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CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension

BACKGROUND: Patients with pulmonary arterial hypertension (PAH) carrying bone morphogenetic protein receptor type 2 (Bmpr2) mutations present earlier with severe hemodynamic compromise and have poorer survival outcomes than those without mutation. The mechanism underlying the worsening clinical phen...

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Autores principales: Kabwe, Jane Chanda, Sawada, Hirofumi, Mitani, Yoshihide, Oshita, Hironori, Tsuboya, Naoki, Zhang, Erquan, Maruyama, Junko, Miyasaka, Yoshiki, Ko, Hideyoshi, Oya, Kazunobu, Ito, Hiromasa, Yodoya, Noriko, Otsuki, Shoichiro, Ohashi, Hiroyuki, Okamoto, Ryuji, Dohi, Kaoru, Nishimura, Yuhei, Mashimo, Tomoji, Hirayama, Masahiro, Maruyama, Kazuo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8994407/
https://www.ncbi.nlm.nih.gov/pubmed/35395852
http://dx.doi.org/10.1186/s12931-022-02005-w
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author Kabwe, Jane Chanda
Sawada, Hirofumi
Mitani, Yoshihide
Oshita, Hironori
Tsuboya, Naoki
Zhang, Erquan
Maruyama, Junko
Miyasaka, Yoshiki
Ko, Hideyoshi
Oya, Kazunobu
Ito, Hiromasa
Yodoya, Noriko
Otsuki, Shoichiro
Ohashi, Hiroyuki
Okamoto, Ryuji
Dohi, Kaoru
Nishimura, Yuhei
Mashimo, Tomoji
Hirayama, Masahiro
Maruyama, Kazuo
author_facet Kabwe, Jane Chanda
Sawada, Hirofumi
Mitani, Yoshihide
Oshita, Hironori
Tsuboya, Naoki
Zhang, Erquan
Maruyama, Junko
Miyasaka, Yoshiki
Ko, Hideyoshi
Oya, Kazunobu
Ito, Hiromasa
Yodoya, Noriko
Otsuki, Shoichiro
Ohashi, Hiroyuki
Okamoto, Ryuji
Dohi, Kaoru
Nishimura, Yuhei
Mashimo, Tomoji
Hirayama, Masahiro
Maruyama, Kazuo
author_sort Kabwe, Jane Chanda
collection PubMed
description BACKGROUND: Patients with pulmonary arterial hypertension (PAH) carrying bone morphogenetic protein receptor type 2 (Bmpr2) mutations present earlier with severe hemodynamic compromise and have poorer survival outcomes than those without mutation. The mechanism underlying the worsening clinical phenotype of PAH with Bmpr2 mutations has been largely unaddressed in rat models of pulmonary hypertension (PH) because of the difficulty in reproducing progressive PH in mice and genetic modification in rats. We tested whether a clinically-relevant Bmpr2 mutation affects the progressive features of monocrotaline (MCT) induced-PH in rats. METHODS: A monoallelic single nucleotide insertion in exon 1 of Bmpr2 (+/44insG) was generated in rats using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9, then PH, pulmonary vascular disease (PVD) and survival after MCT injection with or without a phosphodiesterase type 5 inhibitor, tadalafil, administration were assessed. RESULTS: The +/44insG rats had reduced BMPR2 signalling in the lungs compared with wild-type. PH and PVD assessed at 3-weeks after MCT injection were similar in wild-type and +/44insG rats. However, survival at 4-weeks after MCT injection was significantly reduced in +/44insG rats. Among the rats surviving at 4-weeks after MCT administration, +/44insG rats had increased weight ratio of right ventricle to left ventricle plus septum (RV/[LV + S]) and % medial wall thickness (MWT) in pulmonary arteries (PAs). Immunohistochemical analysis showed increased vessels with Ki67-positive cells in the lungs, decreased mature and increased immature smooth muscle cell phenotype markers in the PAs in +/44insG rats compared with wild-type at 3-weeks after MCT injection. Contraction of PA in response to prostaglandin-F2α and endothelin-1 were significantly reduced in the +/44insG rats. The +/44insG rats that had received tadalafil had a worse survival with a significant increase in RV/(LV + S), %MWT in distal PAs and RV myocardial fibrosis compared with wild-type. CONCLUSIONS: The present study demonstrates that the Bmpr2 mutation promotes dedifferentiation of PA smooth muscle cells, late PVD and RV myocardial fibrosis and adversely impacts both the natural and post-treatment courses of MCT-PH in rats with significant effects only in the late stages and warrants preclinical studies using this new genetic model to optimize treatment outcomes of heritable PAH. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12931-022-02005-w.
