM Frequency (bp) SpCas9 Cas9 AsCas12a Cpf1 (Cas12a) SpCas9 NGG 1 in 8 SaCas9 NNGRRT 1 in 32 SaCas9 KKH NNNRRT 1 in 8 Targets G-rich PAMs Targets T- and C-rich PAMs AsCas12a TTTV 1 in 43 Separate crRNA and trRNA that Natura
Slide 4: Cas12a Applications for Engineered Cell Medicines: Sickle Cell Disease, NK Cell Therapy for Cancer
Cas12a Applications for Engineered Cell Medicines: Sickle Cell Disease NK Cell Therapy for Cancer editasmedicine.com © 2020 Editas Medicine 4
Slide 5: Etiology of Sickle Cell Disease
Etiology of Sickle Cell Disease Amino Hemoglobin Low DNA Acid Tetramer O2 a a a a b-globin GLU GAG b b b b HbA a a a a a a a a bs-globin G T G VAL b s b s b s bs bs bs b s bs HbS HbS Fiber • Sickle cell disease (SCD) is caused by a single mutation E6V of the b-globin chain, leading to polymerization
Slide 6: Harnessing Natural Anti-sickling Hemoglobin to Treat Sickle Cell Disease
Harnessing Natural Anti-sickling Hemoglobin to Treat Sickle Cell Disease Embryo Fetus Adult b-globin locus HBE HBG2 HBG1 HBD HBB Insulator Enhancer Insulator (5’HS) (HS1-4) (3’HS1) LCR a a a a g g HbS b b HbF HbA 100 % Globin Synthesis a 75 g b a a a a 50 g g b b Onset of Globin switch e SCD 25 Symp
Slide 7: Genome Editing to Reverse Hemoglobin Switching for Treating Sickle Cell Disease
Genome Editing to Reverse Hemoglobin Switching for Treating Sickle Cell Disease CRISPR-based disruption of HbF-repressing machinery Engineered patient Patient CD34+ cells CD34+ cells with switching reversed a a a a a a bs bs g g bs bs HbS HbF HbS From - Chang et al. Annual Meeting of the American So
Slide 8: Optimized CRISPR Cas12a-based Editing at the HBG Distal CCAAT box Region – EDIT-301
Optimized CRISPR Cas12a-based Editing at the HBG Distal CCAAT box Region – EDIT-301 From – De Dreuzy et al. Annual Meeting of the American Society of Hematology, Orlando. FL, December 9, 2019 © 2020 Editas Medicine 8
Slide 9: Cas12a Showed Improved Editing in HSCs with Desired Indel Size and Specificity
Cas12a Showed Improved Editing in HSCs with Desired Indel Size and Specificity From – De Dreuzy et al. Annual Meeting of the American Society of Hematology, Orlando. FL, December 9, 2019 © 2020 Editas Medicine 9
Slide 10: Editing HBG1/2 Induces Robust Fetal Hemoglobin In Vivo
Editing HBG1/2 Induces Robust Fetal Hemoglobin In Vivo Robust HbF in human CD34+ cells in vivo High pan-cellular human HbF in red blood cells 100 100% 100 100% HbF+ Red Blood Cells HbF / (HbF + HbA) 80 80 60 60 89% 40 40 52% 20 20 4% 4% 0 0 Unedited EDIT-301 HBG1/2 Unedited HBG1/2 HBG1/2 Edited Edit
Slide 11: Cas12a Editing for SCD
Cas12a Editing for SCD Cas12a produced more long term HbF inducing indels than SpCas9 at the HBG distal CCAAT box region. Greater than 90% indels were achieved after optimization of electroporation conditions and selection of best performing Cas12a variant and gRNA modifications EDIT-301 had no dete
Slide 12: Oncology Cell Therapy Development
Oncology Cell Therapy Development Autologous Start Patient Hospital Apheresis Centralized Production Hospital Patient COGs Scope Journey Manufacturing Time - Weeks Treatment $$$ Allogeneic Multiple Patient Healthy Apheresis/gene edit Expand Cells Hospital Treatments Donor $$ Off-the-shelf Multiple g
Slide 13: Potential Cell Types for Allogeneic Cell Medicines
Potential Cell Types for Allogeneic Cell Medicines Pluripotent Stem Cell CD34+ Cells ab T cells gd T cells NK cells Immune System Adaptive ADAPTIVE Innate INNATE Tumor Recognition • ab T cell receptor • gd T cell receptor • Innate receptors • CAR • Innate receptors • Antibody-directed • CAR • CAR Gr
Slide 14: Multiple Gene Edits Allow for Generation of an Off-the-shelf Allogeneic T Cell Product
Multiple Gene Edits Required for the Generation Multiple Gene Edits Allow for the Generation of an “Off-the-shelf”ofAllogeneic an “Off-the-shelf”TAllogeneic Cell Products T Cell Product MHC TCR Class I Host CIITA Epithelial Host CD8 Cell B2M T Cell Engineered MHC T Cell Class II Host CD4 T Cell Resp
