The human glucocorticoid receptor variant rs6190 increases blood cholesterol and promotes atherosclerosis
Hima Bindu Durumutla, April Haller, Greta Noble, Ashok Daniel Prabakaran, Kevin McFarland, Hannah Latimer, Akanksha Rajput, Olukunle Akinborewa, Bahram Namjou-Khales, David Y. Hui, Mattia Quattrocelli
Hima Bindu Durumutla, April Haller, Greta Noble, Ashok Daniel Prabakaran, Kevin McFarland, Hannah Latimer, Akanksha Rajput, Olukunle Akinborewa, Bahram Namjou-Khales, David Y. Hui, Mattia Quattrocelli
View: Text | PDF
Research Article Endocrinology Genetics Metabolism

The human glucocorticoid receptor variant rs6190 increases blood cholesterol and promotes atherosclerosis

Abstract

Elevated cholesterol poses cardiovascular risks. The glucocorticoid receptor (GR) harbors a still undefined role in cholesterol regulation. Here, we report that a coding SNP in the gene encoding the GR, rs6190, is associated with increased cholesterol in women according to UK Biobank and All of Us (NIH) datasets. In SNP-genocopying mice, we found that the SNP enhanced hepatic GR activity to transactivate Pcsk9 and Bhlhe40, negative regulators of LDL and HDL receptors, respectively. In mice, the SNP was sufficient to elevate circulating cholesterol across all lipoprotein fractions and the risk and severity of atherosclerotic lesions on the proatherogenic hAPOE*2/*2 background. The SNP effect on atherosclerosis was blocked by in vivo liver knockdown of Pcsk9 and Bhlhe40. Also, corticosterone and testosterone were protective against the mutant GR program in cholesterol and atherosclerosis in male mice, while the SNP effect was additive to estrogen loss in females. Remarkably, we found that the mutant GR program was conserved in human hepatocyte-like cells using CRISPR-engineered, SNP-genocopying human induced pluripotent stem cells. Taken together, our study leverages a nonrare human variant to uncover a GR-dependent mechanism contributing to atherogenic risk, particularly in women.

Authors

Hima Bindu Durumutla, April Haller, Greta Noble, Ashok Daniel Prabakaran, Kevin McFarland, Hannah Latimer, Akanksha Rajput, Olukunle Akinborewa, Bahram Namjou-Khales, David Y. Hui, Mattia Quattrocelli

×

Figure 5

Hormonal determinants of sexual dimorphism in the SNP effect.

Options: View larger image (or click on image) Download as PowerPoint
Hormonal determinants of sexual dimorphism in the SNP effect. (A) Serum ...
(A) Serum corticosterone at its typical peak circadian time in mice ZT16 was assessed by ELISA and was higher in females than males of both control and mutant genotypes. (B) Western blot analyses in liver tissue showed that the molecular SNP effect on the identified protein cascade (increased PCSK9 and BHLHE40, decreased LDLR and SR-B1) was present in males, but the effect magnitude was significantly higher in females. (C) Experimental plan for gonadectomy experiment with ELISA-based validation of serum testosterone or estradiol depletion in WT mice on regular chow at 2 weeks after surgery. (D) In male mice, orchiectomy unmasked the SNP effect on increased cholesterol across lipoprotein fractions in either normal (WT background with regular chow) or proatherogenic (hAPOE*2/*2 background with Western diet) conditions (pink arrows). In females, the SNP effect was recapitulated in sham mice (red arrows) and maintained after ovariectomy (red arrows). Scales of y axes are different between normal and atherogenic conditions to adapt to the different cholesterol levels. (E) In atherogenic conditions, orchiectomy unmasked the SNP effect on male plaque/aorta surface ratio (pink arrows); in females the SNP effect was additive to the ovariectomy effect on the aorta plaque profile (red and pink arrows). (F) Male hiPSC-hepatocytes showed higher mRNA expression of the AR than the ER per qPCR. The hepatocyte-autonomous protective effect of the AR agonist Cl-4AS1 on HDL or LDL uptake was modest but significant in vitro in both control and mutant genotypes. We did not detect significant changes induced by estradiol. Scale bars: 100 μm. n = 3–5/sex/group; 2-way ANOVA and Šidák’s test; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. bg, background; CL-4, CL-4AS1, AR agonist; estrad, β-estradiol.