Embryonic lethality and defective mammary gland development of activator-function impaired conditional knock-in Erbb3V943R mice

Abstract

ERBB3 is a pseudokinase domain-containing member of the ERBB family of receptor tyrosine kinases (RTKs). Following ligand binding, ERBB receptors homo- or hetero-dimerize, leading to a head-to-tail arrangement of the intracellular kinase domains, where the “receiver” kinase domain of one ERBB is activated by the “activator” domain of the other ERBB in the dimer. In ERBB3, a conserved valine at codon 943 (V943) in the kinase C-terminal domain has been shown to be important for its function as an “activator” kinase in vitro. Here we report a knock-in mouse model where we have modified the endogenous Erbb3 allele to allow for tissue-specific conditional expression of Erbb3^V943R (Erbb3^CKI-V943R). Additionally, we generated an Erbb3^D850N (Erbb3^CKI-D850N) conditional knock-in mouse model where the conserved aspartate in the DFG motif of the pseudokinase domain was mutated to abolish any potential residual kinase activity. While Erbb3^D850N/D850N animals developed normally, homozygous Erbb3^V943R/V943R expression during development resulted in embryonic lethality. Further, tissue specific expression of Erbb3^V943R/V943R in the mammary gland epithelium following its activation using MMTV-Cre resulted in delayed elongation of the ductal network during puberty. Single-cell RNA-seq analysis of Erbb3^V943R/V943R mammary glands showed a reduction in a specific subset of fibrinogen-producing luminal epithelial cells.