Development of a Novel 3D Tumor-tissue Invasion Model for High-throughput, High-content Phenotypic Drug Screening

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.

Cell Culture Based in vitro Test Systems for Anticancer Drug Screening

“…3D co-cultures of non-small cell lung cancer (NSCLC) and fibroblasts embedded in a Matrigel or encapsulated in alginate are used as models in drug discovery for analysis of immune cell infiltration (Osswald et al, 2019). Also, described is a highpotential tumor spheroid model drug screening, which consists of pancreatic ductal adenocarcinoma (PDAC) cell lines (Panc-1 and BxPC-3) and cancer-associated fibroblasts (CAFs) surrounding by oligomeric type I collagen (Oligomer) for creation of the interstitial ECM supports definition (Puls et al, 2018).…”

The Mini‐Organo: A rapid high‐throughput 3D coculture organotypic assay for oncology screening and drug development

“…One caveat, however, is that many of these 3D in vitro model systems are time‐consuming and work‐intensive with often limited scalability. Thus, the miniaturisation and integration of organotypic culture models into high‐throughput assays can lead to the rapid evaluation of novel drugs in a cost‐effective and time‐effective manner . From a clinical perspective, a cost‐efficient protocol to validate drug efficacy in ex vivo patient‐derived transplants could facilitate personalised decision making in oncology …”

“…Thus, the miniaturisation and integration of organotypic culture models into high-throughput assays can lead to the rapid evaluation of novel drugs in a cost-effective and timeeffective manner. [15][16][17][18] From a clinical perspective, a cost-efficient protocol to validate drug efficacy in ex vivo patient-derived transplants could facilitate personalised decision making in oncology. [19][20][21] Extracellular matrix deposition, contraction, and remodelling are common to diseases such as tissue fibrosis, diabetes, and tissue aging and importantly are salient features of almost all solid tumours.…”

Three‐Dimensional Osteosarcoma Models for Advancing Drug Discovery and Development

“…[67] This co-culture promoted the formation of a well-organized network of tubule-like structures of HUVECs, stimulated by the enhanced expression of angiogenic factors like VEGF, intercellular adhesion molecule 1, and C-X-C chemokine receptor type 4 as a result of the crosstalk Adv between them (Figure 3). [56,[69][70][71] Superhydrophobic surfaces patterned with wettable spots proposed by Oliveira et al are a great example of the effort that have been made in recent years, demonstrating that this robust platform allow the formation of SaOS-2 and L929 cell spheroids and in situ combinatorial high-throughput doxorubicin screening. The in vitro co-culture of stromal cells, such as fibroblasts and bone marrow (BM)-MSCs, as well as the epithelial cells in the bone, osteoblasts, that mimic the in vivo situation, demonstrated to have a crucial effect in tumor behavior and drug sensitivity.…”

“…The in vitro co-culture of stromal cells, such as fibroblasts and bone marrow (BM)-MSCs, as well as the epithelial cells in the bone, osteoblasts, that mimic the in vivo situation, demonstrated to have a crucial effect in tumor behavior and drug sensitivity. [70] Although pancreatic cancer cell lines had been used to the proof-of-concept, this system has the potential to be applied to other tumor types. Therefore, it is imperative to develop more complex and reliable in vitro OS models that can more accurately mimic the in vivo cell function and provide physiologically relevant results.…”