Human Organoids Share Structural and Genetic Features with Primary Pancreatic Adenocarcinoma Tumors

Abstract

Patient-derived pancreatic ductal adenocarcinoma (PDAC) organoid systems show great promise for understanding the biological underpinnings of disease and advancing therapeutic precision medicine. Despite the increased use of organoids, the fidelity of molecular features, genetic heterogeneity, and drug response to the tumor of origin remain important unanswered questions limiting their utility. To address this gap in knowledge, primary tumor- and patient-derived xenograft (PDX)-derived organoids, and 2D cultures for in-depth genomic and histopathologic comparisons with the primary tumor were created. Histopathologic features and PDAC representative protein markers (e.g., claudin 4 and CA19-9) showed strong concordance. DNA- and RNA-sequencing (RNAseq) of single organoids revealed patient-specific genomic and transcriptomic consistency. Single-cell RNAseq demonstrated that organoids are primarily a clonal population. In drug response assays, organoids displayed patient-specific sensitivities. In addition, the PDX response to FOLFIRINOX and gemcitabine/abraxane treatments were examined, which was recapitulated with organoids. This study has demonstrated that organoids are potentially invaluable for precision medicine as well as preclinical drug treatment studies because they maintain distinct patient phenotypes and respond differently to drug combinations and dosage. The patient-specific molecular and histopathologic fidelity of organoids indicate that they can be used to understand the etiology of the patient's tumor and the differential response to therapies and suggests utility for predicting drug responses.

Hippo-YAP1 Is a Prognosis Marker and Potentially Targetable Pathway in Advanced Gallbladder Cancer

“…GBC-PDOs cultures were established from tumor tissue samples of patients diagnosed with moderated differentiated and pT3 N0/N1 Mx stage (Eight edition of American Joint Committee on Cancer (AJCC)) advanced tubular GBC. Gallbladder tumor organoids were generated as previously described [54]. For organoids cultures, dissociated cells were washed in 1× DPBS and embedded in growth factor reduced Matrigel (#356231, Corning, NY, USA) and cultured in complete media (Intesticult (#6005, Stemcell Technologies, Vancouver, BC, Canada), 0.5 µmol/L A83-01 (#SML0788; Sigma-Aldrich), 100 ng/mL, FGF10 (#100-26, PeproTech, Inc., Rocky Hill, NJ, USA), 100 ng/mL HGF (#100-39; Peprotech), 10 nmol/L Gastrin I (#G9145; Sigma), 10 mmol/L N-acetyl-l-cysteine (#A9165; Sigma-Aldrich) 10 mmol/L nicotinamide (#N0636; Sigma-Aldrich), 1× B27 supplement (#17504-044; Gibco, Thermo Fisher Scientific, Waltham, MA, USA), 1× N2 supplement (#175020-048; Gibco), 1 mg/mL Primocin (#ant-pm-1; InvivoGen, San Diego, CA, USA) and 10.5 µmol/L Y-27632 (#Y0503, Sigma-Aldrich).…”

Optical Metabolic Imaging of Heterogeneous Drug Response in Pancreatic Cancer Patient Organoids

“…Alternatively, pancreatic organoids have emerged as an appealing method to tailor treatments by performing high-throughput drug screening directly on a patient's tumor cells (7)(8)(9). In vitro organoids recapitulate the genetic and histopathological characteristics of the original pancreatic tumor, along with its complex 3-dimensional organization (10)(11)(12)(13)(14). Organoid cultures also preserve interactions between tumor cells, immune cells (15), and fibroblasts (16), which can influence tumor drug response and are potential drug targets (17,18).…”

An Organotypic Microcosm for the Pancreatic Tumor Microenvironment

“…Additionally, normal murine pancreatic organoids induced with a Kras mutation, which occurs in 94% of PDACs, displayed typical features of early PDAC, including multilayering epithelial cells, disorganized cells, and overlapping nuclei [14][15][16]. PDAC organoids also harbor pancreatic cancer markers CK7, CK19, P53, and lack of CK20, consistent with paired tumor tissues [12,13]. Furthermore, organoids are representative models of the genetic landscape of the tumor of origin and have been indicated as a beneficial drug sensitivity model for PDAC patients [17,18].…”

“…PDAC tumor organoids represent the morphologic features of the primary tumor ( Figure 1). Compared to normal pancreatic organoids, tumor organoids contain nuclear irregularity, nucleolar prominence, and cell-cell adhesions as seen by hematoxylin and eosin staining [12,13]. Additionally, normal murine pancreatic organoids induced with a Kras mutation, which occurs in 94% of PDACs, displayed typical features of early PDAC, including multilayering epithelial cells, disorganized cells, and overlapping nuclei [14][15][16].…”