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Extracellular matrix and Hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids

BACKGROUND: Uterine fibroids are highly prevalent, collagen‐rich, mechanically stiff, fibrotic tumors for which new therapeutic options are needed. Increased extracellular matrix (ECM) stiffness activates mechanical signaling and Hippo/YAP promoting fibroid growth, but no prior studies have tested e...

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Autores principales: Islam, Md Soriful, Afrin, Sadia, Singh, Bhuchitra, Jayes, Friederike L., Brennan, Joshua T., Borahay, Mostafa A., Leppert, Phyllis C., Segars, James H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8255059/
https://www.ncbi.nlm.nih.gov/pubmed/34323413
http://dx.doi.org/10.1002/ctm2.475
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author Islam, Md Soriful
Afrin, Sadia
Singh, Bhuchitra
Jayes, Friederike L.
Brennan, Joshua T.
Borahay, Mostafa A.
Leppert, Phyllis C.
Segars, James H.
author_facet Islam, Md Soriful
Afrin, Sadia
Singh, Bhuchitra
Jayes, Friederike L.
Brennan, Joshua T.
Borahay, Mostafa A.
Leppert, Phyllis C.
Segars, James H.
author_sort Islam, Md Soriful
collection PubMed
description BACKGROUND: Uterine fibroids are highly prevalent, collagen‐rich, mechanically stiff, fibrotic tumors for which new therapeutic options are needed. Increased extracellular matrix (ECM) stiffness activates mechanical signaling and Hippo/YAP promoting fibroid growth, but no prior studies have tested either as a therapeutic target. We tested the hypothesis that injection of a purified form of collagenase Clostridium histolyticum (CCH) that selectively digests type I and type III collagens would alter ECM stiffness, Hippo signaling, and selectively reduce fibroid cell growth. We also used two FDA‐approved drugs, verteporfin and nintedanib, to elucidate the role of Hippo/YAP signaling in uterine fibroid and myometrial cells. METHODS: The clinical trial was registered (NCT02889848). Stiffness of samples was measured by rheometry. Protein expression in surgical samples was analyzed via immunofluorescence. Protein and gene expression in uterine fibroid or myometrial cell lines were measured by real time PCR and western blot, and immunofluorescence. RESULTS: Injection of CCH at high doses (0.1–0.2 mg/cm(3)) into fibroids resulted in a 46% reduction in stiffness in injected fibroids compared to controls after 60 days. Levels of the cell proliferation marker proliferative cell nuclear antigen (PCNA) were decreased in fibroids 60 days after injection at high doses of CCH. Key Hippo signaling factors, specifically the transcriptionally inactive phosphorylated YAP (p‐YAP), was increased at high CCH doses, supporting the role of YAP in fibroid growth. Furthermore, inhibition of YAP via verteporfin (YAP inhibitor) decreased cell proliferation, gene and protein expression of key factors promoting fibrosis and mechanotransduction in fibroid cells. Additionally, the anti‐fibrotic drug, nintedanib, inhibited YAP and showed anti‐fibrotic effects. CONCLUSIONS: This is the first report that in vivo injection of collagenase into uterine fibroids led to a reduction in Hippo/YAP signaling and crucial genes and pathways involved in fibroid growth. These results indicate that targeting ECM stiffness and Hippo signaling might be an effective strategy for uterine fibroids.
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spelling pubmed-82550592021-07-13 Extracellular matrix and Hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids Islam, Md Soriful Afrin, Sadia Singh, Bhuchitra Jayes, Friederike L. Brennan, Joshua T. Borahay, Mostafa A. Leppert, Phyllis C. Segars, James H. Clin Transl Med Research Articles BACKGROUND: Uterine fibroids are highly prevalent, collagen‐rich, mechanically stiff, fibrotic tumors for which new therapeutic options are needed. Increased extracellular matrix (ECM) stiffness activates mechanical signaling and Hippo/YAP promoting fibroid growth, but no prior studies have tested either as a therapeutic target. We tested the hypothesis that injection of a purified form of collagenase Clostridium histolyticum (CCH) that selectively digests type I and type III collagens would alter ECM stiffness, Hippo signaling, and selectively reduce fibroid cell growth. We also used two FDA‐approved drugs, verteporfin and nintedanib, to elucidate the role of Hippo/YAP signaling in uterine fibroid and myometrial cells. METHODS: The clinical trial was registered (NCT02889848). Stiffness of samples was measured by rheometry. Protein expression in surgical samples was analyzed via immunofluorescence. Protein and gene expression in uterine fibroid or myometrial cell lines were measured by real time PCR and western blot, and immunofluorescence. RESULTS: Injection of CCH at high doses (0.1–0.2 mg/cm(3)) into fibroids resulted in a 46% reduction in stiffness in injected fibroids compared to controls after 60 days. Levels of the cell proliferation marker proliferative cell nuclear antigen (PCNA) were decreased in fibroids 60 days after injection at high doses of CCH. Key Hippo signaling factors, specifically the transcriptionally inactive phosphorylated YAP (p‐YAP), was increased at high CCH doses, supporting the role of YAP in fibroid growth. Furthermore, inhibition of YAP via verteporfin (YAP inhibitor) decreased cell proliferation, gene and protein expression of key factors promoting fibrosis and mechanotransduction in fibroid cells. Additionally, the anti‐fibrotic drug, nintedanib, inhibited YAP and showed anti‐fibrotic effects. CONCLUSIONS: This is the first report that in vivo injection of collagenase into uterine fibroids led to a reduction in Hippo/YAP signaling and crucial genes and pathways involved in fibroid growth. These results indicate that targeting ECM stiffness and Hippo signaling might be an effective strategy for uterine fibroids. John Wiley and Sons Inc. 2021-07-04 /pmc/articles/PMC8255059/ /pubmed/34323413 http://dx.doi.org/10.1002/ctm2.475 Text en © 2021 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Islam, Md Soriful
Afrin, Sadia
Singh, Bhuchitra
Jayes, Friederike L.
Brennan, Joshua T.
Borahay, Mostafa A.
Leppert, Phyllis C.
Segars, James H.
Extracellular matrix and Hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids
title Extracellular matrix and Hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids
title_full Extracellular matrix and Hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids
title_fullStr Extracellular matrix and Hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids
title_full_unstemmed Extracellular matrix and Hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids
title_short Extracellular matrix and Hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids
title_sort extracellular matrix and hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8255059/
https://www.ncbi.nlm.nih.gov/pubmed/34323413
http://dx.doi.org/10.1002/ctm2.475
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