Next-generation sequencing of 32 genes associated with hereditary aortopathies and related disorders of connective tissue in a cohort of 199 patients

Sina Renner, Helke Schüler, Malik Alawi, Verena Kolbe, Meike Rybczynski, Rixa Woitschach, Sara Sheikhzadeh, Veronika C Stark, Jakob Olfe, Elke Roser, Friederike Sophia Seggewies, Adrian Mahlmann, Maja Hempel, Melanie J Hartmann, Mathias Hillebrand, Dagmar Wieczorek, Alexander Erich Volk, Katja Kloth, Margarete Koch-Hogrebe, Rami Abou Jamra, Diana Mitter, Janine Altmüller, Alexandra Wey-Fabrizius, Christine Petersen, Isabella Rau, Guntram Borck, Christian Kubisch, Thomas S Mir, Yskert von Kodolitsch, Kerstin Kutsche, Georg Rosenberger, Sina Renner, Helke Schüler, Malik Alawi, Verena Kolbe, Meike Rybczynski, Rixa Woitschach, Sara Sheikhzadeh, Veronika C Stark, Jakob Olfe, Elke Roser, Friederike Sophia Seggewies, Adrian Mahlmann, Maja Hempel, Melanie J Hartmann, Mathias Hillebrand, Dagmar Wieczorek, Alexander Erich Volk, Katja Kloth, Margarete Koch-Hogrebe, Rami Abou Jamra, Diana Mitter, Janine Altmüller, Alexandra Wey-Fabrizius, Christine Petersen, Isabella Rau, Guntram Borck, Christian Kubisch, Thomas S Mir, Yskert von Kodolitsch, Kerstin Kutsche, Georg Rosenberger

Abstract

Purpose: Heritable factors play an important etiologic role in connective tissue disorders (CTD) with vascular involvement, and a genetic diagnosis is getting increasingly important for gene-tailored, personalized patient management.

Methods: We analyzed 32 disease-associated genes by using targeted next-generation sequencing and exome sequencing in a clinically relevant cohort of 199 individuals. We classified and refined sequence variants according to their likelihood for pathogenicity.

Results: We identified 1 pathogenic variant (PV; in FBN1 or SMAD3) in 15 patients (7.5%) and ≥1 likely pathogenic variant (LPV; in COL3A1, FBN1, FBN2, LOX, MYH11, SMAD3, TGFBR1, or TGFBR2) in 19 individuals (9.6%), together resulting in 17.1% diagnostic yield. Thirteen PV/LPV were novel. Of PV/LPV-negative patients 47 (23.6%) showed ≥1 variant of uncertain significance (VUS). Twenty-five patients had concomitant variants. In-depth evaluation of reported/calculated variant classes resulted in reclassification of 19.8% of variants.

Conclusion: Variant classification and refinement are essential for shaping mutational spectra of disease genes, thereby improving clinical sensitivity. Obligate stringent multigene analysis is a powerful tool for identifying genetic causes of clinically related CTDs. Nonetheless, the relatively high rate of PV/LPV/VUS-negative patients underscores the existence of yet unknown disease loci and/or oligogenic/polygenic inheritance.

Keywords: Marfan syndrome; TAAD; aortopathy; connective tissue disorder; next-generation sequencing.

