Novel positive allosteric modulator of protease‐activated receptor 1 promotes skin wound healing in hairless mice
Background and Purpose Protease‐activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1 activation plays a crucial role in the process of skin wound healing such as thrombosis, inflammation, proliferation and tissue repair. In th...
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| Published in | British journal of pharmacology Vol. 178; no. 17; pp. 3414 - 3427 |
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| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
England
Blackwell Publishing Ltd
01.09.2021
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| Subjects | |
| Online Access | Get full text |
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| Abstract | Background and Purpose
Protease‐activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1 activation plays a crucial role in the process of skin wound healing such as thrombosis, inflammation, proliferation and tissue repair. In the present study, we identified a novel positive allosteric modulator of PAR1, GB83, and investigated its effect on skin wound healing.
Experimental Approach
The enhancement of PAR1 activity by GB83 was measured using Fluo‐4 calcium assay. In silico docking analysis of GB83 in PAR1 was performed using dock ligands method (CDOCKER) with CHARMm force field. Effects of GB83 on cell viability and gene expression were observed using MTS assay and quantitative real‐time PCRs, respectively. SKH‐1 hairless mice were used to investigate the wound healing effect of GB83.
Key Results
We demonstrated that GB83 did not activate PAR1 by itself but strongly enhanced PAR1 activation by thrombin and PAR1‐activating peptide (AP). In silico docking analysis revealed that GB83 can bind to the PAR1 binding site of vorapaxar. GB83 significantly promoted PAR1‐mediated cell viability and migration. In addition, the enhancement of PAR1 activity by GB83 strongly increased gene expression of TGF‐β, fibronectin and type I collagen in vitro and promoted skin wound healing in vivo.
Conclusion and Implications
Our results revealed that GB83 is the first positive allosteric modulator of PAR1 and it can be a useful pharmacological tool for studying PAR1 and a potential therapeutic agent for skin wound healing. |
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| AbstractList | Background and Purpose
Protease‐activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1 activation plays a crucial role in the process of skin wound healing such as thrombosis, inflammation, proliferation and tissue repair. In the present study, we identified a novel positive allosteric modulator of PAR1, GB83, and investigated its effect on skin wound healing.
Experimental Approach
The enhancement of PAR1 activity by GB83 was measured using Fluo‐4 calcium assay. In silico docking analysis of GB83 in PAR1 was performed using dock ligands method (CDOCKER) with CHARMm force field. Effects of GB83 on cell viability and gene expression were observed using MTS assay and quantitative real‐time PCRs, respectively. SKH‐1 hairless mice were used to investigate the wound healing effect of GB83.
Key Results
We demonstrated that GB83 did not activate PAR1 by itself but strongly enhanced PAR1 activation by thrombin and PAR1‐activating peptide (AP). In silico docking analysis revealed that GB83 can bind to the PAR1 binding site of vorapaxar. GB83 significantly promoted PAR1‐mediated cell viability and migration. In addition, the enhancement of PAR1 activity by GB83 strongly increased gene expression of TGF‐β, fibronectin and type I collagen in vitro and promoted skin wound healing in vivo.
Conclusion and Implications
Our results revealed that GB83 is the first positive allosteric modulator of PAR1 and it can be a useful pharmacological tool for studying PAR1 and a potential therapeutic agent for skin wound healing. Protease-activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1 activation plays a crucial role in the process of skin wound healing such as thrombosis, inflammation, proliferation and tissue repair. In the present study, we identified a novel positive allosteric modulator of PAR1, GB83, and investigated its effect on skin wound healing.BACKGROUND AND PURPOSEProtease-activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1 activation plays a crucial role in the process of skin wound healing such as thrombosis, inflammation, proliferation and tissue repair. In the present study, we identified a novel positive allosteric modulator of PAR1, GB83, and investigated its effect on skin wound healing.The enhancement of PAR1 activity by GB83 was measured using Fluo-4 calcium assay. In silico docking analysis of GB83 in PAR1 was performed using dock ligands method (CDOCKER) with CHARMm force field. Effects of GB83 on cell viability and gene expression were observed using MTS assay and quantitative real-time PCRs, respectively. SKH-1 hairless mice were used to investigate the wound healing effect of GB83.EXPERIMENTAL APPROACHThe enhancement of PAR1 activity by GB83 was measured using Fluo-4 calcium assay. In silico docking analysis of GB83 in PAR1 was performed using dock ligands method (CDOCKER) with CHARMm force field. Effects of GB83 on cell viability and gene expression were observed using MTS assay and quantitative real-time PCRs, respectively. SKH-1 hairless mice were used to investigate the wound healing effect of GB83.We demonstrated that GB83 did not activate PAR1 by itself but strongly enhanced PAR1 activation by thrombin and PAR1-activating peptide (AP). In silico docking analysis revealed that GB83 can bind to the PAR1 binding site of vorapaxar. GB83 significantly promoted PAR1-mediated cell viability and migration. In addition, the enhancement of PAR1 activity by GB83 strongly increased gene expression of TGF-β, fibronectin and type I collagen in vitro and promoted skin wound healing in vivo.KEY RESULTSWe demonstrated that GB83 did not activate PAR1 by itself but strongly enhanced PAR1 activation by thrombin and