Top

Published in:

Open Access 31-01-2024 | Hereditary Hemorrhagic Telangiectasia | Original Paper

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

Authors: T. Al Tabosh, H. Liu, D. Koça, M. Al Tarrass, L. Tu, S. Giraud, L. Delagrange, M. Beaudoin, S. Rivière, V. Grobost, M. Rondeau-Lutz, O. Dupuis, N. Ricard, E. Tillet, P. Machillot, A. Salomon, C. Picart, C. Battail, S. Dupuis-Girod, C. Guignabert, A. Desroches-Castan, S. Bailly

Published in: Angiogenesis | Issue 2/2024

Abstract

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.

Available only for authorised users

Shovlin CL (2010) Hereditary haemorrhagic telangiectasia: pathophysiology, diagnosis and treatment. Blood Rev 24:203–219. https://doi.org/10.1016/j.blre.2010.07.001CrossRefPubMed

Gallione CJ, Repetto GM, Legius E et al (2004) A combined syndrome of juvenile polyposis and hereditary haemorrhagic telangiectasia associated with mutations in MADH4 (SMAD4). The Lancet 363:852–859. https://doi.org/10.1016/S0140-6736(04)15732-2CrossRef

Lesca G, Burnichon N, Raux G et al (2006) Distribution of ENG and ACVRL1 (ALK1) mutations in French HHT patients. Hum Mutat 27:598–598. https://doi.org/10.1002/humu.9421CrossRefPubMed

Wooderchak-Donahue WL, McDonald J, O’Fallon B et al (2013) BMP9 mutations cause a vascular-anomaly syndrome with phenotypic overlap with hereditary hemorrhagic telangiectasia. Am J Hum Genet 93:530–537. https://doi.org/10.1016/j.ajhg.2013.07.004CrossRefPubMedPubMedCentral

David L, Mallet C, Mazerbourg S et al (2007) Identification of BMP9 and BMP10 as functional activators of the orphan activin receptor-like kinase 1 (ALK1) in endothelial cells. Blood 109:1953–1961. https://doi.org/10.1182/blood-2006-07-034124CrossRefPubMed

Shi Y, Massagué J (2003) Mechanisms of TGF-β signaling from cell membrane to the nucleus. Cell 113:685–700. https://doi.org/10.1016/S0092-8674(03)00432-XCrossRefPubMed

David L, Feige J-J, Bailly S (2009) Emerging role of bone morphogenetic proteins in angiogenesis. Cytokine Growth Factor Rev 20:203–212. https://doi.org/10.1016/j.cytogfr.2009.05.001CrossRefPubMed

David L, Mallet C, Keramidas M et al (2008) Bone morphogenetic protein-9 is a circulating vascular quiescence factor. Circ Res 102:914–922. https://doi.org/10.1161/CIRCRESAHA.107.165530CrossRefPubMedPubMedCentral

Robert F, Desroches-Castan A, Bailly S et al (2020) Future treatments for hereditary hemorrhagic telangiectasia. Orphanet J Rare Dis 15:4. https://doi.org/10.1186/s13023-019-1281-4CrossRefPubMedPubMedCentral

10.

Dupuis-Girod S, Shovlin CL, Kjeldsen AD et al (2022) European Reference Network for Rare Vascular Diseases (VASCERN): when and how to use intravenous bevacizumab in hereditary haemorrhagic telangiectasia (HHT)? Eur J Med Genet 65:104575. https://doi.org/10.1016/j.ejmg.2022.104575CrossRefPubMed

11.

Mallet C, Lamribet K, Giraud S et al (2015) Functional analysis of endoglin mutations from hereditary hemorrhagic telangiectasia type 1 patients reveals different mechanisms for endoglin loss of function. Hum Mol Genet 24:1142–1154. https://doi.org/10.1093/hmg/ddu531CrossRefPubMed

12.

Ricard N, Bidart M, Mallet C et al (2010) Functional analysis of the BMP9 response of ALK1 mutants from HHT2 patients: a diagnostic tool for novel ACVRL1 mutations. Blood 116:1604–1612. https://doi.org/10.1182/blood-2010-03-276881CrossRefPubMed

13.

Pece-Barbara N, Cymerman U, Vera S et al (1999) Expression analysis of four endoglin missense mutations suggests that haploinsufficiency is the predominant mechanism for hereditary hemorrhagic telangiectasia type 1. Hum Mol Genet 8:2171–2181. https://doi.org/10.1093/hmg/8.12.2171CrossRefPubMed

14.

