Slide 1: Pancreas-Targeted GLP-1 Gene Therapy Enhances β-Cell Function in Preclinical T2D Models
Pancreas-Targeted GLP-1 Gene Therapy Enhances β-Cell Function and Reduces Cellular Stress in Preclinical Type 2 Diabetes Models Joan Sabadell-Basallote, Alice Liou Fitzpatrick, Lin Quek, Jessica Von Stetina, Suya Wang, Lindsay Schulman, Nicole Picard, Abdul Alhamood, Keiko Ishida, Rebecca Reese, Cam
Slide 2: Presenter Disclosure
Presenter Disclosure ▪ Joan Sabadell-Basallote, PhD Type of Support — Categories • Employee: Fractyl Health, Inc. • Stock/Shareholder: Fractyl Health, Inc. • Research Support: Fractyl Health, Inc. (sponsor of the work presented) Co-authors: All co-authors are current or former employees / stockholde
Slide 3: T2D is a Chronic, Progressive Disease Driven By Beta Cell Failure
T2D is a Chronic, Progressive Disease Driven By Beta Cell Failure Despite treatment advancements, a clear unmet need remains • Type 2 diabetes (T2D) and associated co-morbidities affect >30M Americans1,2,3 • Insulin resistance and beta cell dysfunction lead to progressive metabolic failure4 Only 50%
Slide 4: Rejuva: A "Smart GLP-1" AAV Gene Therapy Platform for T2D
Rejuva: A "Smart GLP-1TM" AAV Gene Therapy Platform for T2D Single treatment → nutrient-responsive, adaptive, and durable effect Novel ROA Novel MOA Targeted administration via purpose-built GLP-1 under insulin-based regulatory and secretory endoscopic ultrasound-guided device control for beta cell-
Slide 5: Human Insulin-based Promoter (INSp) Drives Expression in Beta Cells
Human Insulin-based Promoter (INSp) Drives Expression in Beta Cells In Vitro: Transduced Human Cell Lines Beta cells DRG Liver Adrenal gland Gastric epithelial CMV_GFP (ubiquitous) INSp_GFP (specific) • Cells transduced with a GFP reporter surrogate under the engineered human insulin promoter (INSp)
Slide 6: INSp Drives Nutrient-Responsive Transgene Expression
INSp Drives Nutrient-Responsive Transgene Expression Chronic high glucose leads to increased transgene expression In Vitro: Transduced Human Beta Cells Transgene RNA Expression Transgene Protein (GFP) 3×10 6 5.5 mM Adj. transgene RNA copies per 10 ng total RNA 2×10 6 INSp_GFP (72h) 20 mM 1×10 6 0 Ho
Slide 7: AAV-INSp-GLP1 Drives Nutrient-Responsive Secretion of Bioactive GLP-1
AAV-INSp-GLP1 Drives Nutrient-Responsive Secretion of Bioactive GLP-1 With dose-responsive expression observed In Vitro: Transduced Human Beta Cells GLP-1 Content GLP-1 Secretion GLP-1 Bioactivity (lysate) (supernatant) (supernatant) 15000 10000 5000 AAV-INSp-GFP AAV-INSp-GLP1 Low Bioactive GLP-1 [p
Slide 8: AAV-INSp-GLP1 Preserves and Protects Human Beta Cell Health
AAV-INSp-GLP1 Preserves and Protects Human Beta Cell Health Consistent with GLP-1’s known beta cell cytoprotective biology In Vitro: Transduced Human Beta Cells No ROS Generation or Reduction in Cell Viability Reduced Baseline AND TGN-induced ER Stress ROS Generation Cell Viability RNA Expression of
Slide 9: Primary Human Islets Confirm Enhanced GLP-1 Secretion and Preserved Beta cell Health
Primary Human Islets Confirm: Enhanced GLP-1 Secretion and Preserved Beta cell Health Ex Vivo: Transduced Human Primary Islets GLP-1 Content GLP-1 Secretion Reduced ER Stress Gene Expression (lysate) (supernatant) 1.5 ER stress markers Insulin 4000 50 Relative RNA Expression 40 Total GLP-1 [pM] Tota
