Molecular autopsy in sudden unexplained death

Yıl 2022, Cilt: 36 Sayı: 2, 40 - 47, 01.08.2022

Esra Tekcan Sengul Tural

Öz

Molecular autopsy is the process of investigating sudden unexplained deaths through genetic analysis. It is particularly useful in cases where the cause of death is unexplained or shows non-diagnostic features despite macroscopic, histopathological, and toxicological examinations at conventional autopsy. Postmortem genetic testing is a complementary tool to a rigorous autopsy. The ultimate purpose of molecular autopsy is to assist forensic investigations and guide the genetic screening of relatives of the victim who may be at risk for sudden unexplained death. Earlier molecular autopsy attempts relied on Sanger sequencing, which although accurate and easy to use, has a low yield and can only be used to analyze a small panel of genes. The recent adoption of next-generation sequencing (NGS) technologies has allowed for exome/genome-wide examination, resulting in an increase in detection of pathogenic variants and the discovery of newer genotype-phenotype associations.
This review summarizes the scientific evidence for the use of molecular autopsy to investigate sudden unexplained deaths. Unlike other recently published articles dealing with this topic, we aimed to the technical aspects of data processing and interpretation as well as report the latest data, focusing on the use of NGS technologies for molecular autopsy.

Anahtar Kelimeler

Autopsy, sudden death, molecular genetic

Ani açıklanamayan ölümlerde moleküler otopsi

Öz

Moleküler otopsi, ani açıklanamayan ölümlerin genetik analiz yoluyla araştırılması işlemidir. Geleneksel otopside makroskobik, histopatolojik, toksikolojik incelemelere karşın ölüm nedeninin açıklanamadığı veya tanısal olmayan özellikler gösterdiği durumlarda özellikle yararlıdır. Postmortem genetik testler titizlikle yapılan bir otopsi için tamamlayıcı bir araçtır. Moleküler otopsinin nihai amacı, adli tıp araştırmalarına yardımcı olmak ve mağdurun ani açıklanamayan ölümlere karşı risk altında olabilecek akrabalarının genetik taramasına rehberlik etmektir. Eski moleküler otopsi girişimleri, doğru ve kullanımı kolay olmasına rağmen, düşük bir verime sahip olan ve yalnızca küçük bir gen panelini analiz etmek için kullanılabilen Sanger dizilimine dayanıyordu. Yeni nesil dizileme (NGS) teknolojilerinin yakın zamanda benimsenmesi, ekzom/genom çapında incelemeye izin vererek, patojenik varyantların tespitinde bir artış ve daha yeni genotip-fenotip birlikteliklerinin keşfini sağlamıştır.
Bu derlemede, ani açıklanamayan ölümlerin araştırması için moleküler otopsi kullanımına ilişkin bilimsel kanıtlar özetlenmekte ayrıca bu konuyu ele alan yakın zamanda yayınlanan diğer makalelerden farklı olarak moleküler otopsi için NGS teknolojilerinin kullanımına odaklanarak en son verileri, hem de veri işleme ve yorumlamaya ilişkin teknik yönleri raporlamayı amaçladık.

