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Research article
Retinoic acid receptor-related orphan receptor α reduces lipid droplets by upregulating neutral cholesterol ester hydrolase 1 in macrophages
Hiroshi MATSUOKA
Fukuyama University
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9423-5473
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Neutral cholesterol ester hydrolase 1 (NCEH1) catalyzes the hydrolysis of cholesterol ester (CE) in macrophages. Genetic ablation of NCEH1 promotes CE-laden macrophages and the development of atherosclerosis in mice. Dysregulation of NCEH1 levels is involved in the pathogenesis of multiple disorders including metabolic diseases and atherosclerosis; however, relatively little is known regarding the mechanisms regulating NCEH1. Retinoic acid receptor-related orphan receptor α (RORα)-deficient mice exhibit several phenotypes indicative of aberrant lipid metabolism, including dyslipidemia and increased susceptibility to atherosclerosis.
Results: In this study, inhibition of lipid droplet formation by RORα positively regulated NCEH1 expression in macrophages. In mammals, the NCEH1 promoter region was found to harbor putative RORα response elements (ROREs). Electrophoretic mobility shift, chromatin immunoprecipitation, and luciferase reporter assays showed that RORα binds and responds to ROREs in human NCEH1. Moreover, NCEH1 was upregulated through RORα via a phorbol myristate acetate-dependent mechanism during macrophage differentiation from THP1 cells. siRNA-mediated knockdown of RORα significantly downregulated NCEH1 expression and accumulated lipid droplets in human hepatoma cells. In contrast, NCEH1 expression and removal of lipid droplets were induced by RORα agonist treatments and RORα overexpression in macrophages.
Conclusion: These data strongly suggested that NCEH1 is a direct RORα target, defining potential new roles for RORα in the inhibition of lipid droplet formation through NCEH1.
Keywords
Atherosclerosis, Cholesterol metabolism, Lipid droplets, Macrophages, NCEH1, RORα, Transcriptional regulation
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This preprint is under consideration at BMC Molecular and Cell Biology. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
Retinoic acid receptor-related orphan receptor α reduces lipid droplets by upregulating neutral cholesterol ester hydrolase 1 in macrophages
Hiroshi MATSUOKA
Fukuyama University
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9423-5473
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 5
Posted 13 Apr, 2020
No community comments so far
Submission checks complete
On 07 Apr, 2020
No comments provided
Submission checks complete
On 31 Mar, 2020
Editorial decision: Minor revision
On 31 Mar, 2020
No comments provided
Editorial decision: Minor revision
On 26 Mar, 2020
Editor assigned
On 25 Mar, 2020
Submission checks complete
On 24 Mar, 2020
Editor invited
On 24 Mar, 2020
No comments provided
Editorial decision: Minor revision
On 09 Feb, 2020
Review #2 received
Received 07 Feb, 2020
Reviewer #2 agreed
On 25 Jan, 2020
Reviewer #1 agreed
On 22 Jan, 2020
Review #1 received
Received 22 Jan, 2020
Reviewers invited
Invitations sent on 25 Dec, 2019
Editor assigned
On 22 Dec, 2019
Editor invited
On 21 Dec, 2019
Submission checks complete
On 21 Dec, 2019
No comments provided
Editorial decision: Major revision
On 11 Sep, 2019
Review #2 received
Received 10 Sep, 2019
Reviewer #3 agreed
On 27 Aug, 2019
Reviewer #2 agreed
On 27 Aug, 2019
Reviewer #1 agreed
On 14 Aug, 2019
Review #1 received
Received 14 Aug, 2019
Reviewers invited
Invitations sent on 13 Aug, 2019
Editor assigned
On 26 Jul, 2019
Submission checks complete
On 25 Jul, 2019
Editor invited
On 25 Jul, 2019
First submitted
On 17 Jul, 2019
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Neutral cholesterol ester hydrolase 1 (NCEH1) catalyzes the hydrolysis of cholesterol ester (CE) in macrophages. Genetic ablation of NCEH1 promotes CE-laden macrophages and the development of atherosclerosis in mice. Dysregulation of NCEH1 levels is involved in the pathogenesis of multiple disorders including metabolic diseases and atherosclerosis; however, relatively little is known regarding the mechanisms regulating NCEH1. Retinoic acid receptor-related orphan receptor α (RORα)-deficient mice exhibit several phenotypes indicative of aberrant lipid metabolism, including dyslipidemia and increased susceptibility to atherosclerosis.
Results: In this study, inhibition of lipid droplet formation by RORα positively regulated NCEH1 expression in macrophages. In mammals, the NCEH1 promoter region was found to harbor putative RORα response elements (ROREs). Electrophoretic mobility shift, chromatin immunoprecipitation, and luciferase reporter assays showed that RORα binds and responds to ROREs in human NCEH1. Moreover, NCEH1 was upregulated through RORα via a phorbol myristate acetate-dependent mechanism during macrophage differentiation from THP1 cells. siRNA-mediated knockdown of RORα significantly downregulated NCEH1 expression and accumulated lipid droplets in human hepatoma cells. In contrast, NCEH1 expression and removal of lipid droplets were induced by RORα agonist treatments and RORα overexpression in macrophages.
Conclusion: These data strongly suggested that NCEH1 is a direct RORα target, defining potential new roles for RORα in the inhibition of lipid droplet formation through NCEH1.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8