AUTHOR=Takashima Tomoya , Brisset Sophie , Furukawa Asuka , Taniguchi Hirohisa , Takeyasu Rika , Kawamura Akio , Tamura Yuichi 

TITLE=Case Report: BMPR2-Targeted MinION Sequencing as a Tool for Genetic Analysis in Patients With Pulmonary Arterial Hypertension

JOURNAL=Frontiers in Cardiovascular Medicine

VOLUME=8

YEAR=2021

URL=https://www.frontiersin.org/journals/cardiovascular-medicine/articles/10.3389/fcvm.2021.711694

DOI=10.3389/fcvm.2021.711694

ISSN=2297-055X

ABSTRACT=<p><bold>Background:</bold> Mutations in the bone morphogenetic protein receptor type 2 gene (<italic>BMPR2</italic>) represent a major genetic cause of pulmonary arterial hypertension (PAH). Identification of <italic>BMPR2</italic> mutations is crucial for the genetic diagnosis of PAH. MinION nanopore sequencer is a portable third-generation technology that enables long-read sequencing at a low-cost. This nanopore technology-based device has not been used previously for PAH diagnosis. This study aimed to determine the feasibility of using MinION nanopore sequencing for the genetic analysis of PAH patients, focused on <italic>BMPR2</italic>.</p><p><bold>Methods:</bold> We developed a protocol for the custom bioinformatics pipeline analysis of long reads generated by long-PCR. To evaluate the potential of using MinION sequencing in PAH, we analyzed five samples, including those of two idiopathic PAH patients and a family of three members with one affected patient. Sanger sequencing analysis was performed to validate the variants.</p><p><bold>Results:</bold> The median read length was around 3.4 kb and a good mean quality score of approximately 19 was obtained. The total number of reads generated was uniform among the cases and ranged from 2,268,263 to 3,126,719. The coverage was consistent across flow cells in which the average number of reads per base ranged from 80,375 to 135,603. We identified two polymorphic variants and three mutations in four out of five patients. Certain indel variant calling-related errors were observed, mostly outside coding sequences.</p><p><bold>Conclusion:</bold> We have shown the ability of this portable nanopore sequencer to detect <italic>BMPR2</italic> mutations in patients with PAH. The MinION nanopore sequencer is a promising tool for screening <italic>BMPR2</italic> mutations, especially in small laboratories and research groups.</p>