Anahtar Kelimeler

Otopsi, ani ölüm, moleküler genetik

Kaynakça

• Banner J, Basso C, Tolkien Z, Kholova I, Michaud K, Gallaghe PJ. Autopsy examination in sudden cardiac death: a current perspective on behalf of the association for European Cardiovascular Pathology. Virchows Archiv 2021;478(4):687-93.
• Castiglione EV, Modena M, Aimo A, Chiti E, Botto N, Vittorini S, Guidi B, Vergaro G, Barison A, Rossi A, Passino C, Giannoni A, Paolo MD, Emdin M. Molecular autopsy of sudden cardiac death in the genomics. Diagnostics 2021;11(8):1378.
• Vahatalo JH, Holmström LTA, Pylkas K, Skarp S, Porvari K, Pakanen L, Kaikkonen KS, Perkiömaki JS, Kerkela R, Huikuri HV, Myerburg RJ, Junttila MJ. Genetic variants associated with sudden cardiac death in victims with single vessel coronary artery disease and left ventricular hypertrophy with or without fibrosis. Front Cardiovasc Med 2022;8:755062.
• Mazzaccara C, Mirra B, Barretta B, Lombardo B, Scudiero O, Frisso G. Sudden cardiac death in young athletes: Literature review of molecular basis. Cardiogenetics 2020;10:8860.
• Ergül Y, Şahin GT, Kafalı HC, Öztürk E, Özgür S, Haydin S, Güzeltaş A. Clinical and genetic characteristics and course of congenital long QT syndrome in children: A nine-year single-center experience. Anatol J Cardiol 2021;25:250-7.
• Priori SG, Blomstrom-Lundqvist C, Mazzanti A, Bloma N, Borggrefe M, Camm J, et al. 2015 ESC Guidelines for the management of patients with ventricular arrhythmias and the preventionof sudden cardiac death the task force for the management of patients with ventricular arrhythmias and the prevention ofsudden cardiac death of the Europea. Eur Heart J 2015;36(41):2793-867.
• Scheiper-Welling S, Tabunscik M, Gross TE, Jenewein T, Beckmann BM, Niess C, Gradhand E, Wunder C, Schneider PM, Rothschild MA, Verhof MA, Kauferstein S. Variant interpretation in molecular autopsy: a useful dilemma. International Journal of Legal Medicine 2022;136(2):475-82.
• Brohus M, Arsov T, Wallace DA, Jensen HH, Nyegaard M, Crotti L, Adamski M, Zhang Y, Field MA, Athanasopoulos V, Baro I, Ribeiro de Oliveira-Mendes BB, Redon R, Charpentier F, Raju H, DiSilvestre D, Wei J, Wang R, Rafehi H, Kaspi A, Bahlo M, Dick IE, Chen SRW, Cook MC, Vinuesa CG, Overgaard MT, Schwartz PJ. Infanticide vs. inherited cardiac arrhythmias. EP Eur 2021;23:441-50.
• Grassi S, Vidal MC, Campuzano O, Arena V, Alfonsetti A, Rossi SS, Scarnicci F, Iglesias A, Brugada R, Oliva A. Sudden death without a clear cause after comprehensive ınvestigation: An example of forensic approach to atypical/uncertain findings. Diagnostics 2021;11:886.
• Goodwin S, McPherson JD, McCombie WR. Coming of age: Ten years of next-generation sequencing technologies. Nat Rev Genet 2016;17(6):333-51.
• Ackerman MJ, Tester DJ, Porter CJ, Edwards WD. Molecular diagnosis of the inherited Long-QT syndrome in a woman who died after near-drowning. N Engl J Med 1999; 341(15):1121-5.
• Chugh SS, Senashova O, Watts A, Tran PT, Zhou Z, Gong Q, Titus JL, Hayflick SJ. Postmortem molecular screening in unexplained sudden death. J Am Coll Cardiol 2004; 43(9):1625-9.
• Di Paolo M, Luchini D, Bloise R, Priori SG. Postmortem Molecular Analysis in Victims of Sudden Unexplained Death. Am. J. Forensic Med. Pathol. 2004;25(2):182-4.
• Skinner JR, Crawford J, Smith W, Aitken A, Heaven D, Evans CA, Hayes I, Neas, KR, Stables S, Koelmeyer T, Denmark L, Vuletic J, Maxwell F, White K, Yang T, Roden DM, Leren TP, Shelling A, Love DR. Prospective, population-based Long QT molecular autopsy study of postmortem negative sudden death in 1 to 40 year olds. Heart Rhythm 2011;8(3):412-9.
• Winkel BG, Holst AG, Theilade J, Kristensen IB, Thomsen JL, Ottesen GL, Bundgaard H, Svendsen JH, Haunso S, Tfelt-Hansen J. Nationwide study of sudden cardiac death in persons aged 1-35 years. Eur Heart J 2011;32(8):983-90.
• Tester DJ, Medeiros-Domingo A, Will ML, Haglund CM, Ackerman MJ. Cardiac channel molecular autopsy: Insights from 173 consecutive cases of autopsy-negative sudden unexplained death referred for postmortem genetic testing. Mayo Clin Proc 2012;87(6):524-39.
