Project description:To investigate the impact of perturbing key differentiation driver genes on hematopoietic stem cell fate decision, we employed a Dapp1 gene knockout approach. This allowed us to observe changes in the subpopulations of hematopoietic stem cells at the LSK level in mice. Additionally, we assessed alterations in their fate decision and lineage differentiation tendencies
Project description:To investigate the impact of radiation damage on hematopoietic stem cell fate decision, we conducted a longitudinal study, collecting single-cell data on days 2 post-radiation exposure. This approach enabled us to observe changes in the subpopulations of hematopoietic stem cells at the LSK level in mice over time. Additionally, we assessed alterations in their fate determination and lineage differentiation tendencies throughout the recovery process.
Project description:In this study, we aimed to investigate the differentiation dynamics of LSK (Lin-Sca1+c-Kit+) cells following radiation injury. LSK cells, which are a critical component of the hematopoietic stem and progenitor cell population, play a vital role in the regeneration and repair of the hematopoietic system after damage. By utilizing these cells, we sought to understand the mechanisms underlying their response to radiation, focusing on their capacity for self-renewal and differentiation into various blood cell lineages.
Project description:Gene expression of LSK (lin-Sca-Kit+) hematopoietic stem cells from wild type mice was compared with LSK from Cebpa knock-in mutant mice (K/K, K/L, and L/L mutants). Fetal liver cells for each genotype were competitively transplanted into irradiant recipients. Donor-derived LSK cells were isolated by FACS sorting of recipient bone marrow. 3 biological replicates of each were generated and expression profiles were determined by hybridization to Affymetrix Moe430_2 arrays.
Project description:Ezh1 is a protein member of PRC2. Ezh1 has been described as a functional repressor gene, such as its homologous Ezh2. We are investigating the role of Ezh1 in hematopoietic stem cells, aging, self-renewal and differentiation. We used microarrays to detail the global program of gene expression in LSK cells from mice with knocked-down expression of Ezh1. LSK cells from EZH1 knockout and control mice were marked and isolated by fluorescence-activated cell sorting. 2 replicates each.
Project description:Little is known of hematopoietic stem (HSC) and progenitor (HPC) cell self-renewal. The role of Brahma (BRM), a chromatin remodeler, in HSC/HPC function is unknown. Bone marrow (BM) from Brm-/- mice manifested increased numbers of long- and short-term HSCs, GMPs, and increased numbers and cycling of functional HPCs. Brm-/- HSC/HPC show demonstrated functional differences compared to wildtype HSC/HPC in vivo and ex vivo, due in part to increased intracellular valine in lineage negative BM. To determine changes in gene programs associated with loss of BRM, we performed RNA sequencing on Brm-/- LSK cells. Brm-/- LSK exhibited upregulated interferon response/cell cycle gene programs, suggesting important extrinsic effects on primitive HSC/HPC from more mature HPC due to the absence of BRM.
Project description:Ezh2 is a protein member of PRC2 and has described like a functional repressor gene. We are investigating the role of Ezh2 in hematopoietic stem cell, aging and leukemia. In the present study, we generated mouse models that allow gain-of-function of Ezh2 in the hematopoietic system. Activation of Ezh2 expression specifically in self-renewing HSCs alters gene expression programs and severely compromises hematopoietic function, leading to the development of myeloproliferative disease. We used microarrays to detail the global programme of gene expression in LSK that has relationship with the overexpression of Ezh2. LSK were marked and isolated by Fluorescence-activated cell sorting in Trizol.
Project description:We performed RNA sequencing analyses of adult mouse bone marrow lineage-negative, Sca-1-positive, and c-kit-positive (LSK) multipotent progenitor cells in wildtype and Drd2 Drd3 double knockout mice