High-Throughput Screening Identifies MicroRNAs Regulating Human PCSK9 and Hepatic Low-Density Lipoprotein Receptor Expression
Investigations into the regulatory mechanisms controlling cholesterol homeostasis have proven fruitful in identifying low-density lipoprotein (LDL)-lowering therapies to reduce the risk of atherosclerotic cardiovascular disease. A major advance was the discovery of proprotein convertase subtilisin/k...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2021-07-01
|
| Series: | Frontiers in Cardiovascular Medicine |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcvm.2021.667298/full |
| _version_ | 1818738088006909952 |
|---|---|
| author | Coen van Solingen Scott R. Oldebeken Alessandro G. Salerno Amarylis C. B. A. Wanschel Kathryn J. Moore Kathryn J. Moore |
| author_facet | Coen van Solingen Scott R. Oldebeken Alessandro G. Salerno Amarylis C. B. A. Wanschel Kathryn J. Moore Kathryn J. Moore |
| author_sort | Coen van Solingen |
| collection | DOAJ |
| description | Investigations into the regulatory mechanisms controlling cholesterol homeostasis have proven fruitful in identifying low-density lipoprotein (LDL)-lowering therapies to reduce the risk of atherosclerotic cardiovascular disease. A major advance was the discovery of proprotein convertase subtilisin/kexin type 9 (PCSK9), a secreted protein that binds the LDL receptor (LDLR) on the cell surface and internalizes it for degradation, thereby blunting its ability to take up circulating LDL. The discovery that loss-of-function mutations in PCSK9 lead to lower plasma levels of LDL cholesterol and protection from cardiovascular disease led to the therapeutic development of PCSK9 inhibitors at an unprecedented pace. However, there remain many gaps in our understanding of PCSK9 regulation and biology, including its posttranscriptional control by microRNAs. Using a high-throughput region(3′-UTR) of human microRNA library screen, we identified microRNAs targeting the 3′ untranslated region of human PCSK9. The top 35 hits were confirmed by large-format PCSK9 3′-UTR luciferase assays, and 10 microRNAs were then selected for further validation in hepatic cells, including effects on PCSK9 secretion and LDLR cell surface expression. These studies identified seven novel microRNAs that reduce PCSK9 expression, including miR-221-5p, miR-342-5p, miR-363-5p, miR-609, miR-765, and miR-3165. Interestingly, several of these microRNAs were also found to target other genes involved in LDLR regulation and potently upregulate LDLR cell surface expression in hepatic cells. Together, these data enhance our understanding of post-transcriptional regulators of PCSK9 and their potential for therapeutic manipulation of hepatic LDLR expression. |
| first_indexed | 2024-12-18T01:03:22Z |
| format | Article |
| id | doaj.art-d7a3d43ee4e840f0bee2edb841757972 |
| institution | Directory Open Access Journal |
| issn | 2297-055X |
| language | English |
| last_indexed | 2024-12-18T01:03:22Z |
| publishDate | 2021-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cardiovascular Medicine |
| spelling | doaj.art-d7a3d43ee4e840f0bee2edb8417579722022-12-21T21:26:18ZengFrontiers Media S.A.Frontiers in Cardiovascular Medicine2297-055X2021-07-01810.3389/fcvm.2021.667298667298High-Throughput Screening Identifies MicroRNAs Regulating Human PCSK9 and Hepatic Low-Density Lipoprotein Receptor ExpressionCoen van Solingen0Scott R. Oldebeken1Alessandro G. Salerno2Amarylis C. B. A. Wanschel3Kathryn J. Moore4Kathryn J. Moore5Leon H. Charney Division of Cardiology, Department of Medicine, New York University Cardiovascular Research Center, New York University School of Medicine, New York, NY, United StatesLeon H. Charney Division of Cardiology, Department of Medicine, New York University Cardiovascular Research Center, New York University School of Medicine, New York, NY, United StatesLeon H. Charney Division of Cardiology, Department of Medicine, New York University Cardiovascular Research Center, New York University School of Medicine, New York, NY, United StatesLeon H. Charney Division of Cardiology, Department of Medicine, New York University Cardiovascular Research Center, New York University School of Medicine, New York, NY, United StatesLeon H. Charney Division of Cardiology, Department of Medicine, New York University