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RNA sequencing of sorted cells from mouse PDAC

ABSTRACT: RNA sequencing of sorted cells from mouse PDAC

PROVIDER: PRJNA418408 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6292400.fastq.gz	Fastqsanger.gz
	SRR6292410.fastq.gz	Fastqsanger.gz
	SRR6292401.fastq.gz	Fastqsanger.gz
	SRR6292411.fastq.gz	Fastqsanger.gz
	SRR6292402.fastq.gz	Fastqsanger.gz

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RNA sequencing of sorted cells from mouse PDAC

Project description:Pancreatic ductal adenocarcinoma (PDAC) is characterized by the presence of abundant desmoplatic stroma primarily composed of cancer-associated fibroblasts (CAFs). It is generally accepted that CAFs stimulate tumor progression and might be implicated in drug resistance and immunosuppression. Here, we have compared the transcriptional profile of PDGFRα+ CAFs isolated from genetically engineered mouse PDAC tumors with that of normal pancreatic fibroblasts (NPFs) to identify genes potentially implicated in their pro-tumorigenic properties. We report that the most differentially expressed gene, Saa3, a member of the acute-phase Serum Amyloid A (SAA) apolipoprotein family, is a key mediator of the pro-tumorigenic activity of PDGFRα+ CAFs. Whereas Saa3 competent CAFs stimulate the growth of PDAC tumor cells in an orthotopic model, Saa3 null CAFs inhibit tumor growth. Saa3 also plays a role in the cross-talk between CAFs and tumor cells. Ablation of Saa3 in pancreatic tumor cells makes them insensitive to the inhibitory effect of Saa3 null CAFs. As a consequence, germline ablation of Saa3 does not prevent PDAC tumor development in mice. The pro-tumorigenic activity of Saa3 in CAFs is mediated by Mpp6, a member of the palmitoylated membrane protein subfamily of the peripheral membrane-associated guanylate kinases. Finally, we interrogated whether these observations could be translated to a human scenario. Indeed, SAA1, the orthologue of murine Saa3, is overexpressed in human CAFs. Moreover, high levels of SAA1 in the stromal component correlate with worse survival. These findings support the concept that selective inhibition of SAA1 in CAFs may provide potential therapeutic benefit to PDAC patients.

2018-01-22 | GSE106901 | GEO

CUT&Tag sequencing in PDAC cells with Setd2-KO and Setd2-WT sorted from orthotopic PDAC tumor

Project description:To gain mechanistic insight into how Setd2 loss reprogramming tumor cell metabolism, we performed CUT&Tag sequencing in PDAC cells with Setd2-KO and Setd2-WT sorted from orthotopic PDAC tumor. Setd2-WT and -KO murine PDAC cells were sorted with DAPI-CD45.2-Pdpn-Epcam+ or cultured, and freshly collected as soon as possible. Hyperactive pA-Tn5 Transposase for CUT&Tag kit from Vazyme (TD901), and antibodies against H3K36me3, H3K27Ac and H3K27me3 were employed. Trueprep index kit v2 and v3 for illumina were used to establish DNA library.

2024-08-07 | GSE196241 | GEO

RNA-seq of sorted KPC Sf3b1-K700E murine PDAC cells

Project description:RNA-seq of sorted KPC Sf3b1-K700E murine PDAC cells

| PRJNA839859 | ENA

Bulk RNA-seq analysis of PDAC tumor Cells

Project description:Orthotopic PDAC tumor were established with KPC and KPSC cells, respectively. After about 10-14 days, PDAC cells with Setd2-KO and Setd2-WT were sorted from orthotopic PDAC tumor by DPAI-CD45.2-Pdpn-Epcam+, and used for RNA preparation.

2024-08-07 | GSE196752 | GEO

RNA-seq of sorted KPC Sf3b1-K700E murine PDAC cells

Project description:We established a mouse model of KrasG12D, Trp53-/- and Sf3b1-K700E murine pancreatic cancer to elucidate the impact of the SF3B1 mutation found in human PDAC on the KPC mouse model.

2022-06-07 | GSE203339 | GEO

RNA-seq comparison of CD90- myCAFs and CD90+ myCAFs flow-sorted from murine PDAC tumors derived from the orthotopic transplantation of PDAC KPC T-LOH organoids

Project description:We investigated differences between CD90- myCAFs and CD90+ myCAFs flow-sorted from murine PDAC tumors derived from the orthotopic transplantation of PDAC KPC T-LOH organoids

2023-11-30 | GSE243828 | GEO

RNA-seq of epithelial cells flow-sorted from PDAC tumors derived from the orthotopic transplantation of PDAC T-LOH KPC organoids with either Egfr WT PSCs or Egfr KO PSCs in NSG mice

Project description:We investigated differences in epithelial cells flow-sorted from PDAC tumors derived from the orthotopic transplantation of PDAC T-LOH KPC organoids with either Egfr WT PSCs or Egfr KO PSCs

2023-11-30 | GSE243888 | GEO

CUT&Tag sequencing in PDAC cells with Setd2-KO and Setd2-WT sorted from orthotopic PDAC tumor

Project description:CUT&Tag sequencing in PDAC cells with Setd2-KO and Setd2-WT sorted from orthotopic PDAC tumor

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RNA-seq analysis of CAFs and epithelial cells flow-sorted from PDAC tumors derived from the orthotopic transplantation of PDAC T-LOH KPC organoids in C57BL/6J mice following 2-week treatment with Neratinib or vehicle

Project description:We investigated changes in CAFs and epithelial cells flow-sorted from PDAC tumors derived from the orthotopic transplantation of PDAC T-LOH KPC organoids in C57BL/6J mice following 2-week treatment with Neratinib compared to vehicle

2023-11-30 | GSE243889 | GEO

Analysis of oncogenic Kras-induced changes in the RNA-bound proteome of PDAC cells

Project description:In order to reveal the impact of oncogenic KRAS signaling on the RNA-bound proteome of Pancreatic Ductal Adenocarcinoma (PDAC), we carried out quantitative whole Transcriptome RNA Interactome Capture (RIC) analysis in tumour cells from an inducible mouse model of PDAC (iKras PDAC) (Ying et al., 2012). In this model, oncogenic Kras (G12D) expression can be controlled by administration of doxycycline (Dox). For RIC, we employed Orthogonal Organic Phase Separation (OOPS)(Queiroz et al., 2019). In this method, UV-C crosslinking coupled with phenol-chloroform based phase separation is used to isolate RNA-crosslinked proteins, which concentrate in the interface fraction, whilst free proteins concentrate in the organic fraction. SILAC was employed for quantitative analysis of Dox induced (24hrs) changes in the RNA-bound proteome. To assess the impact of ERK signalling, an independent experiment was carried out in which Trametinib (10nM) was added to the cells at the same time as Dox (both with or without Dox cells). A total of 6 biological replicates without Trametinib, and 4 biological replicates with Trametinib, were conducted, with switching the Heavy and Light SILAC labels for +Dox and -Dox conditions. Both the Interface as well as the organic phase fractions were analysed by mass spectrometry. In a separate experiment, the total proteome changes in response to dox (24hrs) were quantified from two reciprocally SILAC labelled mixes of heavy and light iKras PDAC whole cell lysates (See the READ ME.XLS for details of the experimental conditions).

2023-02-12 | PXD030825 | Pride

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