Slide 15: Making an Allo T Cell: Cas12a Comparable to SpCas9
Making an Allo T Cell: Cas12a Comparable to SpCas9 Efficient Multiplexing by Cas12a 100 100 80 80 % Editing by NGS % Editing by NGS 60 Cpf1 Cas12a 60 Cas9 Cas9 40 40 20 20 0 TRAC B2M CIITA 0 TRAC B2M CIITA Zuris et al. Molecular Therapy Vol 27 No 4S1 April 2019 © 2020 Editas Medicine 15
Slide 16: Potential Cell Types for Allogeneic Cell Medicines
Potential Cell Types for Allogeneic Cell Medicines Pluripotent Stem Cell CD34+ Cells ab T cells gd T cells NK cells Immune System Adaptive ADAPTIVE Innate INNATE Tumor Recognition • ab T cell receptor • gd T cell receptor • Innate receptors • CAR • Innate receptors • Antibody-directed • CAR • CAR Gr
Slide 17: NK Cells Can Both Directly Kill Tumors and Stimulate the Endogenous Immune System
NK Cells Can Both Directly Kill Tumors and Stimulate the Endogenous Immune System Souza-Fonseca-Guimaraes, et al., Trends in Immunology, 2019, Vol. 40, 142-158 © 2020 Editas Medicine 17
Slide 18: TGF-β is a Major NK Cell Suppressive Cytokine
TGF-β is a Major NK Cell Suppressive Cytokine TGF-b isTGFβ is an immunosuppressive Overexpressed factorTumors in Many Solid abundant in the solid tumor microenvironment (mRNA expression) TGFB1 Log2 Fc Matched TCGA normal and GTEx database Stomach cancer HNSCC n=408 (pt) n=529 (pt) n=211 (normal) n=4
Slide 19: Potency Comparison of Engineered Cas12a and SpCas9 in NK Cells
Potency Comparison of Engineered Cas12a and SpCas9 in NK Cells at the Same Target Site in the Genome Potency of Cas12a vs. Cas9 in NK cells Control target site for comparing SpCas9 and AsCas12a 100 MS5 80 MS5 chosen from Kleinstiver et al. % Editing by NGS Nat Biotech 2016 vol 34, 869-874 Cas12a 60
Slide 20: Robust Gene Editing at TGFBR2 in Healthy Donor NK Cells with an Engineered Cas12a
Robust Gene Editing at TGFBR2 in Healthy Donor NK Cells with an Engineered Cas12a Rationale Editing Biology NK cell KO NK cells, minimal pSMAD2/3 Editing of TGFBR2 in three different donors in presence of TGF-β 100 D a ta 1 D Daa tt a a 1 1 6 6 6 pSMAD2/3 3 technical replicates lizeedd ppSSMMAADD22/
Slide 21: TGFBR2 Knockout NK Cells Showed Superior Effector Function than Unedited Control
TGFBR2 Knockout (KO) NK Cells Showed Superior Effector Function than Unedited Control NK Cells xCELLigence assay Assay readouts Target - SKOV3 NK cell addition • % Cytolysis by xCELLigence OvCa cells 0hr 24hr • IFNγ production by FACS Attached 2D tumor cell monolayer % Cytolysis %IFNγ+NK cells at 24
Slide 22: TGFBR2 KO Exerted Superior Control of Tumor Spheroids for >125hrs
TGFBR2 KO Exerted Superior Control of Tumor Spheroids for >125hrs in the Presence of TGF-β NK cell addition Spheroid assay Assay readouts TGF-b • Changes in spheroid size over time 0hr 125hr (killing by NK cells) (measurement of red object intensity) Tumor spheroid IL 1 5 v s IL 1 5 + T G F B 20:1 E
Slide 23: NK Therapeutic Strategy for Winning in Solid Tumors
NK Therapeutic Strategy for Winning in Solid Tumors Multiple innate receptors Multiple innate receptors NK CELL CARs Multiple innate receptors NK CELL ADCC NK CELL ADCC CRISPR editing Antibody-directed cellular cytotoxicity (ADCC) CRISPR editing Improved ADCC, persistence, Improved ADCC, persistence
Slide 24: Cas12a Editing for NK Cell-based Therapies
Cas12a Editing for NK Cell-based Therapies The next generation of allo and off-the-shelf cell therapies for cancer will require robust and specific gene editing. Cas12a produces efficient and specific gene editing, comparable or superior to SpCas9 in both primary T cells and NK cells. Greater than 9
Slide 25: Acknowledgements
Acknowledgements Sickle Cell Disease Team KaiHsin Chang, Edouard de Dreuzy, Jack Heath Healthy Donor NK Cell Team Karrie Wong, Chris Borges, John Zuris Editas Discovery, Development, Chemistry, and Operations Partners Bristol-Myers Squibb BlueRock Therapeutics Sandhill Therapeutics Integrated DNA Te
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