Cardiovascular Research Center, New York University School of Medicine, New York, NY, United StatesDepartment of Cell Biology, New York University School of Medicine, New York, NY, United StatesInvestigations into the regulatory mechanisms controlling cholesterol homeostasis have proven fruitful in identifying low-density lipoprotein (LDL)-lowering therapies to reduce the risk of atherosclerotic cardiovascular disease. A major advance was the discovery of proprotein convertase subtilisin/kexin type 9 (PCSK9), a secreted protein that binds the LDL receptor (LDLR) on the cell surface and internalizes it for degradation, thereby blunting its ability to take up circulating LDL. The discovery that loss-of-function mutations in PCSK9 lead to lower plasma levels of LDL cholesterol and protection from cardiovascular disease led to the therapeutic development of PCSK9 inhibitors at an unprecedented pace. However, there remain many gaps in our understanding of PCSK9 regulation and biology, including its posttranscriptional control by microRNAs. Using a high-throughput region(3′-UTR) of human microRNA library screen, we identified microRNAs targeting the 3′ untranslated region of human PCSK9. The top 35 hits were confirmed by large-format PCSK9 3′-UTR luciferase assays, and 10 microRNAs were then selected for further validation in hepatic cells, including effects on PCSK9 secretion and LDLR cell surface expression. These studies identified seven novel microRNAs that reduce PCSK9 expression, including miR-221-5p, miR-342-5p, miR-363-5p, miR-609, miR-765, and miR-3165. Interestingly, several of these microRNAs were also found to target other genes involved in LDLR regulation and potently upregulate LDLR cell surface expression in hepatic cells. Together, these data enhance our understanding of post-transcriptional regulators of PCSK9 and their potential for therapeutic manipulation of hepatic LDLR expression.https://www.frontiersin.org/articles/10.3389/fcvm.2021.667298/fullmicroRNALDL receptorlipoproteinproprotein convertase subtilisin kexin type 9hepatocytes |
| spellingShingle | Coen van Solingen Scott R. Oldebeken Alessandro G. Salerno Amarylis C. B. A. Wanschel Kathryn J. Moore Kathryn J. Moore High-Throughput Screening Identifies MicroRNAs Regulating Human PCSK9 and Hepatic Low-Density Lipoprotein Receptor Expression Frontiers in Cardiovascular Medicine microRNA LDL receptor lipoprotein proprotein convertase subtilisin kexin type 9 hepatocytes |
| title | High-Throughput Screening Identifies MicroRNAs Regulating Human PCSK9 and Hepatic Low-Density Lipoprotein Receptor Expression |
| title_full | High-Throughput Screening Identifies MicroRNAs Regulating Human PCSK9 and Hepatic Low-Density Lipoprotein Receptor Expression |
| title_fullStr | High-Throughput Screening Identifies MicroRNAs Regulating Human PCSK9 and Hepatic Low-Density Lipoprotein Receptor Expression |
| title_full_unstemmed | High-Throughput Screening Identifies MicroRNAs Regulating Human PCSK9 and Hepatic Low-Density Lipoprotein Receptor Expression |
| title_short | High-Throughput Screening Identifies MicroRNAs Regulating Human PCSK9 and Hepatic Low-Density Lipoprotein Receptor Expression |
| title_sort | high throughput screening identifies micrornas regulating human pcsk9 and hepatic low density lipoprotein receptor expression |
| topic | microRNA LDL receptor lipoprotein proprotein convertase subtilisin kexin type 9 hepatocytes |
| url | https://www.frontiersin.org/articles/10.3389/fcvm.2021.667298/full |
| work_keys_str_mv | AT coenvansolingen highthroughputscreeningidentifiesmicrornasregulatinghumanpcsk9andhepaticlowdensitylipoproteinreceptorexpression AT scottroldebeken highthroughputscreeningidentifiesmicrornasregulatinghumanpcsk9andhepaticlowdensitylipoproteinreceptorexpression AT alessandrogsalerno highthroughputscreeningidentifiesmicrornasregulatinghumanpcsk9andhepaticlowdensitylipoproteinreceptorexpression AT amaryliscbawanschel highthroughputscreeningidentifiesmicrornasregulatinghumanpcsk9andhepaticlowdensitylipoproteinreceptorexpression AT kathrynjmoore highthroughputscreeningidentifiesmicrornasregulatinghumanpcsk9andhepaticlowdensitylipoproteinreceptorexpression AT kathrynjmoore highthroughputscreeningidentifiesmicrornasregulatinghumanpcsk9andhepaticlowdensitylipoproteinreceptorexpression |