The role of Jagged1 as a dynamic switch of cancer cell plasticity in PDAC assembloids

Abstract

Background: Pancreatic ductal adenocarcinoma (PDAC), which commonly relapses due to chemotherapy resistance, has a poor 5-year survival rate (< 10%). The ability of PDAC to dynamically switch between cancer-initiating cell (CIC) and non-CIC states, which is influenced by both internal and external events, has been suggested as a reason for the low drug efficacy. However, cancer cell plasticity using patient-derived PDAC organoids remains poorly understood. Methods: First, we successfully differentiated CICs, which were the main components of PDAC organoids, toward epithelial ductal carcinomas. We further established PDAC assembloids of organoid-derived differentiated ductal cancer cells with endothelial cells (ECs) and autologous immune cells. To investigate the mechanism for PDAC plasticity, we performed single-cell RNA sequencing analysis after culturing the assembloids for 7 days. Results: In the PDAC assembloids, the ECs and immune cells acted as tumor-supporting cells and induced plasticity in the differentiated ductal carcinomas. We also observed that the transcriptome dynamics during PDAC re-programming were related to the WNT/beta-catenin pathway and apoptotic process. Interestingly, we found that WNT5B in the ECs was highly expressed by trans interaction with a JAG1. Furthermore, JAG1 was highly expressed on PDAC during differentiation, and NOTCH1/NOTCH2 were expressed on the ECs at the same time. The WNT5B expression level correlated positively with those of JAG1, NOTCH1, and NOTCH2, and high JAG1 expression correlated with poor survival. Additionally, we experimentally demonstrated that neutralizing JAG1 inhibited cancer cell plasticity. Conclusions: Our results indicate that JAG1 on PDAC plays a critical role in cancer cell plasticity and maintenance of tumor heterogeneity.

Gene expression in organoids: an expanding horizon

“…Lung organoids have been used to model early-stage lung adenocarcinoma [ 149 ] and to identify differentially expressed genes in alveolar epithelial progenitor cells of patients affected by smoking-associated disease [ 150 ]. Organoid model of PDAC allowed identification of new intermediate pancreatic ductal adenocarcinoma (PDAC) transcriptional cell states [ 151 ], while co-culture of PDAC organoids with endothelial cells shed light into the role of JAG1 and NOTCH pathways in cancer cell plasticity [ 152 ]. Single-cell transcriptomics of patient-derived prostate organoids unveiled presence of heterogeneous populations of prostate epithelial cells characterised by enhanced androgen signalling along with a cluster of tumour-associated club cells that may be linked to prostate carcinogenesis [ 153 ].…”

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

“…This model allowed to identify JAG1 as an essential player in this process and as a potential therapeutic target. 48…”

“…This model allowed to identify JAG1 as an essential player in this process and as a potential therapeutic target. 48 To reflect the complexity of processes in the human tissues, the complexity of organoids must be increased and assembloids are considered to be the nearest next step in this long journey. Different approaches in their assembly should be mixed and used to create highly complex, tissue-mimetic organoid models that could serve as very realistic laboratory models.…”

Exploring New Dimensions of Tumor Heterogeneity: The Application of Single Cell Analysis to Organoid‐Based 3D In Vitro Models

“…This analysis implicated Notch pathway activation in endothelial cells by tumor cells as an important player in maintaining tumor cell stemness and metastasis. [ 210 ] Schuth et al. characterized a PDAC PDO co‐culture model with patient matched CAFs using scRNA‐seq and showed changes in CAF and PDAC PDO cell phenotypes in co‐culture compared to monoculture.…”

“…This analysis implicated Notch pathway activation in endothelial cells by tumor cells as an important player in maintaining tumor cell stemness and metastasis. [210] Schuth et al characterized a PDAC PDO co-culture model with patient matched CAFs using scRNA-seq and showed changes in CAF and PDAC PDO cell phenotypes in co-culture compared to monoculture. Furthermore, ligand-receptor analysis identified several possible interactions between PDO cells and CAFs that may contribute to the upregulation of EMT gene expression observed in PDOs co-cultured with CAFs.…”