A bioengineered heterotypic stroma-cancer microenvironment model to study pancreatic ductal adenocarcinoma.

Cole R Drifka,Kevin W Eliceiri,W John Kao,Sharon M Weber

doi:10.1039/c3lc50487e

Abstract

Interactions between neoplastic epithelial cells and components of a reactive stroma in pancreatic ductal adenocarcinoma (PDAC) are of key significance behind the disease's dismal prognosis. Despite extensive published research in the importance of stroma-cancer interactions in other cancers and experimental evidence supporting the importance of the microenvironment in PDAC progression, a reproducible three-dimensional (3D) in vitro model for exploring stroma-cancer interplay and evaluating therapeutics in a physiologically relevant context has been lacking. We introduce a humanized microfluidic model of the PDAC microenvironment incorporating multicellularity, extracellular matrix (ECM) components, and a spatially defined 3D microarchitecture. Pancreatic stellate cells (PSCs) isolated from clinically-evaluated human tissue specimens were co-cultured with pancreatic ductal adenocarcinoma cells as an accessible 3D construct that maintained important tissue features and disease behavior. Multiphoton excitation (MPE) and Second Harmonic Generation (SHG) imaging techniques were utilized to image the intrinsic signal of stromal collagen in human pancreatic tissues and live cell-collagen interactions within the optically-accessible microfluidic tissue model. We further evaluated the dose-response of the model with the anticancer agent paclitaxel. This bioengineered model of the PDAC stroma-cancer microenvironment provides a complementary platform to elucidate the complex stroma-cancer interrelationship and to evaluate the efficacy of potential therapeutics in a humanized system that closely recapitulates key PDAC microenvironment characteristics.

Highlights

Pancreatic ductal adenocarcinoma (PDAC) is the fourth most common cause of cancer death in the United States
As air is drawn from the outlet port, the cell–extracellular matrix (ECM) solution droplets flow into the device and pattern as a continuous trilayer over the length of the central culture channel due to laminar flow (Fig. 1B)
We have routinely shown that Pancreatic stellate cells (PSCs) contract the ECM rapidly within the first 24 h following culture loading before reaching a state of tensional homeostasis when using culture parameters of 2 mg mL21 ColI, 2 mg mL21 HA, 1 h incubation at 4 uC, 1 6 106 PANC-1 cells mL21, and 2.5 6 105 PSCs/mL

Summary

Introduction

Pancreatic ductal adenocarcinoma (PDAC) is the fourth most common cause of cancer death in the United States. PDAC presents itself as one of the most dismal human cancers It is one of the few human malignancies with nearly 100% mortality with a median survival time of less than 6 months and a 5-year survival rate of less than 4%.1. This is a result of the inherently aggressive disease biology and a pronounced resistance to conventional c3lc50487e therapeutic regimens.[2,3,4] a number of genetic and environmental factors have been implicated in PDAC carcinogenesis, exact causes and pathological mechanisms remain incompletely understood It is one of the few human malignancies with nearly 100% mortality with a median survival time of less than 6 months and a 5-year survival rate of less than 4%.1 This is a result of the inherently aggressive disease biology and a pronounced resistance to conventional c3lc50487e therapeutic regimens.[2,3,4] a number of genetic and environmental factors have been implicated in PDAC carcinogenesis, exact causes and pathological mechanisms remain incompletely understood

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Conclusion