Precision Biosciences, Inc. · NASDAQ:DTIL
Precision Biosciences — American Society of Gene and Cell Therapy 2023, Late-Breaking Abstracts 2
16 slides from Precision Biosciences's conference presentation, May 19, 2023
Slide 1: ARCUS nuclease-mediated excision of the Hot Spot region of the human dystrophin gene for DMD
ARCUS nuclease-mediated excision of the “Hot Spot” region of the human dystrophin gene for the treatment of Duchenne Muscular Dystrophy (DMD) American Society of Gene and Cell Therapy Session: Late-Breaking Abstracts 2 Gary Owens May 19, 2023
Slide 2: Forward-Looking Statements
Forward-Looking Statements This presentation contains forward-looking statements, as may any related presentations, within the meaning of the Private Securities Litigation Reform Act of 1995. The Company intends such forward-looking statements to be covered by the safe harbor provisions for forward-
Slide 3: Disclosures
Disclosures • I am an employee of Precision BioSciences, Inc. (Nasdaq: DTIL) 3
Slide 4: Duchenne Muscular Dystrophy Currently Lacks a Curative Treatment
Duchenne Muscular Dystrophy Currently Lacks a Curative Treatment Mutation on the X chromosome interferes On average, with dystrophin protein production, which is needed to form and maintain healthy children lose muscle their ability to walk by Affects approximately age 12 1 in 3,500 live male births
Slide 5: ARCUS: Engineering Nature's Genome Editing System
ARCUS: Engineering Nature’s Genome Editing System ARCUS • Derived from I-CreI, a naturally-occurring green algae homing endonuclease • Target site recognition and cleavage rely solely on an extensive DNA-protein interface • DNA cleavage results in 4 bp 3’ sticky ends • Small size (364 aa) facilitate
Slide 6: Utilizing ARCUS Nucleases to Restore Dystrophin Expression for DMD
Utilizing ARCUS Nucleases to Restore Dystrophin Expression for DMD Strategy to restore dystrophin Delete exons 45-55 using a pair of ARCUS nucleases intended to remove a expression mutation hotspot responsible for approximately 50% of DMD cases ARCUS 1 ARCUS 2 GOAL Exon 44 Exons 45–55 Exon 56 Exon 5
Slide 7: Dystrophin Gene Correction Observed in DMD Patient Myoblasts
Dystrophin Gene Correction Observed in DMD Patient Myoblasts Genomic Editing Dystrophin Protein Expression 40 30 Excision (%) 20 10 Dystrophin 0 mock 10ng 20ng 40ng 80ng 160ng ARCUS mRNA delivered ARCUS mRNA delivered Exons 45-55 deleted Exon 44 spliced to Exon 56 Dystrophin protein expressed Trunca
Slide 8: In Vivo POC Study Design
In Vivo POC Study Design • Objective: Assess muscle function in a humanized, murine model of DMD • Test Article: A single AAV with 2 early generation ARCUS nucleases, expression driven by a muscle specific promoter • Mouse Models: Group Mouse Strain N - hDMD/mdx * • Contains human dystrophin gene an
Slide 9: Excision of the Hot Spot Region of the Dystrophin Gene in Target Tissues
Excision of the “Hot Spot Region” of the Dystrophin Gene in Target Tissues Quadricep Gastrocnemius Heart Diaphragm 25 25 8 6 20 20 6 Excision (%) Excision (%) Excision (%) Excision (%) 4 15 15 4 10 10 2 2 5 5 0 0 0 0 d d d d d d d d te te te te te te te te ea ea ea ea ea ea ea ea Tr r Tr r Tr Tr r r
Slide 10: Edited Dystrophin Protein Variant Expressed in Target Tissues
Edited Dystrophin Protein Variant Expressed in Target Tissues Quadricep Gastrocnemius Heart Diaphragm 20 8 20 10 % Dystrophin Restoration % Dystrophin Restoration % Dystrophin Restoration % Dystrophin Restoration 8 15 6 15 6 10 4 10 4 5 2 5 2 0 0 d 0 0 d d d d d d d te te te te te te te te ea ea ea
Slide 11: Maximum Force Output of Gastrocnemius Muscle in ARCUS-Treated Mice was Significantly Improved
Maximum Force Output of Gastrocnemius Muscle in ARCUS-Treated Mice was Significantly Improved 400 hDMD/mdx (#3) *# *# 300 *# treated hDMDdel52/mdx (#1) Force (mN) untreated hDMDdel52/mdx (#2) *# 200 *# *# 100 *# *# *p<0.05 Group 1 vs 2 by Two-Way RM ANOVA #p<0.05 Group 3 vs 2 by Two-Way RM ANOVA 0 0
Slide 12: Restoration of Dystrophin in Gastrocnemius Muscle
Restoration of Dystrophin in Gastrocnemius Muscle Dystrophin Laminin Untreated Green = Dystrophin Non-Disease Model Purple = Laminin (hDMD/mdx) Untreated Disease Model (hDMDdel52/mdx) Treated Disease Model (hDMDdel52/mdx) 12
Slide 13: BaseScope Duplex Assay
BaseScopeTM Duplex Assay • BaseScope TM Duplex Assay can be used for simultaneous visualization of two RNA targets while maintaining single cell resolution • Detection of short RNA target sequences and exon junctions in cells and tissues with morphological context • Highly specific and sensitive det
Slide 14: Edited Dystrophin Transcript in PAX7+ cells, a Marker for Muscle Satellite Cells
Edited Dystrophin Transcript in PAX7+ cells, a Marker for Muscle Satellite Cells • 44-56 Dystrophin mRNA splice edit by Pax7+ 40X BaseScope Edited DMD 44-56 • Following AAV delivery, we observed evidence of the edited dystrophin transcript in PAX7+ cells, a marker for muscle satellite cells 40X 44-5
Slide 15: Conclusions
Conclusions • Here, we report in vivo proof of concept of a dual ARCUS nuclease approach in a humanized DMD mouse model • Following AAV delivery, we observed the edited dystrophin protein variant in multiple tissue types including heart, diaphragm, and skeletal muscle • The maximum force output (MFO
Slide 16: Acknowledgements
Acknowledgements Precision BioSciences Myologica LLC – Cassie Gorsuch – Ramzi J. Khairallah – Nicole Heard – Jennifer Martin – Jason Holt – Christopher Ward – Derek Jantz – Matt Jordan-Steele – Janel Lape – Whitney Lewis – Emily Manzon – Daniel Martin – Ben Morris – Dave Morris – Gary Owens – Traci