Shovlin CL, Guttmacher AE, Buscarini E et al (2000) Diagnostic criteria for hereditary hemorrhagic telangiectasia (Rendu-Osler-Weber syndrome). Am J Med Genet 91:66–67. https://doi.org/10.1002/(sici)1096-8628(20000306)91:1%3c66::aid-ajmg12%3e3.0.co;2-pCrossRefPubMed

15.

Aldred MA, Morrell NW, Guignabert C (2022) New mutations and pathogenesis of pulmonary hypertension: progress and puzzles in disease pathogenesis. Circ Res 130:1365–1381. https://doi.org/10.1161/CIRCRESAHA.122.320084CrossRefPubMedPubMedCentral

16.

Simonneau G, Montani D, Celermajer DS et al (2019) Haemodynamic definitions and updated clinical classification of pulmonary hypertension. Eur Respir J 53:1801913. https://doi.org/10.1183/13993003.01913-2018CrossRefPubMedPubMedCentral

17.

Humbert M, Kovacs G, Hoeper MM et al (2022) 2022 ESC/ERS guidelines for the diagnosis and treatment of pulmonary hypertension. Eur Heart J 43:3618–3731. https://doi.org/10.1093/eurheartj/ehac237CrossRefPubMed

18.

Walsh LJ, Collins C, Ibrahim H et al (2022) Pulmonary arterial hypertension in hereditary hemorrhagic telangiectasia associated with ACVRL1 mutation: a case report. J Med Case Rep 16:99. https://doi.org/10.1186/s13256-022-03296-9CrossRefPubMedPubMedCentral

19.

Yokokawa T, Sugimoto K, Kimishima Y et al (2020) Pulmonary hypertension and hereditary hemorrhagic telangiectasia related to an ACVRL1 mutation. Intern Med 59:221–227. https://doi.org/10.2169/internalmedicine.3625-19CrossRefPubMed

20.

Girerd B, Montani D, Coulet F et al (2010) Clinical outcomes of pulmonary arterial hypertension in patients carrying an ACVRL1 (ALK1) mutation. Am J Respir Crit Care Med 181:851–861. https://doi.org/10.1164/rccm.200908-1284OCCrossRefPubMed

21.

Roman BL, Hinck AP (2017) ALK1 signaling in development and disease: new paradigms. Cell Mol Life Sci 74:4539–4560. https://doi.org/10.1007/s00018-017-2636-4CrossRefPubMedPubMedCentral

22.

Paschalaki KE, Randi AM (2018) Recent advances in endothelial colony forming cells toward their use in clinical translation. Front Med 5:295. https://doi.org/10.3389/fmed.2018.00295CrossRef

23.

Dumortier J, Dupuis-Girod S, Valette P-J et al (2019) Recurrence of hereditary hemorrhagic telangiectasia after liver transplantation: clinical implications and physiopathological insights. Hepatology 69:9CrossRef

24.

Smadja DM, Melero-Martin JM, Eikenboom J et al (2019) Standardization of methods to quantify and culture endothelial colony-forming cells derived from peripheral blood: position paper from the International Society on Thrombosis and Haemostasis SSC. J Thromb Haemost 17:1190–1194. https://doi.org/10.1111/jth.14462CrossRefPubMedPubMedCentral

25.

Ricard N, Ciais D, Levet S et al (2012) BMP9 and BMP10 are critical for postnatal retinal vascular remodeling. Blood 119:6162–6171. https://doi.org/10.1182/blood-2012-01-407593CrossRefPubMedPubMedCentral

26.

Panov J, Simchi L, Feuermann Y, Kaphzan H (2020) Bioinformatics analyses of the transcriptome reveal Ube3a-dependent effects on mitochondrial-related pathways. Int J Mol Sci 21:4156. https://doi.org/10.3390/ijms21114156CrossRefPubMedPubMedCentral

27.

Scharpfenecker M, van Dinther M, Liu Z et al (2007) BMP-9 signals via ALK1 and inhibits bFGF-induced endothelial cell proliferation and VEGF-stimulated angiogenesis. J Cell Sci 120:964–972. https://doi.org/10.1242/jcs.002949CrossRefPubMed

28.

Townson SA, Martinez-Hackert E, Greppi C et al (2012) Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex. J Biol Chem 287:27313–27325. https://doi.org/10.1074/jbc.M112.377960CrossRefPubMedPubMedCentral

29.

Salmon RM, Guo J, Wood JH et al (2020) Molecular basis of ALK1-mediated signalling by BMP9/BMP10 and their prodomain-bound forms. Nat Commun 11:1621. https://doi.org/10.1038/s41467-020-15425-3CrossRefPubMedPubMedCentral

30.

