Three novel BMPR2 mutations associated with advanced pulmonary arterial hypertension

Hara, Hironori; Takeda, Norifumi; Morita, Hiroyuki; Hatano, Masaru; Amiya, Eisuke; Maki, Hisataka; Minatsuki, Shun; Taki, Mizuri; Fujiwara, Takayuki; Maemura, Sonoko; Komuro, Issei

doi:10.1038/hgv.2017.10

Cited by 7 publications

(4 citation statements)

References 25 publications

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“…Total RNA was extracted from white blood cells, and cDNA synthesis was performed, as previously described [9]. Splicing patterns were analyzed using PCR with the following primers: forward, 5′-ggggagaagaagttgctgtt-3′ and reverse, 5′-tgtcttactgccagtcctaagt-3′, designed according to exons 4 and 6 sequences, respectively.…”

Section: Splice Analysis Of Tgfbr1 Exonmentioning

confidence: 99%

Distinct variants affecting differential splicing of TGFBR1 exon 5 cause either Loeys–Dietz syndrome or multiple self-healing squamous epithelioma

Fujiwara¹,

Takeda

Hara

et al. 2018

Eur J Hum Genet

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Variants in TGFBR1 have been reported to induce two completely distinct diseases, namely Loeys-Dietz syndrome (LDS) and multiple self-healing squamous epithelioma (MSSE). However, detailed mechanisms underlying this effect remain unknown. We report a Japanese familial case of LDS with a novel splice donor site variant in TGFBR1 gene (c.973 + 1 G > A; NG_007461.1). The intronic variant was predicted to mediate in-frame exon 5 skipping within the serine/threonine kinase (STK) domain, which may also be mediated by a similar TGFBR1 variant of a splice acceptor site in intron 4 (c.806-2 A > C), identified in a British familial case of MSSE. Therefore, ex vivo splicing and functional assays were performed in mammalian cells to evaluate the effect of these sequence variants. The MSSE variant activated a cryptic acceptor site at 76 bp downstream of the 3' natural splice acceptor site, which produced an out-of-frame transcript (r.807_882del, p.Asn270Thrfs*8). In contrast, the LDS variant generated two types of in-frame transcription products, r.[806_973del, 965_973 del], and produced two functionally inactivated proteins, p.[Asp269_Gln324del, Thr323_Gly325del], as a result of exon 5 skipping and the activation of a cryptic donor splice site at 9 bp upstream of the 5' natural splice donor site, respectively. Our results support the previously proposed but not yet approved mechanism that dominant-negative and truncating variants in STK domain induce LDS and MSSE, respectively.

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Section: Splice Analysis Of Tgfbr1 Exonmentioning

confidence: 99%

Distinct variants affecting differential splicing of TGFBR1 exon 5 cause either Loeys–Dietz syndrome or multiple self-healing squamous epithelioma

Fujiwara¹,

Takeda

Hara

et al. 2018

Eur J Hum Genet

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“…BMPR2 expression and activity are crucial in the pulmonary vasculature and mutations in BMPR2 underlie the vast majority of heritable forms of PAH. 32 , 33 Furthermore, many idiopathic cases of PAH are a result of dysfunctional BMPR2 signaling. 34 Therefore, increased BMPR2 signaling may have contributed to the therapeutic effects of miR96.…”

Section: Discussionmentioning

confidence: 99%

Direct Delivery of MicroRNA96 to the Lungs Reduces Progression of Sugen/Hypoxia-Induced Pulmonary Hypertension in the Rat

