Impact of heterozygous ALK1 mutations on the transcriptomic response to BMP9 and BMP10 in endothelial cells from hereditary hemorrhagic telangiectasia and pulmonary arterial hypertension donors

• Published in: Angiogenesis (London) 2024-05, Vol.27 (2), p.211-227
• Main Authors: Al Tabosh, T., Liu, H., Koça, D., Al Tarrass, M., Tu, L., Giraud, S., Delagrange, L., Beaudoin, M., Rivière, S., Grobost, V., Rondeau-Lutz, M., Dupuis, O., Ricard, N., Tillet, E., Machillot, P., Salomon, A., Picart, C., Battail, C., Dupuis-Girod, S., Guignabert, C., Desroches-Castan, A., Bailly, S.
• Format: Article
• Language: English
• Subjects: Activin; Biomedical and Life Sciences; Biomedicine; Bone morphogenetic protein 9; Cancer Research; Cardiology; Cell Biology; Cell surface; Colony-forming cells; Endothelial cells; Gene expression; Gene regulation; Gene sequencing; Genes; Genotypes; Hemorrhage; Hereditary hemorrhagic telangiectasia; Heterozygosity; Hypertension; Kinases; Life Sciences; Microvasculature; Mutation; Oncology; Ophthalmology; Original Paper; Pathogenesis; Pulmonary hypertension; Stimulation; Therapeutic targets; Transcriptomics; Vascular diseases
• ISSN: 0969-6970; 1573-7209
• DOI: 10.1007/s10456-023-09902-8

Heterozygous activin receptor-like kinase 1 ( ALK1 ) mutations are associated with two vascular diseases: hereditary hemorrhagic telangiectasia (HHT) and more rarely pulmonary arterial hypertension (PAH). Here, we aimed to understand the impact of ALK1 mutations on BMP9 and BMP10 transcriptomic responses in endothelial cells. Endothelial colony-forming cells (ECFCs) and microvascular endothelial cells (HMVECs) carrying loss of function ALK1 mutations were isolated from newborn HHT and adult PAH donors, respectively. RNA-sequencing was performed on each type of cells compared to controls following an 18 h stimulation with BMP9 or BMP10. In control ECFCs, BMP9 and BMP10 stimulations induced similar transcriptomic responses with around 800 differentially expressed genes (DEGs). ALK1 -mutated ECFCs unexpectedly revealed highly similar transcriptomic profiles to controls, both at the baseline and upon stimulation, and normal activation of Smad1/5 that could not be explained by a compensation in cell-surface ALK1 level. Conversely, PAH HMVECs revealed strong transcriptional dysregulations compared to controls with > 1200 DEGs at the baseline. Consequently, because our study involved two variables, ALK1 genotype and BMP stimulation, we performed two-factor differential expression analysis and identified 44 BMP9-dysregulated genes in mutated HMVECs, but none in ECFCs. Yet, the impaired regulation of at least one hit, namely lunatic fringe ( LFNG ), was validated by RT-qPCR in three different ALK1 -mutated endothelial models. In conclusion, ALK1 heterozygosity only modified the BMP9/BMP10 regulation of few genes, including LFNG involved in NOTCH signaling. Future studies will uncover whether dysregulations in such hits are enough to promote HHT/PAH pathogenesis, making them potential therapeutic targets, or if second hits are necessary.