Fig 2: Optimization of Retrograde Intraductal Infusion via Catheterizing the Common Bile Duct through the Gallbladder and Cystic Duct(A) Uniform Evans Blue dye delivery to the entire pancreas via the retrograde intraductal infusion procedure. (B) A C57BL/6 mouse was dosed with 1 × 1011 vg of scAAV6.GFP via retrograde intraductal infusion, and pancreatic GFP expression was observed via direct fluorescence. (C–E) A cohort of C57BL/6 mice was dosed with scAAV6.GFP, 8, or 9 at 1 × 1011 vg/animal (n = 3–4/group). Serial sections of frozen (6 µm) pancreatic tissue collected 3 weeks post-vector infusion were stained for (C) amylase (AMY2, acinar cells), (D) Cytokeratin 19 (CK19, ductal cells), or (E) insulin (INS, islets) or with H&E to quantify cell-specific GFP transduction percentages. GFP, green; AMY2, CK19, or INS, red; DAPI, blue; 20× magnification; scale bar, 50 µm; representative images are presented. (F) A cohort of C57BL/6 mice was intraductally infused with different doses of scAAV6.GFP (n = 4–6/group), and the percentage of GFP+ acinar cells was determined. Data represent mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001.

Fig 3: Retrograde Pancreatic Intraductal Delivery of scAAV6 Targets Acinar, Epithelial, and Stromal Cells and Shows Long-Term Gene Expression in PDAC Mice5 × 1011vg of scAAV6.GFP was dosed in 1-month-old KC mice, and pancreata were collected 5 months post-delivery for the late time point and stained for amylase (AMY2), CK19, aSMA, or insulin (INS) to identify acinar cells, ductal cells, PSCs, and islet cells, respectively. Serial sections were stained with H&E, and representative images are presented. GFP, green; AMY2, CK19, aSMA, or INS, red; DAPI, blue; 20× magnification; scale bars, 50 µm; inset, 40×; arrows indicate aSMA+/GFP+ PSCs.