Dataset Information

RNA sequencing of WT and Srp72+/- LT-HSCs from aged mice

ABSTRACT: To investigat the role of Srp72 in hematopoiesis and the development of myelodysplasia and bone marrow aplasia, we generated a transgenic mouse strain with a Srp72 KO strand. Heterozygous loss of SRP72 in mice is not associated with major changes in steady-state hematopoiesis, although we did observe mild reductions in peripheral blood and BM cellularity. Moreover, we also detected minor changes within the stem/progenitor compartment as well as in their propensity to generate a balanced lineage output. We did not observe the development of any haematological disorder, thus the Srp72+/- mouse model only partially recapitulates the more severe aplasia and myelodysplasia observed in families with inherited SRP72 lesions. We thereafter extracted RNA from LT-HSCs of old mice to observe transcriptional changes in those mice. Interestingly, gene expression analysis demonstrated that genes encoding secreted factors, including cytokines and cell-surface receptors, were transcriptionally down-regulated in Srp72+/- animals. This can potentially provide mechanistic insights into why SRP72 lesions are associated with the development of severe aplasia and myelodysplasia in humans.

ORGANISM(S): Mus musculus

PROVIDER: GSE132900 | GEO | 2019/07/03

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Tight regulation of hematopoietic stem cell (HSC) homeostasis is essential for life-long hematopoiesis, for preventing blood cancers and for averting bone marrow failure. The underlying mechanisms are incompletely understood. Here, we identify production of inositol-tetrakisphosphate (IP4) by inositoltrisphosphate 3-kinase B (ItpkB) as essential for HSC quiescence and function. Young ItpkB-/- mice accumulated phenotypic HSC and showed extramedullary hematopoiesis. ItpkB-/- HSC were less quiescent and proliferated more than wildtype controls. They downregulated quiescence and stemness associated mRNAs, but upregulated activation, oxidative metabolism, protein synthesis and lineage associated transcripts. Although they showed no significant homing defects, ItpkB-/- HSC had a severely reduced competitive long-term repopulating potential. Aging ItpkB-/- mice lost hematopoietic stem and progenitor cells and died with severe anemia. Wildtype HSC normally repopulated ItpkB-/- hosts, incidating a HSC-intrinsic ItpkB requirement. ItpkB-/- HSC had reduced cobblestone-area forming cell activity in vitro and showed increased stem-cell-factor activation of the phosphoinositide 3-kinase (PI3K) effector Akt, reversed by exogenous provision of the known PI3K/Akt antagonist IP4. They also showed transcriptome changes consistent with hyperactive Akt/mTOR signaling. Thus, we propose that ItpkB ensures HSC quiescence by limiting cytokine-induced PI3K signaling in HSC. For each of 3 replicate ItpkB-/- or wt samples, we enriched Lin- cells from BM of 4 pooled age-matched mice with Rapidspheres (Stemcell Technologies), FACS-sorted ?10,000 LSK CD34-CD150+CD48-Flk2- LT-HSC into lysis buffer and prepared RNA with RNeasy Micro kits (Quiagen). RNA sequencing was done using an Illumina HISeq Analyzer 2000, Casava v1.8.2 genome analyzer pipeline, TopHat v1.4.1/Bowtie2 genome alignment and Partek v6.6 mRNA annotation software. Statistical analyses were done with edgeR (Bioconductor package), excluding genes with false discovery rates >0.15, fold-change magnitudes ?1.4 and log2(counts per million) ?4 to avoid undefined values and the poorly defined log fold-changes for low counts close to 0. Unsupervised clustering of 441 significantly changed genes was done with dChip using rank correlation and a centroid linkage method. Scatter plots were generated in Spotfire. GSEA was performed with gene set permutation, using gene sets from MSigDB (www.broadinstitute.org/gsea/msigdb/index.jsp) or manually curated from, excluding genes without HUGO approved symbols

Dataset Information

RNA sequencing of WT and Srp72+/- LT-HSCs from aged mice

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets