Customized Massive Parallel Sequencing Panel for Diagnosis of Pulmonary Arterial Hypertension

Figures

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  Figure 1. Filtering pipeline for NGS panel HAP v1.2. A set of filters applied for variant prioritization. Only variants does not fulfil the criteria for variant quality were validated with Sanger sequencing [25].

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  Figure 2. Frequency of gene mutation detected after the analysis of 21 genes. (A) A total of 14% of pathogenic and likely pathogenic variants were identified, with a 15% of variants of unknown significance (VUS) and in approximately 65% of individuals no candidate gene variants were identified. After quality check, 6% of the analyzed samples were discarded due to low quality. (B) The gene with the highest pathogenic variants detected was BMPR2, followed by EIF2AK4, because many patients with PVOD were also included in the analysis.

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  Figure 3. Pedigree and in silico analysis of the families with more than one variant detected. (A) Genealogy of the families in which segregation analysis was performed (when available). (B) Table of in sillico analysis of the candidate variants, including the ACMG classification as well as the reference in which the variant was described. n/a: not applicable, P: pathogenic, VUS: Variant of unknown significance. α: In silico analysis was performed by dbNSFP counting the ones that predict a deleterious effect over the total available.

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  Figure 4. Variants detected in ABCC8. (A) Schematic representation of SUR1 (encoded by ABCC8) protein with its 17 transmembrane domains, and the variants detected in our cohort (red) and in Bohnen et al series (green). (B) Distribution of variants detected in ABCC8 in our cohort based on the type of variant.

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  Figure 5. Survival analysis between carriers’ vs. non-carriers. Kaplan-Meier analysis did not reflect statistical differences between individuals with PAH and pathogenic or likely pathogenic variants and individuals without any pathogenic, likely pathogenic variant detected.