References

    1. Murphy-Ryan M, Psychogios A, Lindor NM. Hereditary disorders of connective tissue: a guide to the emerging differential diagnosis. Genet Med. 2010;12:344–354.
    1. Bateman JF, Boot-Handford RP, Lamande SR. Genetic diseases of connective tissues: cellular and extracellular effects of ECM mutations. Nat Rev Genet. 2009;10:173–183.
    1. Verstraeten A, Luyckx I, Loeys B. Aetiology and management of hereditary aortopathy. Nat Rev Cardiol. 2017;14:197–208.
    1. Goyal A, Keramati AR, Czarny MJ, Resar JR, Mani A. The genetics of aortopathies in clinical cardiology. Clin Med Insights Cardiol. 2017;11:1179546817709787.
    1. Andelfinger G, Loeys B, Dietz H. A decade of discovery in the genetic understanding of thoracic aortic disease. Can J Cardiol. 2016;32:13–25.
    1. Morisaki T, Morisaki H. Genetics of hereditary large vessel diseases. J Hum Genet. 2016;61:21–26.
    1. Erbel R, Aboyans V, Boileau C, et al. 2014 ESC Guidelines on the diagnosis and treatment of aortic diseases: Document covering acute and chronic aortic diseases of the thoracic and abdominal aorta of the adult. The Task Force for the Diagnosis and Treatment of Aortic Diseases of the European Society of Cardiology (ESC). Eur Heart J. 2014;35:2873–2926.
    1. Hiratzka LF, Bakris GL, Beckman JA, et al. 2010 ACCF/AHA/AATS/ACR/ASA/SCA/SCAI/SIR/STS/SVM guidelines for the diagnosis and management of patients with thoracic aortic disease: a report of the American College of Cardiology Foundation/American Heart Association Task Force on Practice Guidelines, American Association for Thoracic Surgery, American College of Radiology, American Stroke Association, Society of Cardiovascular Anesthesiologists, Society for Cardiovascular Angiography and Interventions, Society of Interventional Radiology, Society of Thoracic Surgeons, and Society for Vascular Medicine. Circulation. 2010;121:e266–369.
    1. Poninska JK, Bilinska ZT, Franaszczyk M, et al. Next-generation sequencing for diagnosis of thoracic aortic aneurysms and dissections: diagnostic yield, novel mutations and genotype phenotype correlations. J Transl Med. 2016;14:115.
    1. Proost D, Vandeweyer G, Meester JA, et al. Performant mutation identification using targeted next-generation sequencing of 14 thoracic aortic aneurysm genes. Hum Mutat. 2015;36:808–814.
    1. Wooderchak-Donahue W, VanSant-Webb C, Tvrdik T, et al. Clinical utility of a next generation sequencing panel assay for Marfan and Marfan-like syndromes featuring aortopathy. Am J Med Genet A. 2015;167A:1747–1757.
    1. Ziganshin BA, Bailey AE, Coons C, et al. Routine genetic testing for thoracic aortic aneurysm and dissection in a clinical setting. Ann Thorac Surg. 2015;100:1604–1611.
    1. Weerakkody R, Ross D, Parry DA. et al. Targeted genetic analysis in a large cohort of familial and sporadic cases of aneurysm or dissection of the thoracic aorta. Genet Med. 2018;20:1414–1422.
    1. Fang M, Yu C, Chen S, et al. Identification of novel clinically relevant variants in 70 Southern Chinese patients with thoracic aortic aneurysm and dissection by next-generation sequencing. Sci Rep. 2017;7:10035.
    1. Campens L, Callewaert B, Muino Mosquera L, et al. Gene panel sequencing in heritable thoracic aortic disorders and related entities—results of comprehensive testing in a cohort of 264 patients. Orphanet J Rare Dis. 2015;10:9.
    1. Overwater E, Marsili L, Baars MJH, et al. Results of next-generation sequencing gene panel diagnostics including copy-number variation analysis in 810 patients suspected of heritable thoracic aortic disorders. Hum Mutat. 2018;39:1173–1192.
    1. Richards S, Aziz N, Bale S, et al. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med. 2015;17:405–424.
    1. Biesecker LG, Harrison SM. The ACMG/AMP reputable source criteria for the interpretation of sequence variants. Genet Med. 2018;20:1687–1688.
    1. Landrum MJ, Lee JM, Benson M, et al. ClinVar: improving access to variant interpretations and supporting evidence. Nucleic Acids Res. 2018;46(D1):D1062–D1067.
    1. Stenson PD, Ball EV, Mort M, et al. Human Gene Mutation Database (HGMD): 2003 update. Hum Mutat. 2003;21:577–581.
    1. Guo DC, Regalado ES, Gong L, et al. LOX mutations predispose to thoracic aortic aneurysms and dissections. Circ Res. 2016;118:928–934.
    1. Amendola LM, Jarvik GP, Leo MC, et al. Performance of ACMG-AMP variant-interpretation guidelines among nine laboratories in the Clinical Sequencing Exploratory Research Consortium. Am J Hum Genet. 2016;99:247.
    1. Ghosh R, Oak N, Plon SE. Evaluation of in silico algorithms for use with ACMG/AMP clinical variant interpretation guidelines. Genome Biol. 2017;18:225.
    1. Ioannidis NM, Rothstein JH, Pejaver V, et al. REVEL: an ensemble method for predicting the pathogenicity of rare missense variants. Am J Hum Genet. 2016;99:877–885.
    1. Jagadeesh KA, Wenger AM, Berger MJ, et al. M-CAP eliminates a majority of variants of uncertain significance in clinical exomes at high sensitivity. Nat Genet. 2016;48:1581–1586.
    1. Alirezaie N, Kernohan KD, Hartley T, Majewski J, Hocking TD. ClinPred: prediction tool to identify disease-relevant nonsynonymous single-nucleotide variants. Am J Hum Genet. 2018;103:474–483.
    1. Vanakker O, Callewaert B, Malfait F, Coucke P. The genetics of soft connective tissue disorders. Annu Rev Genomics Hum Genet. 2015;16:229–255.
    1. Isselbacher EM, Lino Cardenas CL, Lindsay ME. Hereditary influence in thoracic aortic aneurysm and dissection. Circulation. 2016;133:2516–2528.
    1. Brownstein AJ, Ziganshin BA, Kuivaniemi H, Body SC, Bale AE, Elefteriades JA. Genes associated with thoracic aortic aneurysm and dissection: an update and clinical implications. Aorta (Stamford). 2017;5:11–20.
    1. Lee VS, Halabi CM, Hoffman EP, et al. Loss of function mutation in LOX causes thoracic aortic aneurysm and dissection in humans. Proc Natl Acad Sci U S A. 2016;113:8759–8764.
    1. Chen X, Greenaway FT. Identification of the disulfide bonds of lysyl oxidase. J Neural Transm (Vienna). 2011;118:1111–1114.
    1. Baetens M, Van Laer L, De Leeneer K, et al. Applying massive parallel sequencing to molecular diagnosis of Marfan and Loeys-Dietz syndromes. Hum Mutat. 2011;32:1053–1062.
    1. Loeys B, De Backer J, Van Acker P, et al. Comprehensive molecular screening of the FBN1 gene favors locus homogeneity of classical Marfan syndrome. Hum Mutat. 2004;24:140–146.
    1. Sakai H, Suzuki S, Mizuguchi T, et al. Rapid detection of gene mutations responsible for non-syndromic aortic aneurysm and dissection using two different methods: resequencing microarray technology and next-generation sequencing. Hum Genet. 2012;131:591–599.
    1. Frank M, Albuisson J, Ranque B, et al. The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome. Eur J Hum Genet. 2015;23:1657–1664.
    1. Pepin MG, Schwarze U, Rice KM, Liu M, Leistritz D, Byers PH. Survival is affected by mutation type and molecular mechanism in vascular Ehlers-Danlos syndrome (EDS type IV). Genet Med. 2014;16:881–888.
    1. Buchan JG, Alvarado DM, Haller GE, et al. Rare variants in FBN1 and FBN2 are associated with severe adolescent idiopathic scoliosis. Hum Mol Genet. 2014;23:5271–5282.

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