Aberrant Alternative Splicing In U2af1/Tet2 Double Mutant Mice Contributes To Major Hematological Phenotypes

Mutations in splicing factors are recurrent somatic alterations identified in myelodysplastic syndromes (MDS) and they frequently coincide with mutations in epigenetic factors. About 25% of patients present concurrent mutations in such pathways, suggesting a cooperative role in the pathogenesis of MDS. We focused on the splicing factor U2AF1 involved in the recognition of the 3 ' splice site during pre-mRNA splicing. Using a CRISPR/Cas9 system, we created heterozygous mice with a carboxy-terminal truncated U2af1 allele (U2af1(mut/+)), studied the U2af1(mut/+) hematopoietic system, and did not observe any gross differences in both young (12-13 weeks) and old (23 months) U2af1(mut/+) mice, except for a reduction in size of approximately 20%. However, hematopoietic stem/progenitor cells lacked reconstitution capacity in transplantation assays and displayed an aberrant RNA splicing by RNA sequencing. We also evaluated U2af1(mut/+) in conjunction with Tet2-deficiency. Novel double mutant U2af1(mut/+) Tet2(-/-) mice showed increased monogranulocytic precursors. Hematopoietic stem/progenitor cells were also enhanced and presented functional and transcriptomic alterations. Nonetheless, U2af1(mut/+) Tet2(-/-) mice did not succumb to MDS disease over a 6-month observation period. Collectively, our data suggest that cooperation between mutant U2af1 and Tet2 loss is not sufficient for MDS initiation in mice.