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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.
Afiliación
  • Kabwe JC; The Department of Anesthesiology and Critical Care Medicine, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu city, Mie, 514-8507, Japan.
  • Sawada H; The Department of Anesthesiology and Critical Care Medicine, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu city, Mie, 514-8507, Japan. hisawada@med.mie-u.ac.jp.
  • Mitani Y; The Department of Pediatrics, Mie University Graduate School of Medicine, Mie, Japan. hisawada@med.mie-u.ac.jp.
  • Oshita H; The Department of Pediatrics, Mie University Graduate School of Medicine, Mie, Japan.
  • Tsuboya N; The Department of Pediatrics, Mie University Graduate School of Medicine, Mie, Japan.
  • Zhang E; The Department of Pediatrics, Nagoya City University School of Medicine, Aichi, Japan.
  • Maruyama J; The Department of Pediatrics, Mie University Graduate School of Medicine, Mie, Japan.
  • Miyasaka Y; The Department of Anesthesiology and Critical Care Medicine, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu city, Mie, 514-8507, Japan.
  • Ko H; The Department of Neonatology, Fuzhou Children's Hospital of Fujian Province, Fujian Medical University, Fujian, China.
  • Oya K; The Department of Clinical Engineering, Suzuka University of Medical Science, Mie, Japan.
  • Ito H; Institute of Experimental Animal Sciences, Osaka University Graduate School of Medicine, Osaka, Japan.
  • Yodoya N; The Department of Clinical Engineering, Suzuka University of Medical Science, Mie, Japan.
  • Otsuki S; The Department of Pediatrics, Mie University Graduate School of Medicine, Mie, Japan.
  • Ohashi H; The Department of Cardiology and Nephrology, Mie University Graduate School of Medicine, Mie, Japan.
  • Okamoto R; The Department of Pediatrics, Mie University Graduate School of Medicine, Mie, Japan.
  • Dohi K; The Department of Pediatrics, Mie University Graduate School of Medicine, Mie, Japan.
  • Nishimura Y; The Department of Pediatrics, Mie University Graduate School of Medicine, Mie, Japan.
  • Mashimo T; The Department of Cardiology and Nephrology, Mie University Graduate School of Medicine, Mie, Japan.
  • Hirayama M; The Department of Cardiology and Nephrology, Mie University Graduate School of Medicine, Mie, Japan.
  • Maruyama K; The Department of Integrative Pharmacology, Mie University Graduate School of Medicine, Mie, Japan.
Respir Res ; 23(1): 87, 2022 Apr 08.
Article en En | MEDLINE | ID: mdl-35395852
ABSTRACT

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.
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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Hipertensión Arterial Pulmonar / Hipertensión Pulmonar Límite: Animals / Humans Idioma: En Revista: Respir Res Año: 2022 Tipo del documento: Article País de afiliación: Japón

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Hipertensión Arterial Pulmonar / Hipertensión Pulmonar Límite: Animals / Humans Idioma: En Revista: Respir Res Año: 2022 Tipo del documento: Article País de afiliación: Japón