Compound haploinsufficiencies of Ebf1 and Runx1 genes impede B cell lineage progression

Early B cell factor (EBF)1 is essential for B lineage specification. Previously, we demonstrated the synergistic activation of Cd79a (mb-1) genes by EBF1 and its functional partner, RUNX1. Here, we identified consequences of Ebf1 haploinsufficiency together with haploinsufficiency of Runx1 genes in mice. Although numbers of committed pro-B cells were maintained in Ebf1(+/-)Runx1(+/-)(ERhet) mice, activation of B cell-specific gene transcription was depressed in these cells. Expression of genes encoding Aiolos, kappa 0 sterile transcripts, CD2 and CD25 were reduced and delayed in ERhet pro-B cells, whereas surface expression of BP-1 was increased on late pro-B cells in ERhet mice. Late pre-B and immature and mature B cells were decreased in the bone marrow of Ebf1(+/-)(E-het) mice and were nearly absent in ERhet mice. Althoughwedid not observe significant effects of haploinsuficiencies on IgH or Ig. rearrangements, a relative lack of Ig. rearrangements was detected in Ehet and ERhet pre-B cells. Together, these observations suggest that Bcell lineage progression is impaired at multiple stages in the bone marrow of Ehet and ERhet mice. Furthermore, enforced expression of EBF1 and RUNX1 in terminally differentiated plasmacytoma cells activated multiple early B cell-specific genes synergistically. Collectively, these studies illuminate the effects of reduced Ebf1 dosage and the compounding effects of reduced Runx1 dosage. Our data confirm and extend the importance of EBF1 in regulating target genes and Ig gene rearrangements necessary for B cell lineage specification, developmental progression, and homeostasis.