Utilizing cell line-derived organoids to evaluate the efficacy of a novel LIFR-inhibitor, EC359 in targeting pancreatic tumor stroma
Bradley R. Hall1,2, Andrew Cannon1, Christopher Thompson1, Bindu Santhamma4, Alejandra Chavez-Riveros4, Rakesh Bhatia1, Hareesh B. Nair4, Klaus Nickisch4, Surinder K. Batra1,3 and Sushil Kumar1
1 Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, USA
2 Department of General Surgery, University of Nebraska Medical Center, Omaha, NE, USA
3 The Fred and Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE, USA
4 Evestra Inc., San Antonio, Texas, USA
Correspondence:
Sushil Kumar, email:
Correspondence:
Surinder K Batra, email:
Keywords: pancreatic ductal adenocarcinoma; 3D model; pancreatic stroma; LIF; LIFR
Received: September 20, 2018 Accepted: November 27, 2018 Published: December 08, 2018
Abstract
Survival of pancreatic cancer (PC) patient is poor due to lack of effective treatment modalities, which is partly due to the presence of dense desmoplasia that impedes the delivery of chemotherapeutics. Therefore, PC stroma-targeting therapies are expected to improve the efficacy of chemotherapeutics. However, in vitro evaluation of stromal-targeted therapies requires a culture system which includes components of both tumor stroma and parenchyma. We aim to generate a cell line-derived 3D organoids to test the efficacy of stromal-targeted, LIFR-inhibitor EC359. Murine PC (FC1245) and stellate (ImPaSC) cells were cultured to generate organoids that recapitulated the histological organization of PC with the formation of ducts by epithelial cells surrounded by activated fibroblasts, as indicated by CK19 and α-SMA staining, respectively. Analysis by qRT-PCR demonstrated a significant downregulation of markers of activated stroma, POSTN, FN1, MMP9, and SPARC (p<0.0001), when treated with gemcitabine in combination with EC359. Concurrently, collagen proteins including COL1A1, COL1A2, COL3A1, and COL5A1 were significantly downregulated (p<0.0001) after treatment with gemcitabine in combination with EC359. Overall, our study demonstrates the utility of cell lines-derived 3D organoids to evaluate the efficacy of stroma-targeted therapies as well as the potential of EC359 to target activated stroma in PC.
