Dysregulated microRNA Activity in Shwachman-Diamond Syndrome
Annual rept. 15 Aug 2014-14 Aug 2015
DANA-FARBER CANCER INST BOSTON MA
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Shwachman-Diamond Syndrome SDS is an inherited bone marrow failure primarily affecting myeloid development. Because the affected cells are rare and heterogeneous, the altered genetic networks in vivo remain unknown. The central goal of this grant is to define transcriptional signatures of bone marrow failure in SDS using single cell RNA-seq of patient cells. We will analyze these datasets to test the novel hypothesis that reduced microRNA activity contributes to hematopoietic dysfunction in SDS. To date, we have sequenced 300 hematopoietic stem and progenitor cells HSPC from normal donors and SDS patients and established, to our knowledge, the first hematopoietic ontogeny at single cell resolution. Differential gene expression analyses between normal and SDS cells revealed cell-type restricted gene expression changes in every subpopulation of HSPC. Our preliminary results suggest that closely related HSPC subpopulations are variably affected by SBDS mutations, which may contribute to complex and unstable hematopoietic symptoms in patients. Ongoing and future work includes 1 annotation of differentially expressed genes to hematopoietic phenotypes in cellular and animal models of SDS, 2 targeted single cell RNA-seq to generate quantitative expression data for a panel of low abundance, disease-relevant genes that were impossible to detect using traditional RNAseq and 3 generation of microRNA expression profiles from HSPCs to be overlaid onto mRNA profiles.
- Genetic Engineering and Molecular Biology
- Anatomy and Physiology
- Medicine and Medical Research