Hematopoiesis in Murine Fancd2-/- vs WT Fetal Liver

Fanconi anemia (FA) is a rare inherited bone marrow failure (BMF) syndrome, caused by mutation in one of at least 23 proteins in the FA complementation group. The mechanism of BMF in FA is currently unknown. Previous studies have shown that functional defects in hematopoeitic stem and progenitor cells (HSPC) arise spontaneously in response to developmental replication stress in murine FA fetal livers (FL) as early as 13.5 dpc. In this study, we harvested E13.5 wild-type (WT) and Fancd2-/- FL from three pregnancies, depleted samples of erythroid progenitors via FACS against Ter-119, and performed single-cell RNA sequencing. Our goal was to identify which popoulations in the fetal liver at this time point show severe transcriptional dysregulation, as well as to identify what cellular processes in these populations are most affected, in order to gain insight into the mechanism of BMF in FA and guide future studies. Overall design: single-cell RNA sequencing of live TER119- E13.5 WT and Fancd2-/- FL cells libraries generated from freshly isolated cells (same day) of sex-matched littermates from three pregnancies pregnancy #1: male WT (1.1), male Fancd2-/- (1.1) pregnancy #2: female WT (2.1), female Fancd2-/- (2.1) pregnancy #3: male WT (3.1), male WT (3.2), female Fancd2-/- (3.1), female Fancd2-/- (3.2)