|
| Status |
Public on May 01, 2024 |
| Title |
BRD9 regulates normal human hematopoietic stem cell function and lineage differentiation [RNA-Seq] |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Bromodomain containing protein 9 (BRD9), a member of the non-canonical BRG1/BRM-associated factor (ncBAF) chromatin remodeling complex, has been implicated as a synthetic lethal target in AML but its function in normal human hematopoiesis is unknown. In hematopoietic stem and progenitor cells (HSPC) genomic or chemical inhibition of BRD9 led to a proliferative disadvantage and loss of stem cells in vitro. Human HSPCs with reduced BRD9 protein levels produced lower numbers of immature mixed multipotent GEMM colonies in semi-solid media. In lineage-promoting culture conditions, cells with reduced BRD9 levels failed to differentiate into the megakaryocytic lineage and showed delayed differentiation into erythroid cells but enhanced terminal myeloid differentiation. HSPCs with BRD9 knock down (KD) had reduced long-term multilineage engraftment in a xenotransplantation assay. An increased number of downregulated genes in RNAseq analysis after BRD9 KD coupled with a gain in chromatin accessibility at the promoters of several repressive transcription factors (TF) suggest that BRD9 functions in the maintenance of active transcription during HSC differentiation. In particular, the hematopoietic master regulator GATA1 was identified as one of the core TFs regulating the gene networks modulated by BRD9 loss in HSPCs. BRD9 inhibition reduced a GATA1-luciferase reporter signal, further suggesting a role for BRD9 in regulating GATA1 activity. BRD9 is therefore an additional example of epigenetic regulation of human hematopoiesis.
|
| |
|
| Overall design |
To investigate the function of BRD9 in human CD34+ hematopoietic stem and progenitor cells, cord blood derived CD34 cells were lentivirally infected in triplicates with GFP tagged vector containing either control (shNT) or hairpin against BRD9 (shBRD9#33 and shBRD9#81), sorted for GFP at 72 hours post infection and collected for RNA sequencing. Differential gene expression analysis was performed on CD34 cells with BRD9 knockdown compared to control.
|
| |
|
| Contributor(s) |
Garg S, Griffin JD |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
Apr 15, 2024 |
| Last update date |
May 01, 2024 |
| Contact name |
Swati Garg |
| E-mail(s) |
swati.garg15@gmail.com
|
| Organization name |
Dana Farber Cancer Institute
|
| Street address |
450 Brookline Ave
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02215 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL20301 |
Illumina HiSeq 4000 (Homo sapiens) |
|
| Samples (9)
|
|
| Relations |
| BioProject |
PRJNA1100540 |
Less...
More...