Unknown,Transcriptomics,Genomics,Proteomics

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Single cell RNA-seq of Stmn1+ isthmal cells and Pgc+ chief cells to compare and characterize distinct gastric corpus stem cells at molecular level

ABSTRACT: The purpose of this single cell experiment is to compare and characterize at molecular level actively cycling stem cells in the isthmus and quiescent stem cells in the base of the mouse stomach corpus. The lineage tracing data from Stmn1-CreERT2 shows that Stmn1+ cells in the corpus isthmus include fast dividing isthmus stem cells with long-term potency. On the other hand, chief cells including Lgr5+ subpopulation can play as quiescent stem cells in the base that are largely quiescent in homeostasis, but are activated upon injury. The isthmus stem cells and chief cells are isolated by Stmn1 and Pgc, respectively, and were subject to single cell RNA-seq experiment.

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Mus musculus

SUBMITTER: Seungmin Han

PROVIDER: E-MTAB-6879 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Carcinoma-associated fibroblasts (CAFs) that express ?-smooth-muscle-actin (?SMA+) contribute to cancer progression, but their precise origin and role in tumorigenesis is not established. Using mouse models of inflammation-induced gastric cancer, we show that at least 20% of CAFs originate from bone marrow and derive from mesenchymal stem cells (MSCs). Surprisingly, we find that ?SMA+ myofibroblasts (MF) are niche cells normally present in bone marrow and increase markedly in the bone marrow and blood during progression to dysplasia. MSC-derived CAFs that are recruited to the dysplastic stomach express IL-6, Wnt5? and BMP4 and show DNA hypomethylation. Bone marrow (BM)-derived CAFs strongly promote tumor growth in organotypic and xenograft models. In addition, CAFs are generated from MSCs and are recruited to distant tumor sites in a TGF-?- and SDF-1?-dependent manner. Carcinogenesis therefore involves the expansion and relocation of normal bone marrow niche cells to the tumor site where they create a new niche to sustain cancer progression. Since resident (non-BM-derived) CAFs could not be cultured and directly compared to BM-derived CAFs, we additionally isolated total RFP(+) gastric CAFs from aSMA-RFP mice with Helicobacter felis-induced dysplasia, and compared them to GFP(+) BM-derived gastric CAFs from mice with H. felis-induced dysplasia mice that had been transplanted with UBC-EGFP bone marrow. The RFP+ CAFs (HF CAF) represent total CAFs (of which only 20% were BM-derived), while the latter represented only BM-derived CAFs (BM CAF). We compared their gene expression using the Illumina array (MouseWG-6v2) directly after FACS sorting. Interestingly, the GFP+ BM-derived CAFs expressed higher levels of inflammatory genes (IL-6, IL-1?, IL-33) and a number of tumor and stem cell associated factors (CCL5, SPP1, Notch3, MMP9, CD47, CXCR4, PARP10,) compared to the total (RFP+) population of gastric CAFs. Comparison of bone marrow-derived GFP-labeled gastric CAFs versus all gastric CAFs.

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Single cell RNA-seq of Stmn1+ isthmal cells and Pgc+ chief cells to compare and characterize distinct gastric corpus stem cells at molecular level

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