Desroches-Castan A, Tillet E, Bouvard C, Bailly S (2022) BMP9 and BMP10: two close vascular quiescence partners that stand out. Dev Dyn 251:158–177. https://doi.org/10.1002/dvdy.395CrossRef

31.

Morikawa M, Koinuma D, Tsutsumi S et al (2011) ChIP-seq reveals cell type-specific binding patterns of BMP-specific Smads and a novel binding motif. Nucleic Acids Res 39:8712–8727. https://doi.org/10.1093/nar/gkr572CrossRefPubMedPubMedCentral

32.

Vadasz Z, Balbir Gurman A, Meroni P et al (2019) Lysyl oxidase—a possible role in systemic sclerosis–associated pulmonary hypertension: a multicentre study. Rheumatology 58:1547–1555. https://doi.org/10.1093/rheumatology/kez035CrossRefPubMed

33.

Zimmerman LB, De Jesús-Escobar JM, Harland RM (1996) The spemann organizer signal noggin binds and inactivates bone morphogenetic protein 4. Cell 86:599–606. https://doi.org/10.1016/S0092-8674(00)80133-6CrossRefPubMed

34.

Gazzerro E, Gangji V, Canalis E (1998) Bone morphogenetic proteins induce the expression of noggin, which limits their activity in cultured rat osteoblasts. J Clin Investig 102:2106–2114. https://doi.org/10.1172/JCI3459CrossRefPubMedPubMedCentral

35.

Snellings DA, Gallione CJ, Clark DS et al (2019) Somatic mutations in vascular malformations of hereditary hemorrhagic telangiectasia result in bi-allelic loss of ENG or ACVRL1. Am J Hum Genet 105:894–906. https://doi.org/10.1016/j.ajhg.2019.09.010CrossRefPubMedPubMedCentral

36.

Brouillard P, Vikkula M (2007) Genetic causes of vascular malformations. Hum Mol Genet 16:R140–R149. https://doi.org/10.1093/hmg/ddm211CrossRefPubMed

37.

Snellings DA, Girard R, Lightle R et al (2022) Developmental venous anomalies are a genetic primer for cerebral cavernous malformations. Nat Cardiovasc Res 1:246–252. https://doi.org/10.1038/s44161-022-00035-7CrossRefPubMedPubMedCentral

38.

Rabinovitch M, Guignabert C, Humbert M, Nicolls MR (2014) Inflammation and immunity in the pathogenesis of pulmonary arterial hypertension. Circ Res 115:165–175. https://doi.org/10.1161/CIRCRESAHA.113.301141CrossRefPubMedPubMedCentral

39.

Schmid CD, Olsavszky V, Reinhart M et al (2023) ALK1 controls hepatic vessel formation, angiodiversity, and angiocrine functions in hereditary hemorrhagic telangiectasia of the liver. Hepatol Baltim Md 77:1211–1227. https://doi.org/10.1002/hep.32641CrossRef

40.

Crist AM, Zhou X, Garai J et al (2019) Angiopoietin-2 inhibition rescues arteriovenous malformation in a Smad4 hereditary hemorrhagic telangiectasia mouse model. Circulation 139:2049–2063. https://doi.org/10.1161/CIRCULATIONAHA.118.036952CrossRefPubMedPubMedCentral

41.

Fernández-Chacón M, García-González I, Mühleder S, Benedito R (2021) Role of Notch in endothelial biology. Angiogenesis 24:237–250. https://doi.org/10.1007/s10456-021-09793-7CrossRefPubMed

42.

Carlson TR, Yan Y, Wu X et al (2005) Endothelial expression of constitutively active Notch4 elicits reversible arteriovenous malformations in adult mice. Proc Natl Acad Sci USA 102:9884–9889. https://doi.org/10.1073/pnas.0504391102CrossRefPubMedPubMedCentral

43.

Krebs LT, Starling C, Chervonsky AV, Gridley T (2010) Notch1 activation in mice causes arteriovenous malformations phenocopied by ephrinB2 and EphB4 mutants. Genesis. https://doi.org/10.1002/dvg.20599CrossRefPubMedPubMedCentral

44.

Krebs LT, Shutter JR, Tanigaki K et al (2004) Haploinsufficient lethality and formation of arteriovenous malformations in Notch pathway mutants. Genes Dev 18:2469–2473. https://doi.org/10.1101/gad.1239204CrossRefPubMedPubMedCentral

45.