PAR1-activating peptide (AP). In silico docking analysis revealed that GB83 can bind to the PAR1 binding site of vorapaxar. GB83 significantly promoted PAR1-mediated cell viability and migration. In addition, the enhancement of PAR1 activity by GB83 strongly increased gene expression of TGF-β, fibronectin and type I collagen in vitro and promoted skin wound healing in vivo.Our results revealed that GB83 is the first positive allosteric modulator of PAR1 and it can be a useful pharmacological tool for studying PAR1 and a potential therapeutic agent for skin wound healing.CONCLUSION AND IMPLICATIONSOur results revealed that GB83 is the first positive allosteric modulator of PAR1 and it can be a useful pharmacological tool for studying PAR1 and a potential therapeutic agent for skin wound healing. Background and PurposeProtease‐activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1 activation plays a crucial role in the process of skin wound healing such as thrombosis, inflammation, proliferation and tissue repair. In the present study, we identified a novel positive allosteric modulator of PAR1, GB83, and investigated its effect on skin wound healing.Experimental ApproachThe enhancement of PAR1 activity by GB83 was measured using Fluo‐4 calcium assay. In silico docking analysis of GB83 in PAR1 was performed using dock ligands method (CDOCKER) with CHARMm force field. Effects of GB83 on cell viability and gene expression were observed using MTS assay and quantitative real‐time PCRs, respectively. SKH‐1 hairless mice were used to investigate the wound healing effect of GB83.Key ResultsWe demonstrated that GB83 did not activate PAR1 by itself but strongly enhanced PAR1 activation by thrombin and PAR1‐activating peptide (AP). In silico docking analysis revealed that GB83 can bind to the PAR1 binding site of vorapaxar. GB83 significantly promoted PAR1‐mediated cell viability and migration. In addition, the enhancement of PAR1 activity by GB83 strongly increased gene expression of TGF‐β, fibronectin and type I collagen in vitro and promoted skin wound healing in vivo.Conclusion and ImplicationsOur results revealed that GB83 is the first positive allosteric modulator of PAR1 and it can be a useful pharmacological tool for studying PAR1 and a potential therapeutic agent for skin wound healing. Protease-activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1 activation plays a crucial role in the process of skin wound healing such as thrombosis, inflammation, proliferation and tissue repair. In the present study, we identified a novel positive allosteric modulator of PAR1, GB83, and investigated its effect on skin wound healing. The enhancement of PAR1 activity by GB83 was measured using Fluo-4 calcium assay. In silico docking analysis of GB83 in PAR1 was performed using dock ligands method (CDOCKER) with CHARMm force field. Effects of GB83 on cell viability and gene expression were observed using MTS assay and quantitative real-time PCRs, respectively. SKH-1 hairless mice were used to investigate the wound healing effect of GB83. We demonstrated that GB83 did not activate PAR1 by itself but strongly enhanced PAR1 activation by thrombin and PAR1-activating peptide (AP). In silico docking analysis revealed that GB83 can bind to the PAR1 binding site of vorapaxar. GB83 significantly promoted PAR1-mediated cell viability and migration. In addition, the enhancement of PAR1 activity by GB83 strongly increased gene expression of TGF-β, fibronectin and type I collagen in vitro and promoted skin wound healing in vivo. Our results revealed that GB83 is the first positive allosteric modulator of PAR1 and it can be a useful pharmacological tool for studying PAR1 and a potential therapeutic agent for skin wound healing. |
| Author | Kim, Tae Gun Chang, Jiwon Jo, Sungwoo Park, So‐Hyeon Jeon, Dong‐Kyu Seo, Yohan Lee, Chulho Heo, Yunkyung Namkung, Wan Han, Gyoonhee |
| Author_xml | – sequence: 1 givenname: Yohan surname: Seo fullname: Seo, Yohan organization: Daegu‐Gyeongbuk Medical Innovation Foundation – sequence: 2 givenname: Yunkyung surname: Heo fullname: Heo, Yunkyung organization: Yonsei University – sequence: 3 givenname: Sungwoo surname: Jo fullname: Jo, Sungwoo organization: Yonsei University – sequence: 4 givenname: So‐Hyeon surname: Park fullname: Park, So‐Hyeon organization: Yonsei University – sequence: 5 givenname: Chulho surname: Lee fullname: Lee, Chulho organization: Yonsei University – sequence: 6 givenname: Jiwon surname: Chang fullname: Chang, Jiwon organization: Yonsei University – sequence: 7 givenname: Dong‐Kyu surname: Jeon fullname: Jeon, Dong‐Kyu organization: Yonsei University – sequence: 8 givenname: Tae Gun surname: Kim fullname: Kim, Tae Gun organization: Yonsei University – sequence: 9 givenname: Gyoonhee surname: Han fullname: Han, Gyoonhee organization: Yonsei University – sequence: 10 givenname: Wan orcidid: 0000-0002-1480-7433 surname: Namkung fullname: Namkung, Wan email: wnamkung@yonsei.ac.kr organization: Yonsei University |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/33837955$$D View this record in MEDLINE/PubMed |
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Protease‐activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1... Protease-activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1 activation plays a... Background and PurposeProtease‐activated receptor 1 (PAR1) is a GPCR expressed in several skin cell types, including keratinocyte and dermal fibroblast. PAR1... |
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| SubjectTerms | Allosteric properties Animals Binding sites Calcium Cell migration Cell viability Collagen (type I) Fibroblasts Fibronectin G protein-coupled receptors GB83 Gene expression Hairless Keratinocytes Mice Mice, Hairless PAR1 positive allosteric modulator Proteinase Receptor, PAR-1 Skin skin wound healing Thrombin Thrombosis Wound Healing |
| Title | Novel positive allosteric modulator of protease‐activated receptor 1 promotes skin wound healing in hairless mice |
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