Slide 10: AAV-INSp-GLP1 Improves Metabolic Control in db/db Mice
AAV-INSp-GLP1 Improves Metabolic Control in db/db Mice Dose-responsive improvement in blood glucose, plasma insulin, and body weight In Vivo: Treated 8-week-old BKS db/db mice, 46 days post single IP AAV injection Fasting Blood Glucose Fasting Plasma Insulin Body Weight ✱✱✱ Fasting Blood Glucose [mg
Slide 11: Isolated Islets From Treated db/db Mice Confirm Enhanced GLP-1 and Insulin Content
Isolated Islets From Treated db/db Mice Confirm: Enhanced GLP-1 and Insulin Content and Co-secretion GLP-1 Content GLP-1 Secretion ✱✱✱✱ 8000 80 ✱✱✱✱ 49 days ✱✱ Strong positive correlation between GLP-1 Total GLP-1 [pM] Total GLP-1 [pM] 8-week old BKS db/db 6000 ✱✱ 60 ✱✱✱✱ and insulin secretion 4000
Slide 12: AAV-INSp-GLP1 "Smart GLP-1" Mimics Endogenous Physiology
AAV-INSp-GLP1“Smart GLP-1” Mimics Endogenous Physiology Potent efficacy with near physiologic circulating levels BKS db/db mice Comparison Chart Active GLP-1 Levels1 Circulating GLP-1 Modality Levels Tolerability • “Smart” GLP-1 levels are significantly ✱ lower than pharmacologic GLP-1RA 35 drug lev
Slide 13: Purpose-built Rejuva Device For Targeted Pancreatic AAV Delivery
Purpose-built Rejuva Device For Targeted Pancreatic AAV Delivery Large-animal safety model – Yucatan pig • Rejuva catheter introduced directly to pancreas parenchyma via standard EUS techniques • Upper endoscopic procedure conducted in ~ 20 minutes Novel ROA for AAV Delivery Rejuva Device Rejuva Nee
Slide 14: Effective and Precise Delivery to Pancreas in Yucatan Pigs
Effective and Precise Delivery to Pancreas in Yucatan Pigs AAV DNA, transgene RNA, and active GLP-1 protein all enriched at the targeted infusion site In Vivo: Treated Yucatan Pigs AAV DNA Transgene RNA GLP-1 Protein 10 3E4 1.0 [copies per 10 ng RNA] Infusion site Active GLP-1 [pg/μg] 8 0.8 Transgen
Slide 15: EUS-Guided AAV Delivery is Safe and Well-Tolerated in Yucatan Pigs
EUS-Guided AAV Delivery is Safe and Well-Tolerated in Yucatan Pigs Constrained biodistribution and no procedural or vector-related safety signals In Vivo: Treated Yucatan Pigs Pancreatitis Neurotoxicity (lipase) (Nf-L) 50 50 3xULN Fasting lipase [U/L] 40 40 NF-L [pg/mL] Biodistribution (AAV DNA) 30
Slide 16: GLP-1R Antagonism Provides Reversible Pharmacological Inhibition
GLP-1R Antagonism Provides Reversible Pharmacological Inhibition Blocks GLP-1 secretion and GLP-1R signaling In Vitro: Transduced Human Beta Cells + Avexitide GLP-1 Secretion GLP-1R Signaling (Beta Cells) (cAMP Reporter Assay) 76% reduction High Glucose Avexitide 125 68% reduction 125 AAV-INSp-GFP G
Slide 17: Rejuva: A First-in-Class "Smart GLP-1" AAV Gene Therapy for T2D
Rejuva: A First-in-Class “Smart GLP-1” AAV Gene Therapy for T2D Preclinical foundation complete, clinical translation underway KEY TAKEAWAYS ● Insulin promoter and regulatory features drive beta cell–specific, nutrient-regulated GLP-1 expression and secretion ● Restoration of beta cell function in d
Slide 18: Acknowledgements
Acknowledgements Shimyn Slomovic, PhD In vitro Discovery In/ex vivo Studies Scientific Advisory Board Exec. Director, Head of R&D Lin Quek, PhD Alice Fitzpatrick, DVM, PhD Director Director Jacques Bergman, MD, PhD Amsterdam UMC Tech Ops and Research Ops Junghun Lee, PhD Jessica von Stetina, PhD Mar