Introduction: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive and lethal malignancies, ranking as the seventh leading cause of cancer-related deaths globally. Despite extensive research efforts, its prognosis remains dismal, with a 5-year survival rate of approximately 12.5%. The lack of effective targeted therapies and the reliance on cytotoxic chemotherapy underline the urgent need for innovative preclinical models. Patient-derived organoids (PDOs) have emerged as promising three-dimensional culture systems that replicate the histological and molecular features of primary tumors. Concurrently, liquid biopsy techniques, including bile-derived cell-free DNA (cfDNA), provide a minimally invasive approach to studying tumor dynamics, though their potential in biliopancreatic cancers remains underexplored. Aims: This study aimed to evaluate the feasibility of generating PDOs from endoscopic fine-needle biopsy (EUS-FNB) samples and from bile and to compare their histological concordance with native PDAC tissue. A secondary objective was to assess the potential of bile as a source of cfDNA and analyze its molecular profile compared to plasma-derived cfDNA. Methods: A prospective observational study enrolled patients with suspected or recently diagnosed PDAC presenting with obstructive jaundice. EUS-FNB was performed for histological diagnosis and PDO generation. Bile and plasma samples were collected during biliary drainage for cfDNA extraction and analysis using next-generation sequencing (NGS). Morphological comparisons between PDOs and native tumors were performed, and cfDNA concentrations from bile and plasma were analyzed. Results: Between May 2023 and November 2024, 72 patients were enrolled, of whom 44 were diagnosed with PDAC. PDOs were successfully established in 73% of tissue-derived samples and 58% of bile-derived samples. Histopathological analysis revealed high morphological concordance between PDOs and native PDAC tissue, including cellular morphology, nuclear atypia and glandular formation. Bile-derived cfDNA concentrations were significantly higher than plasma-derived cfDNA (47.26 ng/µL vs. 2.67 ng/µL, p = 0.0001), with longer DNA fragments observed in bile. Conclusions: PDO generation from EUS-FNB is feasible and provides robust models for studying PDAC biology. Bile-derived cfDNA offers superior molecular insights compared to plasma, highlighting its potential as a valuable tool for NGS analysis and tumor mutation profiling. The integration of PDOs and bile liquid biopsy represents a promising strategy for advancing diagnostic and therapeutic approaches in PDAC management.

Establishment of biliopancreatic cancer patient-derived organoids and propagation from biliary cultures: preliminary results from the Bile Biopsy study

PALANO, GIORGIO
2022/2023

Abstract

Introduction: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive and lethal malignancies, ranking as the seventh leading cause of cancer-related deaths globally. Despite extensive research efforts, its prognosis remains dismal, with a 5-year survival rate of approximately 12.5%. The lack of effective targeted therapies and the reliance on cytotoxic chemotherapy underline the urgent need for innovative preclinical models. Patient-derived organoids (PDOs) have emerged as promising three-dimensional culture systems that replicate the histological and molecular features of primary tumors. Concurrently, liquid biopsy techniques, including bile-derived cell-free DNA (cfDNA), provide a minimally invasive approach to studying tumor dynamics, though their potential in biliopancreatic cancers remains underexplored. Aims: This study aimed to evaluate the feasibility of generating PDOs from endoscopic fine-needle biopsy (EUS-FNB) samples and from bile and to compare their histological concordance with native PDAC tissue. A secondary objective was to assess the potential of bile as a source of cfDNA and analyze its molecular profile compared to plasma-derived cfDNA. Methods: A prospective observational study enrolled patients with suspected or recently diagnosed PDAC presenting with obstructive jaundice. EUS-FNB was performed for histological diagnosis and PDO generation. Bile and plasma samples were collected during biliary drainage for cfDNA extraction and analysis using next-generation sequencing (NGS). Morphological comparisons between PDOs and native tumors were performed, and cfDNA concentrations from bile and plasma were analyzed. Results: Between May 2023 and November 2024, 72 patients were enrolled, of whom 44 were diagnosed with PDAC. PDOs were successfully established in 73% of tissue-derived samples and 58% of bile-derived samples. Histopathological analysis revealed high morphological concordance between PDOs and native PDAC tissue, including cellular morphology, nuclear atypia and glandular formation. Bile-derived cfDNA concentrations were significantly higher than plasma-derived cfDNA (47.26 ng/µL vs. 2.67 ng/µL, p = 0.0001), with longer DNA fragments observed in bile. Conclusions: PDO generation from EUS-FNB is feasible and provides robust models for studying PDAC biology. Bile-derived cfDNA offers superior molecular insights compared to plasma, highlighting its potential as a valuable tool for NGS analysis and tumor mutation profiling. The integration of PDOs and bile liquid biopsy represents a promising strategy for advancing diagnostic and therapeutic approaches in PDAC management.
2022
Establishment of biliopancreatic cancer patient-derived organoids and propagation from biliary cultures: preliminary results from the Bile Biopsy study
organoids
biliopancreatic
Bile Biopsy
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12608/81465