Project description:RNA sequencing of hematopoietic stem cells isolated from young (8-12 week) and old (24-27 month) BL6 mice from experimental groups that are intended to describe the transcriptomic consequences of parabiosis on the function of old hematopoietic stem cells.
Project description:Identification of differentially expressed genes in young (3 month old) versus aged (24 month old) mouse hematopoietic stem cells. Comparison of genes differentially expressed in hematopoietic stem cells of young mice with conditional deletion of mTOR within vascular endothelium.
Project description:Purpose: Compare the transcriptome of hematopoietic stem cells (HSCs) that were aged in old and young niches Methods: barcoded GFP+ HSCs were FACS-sorted from a) three recipient mice 15 months post transplantation, and b) six serial transplantation recipient mice 5 months after the 8th transplantation, then subjected to processed using the Chromium Single-cell 3′ v2 Library Kit (10× Genomics, Pleasanton, CA) following the manufacturer’s instructions Results: we obtained transcriptomes of about 12k HSCs aged in young niche, and about 10k HSCs aged in old niche, with the average sequencing depth at close to 50k reads per cell Conclusions: we identified striking differences in gene expression profiles 1) between HSCs aged in young niches from mice with early aging and from mice with delayed aging, and 2) between HSCs aged in old niches and young niches when mice exhibited hematopoietic aging phenotype
Project description:Mass spectrometry was performed with an Orbitrap Fusion Tribrid mass spectrometer (Thermo Scientific) interfaced with an UltiMate 3000 Binary RSLCnano System (Dionex). Proteome Discoverer v.1.4 (Thermo Scientific) with SEQUEST HT search engines was used for the spectra-preprocessing and HCD MS2 spectra were used for peptide identification and quantitation based on TMT reporter ions. TMT isobaric comparison of old versus young haematopoietic stem and progenitor cells. Young 1 and Young 2 are samples 126 and 128 of dataset UTH_1. Old 1 and Old 2 are samples 129 and 130 of UTH_1. Young 3 is sample 131 and Old 3 is sample 130 of dataset UTH_4.
Project description:Hematopoietic stem cells (HSCs) represent a rare population of cells residing in the Bone Marrow (BM) at the top of hematopoietic hierarchy. A critical balance is maintained between self-renewal and lineage differentiation of HSCs to maintain hematopoietic homeostasis. With aging, this balance is altered with an increase of self-renewal long term HSCs and a myeloid biased differentiation, which favors the appearance of myeloid leukemias and anemias. This experiment aims to understand molecular mechanisms that cause this aged-related disequilibrium in the mouse. To this end, we generated single cell RNA-seq data from pools of young and old hematopoietic stem and progenitor cells (HSPCs), isolated from mouse BMs.
Project description:Loss of immune function and an increased incidence of myeloid leukemia are two of the most clinically significant consequences of aging of the hematopoietic system. To better understand the mechanisms underlying hematopoietic aging, we evaluated the cell intrinsic functional and molecular properties of highly purified long-term hematopoietic stem cells (LT-HSCs) from young and old mice. We found that LT-HSC aging was accompanied by cell autonomous changes, including increased stem cell self-renewal, differential capacity to generate committed myeloid and lymphoid progenitors, and diminished lymphoid potential. Expression profiling revealed that LT-HSC aging was accompanied by the systemic down-regulation of genes mediating lymphoid specification and function and up-regulation of genes involved in specifying myeloid fate and function. Moreover, LT-HSCs from old mice expressed elevated levels of many genes involved in leukemic transformation. These data support a model in which age-dependent alterations in gene expression at the stem cell level presage downstream developmental potential and thereby contribute to age-dependent immune decline, and perhaps also to the increased incidence of leukemia in the elderly. 3 old mice and 5 young mice were assayed
Project description:Bulk RNA sequencing (RNAseq) analysis of hematopoietic stem/progenitor cells purifiied from young and longevity mice, both male and female.
Project description:As part of a study of the role of the aryl hydrocarbon receptor (Ahr) in maintenance and senescence of hematopoietic stem cells (HSC), global gene expression profiling was done with HSC isolated from bone marrow restricted conditional Ahr-knockout and AhR floxed mice. HSC from young-adult (8 wk old) cAhR-KO mice had changes in expression of many genes related to HSC maintenance, consistent with the phenotype observed in Ahr-KO mice. Aged cAhR-KO mice (18 months old) also displayed alterations in peripheral white blood cell counts, serial repopulation potential and levels of ROS in bone marrow cells, consistent with previous observations on the role of AhR in the hematopoietic system. 22 samples: 5 young Ahr knockout, 6 old Ahr knockout, 5 young floxed Ahr, 6 old floxed Ahr