Supplemental material from: ESR1 mutations associated with EIS change conformation of ligand receptor complex and alter transcriptome profile

Estrogen insensitivity syndrome (EIS) arises from rare mutations in
estrogen receptor α (ERα, encoded by ESR1 gene) resulting in the inability
of estrogen to exert its biological effects. Due to the rarity, mutations
in ESR1 gene and the underlying molecular mechanisms of EIS have not been
thoroughly studied. Here, we investigate known ESR1 mutants, Q375H and
R394H, associated with EIS patients using in vitro and in vivo systems.
Comparison of the transcriptome and DNA methylome from stable cell lines
of both Q375H and R394H clinical mutants show a differential profile
compared to WT ERα resulting in loss of estrogen-responsiveness. Molecular
dynamic simulation shows that both ESR1 mutations change the ERα
conformation of the ligand receptor complexes. Furthermore, we generated a
mouse model Esr1-Q, harboring the human mutation using CRISPR/Cas9 genome
editing. Female and male Esr1-Q mice are infertile and have similar
phenotypes to αERKO mice. Overall phenotypes of the Esr1-Q mice correspond
to those observed in the Q375H patient. Finally, we explore the effects of
a synthetic progestogen and a GnRH inhibitor in the Esr1-Q mice for
potentially reversing the impaired female reproductive tract function.
These findings provide an important basis for understanding the molecular
mechanistic consequences associated with EIS.