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Single-cell RNA-sequencing of metastatic pancreatic neuroendocrine tumor

ABSTRACT: We report the application of single-cell RNA sequencing (scRNA-seq) technology for single-cell level profiling of the transcriptional features of cells from two primary lesions, one liver metastasis, one normal liver tissue, and peripheral blood monocuclear cells in a patient with metastatic pNET. The transcriptomic profiles of a total of 24.544 cells were captured.

ORGANISM(S): Homo sapiens

PROVIDER: GSE162708 | GEO | 2021/08/24

REPOSITORIES: GEO

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CD73 as a therapeutic target of pancreatic neuroendocrine tumor stem cells

Project description:Background: Identification and purification of cancer stem cells (CSCs) lead to new therapeutic targets; however, there has been no study to identify and isolated pancreatic neuroendocrine tumor (pNET) CSC. Therefore the clinical significance and its target remain unknown. This study aimed to identify pNET CSCs and characterize therapeutic candidate for pNET CSCs. Methods: We isolated CSCs sorting by ALDH activity in pNET surgical section and cell lines. We verified whether these cells have the property of stemness in vivo and in vitro. Additionally in order to acquire CSC gene profile, genome-wide gene expression profiles were investigated using a microarray technique. Results: ALDHhigh cells, but not control bulk cells, formed spheres, proliferated in hypoxia as well as normoxia and promoted cell motility, which are features of CSCs. Injection of as few as 10 ALDHhigh cells led to subcutaneous tumor formation, and 105 ALDHhigh cells established metastases but not control bulk cells in mice. Comprehensive gene expression analysis revealed that genes associated with mesenchymal stem cell, including CD73, and epithelial-mesenchymal transition (EMT) were overexpressed in ALDHhigh cells. APCP, which is CD73 inhibitor, inhibited sphere formation and cell motility in ALDHhigh cells in vitro, and tumor growth inhibition were observed in ALDHhigh cells in vivo. Conclusions: We identified ALDHhigh cells of pNET and elucidated that they have stemness property. Furthermore we identified CD73 as a target of ALDHhigh cells. CD73 is a promising novel target of pNET CSCs.

2016-05-23 | GSE62079 | GEO

RNA-seq analysis of Metastatic and Localized human Pancreatic Neuroendocrine Tumors

Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of RNAs in human pancreatic neuroendocrine tumor (PNET) tissues. We analyzed the transcriptome maps of primary tumor sites from patients that presented metastasis or not (localized). This analysis revelaed that the immune cell and pathway-related molecular signatures were enriched in metastatic compared to localized tumors. Given the wide change of gene expression in metastatic PNETs, we analyzed the RNA-seq data using a L1000 drug-induced signature to identify pharmacologic agents that might preferentially target metastatic disease. In a separate study, we analyzed our RNA-seq data to understand the molecular basis of sexual dimorphisms in PNETs. We found that PNETs are associated with the emergence of unique sex-specific transcriptomic differences that are not observed in non-neoplastic pancreatic islet tissues.

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Project description:We aimed to study the interaction of PNETs and pancreatic stellate cells (PSCs) in PNET progression through exploring specific gene expression of PNETs cultured with conditional media of PSCs.

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RABL6A promotes G1-S phase progression and pancreatic neuroendocrine tumor cell proliferation in an Rb1-dependent manner

Project description:Mechanisms of neuroendocrine tumor (NET) proliferation are poorly understood and therapies that effectively control NET progression and metastatic disease are limited. We found amplification of a putative oncogene, RABL6A, in primary human pancreatic NETs (PNETs) that correlated with high level RABL6A protein expression. Consistent with those results, stable silencing of RABL6A in cultured BON-1 PNET cells revealed that it is essential for their proliferation and survival. Cells lacking RABL6A predominantly arrested in G1 phase with a moderate mitotic block. Pathway analysis of microarray data suggested activation of the p53 and retinoblastoma (Rb1) tumor suppressor pathways in the arrested cells. Specific inactivation of p53 had no effect on the RABL6A knockdown phenotype, indicating RABL6A functions independent of p53 in this setting. By comparison, Rb1 inactivation restored G1 to S phase progression in RABL6A knockdown cells although it was insufficient to override the mitotic arrest and cell death caused by RABL6A loss. Thus, RABL6A promotes G1 progression in PNET cells by inactivating Rb1, an established suppressor of PNET proliferation and development. This work identifies RABL6A as a new negative regulator of Rb1 that is essential for PNET proliferation and survival. We suggest RABL6A is a new potential biomarker and target for anticancer therapy in PNET patients. Total RNA obtained from human BON-1 PNET cells with RABL6A shRNA knockdown compared to BON-1 cells expressing control vector.

2014-07-01 | E-GEOD-53778 | biostudies-arrayexpress

RABL6A promotes G1-S phase progression and pancreatic neuroendocrine tumor cell proliferation in an Rb1-dependent manner

2014-07-01 | GSE53778 | GEO

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Project description:Proteomic and signaling alterations among cells isolated from three sites of metastasis (liver, lung and peritoneum) from a single patient were studied using a SILAC-based quantitative proteomic strategy combined with a high resolution mass spectrometry.

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Single-Cell RNA-Seq Analysis of Metastasis-Primary Tumor Dissimilarity in a Patient with Gastrointestinal Neuroendocrine Cancer

Project description:A 58 year old female diagnosed with a metastic GI-NET underwent resection of primary tumor as well as several metastatic lesions. We performed scRNA-seq of the primary tumor and one lesion from the liver metastasis to perform comparitive analysis between the primary and metastic tumor compositions.

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