Murphy PA, Kim TN, Lu G et al (2012) Notch4 normalization reduces blood vessel size in arteriovenous malformations. Sci Transl Med. https://doi.org/10.1126/scitranslmed.3002670CrossRefPubMedPubMedCentral

46.

Larrivée B, Prahst C, Gordon E et al (2012) ALK1 signaling inhibits angiogenesis by cooperating with the Notch pathway. Dev Cell 22:489–500. https://doi.org/10.1016/j.devcel.2012.02.005CrossRefPubMedPubMedCentral

47.

Moya IM, Umans L, Maas E et al (2012) Stalk cell phenotype depends on integration of Notch and Smad1/5 signaling cascades. Dev Cell 22:501–514. https://doi.org/10.1016/j.devcel.2012.01.007CrossRefPubMedPubMedCentral

48.

Kakuda S, Haltiwanger RS (2017) Deciphering the Fringe-mediated Notch code: identification of activating and inhibiting sites allowing discrimination between ligands. Dev Cell 40:193–201. https://doi.org/10.1016/j.devcel.2016.12.013CrossRefPubMedPubMedCentral

49.

LeBon L, Lee TV, Sprinzak D et al (2014) Fringe proteins modulate Notch-ligand cis and trans interactions to specify signaling states. eLife 3:e02950. https://doi.org/10.7554/eLife.02950CrossRefPubMedPubMedCentral

50.

Brückner K, Perez L, Clausen H, Cohen S (2000) Glycosyltransferase activity of Fringe modulates Notch-Delta interactions. Nature 406:411–415. https://doi.org/10.1038/35019075CrossRefPubMed

51.

Benedito R, Roca C, Sörensen I et al (2009) The Notch ligands Dll4 and Jagged1 have opposing effects on angiogenesis. Cell 137:1124–1135. https://doi.org/10.1016/j.cell.2009.03.025CrossRefPubMed

52.

Ehling M, Adams S, Benedito R, Adams RH (2013) Notch controls retinal blood vessel maturation and quiescence. Development 140:3051–3061. https://doi.org/10.1242/dev.093351CrossRefPubMed

53.

Tual-Chalot S, Mahmoud M, Allinson KR et al (2014) Endothelial depletion of Acvrl1 in mice leads to arteriovenous malformations associated with reduced endoglin expression. PLoS ONE 9:e98646. https://doi.org/10.1371/journal.pone.0098646CrossRefPubMedPubMedCentral

54.

Ristori T, Thuret R, Hooker E et al (2023) Bmp9 regulates Notch signaling and the temporal dynamics of angiogenesis via Lunatic Fringe. BioRxiv. https://doi.org/10.1101/2023.09.25.557123CrossRefPubMedPubMedCentral

55.

Baeyens N (2018) Fluid shear stress sensing in vascular homeostasis and remodeling: towards the development of innovative pharmacological approaches to treat vascular dysfunction. Biochem Pharmacol 158:185–191. https://doi.org/10.1016/j.bcp.2018.10.023CrossRefPubMed

Title: 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
Authors: T. Al Tabosh
H. Liu
D. Koça
M. Al Tarrass
L. Tu
S. Giraud
L. Delagrange
M. Beaudoin
S. Rivière
V. Grobost
M. Rondeau-Lutz
O. Dupuis
N. Ricard
E. Tillet
P. Machillot
A. Salomon
C. Picart
C. Battail
S. Dupuis-Girod
C. Guignabert
A. Desroches-Castan
S. Bailly
Publication date: 31-01-2024
Publisher: Springer Netherlands
Keywords: Hereditary Hemorrhagic Telangiectasia
Arterial Hypertension
Arterial Hypertension
Published in: Angiogenesis / Issue 2/2024
Print ISSN: 0969-6970
Electronic ISSN: 1573-7209
DOI: https://doi.org/10.1007/s10456-023-09902-8

Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin

Prof. Florian Limbourg

Prof. Anoop Chauhan

Developed by: Springer Medicine

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 2/2024

People with a connective tissue disorder may be especially vulnerable to the endothelial damage that characterizes long COVID due to the fragility of their vasculature and slow wound healing

Connective tissue disorders in COVID-19: Reply to “People with a connective tissue disorder may be especially vulnerable to the endothelial damage that characterizes long COVID due to the fragility of their vasculature and slow wound healing”

Role of endothelial PDGFB in arterio-venous malformations pathogenesis

Group XIV C-type lectins: emerging targets in tumor angiogenesis

A high-resolution view of the heterogeneous aging endothelium

Evidence of premature vascular dysfunction in young adults who regularly use e-cigarettes and the impact of usage length

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

At a glance: The STEP trials