Patient-specific modeling of stroma-mediated chemoresistance of pancreatic cancer using a three-dimensional organoid-fibroblast co-culture system

Abstract

Background Cancer-associated fibroblasts (CAFs) are considered to play a fundamental role in pancreatic ductal adenocarcinoma (PDAC) progression and chemoresistance. Patient-derived organoids have demonstrated great potential as tumor avatars for drug response prediction in PDAC, yet they disregard the influence of stromal components on chemosensitivity. Methods We established direct three-dimensional (3D) co-cultures of primary PDAC organoids and patient-matched CAFs to investigate the effect of the fibroblastic compartment on sensitivity to gemcitabine, 5-fluorouracil and paclitaxel treatments using an image-based drug assay. Single-cell RNA sequencing was performed for three organoid/CAF pairs in mono- and co-culture to uncover transcriptional changes induced by tumor-stroma interaction. Results Upon co-culture with CAFs, we observed increased proliferation and reduced chemotherapy-induced cell death of PDAC organoids. Single-cell RNA sequencing data evidenced induction of a pro-inflammatory phenotype in CAFs in co-cultures. Organoids showed increased expression of genes associated with epithelial-to-mesenchymal transition (EMT) in co-cultures and several potential receptor-ligand interactions related to EMT were identified, supporting a key role of CAF-driven induction of EMT in PDAC chemoresistance. Conclusions Our results demonstrate the potential of personalized PDAC co-cultures models not only for drug response profiling but also for unraveling the molecular mechanisms involved in the chemoresistance-supporting role of the tumor stroma.

Organoids: The current status and biomedical applications

“…indicated that knockdown L1CM human colorectal tumoroids presented improved sensitivity to irinotecan. The coculture system of pancreatic ductal adenocarcinoma (PDAC) organoids and CAFs showed resistance to gemcitabine, 5‐FU, and paclitaxel, with increased epithelial‐to‐mesenchymal transition (EMT) gene expression in these organoid cells 231 . Colorectal tumoroids were used to perform drug sensitivity assays and pharmacogenomics analysis, and results showed that the paclitaxel sensitivity was linked to the checkpoint with forkhead and ring finger domains (CHFR) silencing in organoid as well as in xenograft models 232 .…”

Acoustic Printing of Patient‐Derived Organoids That Preserve Tumor Microenvironment for Personalized Drug Screening

“…(Figure 4b) CAFs, as important components of the tumor microenvironment, have been extensively shown to support the development and progression of cancer and to promote therapeutic resistance. [5,8,13] The clinical importance of CAFs in disease progression, treatment response, and patient prognosis has been widely reported in diverse types of cancer. [7] CAFs secrete various chemokines, cytokines, and growth factors to produce a tumorpermissive microenvironment.…”

“…It promotes tumor development, invasion, and metastasis. [8] CAFs also have the capacity to shape the www.advmattechnol.de extra-tumor stroma, forming a barrier to drug or therapeutic immune cell penetration by blocking deeper penetration of drugs and immune cells into tumor tissue, thereby reducing the effectiveness of tumor therapy. [7,9] Histopathological analysis has shown that the abundance of CAFs correlates with the prognosis of different human cancers.…”

The third dimension of tumor microenvironment—The importance of tumor stroma in 3D cancer models

“…Many publications showed that organoid models could accurately recapitulate this biological heterogeneity and maintain phenotypic and genetic characteristics of original tumor epithelium. Moreover, tumor organoid cultures seem to be suitable for modeling the TME heterogeneity and cell interactions using co-culture with different type of cells, such as cancer-associated fibroblasts (CAFs) 37,38 or immune cells. 39,40 As mentioned above, hypoxia, which greatly contributes to malignant behavior and chemo- and radio-resistance, does develop in organoids once they reach a certain size.…”