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spelling pubmed-89944072022-04-10 CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension Kabwe, Jane Chanda Sawada, Hirofumi Mitani, Yoshihide Oshita, Hironori Tsuboya, Naoki Zhang, Erquan Maruyama, Junko Miyasaka, Yoshiki Ko, Hideyoshi Oya, Kazunobu Ito, Hiromasa Yodoya, Noriko Otsuki, Shoichiro Ohashi, Hiroyuki Okamoto, Ryuji Dohi, Kaoru Nishimura, Yuhei Mashimo, Tomoji Hirayama, Masahiro Maruyama, Kazuo Respir Res Research BACKGROUND: Patients with pulmonary arterial hypertension (PAH) carrying bone morphogenetic protein receptor type 2 (Bmpr2) mutations present earlier with severe hemodynamic compromise and have poorer survival outcomes than those without mutation. The mechanism underlying the worsening clinical phenotype of PAH with Bmpr2 mutations has been largely unaddressed in rat models of pulmonary hypertension (PH) because of the difficulty in reproducing progressive PH in mice and genetic modification in rats. We tested whether a clinically-relevant Bmpr2 mutation affects the progressive features of monocrotaline (MCT) induced-PH in rats. METHODS: A monoallelic single nucleotide insertion in exon 1 of Bmpr2 (+/44insG) was generated in rats using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9, then PH, pulmonary vascular disease (PVD) and survival after MCT injection with or without a phosphodiesterase type 5 inhibitor, tadalafil, administration were assessed. RESULTS: The +/44insG rats had reduced BMPR2 signalling in the lungs compared with wild-type. PH and PVD assessed at 3-weeks after MCT injection were similar in wild-type and +/44insG rats. However, survival at 4-weeks after MCT injection was significantly reduced in +/44insG rats. Among the rats surviving at 4-weeks after MCT administration, +/44insG rats had increased weight ratio of right ventricle to left ventricle plus septum (RV/[LV + S]) and % medial wall thickness (MWT) in pulmonary arteries (PAs). Immunohistochemical analysis showed increased vessels with Ki67-positive cells in the lungs, decreased mature and increased immature smooth muscle cell phenotype markers in the PAs in +/44insG rats compared with wild-type at 3-weeks after MCT injection. Contraction of PA in response to prostaglandin-F2α and endothelin-1 were significantly reduced in the +/44insG rats. The +/44insG rats that had received tadalafil had a worse survival with a significant increase in RV/(LV + S), %MWT in distal PAs and RV myocardial fibrosis compared with wild-type. CONCLUSIONS: The present study demonstrates that the Bmpr2 mutation promotes dedifferentiation of PA smooth muscle cells, late PVD and RV myocardial fibrosis and adversely impacts both the natural and post-treatment courses of MCT-PH in rats with significant effects only in the late stages and warrants preclinical studies using this new genetic model to optimize treatment outcomes of heritable PAH. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12931-022-02005-w. BioMed Central 2022-04-08 2022 /pmc/articles/PMC8994407/ /pubmed/35395852 http://dx.doi.org/10.1186/s12931-022-02005-w Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Kabwe, Jane Chanda
Sawada, Hirofumi
Mitani, Yoshihide
Oshita, Hironori
Tsuboya, Naoki
Zhang, Erquan
Maruyama, Junko
Miyasaka, Yoshiki
Ko, Hideyoshi
Oya, Kazunobu
Ito, Hiromasa
Yodoya, Noriko
Otsuki, Shoichiro
Ohashi, Hiroyuki
Okamoto, Ryuji
Dohi, Kaoru
Nishimura, Yuhei
Mashimo, Tomoji
Hirayama, Masahiro
Maruyama, Kazuo
CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension
title CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension
title_full CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension
title_fullStr CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension
title_full_unstemmed CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension
title_short CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension
title_sort crispr-mediated bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8994407/
https://www.ncbi.nlm.nih.gov/pubmed/35395852
http://dx.doi.org/10.1186/s12931-022-02005-w
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