Supplementary Table 1: Raw read counts per gene for bulk RNA-seq IPI-926 intervention study.
Raw read counts of matching biopsy (Bx) and necropsy (Nx) bulk RNA-seq samples after two days of treatment with vehicle or IPI-926.
Supplementary Table 2: Digestion buffer for single cell preparation.
List of reagents optimized for single cell digestion of KPC-derived tumors.
Supplementary Table 3: Single Cell RNA-seq Quality Control.
Overview of KPC-derived scRNA-seq samples, containing treatment information (vehicle or IPI-926; n=3 each), sequencing depth, detected genes, and detection of mitochondrial genes (MT%).
Supplementary Table 4: Network sizes in murine scRNA-seq data.
Number of regulatory proteins for which regulons were generated in each network type, in each cell type, from KPC mouse pancreatic tumors.
Funding
National Cancer Institute (NCI)
United States Department of Health and Human Services
Find out more...Lustgarten Foundation (The Lustgarten Foundation)
Deutsche Forschungsgemeinschaft (DFG)
Charles H. Revson Foundation (Revson Foundation)
Mayo Clinic (The Mayo Clinic)
National Institutes of Health (NIH)
ARTICLE ABSTRACT
The sparse vascularity of pancreatic ductal adenocarcinoma (PDAC) presents a mystery: What prevents this aggressive malignancy from undergoing neoangiogenesis to counteract hypoxia and better support growth? An incidental finding from prior work on paracrine communication between malignant PDAC cells and fibroblasts revealed that inhibition of the Hedgehog (HH) pathway partially relieved angiosuppression, increasing tumor vascularity through unknown mechanisms. Initial efforts to study this phenotype were hindered by difficulties replicating the complex interactions of multiple cell types in vitro. Here we identify a cascade of paracrine signals between multiple cell types that act sequentially to suppress angiogenesis in PDAC. Malignant epithelial cells promote HH signaling in fibroblasts, leading to inhibition of noncanonical WNT signaling in fibroblasts and epithelial cells, thereby limiting VEGFR2-dependent activation of endothelial hypersprouting. This cascade was elucidated using human and murine PDAC explant models, which effectively retain the complex cellular interactions of native tumor tissues.
We present a key mechanism of tumor angiosuppression, a process that sculpts the physiologic, cellular, and metabolic environment of PDAC. We further present a computational and experimental framework for the dissection of complex signaling cascades that propagate among multiple cell types in the tissue environment.This article is featured in Selected Articles from This Issue, p. 201
