Silence Therapeutics PLC · NASDAQ:SLN
Silence Therapeutics — AsiaTIDES Kyoto 2017
17 slides from Silence Therapeutics's conference presentation, Feb 23, 2017
Slide 1: Therapeutic RNA delivery: into hepatocytes and beyond
Therapeutic RNA delivery: into hepatocytes and beyond February 23rd, 2017 Claudine M. Dojahn, PhD © Silence Therapeutics 2017
Slide 2: Disclaimer
Disclaimer The information contained in this presentation is being supplied and communicated to you on a confidential basis solely for your information and may not be reproduced, further distributed to any other person or published, in whole or in part, for any purpose. In accordance with the prohib
Slide 3: Our mission: Establish a clinically viable technology platform for therapeutic RNAs
Our mission: Establish a clinically viable technology platform for therapeutic RNAs Particle Conjugation approach approach mRNA siRNA w/ siRNA Gene editing payloads GalNAc cluster (CRISPR/Cas9) Vasculature Liver Macrophages Hepatocytes 3 © Silence Therapeutics 2017
Slide 4: Technology platform to generate novel delivery systems
Technology platform to generate novel delivery systems Cationic lipids Physicochemical characterization fluores c enc e Helper/ PEGylation TNS co lipids 4 5 6 7 8 pH Zeta potential Particle size Intensity Surface charge +/- Ratio 0 -200 -100 0 100 200 30 60 120 240 480 Size (nm) Formulation proce
Slide 5: Screening for biodistribution of functional mRNA delivery in vivo
lu c ife r a s e a c t iv ity lu c ife r a s e a c t iv ity [ p g /g t is s u e ] [ p g /g t is s u e ] 5 1 10 100 1 0 0 0 1 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 2 0 0 0 3 0 0 0 CD7907 CD7907 CD7910 CD7910 CD7911 CD7911 CD8902 CD8902 CD8903 CD8903 CD8904 CD8904 CD8906 CD8906 CD8908 C57BL/6 (n
Slide 6: Time course and biodistribution of GN2205 functional luciferase mRNA delivery
Time course and biodistribution of GN2205 functional luciferase mRNA delivery -19 days 0h 2h 4h 6h 8h 12h 24h 48h IV administration of scid beige 1mg/kg mRNA Hep3B (5x106 cells s.c.) (FLuc;; ᴪ;; 5MeC;; TriLink) ex vivo quantification of luciferase activity (tumor size ~350mm³, in tissue lysates n=6
Slide 7: Dose response to GN2205 with hEPO and FLuc mRNA
Dose response to GN2205 with hEPO and FLuc mRNA 0h 4h IV administration of 1mg/kg mRNA ex vivo quantification of luciferase (FLuc or hEPO;; ᴪ & 5MeC;; TriLink) activity in tissue lysates or hEPO C57BL/6 (n=6) in EDTA-plasma Luciferase activity hEPO 0.03 mg/kg 100,000 0.1 mg/kg Luciferase activity 1
Slide 8: GN2205 siRNA delivery in the liver
GN2205 siRNA delivery in the liver 0h 48h h e p a to c y te e n d o t h e lia l c e lls ( T T R - liv e r) ( T ie 2 - liv e r) 1 .5 1 .5 IV administration RT-PCR T T R /a c t i n T ie 2 /a c tin 1 .0 1 .0 0.5 or 1.0 mg/kg quantification 0 .5 0 .5 siRNA (Silence) of transcripts C57BL/6 (n=4-6) 0
Slide 9: GN2205 targets vascular cells in liver
GN2205 targets vascular cells in liver 0h 48h L iv e r S p le e n IV administration RT-PCR T ie 2 ( e n d o th e lia l c e lls ) T ie 2 ( e n d o th e lia l c e lls ) 1 .5 2 .0 0.5 mg/kg siRNA quantification (Silence) of transcripts 1 .5 T ie 2 /a c tin T ie 2 /a c tin C57BL/6 (n=4-6) 1 .0 1 .0 0
Slide 10: GN2205 RNA delivery platform
GN2205 RNA delivery platform Liver: ü Hepatocytes & hepatocyte precursors ü Endothelial cells ü Stellate cells ü Macrophages ü Pericytes x Cholangiocytes Spleen, Lung, heart & kidney: x Endothelial cells Liver 10 © Silence Therapeutics 2017
Slide 11: GN2205 co-delivery of guide RNA & spCas9-mRNA
GN2205 co-delivery of guide RNA & spCas9-mRNA day 1 day 35 day 63 day 91 day 119 day 147 day 175 day 203 IV administration of 1mg/kg and quantification of serum protein at indicated 0.5 mg/kg mRNA & guide RNA time points by standardized ELISA (Cas9-mRNA: ᴪ & 5MeC, TriLink;; guide RNA: Silence) c5
Slide 12: What are GalNAc-siRNA conjugates?
What are GalNAc-siRNA conjugates? Scheme based on information provided by Essentials of Glycobiology. 2nd edition. Varki A, Cummings RD, Esko JD, et al., editors. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2009. GalNAc-siRNA conjugate is a delivery platform • contains GalNAc clu
Slide 13: Knockdown of ubiquitous Target1 using GalNAc siRNA conjugates
Knockdown of ubiquitous Target1 using GalNAc siRNA conjugates 0h 24h 0h 48h Murine 1°Hepatocytes lysis and RNA extraction SC administration RT-PCR GalNAc-siRNA conjugate RT-PCR quantification of GalNAc-siRNA conjugate quantification transcripts c57BL/6 (n=4) of transcripts Target1 gene expressio
Slide 14: Knockdown of Target2 using GalNAc siRNA conjugates
Knockdown of Target2 using GalNAc siRNA conjugates 0h 24h 0h 48h Murine 1°Hepatocytes lysis and RNA extraction SC administration RT-PCR GalNAc-siRNA conjugate RT-PCR quantification of GalNAc-siRNA conjugate quantification transcripts c57BL/6 (n=4) of transcripts Target2 gene expression 24h post
Slide 15: Summary
Summary Experience to transfer siRNA formulation into mRNA delivery system Identified LNP with 100 × fold activity in liver Targeting multiple cell types in the liver co-delivery of Cas9 mRNA and guide RNA results in long lasting gene disruption Proprietary ST siRNA payload and ST GalNAc chemistry
Slide 16: Acknowledgement
Acknowledgement Formulation team: Gemma Navarro Biology Team: Ute Schaeper Adrien Weingärtner Vivarium: Birgit Durieux 16 © Silence Therapeutics 2017
Slide 17: Thank you
Thank you © Silence Therapeutics 2017