Abstract
While much progress has been made in the war on cancer, highly invasive cancers such as pancreatic cancer remain difficult to treat and anti-cancer clinical trial success rates remain low. One shortcoming of the drug development process that underlies these problems is the lack of predictive, pathophysiologically relevant preclinical models of invasive tumor phenotypes. While present-day 3D spheroid invasion models more accurately recreate tumor invasion than traditional 2D models, their shortcomings include poor reproducibility and inability to interface with automated, high-throughput systems. To address this gap, a novel 3D tumor-tissue invasion model which supports rapid, reproducible setup and user-definition of tumor and surrounding tissue compartments was developed. High-cell density tumor compartments were created using a custom-designed fabrication system and standardized oligomeric type I collagen to define and modulate ECM physical properties. Pancreatic cancer cell lines used within this model showed expected differential invasive phenotypes. Low-passage, patient-derived pancreatic cancer cells and cancer-associated fibroblasts were used to increase model pathophysiologic relevance, yielding fibroblast-mediated tumor invasion and matrix alignment. Additionally, a proof-of-concept multiplex drug screening assay was applied to highlight this model’s ability to interface with automated imaging systems and showcase its potential as a predictive tool for high-throughput, high-content drug screening.
Highlights
Cell monolayer on 2D transwell insert thin layer or coating thin layer or coating no migration minutes yes none yes yes migration or invasion minutes yes limited tunablity of surroundings no no no yes invasion hours to days no moderate tunablity of surroundings no no
User-defined tumor compartment embedded in 3D matrix
Much work has been done with traditional 3D multicellular spheroid models to demonstrate and improve the relevance of these 3D models over 2D culture[12,13,14], to assess the delivery and efficacy of therapeutics[15,16], and to enable HT-HC screening through standardization and automation[17,18,19]
Summary
Cell monolayer on 2D transwell insert thin layer or coating thin layer or coating no migration minutes yes none yes yes migration or invasion minutes yes limited tunablity of surroundings no no no yes invasion hours to days no moderate tunablity of surroundings no no. While there is advocacy that added model complexity (inclusion of vasculature, various stromal and immune cells) may increase pathophysiologic relevance and predictive power, such approaches fall short with respect to practical logistics[7,8,9] For such models to gain traction and widespread use within both pharmaceutical and academic environments, they must be user-friendly, time-efficient, reproducible within and between laboratories, standardizable, scalable, and ideally, amenable to high-throughput (HT) automation (e.g. automated imaging systems, liquid handling robots)[7,10]. To address the above-mentioned gaps, the goal of this work was to develop a HT-HC phenotypic screening model of PDAC invasion that supports user specification and control of both the tumor and the surrounding tissue compartment, improved pathophysiologic relevance in terms of cell-ECM interactions, and rapid, low-cost implementation. This work presents initial development and validation of this new 3D tumor-tissue invasion model which includes demonstrating pathophysiologically relevant modes of invasion by established PDAC cell lines, application of patient-derived PDAC cells and CAFs to recreate more complex heterogeneous cell-cell interactions and invasion mechanisms, and proof-of-concept (POC) drug dosing with image-based multiplex analysis of tumor cell proliferation, metabolic activity, and invasion
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