Docherty

Denver

Fisher

et al. 2020

Molecular Therapy - Nucleic Acids

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The 5HT1B receptor (5HT1BR) contributes to the pathogenic effects of serotonin in pulmonary arterial hypertension. Here, we determine the effect of a microRNA96 (miR96) mimic delivered directly to the lungs on development of severe pulmonary hypertension in rats. Female rats were dosed with sugen (30 mg/kg) and subjected to 3 weeks of hypobaric hypoxia. In normoxia, rats were dosed with either a 5HT1BR antagonist SB216641 (7.5 mg/kg/day for 3 weeks), miR96, or scramble sequence (50 μg per rat), delivered by intratracheal (i.t) administration, once a week for 3 weeks. Cardiac hemodynamics were determined, pulmonary vascular remodeling was assessed, and gene expression was assessed by qRT-PCR, and in situ hybridization and protein expression were assessed by western blot and ELISA. miR96 expression was increased in pulmonary arteries and associated with a downregulation of the 5HT1BR protein in the lung. miR96 reduced progression of right ventricular systolic pressure, pulmonary arterial remodeling, right ventricular hypertrophy, and the occurrence of occlusive pulmonary lesions. Importantly, miR96 had no off-target effects and did not affect fibrotic markers of liver and kidney function. In conclusion, direct delivery of miR96 to the lungs was effective, reducing progression of sugen/hypoxia-induced pulmonary hypertension with no measured off-target effects. miR96 may be a novel therapy for pulmonary arterial hypertension, acting through downregulation of 5HT1BR.

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“…To confirm the previously reported high-efficacy of Cre-mediated recombination in Tgfbr2 MyeKO mice [13], macrophages were obtained from the peritoneal cavity of 6-week-old mice 4 days after injection of 4% thioglycollate (211260; Becton Dickinson and Company). Total RNA isolation, cDNA conversion, and semi-quantitative reverse transcription polymerase chain reaction (PCR) were performed as previously reported [14,15]. The PCR primers used were as follows (forward and reverse, respectively): Tgfbr2, 5'-GCAAGTTTTGCGATG TGAGA-3'and 5'-GGCATCTTCCAGAGTGAAGC-3'; 18S rRNA, 5'-CGGCTACCACATC CAAGGAA-3' , and 5'-AGCTGGAATTACCGCGGC-3'.…”

Section: Efficacy Of Cre-mediated Recombination In Tgfbr2 Myeko Micementioning

confidence: 99%

Inhibition of transforming growth factor-β signaling in myeloid cells ameliorates aortic aneurysmal formation in Marfan syndrome

et al. 2020

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Increased transforming growth factor-β (TGF-β) signaling contributes to the pathophysiology of aortic aneurysm in Marfan syndrome (MFS). Recent reports indicate that a small but significant number of inflammatory cells are infiltrated into the aortic media and adventitia in MFS. However, little is known about the contribution of myeloid cells to aortic aneurysmal formation. In this study, we ablated the TGF-β type II receptor gene Tgfbr2 in myeloid cells of Fbn1C1039G/+ MFS mice (Fbn1C1039G/+;LysM-Cre/+;Tgfbr2fl/fl mice, hereinafter called Fbn1C1039G/+;Tgfbr2MyeKO) and evaluated macrophage infiltration and TGF-β signaling in the aorta. Aneurysmal formation with fragmentation and disarray of medial elastic fibers observed in MFS mice was significantly ameliorated in Fbn1C1039G/+;Tgfbr2MyeKO mice. In the aorta of Fbn1C1039G/+;Tgfbr2MyeKO mice, both canonical and noncanonical TGF-β signals were attenuated and the number of infiltrated F4/80-positive macrophages was significantly reduced. In vitro, TGF-β enhanced the migration capacity of RAW264.7 macrophages. These findings suggest that TGF-β signaling in myeloid cells promotes aortic aneurysmal formation and its inhibition might be a novel therapeutic target in MFS.

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Three novel BMPR2 mutations associated with advanced pulmonary arterial hypertension

Cited by 7 publications

References 25 publications

Distinct variants affecting differential splicing of TGFBR1 exon 5 cause either Loeys–Dietz syndrome or multiple self-healing squamous epithelioma

Distinct variants affecting differential splicing of TGFBR1 exon 5 cause either Loeys–Dietz syndrome or multiple self-healing squamous epithelioma

Direct Delivery of MicroRNA96 to the Lungs Reduces Progression of Sugen/Hypoxia-Induced Pulmonary Hypertension in the Rat

Inhibition of transforming growth factor-β signaling in myeloid cells ameliorates aortic aneurysmal formation in Marfan syndrome

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