Novel genetic regulators of fibrinogen synthesis identified by an in vitro experimental platform
Copyright © 2022 International Society on Thrombosis and Haemostasis. All rights reserved..
BACKGROUND: Fibrinogen has an established, essential role in both coagulation and inflammatory pathways, and these processes are deeply intertwined in the development of thrombotic and atherosclerotic diseases. Previous studies aimed to better understand the (patho) physiological actions of fibrinogen by characterizing the genomic contribution to circulating fibrinogen levels.
OBJECTIVES: Establish an in vitro approach to define functional roles between genes within these loci and fibrinogen synthesis.
METHODS: Candidate genes were selected on the basis of their proximity to genetic variants associated with fibrinogen levels and expression in hepatocytes and HepG2 cells. HepG2 cells were transfected with small interfering RNAs targeting candidate genes and cultured in the absence or presence of the proinflammatory cytokine interleukin-6. Effects on fibrinogen protein production, gene expression, and cell growth were assessed by immunoblotting, real-time polymerase chain reaction, and cell counts, respectively.
RESULTS: HepG2 cells secreted fibrinogen, and stimulation with interleukin-6 increased fibrinogen production by 3.4 ± 1.2 fold. In the absence of interleukin-6, small interfering RNA knockdown of FGA, IL6R, or EEPD1 decreased fibrinogen production, and knockdown of LEPR, PDIA5, PLEC, SHANK3, or CPS1 increased production. In the presence of interleukin-6, knockdown of FGA, IL6R, or ATXN2L decreased fibrinogen production. Knockdown of FGA, IL6R, EEPD1, LEPR, PDIA5, PLEC, or CPS1 altered transcription of one or more fibrinogen genes. Knocking down ATXN2L suppressed inducible but not basal fibrinogen production via a post-transcriptional mechanism.
CONCLUSIONS: We established an in vitro platform to define the impact of select gene products on fibrinogen production. Genes identified in our screen may reveal cellular mechanisms that drive fibrinogen production as well as fibrin(ogen)-mediated (patho)physiological mechanisms.
| Errataetall: |
CommentIn: J Thromb Haemost. 2023 Mar;21(3):463-464. - PMID 36858793 |
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| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:21 |
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| Enthalten in: |
Journal of thrombosis and haemostasis : JTH - 21(2023), 3 vom: 25. März, Seite 522-533 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Dobson, Dre'Von A [VerfasserIn] |
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| Links: |
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| Themen: |
9001-32-5 |
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| Anmerkungen: |
Date Completed 03.03.2023 Date Revised 02.03.2024 published: Print-Electronic CommentIn: J Thromb Haemost. 2023 Mar;21(3):463-464. - PMID 36858793 Citation Status MEDLINE |
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| doi: |
10.1016/j.jtha.2022.10.027 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM352067306 |
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| 245 | 1 | 0 | |a Novel genetic regulators of fibrinogen synthesis identified by an in vitro experimental platform |
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| 500 | |a Date Revised 02.03.2024 | ||
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| 500 | |a CommentIn: J Thromb Haemost. 2023 Mar;21(3):463-464. - PMID 36858793 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2022 International Society on Thrombosis and Haemostasis. All rights reserved. | ||
| 520 | |a BACKGROUND: Fibrinogen has an established, essential role in both coagulation and inflammatory pathways, and these processes are deeply intertwined in the development of thrombotic and atherosclerotic diseases. Previous studies aimed to better understand the (patho) physiological actions of fibrinogen by characterizing the genomic contribution to circulating fibrinogen levels | ||
| 520 | |a OBJECTIVES: Establish an in vitro approach to define functional roles between genes within these loci and fibrinogen synthesis | ||
| 520 | |a METHODS: Candidate genes were selected on the basis of their proximity to genetic variants associated with fibrinogen levels and expression in hepatocytes and HepG2 cells. HepG2 cells were transfected with small interfering RNAs targeting candidate genes and cultured in the absence or presence of the proinflammatory cytokine interleukin-6. Effects on fibrinogen protein production, gene expression, and cell growth were assessed by immunoblotting, real-time polymerase chain reaction, and cell counts, respectively | ||
| 520 | |a RESULTS: HepG2 cells secreted fibrinogen, and stimulation with interleukin-6 increased fibrinogen production by 3.4 ± 1.2 fold. In the absence of interleukin-6, small interfering RNA knockdown of FGA, IL6R, or EEPD1 decreased fibrinogen production, and knockdown of LEPR, PDIA5, PLEC, SHANK3, or CPS1 increased production. In the presence of interleukin-6, knockdown of FGA, IL6R, or ATXN2L decreased fibrinogen production. Knockdown of FGA, IL6R, EEPD1, LEPR, PDIA5, PLEC, or CPS1 altered transcription of one or more fibrinogen genes. Knocking down ATXN2L suppressed inducible but not basal fibrinogen production via a post-transcriptional mechanism | ||
| 520 | |a CONCLUSIONS: We established an in vitro platform to define the impact of select gene products on fibrinogen production. Genes identified in our screen may reveal cellular mechanisms that drive fibrinogen production as well as fibrin(ogen)-mediated (patho)physiological mechanisms | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
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| 650 | 7 | |a Hemostatics |2 NLM | |
| 700 | 1 | |a Holle, Lori A |e verfasserin |4 aut | |
| 700 | 1 | |a Lin, Feng-Chang |e verfasserin |4 aut | |
| 700 | 1 | |a Huffman, Jennifer E |e verfasserin |4 aut | |
| 700 | 1 | |a Luyendyk, James P |e verfasserin |4 aut | |
| 700 | 1 | |a Flick, Matthew J |e verfasserin |4 aut | |
| 700 | 1 | |a Smith, Nicholas L |e verfasserin |4 aut | |
| 700 | 1 | |a de Vries, Paul S |e verfasserin |4 aut | |
| 700 | 1 | |a Morrison, Alanna C |e verfasserin |4 aut | |
| 700 | 1 | |a Wolberg, Alisa S |e verfasserin |4 aut | |
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