Precision Biosciences, Inc. · NASDAQ:DTIL

Precision Biosciences — ESGCT 2022

19 slides from Precision Biosciences's conference presentation, Nov 8, 2022

  1. Slide 1: Targeting Hepatitis B cccDNA with a Sequence-Specific ARCUS Nuclease to Eliminate HBV In Vivo

    Targeting Hepatitis B cccDNA with a Sequence- Specific ARCUS Nuclease to Eliminate Hepatitis B Virus In Vivo Derek Jantz ESGCT 2022 © 2022 Precision BioSciences, Inc.

  2. 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. All statements contained in this herein and in any related presentation that do not relate to matters of his

  3. Slide 3: Disclosures

    Disclosures • I am an employee of Precision BioSciences © 2022 Precision BioSciences, Inc. 3

  4. Slide 4: Chronic Hepatitis B affects ~250 million people

    ARCUS HBV Infected Hepatocyte Chronic Hepatitis B affects inactivate ~250 million people cccDNA cccDNA and integrated HBV have complicated efforts to inactivate cure the disease integrated HBV Durable Antigen Loss Functional Cure? © 2022 Precision BioSciences, Inc. 4

  5. Slide 5: ARCUS is derived from I-CreI, a homing endonuclease from algae

    I-CreI is 1 expressed 23S rRNA gene ARCUS is derived 5’-CAAACTGTCGTGAGACGTTTTG-3’ 2 I-CreI recognizes its target in the from I-CreI, a homing 3’-GTTTGACAGCACTCTGCAAAAC-5’ 23S rRNA gene endonuclease from algae. I-CreI cuts and 3 produces 3’ 5’-CAAACTGTCGTGA GACGTTTTG-3’ overhangs 3’-GTTTGACAG CACTCTG

  6. Slide 6: Creating and Optimizing ARCUS Nucleases

    Creating and Optimizing ARCUS Nucleases The DNA-binding surface of I-CreI must be extensively re-engineered to produce each new ARCUS nuclease Controls Controls Controls Efficiency Specificity Affinity © 2022 Precision BioSciences, Inc. 6

  7. Slide 7: Engineering ARCUS to Recognize HBV

    Engineering ARCUS to Recognize HBV An ARCUS enzyme was designed to cut a conserved sequence in the HBV polymerase (POL) gene © 2022 Precision BioSciences, Inc. 7

  8. Slide 8: cccDNA Fate after ARCUS Cleavage

    cccDNA Fate after ARCUS Cleavage © 2022 Precision BioSciences, Inc. 8

  9. Slide 9: ARCUS Activity in HBV-Infected Primary Human Hepatocytes

    ARCUS Activity in HBV-Infected Primary Human Hepatocytes • In HBV-infected primary human hepatocytes (PHH), ARCUS delivered via LNP reduced cccDNA by 86% and created indel mutations in 30% of the remaining cccDNA. cccDNA Degradation cccDNA Indels ARCUS LNP 150 ARCUS 30 30% (cccDNA/non-targeting) % T

  10. Slide 10: ARCUS Activity in HBV-Infected Primary Human Hepatocytes

    ARCUS Activity in HBV-Infected Primary Human Hepatocytes • ARCUS treated cells demonstrated an 80% reduction in extracellular HBV DNA and a 77% reduction in secreted sAg with no change in albumin. Extracellular HBV DNA Extracellular HBsAg Extracellular Albumin 150 150 250 Non-targeting ARCUS nucleas

  11. Slide 11: HBV Episomal In Vivo Model

    HBV Episomal In Vivo Model Natural Infection Episomal Model • AAV with a partial HBV sequence expressing HBsAg is administered and is present as the cccDNA surrogate. • An HBV nuclease is subsequently delivered via a lipid nanoparticle (LNP) which can then cut the cccDNA surrogate, AAV-HBsAg. © 2022

  12. Slide 12: Episomal Mouse Model Study Design

    Episomal Mouse Model Study Design • AAV8 HBsAg Dose: 5e11vg • ARCUS LNP Dose: 2 mg/kg • Weekly blood draws for HBsAg © 2022 Precision BioSciences, Inc. 12

  13. Slide 13: Episomal Mouse Model—Molecular Analyses

    Episomal Mouse Model—Molecular Analyses • The ARCUS-HBV nuclease significantly reduced AAV copies in the liver compared to the PBS control group. • The remaining AAV had an average of 86% indels in the ARCUS-HBV treated group. • The AAV degradation and indel formation resulting from ARCUS-HBV cuttin

  14. Slide 14: Episomal Mouse Study—Liver HBsAg Immunohistochemistry

    Episomal Mouse Study—Liver HBsAg Immunohistochemistry • Mice treated with ARCUS show a significant loss in HBsAg in the liver compared to untreated mice. Group 1—No Nuclease Group 2—ARCUS-HBV Blue = Nucleus Brown = HBsAg © 2022 Precision BioSciences, Inc. 14

  15. Slide 15: cccDNA Integration into the Genome of Infected Hepatocytes

    cccDNA Integration into the Genome of Infected Hepatocytes isolate and sequence junctions HBV HBV integrates fragment DNA Chromosomes Chromosomes with Chromosome Fragments Integrated HBV © 2022 Precision BioSciences, Inc. 15

  16. Slide 16: cccDNA Integration into the Genome of Infected Hepatocytes

    cccDNA Integration into the Genome of Infected Hepatocytes Mock (HBV only) ARCUS Cut Site known off- target site? Early Generation ARCUS Treated © 2022 Precision BioSciences, Inc. 16

  17. Slide 17: cccDNA Integration into the Genome of Infected Hepatocytes

    cccDNA Integration into the Genome of Infected Hepatocytes Mock (HBV only) ARCUS Cut Site known off- target site? Optimized ARCUS Treated © 2022 Precision BioSciences, Inc. 17

  18. Slide 18: Conclusions

    Conclusions • ARCUS demonstrates high levels of editing against cccDNA with subsequent reduction of HBsAg levels in PHHs. • We have developed a novel HBV model suitable for mice and NHPs which demonstrated high levels of editing and HBsAg reductions. • Our gene editing approach demonstrates high on-

  19. Slide 19: Acknowledgements

    Acknowledgements Precision BioSciences – Janel Lape – Forrest Karriker – Cassie Gorsuch – Greg Falls – Amod Joshi – Paige Nemec – Jason Holt – Tyler Goodwin – Amy Rhoden Smith – Wendy Shoop – Neil Leatherbury – Jeff Smith – Emma Sevigny – Rob Brown Gilead Sciences – Meghan Holdorf – Nick van Buuren

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