• Adams DR, Eng CM. Next-generation sequencing to diagnose suspected genetic disorders. N Engl J Med 2019;380(2):1353-62.
• Miles CJ, Behr ER. The role of genetic testing in unexplained sudden death. Transl Res J Lab Clin Med 2016;168:59-73.
• Biesecker LG, Harrison SM, ClinGen sequence variant ınterpretationworking group. The ACMG/AMP reputable source criteria for the ınterpretation of sequence variants. Genet Med Off J Am Coll Med Genet 2018;20(12):1687-8.
• Campuzano O, Sarquella-Brugada G, Fernandez-Falgueras A, Coll M, Iglesias A, Ferrer-Costa C, Cesar S, Arbelo E, Garcia-Alvarez A, Jorda P, Toro R, Tiron de Liano C, Grassi S, Oliva A, Brugada J, Brugada R. Reanalysis and reclassification of rare genetic variants associated with ınheritedarrhythmogenic syndromes. E BioMedicine 2020;54:102732.
• Richards S, Aziz N, Bale S, Bick D, Das S, Gastier-Foster J, Grody WW, Hegde M, Lyon E, Spector E, Spector E, Voelkerding K, Rehm HL, ACMG laboratory quality assurance committee. Standards and guidelines for the ınterpretation of sequence variants: A joint consensus recommendation of the American College of Medical Genetics and Genomics and the association for molecular pathology. Genet. Med. Off. J Am Coll Med Genet 2015; 17(5):405-24.
• Stiles MK, Wilde AAM, Abrams DJ, Ackerman MJ, Albert CM, Behr ER, Chugh, SS, Cornel MC, Gardner K, Ingles J, James CA, Juang JMJ, Kaab S, Kaufman ES, Krahn AD, Lubitz SA, MacLeod H, Morillo CA, Nademanee K, Probst V, Saarel EV, Sacilotto L, Semsarian C, Sheppard MN, Shimizu W, Skinner JR, Tfelt-Hansen J, Wang DW. 2020 APHRS/HRS Expert consensus statement on the investigation of decedents with sudden unexplained death and patients with sudden cardiac arrest, and of their families. Heart Rhythm 2021;37(3):481-534.
• Baudhuin LM, Leduc C, Train LJ, Avula R, Kluge ML, Kotzer KE, Lin PT, Ackerman MJ, Maleszewski JJ. Technical advances for the clinical genomic evaluation of sudden cardiac death. Circ Cardiovasc Genet 2017;10:e001844.
• Bagnall RD, Ingles J, Yeates L, Berkovic SF, Semsarian C. Exome sequencing-based molecular autopsy of formalin-fixed paraffin-embedded tissue after sudden death. Genet Med 2017;19(10):1127-33.
• Lin Y, Gryazeva T, Wang D, Zhou B, Um SY, Eng LS, Ruiter K, Rojas L, Williams N, Sampson BA, Tang Y. Using postmortem formalin fixed paraffin-embedded tissues for molecular testing of sudden cardiac death: a cautionary tale of utility and limitations. Forensic Sci Int 2020;308:110177.
• Priest, J.R. A Primer to clinical genome sequencing. Curr Opin Pediatr 2017;29(5):513-9.
• Hata Y, Kinoshita K, Mizumak K, Yamaguchi Y, Hirono K, Ichida F, Takasaki A, Mori H, Nishida N. Postmortem genetic analysis of sudden unexplained death syndrome under 50 years of age: A next-generation sequencing study. Heart Rhythm 2016;13(7):1544-51.
• Modena M, Castiglione V, Aretini P, Mazzanti CM, Chiti E, Giannoni A, Emdin M, Di Paolo M. Unveiling a sudden unexplained death case by whole exome sequencing and bioinformatic analysis. Mol Genet Genomic Med 2020;8(4):e1182.
• Hertz CL, Christiansen SL, Ferrero-Miliani L, Dahl M, Weeke PE, Ottesen GL, Frank-Hansen R, Bundgaard H, Morling N. Next-generation sequencing of 100 candidate genes in young victims of suspected sudden cardiac death with structural abnormalities of the heart. Int J Leg Med 2016;130(1):91-02.
• Ripoll-Vera T, Perez Luengo C, Borondo Alcazar JC, Garcia Ruiz AB, Sanchez Del Valle N, Barcelo Martin B, et al. Sudden cardiac death in persons aged 50 years or younger: diagnostic yield of a regional molecular autopsy program using massive sequencing. Rev Esp Cardiol Engl Ed 2021;74(5):402-13.
• Dewar LJ, Alcaide M, Fornika D, D’Amato L, Shafaatalab S, Stevens CM,et al. Investigating the genetic causes of sudden unexpected death in children through targeted next-generation sequencing analysis. Circ Cardiovasc Genet 2017;10(4):e001738.
• Lahrouchi N, Raju H, Lodder EM, Papatheodorou E, Ware JS, Papadakis M Tadros R, et al. Utility of post-mortem genetic testing in cases of sudden arrhythmic death syndrome. J Am Coll Cardiol 2017; 69(17):2134-145.