ESR1 Mutations Associated With Estrogen Insensitivity Syndrome Change Conformation of Ligand-Receptor Complex and Altered Transcriptome Profile

Abstract

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 its 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 deoxyribonucleic acid methylome from stable cell lines of both Q375H and R394H clinical mutants shows a differential profile compared with wild-type 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 patient with Q375H. Finally, we explore the effects of a synthetic progestogen and a gonadotropin-releasing hormone 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.

A mutant form of ERα associated with estrogen insensitivity affects the coupling between ligand binding and coactivator recruitment

“…3). The effect of differing expression levels of ER-Q375H versus WT ER on gene responsiveness has no influence, as shown by a previous study in which we made stable cell lines in MDA-MB-231 cells, some having considerably higher levels of ER-Q375H than WT ER (25). Even with the high ER-Q375H levels, there was no evidence that the pattern and level of gene expression was altered in any substantial way in the cell lines with different ER levels, suggesting to us that squelching due to different ER expression levels is not likely to be an issue.…”

“…A different homozygous missense mutation in ER, this one located at a critical site of ligand binding stabilization (R394H), was recently identified in three siblings (two females and a male), originating from a consanguineous Algerian family (47). Recently, we have investigated the whole transcriptome profiles of these two clinical ER mutations, Q375H and R394H, in stably expressing the MDA-MB-231 cells (25). Both clinical mutants show a different gene expression profile compared to WT ER, consistent with a loss of estrogen responsiveness.…”

Circulating miR-221/222 expression as microRNA biomarker predicting tamoxifen treatment outcome: a case–control study

“…One important mechanism is the role of miR-221 in degrading ERα messenger RNA, preventing ERα protein translation 52 , 53 . Mutations in the ESR1 gene decrease ERα synthesis 54 . Alternatively, post-translational variation in ERα and crosstalk with growth factor receptors may cause increased proliferation and survival of cancer cells 55 .…”

GMQN: A Reference-Based Method for Correcting Batch Effects and Probe Bias in HumanMethylation BeadChip

“…As a result, we chose two different sets of technical replicates. The first set (EPIC, GSE139687) has nine samples that are repeated three times each, while the second set (450 k, GSE52731) has 56 repetitions of one sample (Aryee et al, 2014;Li et al, 2020). The variances of the probes of the two sets of samples were determined separately.…”