Validation of a Novel EUS-FNB-Derived Organoid Co-Culture System for Drug Screening in Patients with Pancreatic Cancer

Abstract

Cancer-associated fibroblasts (CAFs) have been shown to impact the chemosensitivity of patient-derived tumor organoids (PDTOs). However, the published literature comparing PDTO response to clinical outcome does not include CAFs in the models. Here, a co-culture model was created using PDTOs and CAFs derived from endoscopic ultrasound-guided fine-needle biopsies (EUS-FNBs) for potential use in drug screening applications. Co-cultures were established, and growth was compared to monocultures using image metrics and a commercially available assay. We were able to establish and expand validated malignant PDTOs from 19.2% of adenocarcinomas from EUS-FNBs. CAFs could be established from 25% of the samples. The viability of PDTOs in the mixed cell co-culture could be isolated using image metrics. The addition of CAFs promoted PDTO growth in half of the established co-cultures. These results show that co-cultures can be established from tiny amounts of tissue provided by EUS-FNB. An increased growth of PDTOs was shown in co-cultures, suggesting that the present setup successfully models CAF–PDTO interaction. Furthermore, we demonstrated that standard validation techniques may be insufficient to detect contamination with normal cells in PDTO cultures established from primary tumor core biopsies.

Human patient derived organoids: an emerging precision medicine model for gastrointestinal cancer research

“…Knoblauch M et al established direct 3D co-cultures of primary PDAC organoids and patient-matched CAFs to investigate the influence of stromal components on chemosensitivity ( Knoblauch et al, 2023 ). Similarly, Grützmeier SE et al developed a co-culture model using PDTOs and CAFs derived from endoscopic ultrasound-guided fine-needle biopsies (EUS-FNBs) for potential use in drug screening and molecular mechanisms unraveling involved in the chemoresistance-supporting role of the tumor stroma ( Grützmeier et al, 2023 ). In research reported by Zou et al, HCC-PDOs and PBMC are co-cultured with MSC and CAFs to construct HCC organoid-on-a-chip mimicking the original TME ( Zou et al, 2023 ).…”

Establishment and Advancement of Pancreatic Organoids

“…In particular, organoids can be established in resected tissues before chemotherapy or after neoadjuvant chemotherapy to measure the response to anticancer drugs, which is useful for precision therapy, drug screening, and tumor biology research [48]. Recently, PDOs were generated using endoscopic ultrasound-guided fine-needle aspiration biopsies [46,[49][50][51][52] and utilized in co-culture systems for drug screening [51,52].…”

Role of Endoscopic Ultrasound in Diagnosis of Pancreatic Ductal Adenocarcinoma

“…In a recent feasibility study, authors were able to establish a co-culture using PDOs from a small amount of EUS FNB samples and cancer-associated fibroblasts (CAFs). Limitations like a high risk of contamination (blood as well as benign cells), compromising the yield of tissue available, the suboptimal number of needle passes, and unknown histopathological diagnoses before EUS TA can be obviated in future studies to improve the success rate [ 146 ].…”