Project description:Hematopoietic stem cells (HSCs) constitute a rare cell population in bone-marrow and are capable of live-long self-renewal and production of all mature blood cell types. Cell differentiation processes are governed by epigenetic mechanisms whose study during early differentiation steps will provide insights into stem cell function and differentiation. We performed whole-genome bisulfite sequencing on HSCs and their immediate progeny, namely three different multipotent progenitor subpopulations (MPP1, MPP2, and MPP). Whole-genome bisulfite sequencing of hematopoietic stem cells (HSCs) and 3 different multipotent progenitor subpopulations (MPP). Three independent biological replicates each were analyzed.
Project description:Hematopoietic stem cells (HSCs) constitute a rare cell population in bone-marrow and are capable of live-long self-renewal and production of all mature blood cell types. Cell differentiation processes are governed by epigenetic mechanisms whose study during early differentiation steps will provide insights into stem cell function and differentiation. We performed whole-genome bisulfite sequencing on HSCs and their immediate progeny, namely three different multipotent progenitor subpopulations (MPP1, MPP2, and MPP).
Project description:Hematopoietic stem cells (HSC) continuously regenerate a complete hematologic and immune system. Very few genes that regulate this process have yet been identified. In order to identify factors governing differentiation, we have compared the transcriptome of highly purified HSC with their differentiated progeny, including erythrocytes, granulocytes, monocytes, NK cells, activated and naïve T-cells, and B-cells. Chromosomal analysis revealed that HSC were more transcriptionally active than other cell types across most chromosomes. Each lineage expressed ~100 to 400 genes uniquely, including many previously uncharacterized genes. Overexpression of two fingerprint genes resulted in a significant bias in differentiation indicating a role in cell fate determination, demonstrating the utility of these data for modulation of specific cell types. Keywords: Hematopoietic cells, stem cells, wild type cells, Mus musculus
Project description:Hematopoietic stem cells (HSC) continuously regenerate a complete hematologic and immune system. Very few genes that regulate this process have yet been identified. In order to identify factors governing differentiation, we have compared the transcriptome of highly purified HSC with their differentiated progeny, including erythrocytes, granulocytes, monocytes, NK cells, activated and naïve T-cells, and B-cells. Chromosomal analysis revealed that HSC were more transcriptionally active than other cell types across most chromosomes. Each lineage expressed ~100 to 400 genes uniquely, including many previously uncharacterized genes. Overexpression of two fingerprint genes resulted in a significant bias in differentiation indicating a role in cell fate determination, demonstrating the utility of these data for modulation of specific cell types. Experiment Overall Design: Goal - Expression profiling of hematopoietic cells Experiment Overall Design: Description - Biological replicates of 10 hematopoietic cell types from female C57Bl/6 mice were purified for microarray analysis Experiment Overall Design: Experimental Factors: wild-type female C57Bl/6 CD45.1 Mus musculus Experiment Overall Design: Experimental Design: 25,000- 50,000 cells were FACS sorted, mRNA was purified by RNAqueous (Ambion), DNAse treated with DNAse1, phenol:chloroform:isoamyl alcohol extracted, and amplified through two rounds of in vitro transcription using MessageAmp (Ambion). Second round was biotinylated with bio-CTP and bio-UTP. Experiment Overall Design: Microarrays: Oligonucleotide, Affymetrix, Moe430 v 2.0 Experiment Overall Design: QC: Biological replicates, correlation coefficient > .95; scale factor <10; 5'/3' ratio < 20
Project description:Induction of trained immunity by human Bacille-Calmette-Guérin (BCG) vaccination is implicated in the beneficial heterologous effects of the vaccine, but the underlying mechanisms remain elusive. We performed global transcriptome analysis of sorted progenitors from bone marrow before (D0) and 90 days after vaccination (D90). BCG vaccination induced transcriptomic myeloid priming of the hematopoietic stem and progenitor cell (HSPC) compartment marked by the upregulation of myeloid and granulocytic pathways alongside the induction of transcription factors connected to myeloid cell function, namely Hepatocyte Nuclear Factors (HNF). These findings are corroborated by higher granulocyte numbers in BCG-vaccinated infants, HNF1-related SNP variants correlating with immune training and elevated serum levels of the HNF1 target gene SERPINA1. Taken together, we reveal a transcriptomic reprograming of HSPCs and peripheral monocytes as a trait of in vivo BCG-induced trained immunity.