{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T17:10:51Z","timestamp":1785431451378,"version":"3.56.0"},"reference-count":50,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000780","name":"European Union","doi-asserted-by":"crossref","award":["PE00000019"],"award-info":[{"award-number":["PE00000019"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000780","name":"European Union","doi-asserted-by":"crossref","award":["CUP I33C22006900006"],"award-info":[{"award-number":["CUP I33C22006900006"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"crossref"}]},{"award":["PE00000019"],"award-info":[{"award-number":["PE00000019"]}],"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"publisher"}]},{"award":["CUP I33C22006900006"],"award-info":[{"award-number":["CUP I33C22006900006"]}],"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background\/Objectives: Sudden cardiac death (SCD) is a natural death of cardiac origin, accounting for an estimated 6\u20139 million deaths worldwide each year and representing a major public health challenge. Despite its clinical and forensic relevance, the molecular investigation of SCD remains limited. Peripheral blood or fresh tissue, the preferred specimens for post-mortem genetic testing, are not always available, and DNA extracted from formalin-fixed paraffin-embedded (FFPE) tissues is often suboptimal for conventional genetic analyses. This review evaluates the potential of archived FFPE cardiac tissue as a source for microRNA (miRNA) analysis in molecular autopsy. Methods: A narrative review was conducted by collecting studies investigating miRNA expression in FFPE cardiac tissue relevant to SCD. Ten studies met the inclusion criteria and were critically analyzed. Results: Although the available evidence remains limited, recent studies have identified several differentially expressed miRNAs associated with cardiac diseases relevant to SCD. Owing to their small size and remarkable stability, miRNAs remain detectable in FFPE tissues despite fixation and long-term storage, making them attractive molecular biomarkers. While most available studies were conducted in clinical rather than forensic settings, they demonstrate the feasibility and analytical robustness of miRNA profiling in archived FFPE cardiac specimens. Conclusions: This review underlines the importance of reconsidering archived FFPE tissues not merely as historical or morphological resources, but as promising matrices for molecular autopsy of SCD supporting the identification and validation of novel molecular biomarkers.<\/jats:p>","DOI":"10.3390\/genes17070819","type":"journal-article","created":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T12:00:17Z","timestamp":1784548817000},"page":"819","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["From Old to Bold: Advancing microRNA Studies in Sudden Cardiac Death Through Molecular Analysis of FFPE Heart Tissue"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-9412-9572","authenticated-orcid":false,"given":"Alessia","family":"Bernini Di Michele","sequence":"first","affiliation":[{"name":"Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Polytechnic University of Marche, Via Tronto, 60126 Ancona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9211-810X","authenticated-orcid":false,"given":"Chiara","family":"Turchi","sequence":"additional","affiliation":[{"name":"Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Polytechnic University of Marche, Via Tronto, 60126 Ancona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5721-0912","authenticated-orcid":false,"given":"Mauro","family":"Pesaresi","sequence":"additional","affiliation":[{"name":"Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Polytechnic University of Marche, Via Tronto, 60126 Ancona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1016\/0002-8703(82)90552-X","article-title":"The Necessity of a Uniform Definition of Sudden Coronary Death: Witnessed Death within 1 Hour of the Onset of Acute Symptoms","volume":"103","author":"Goldstein","year":"1982","journal-title":"Am. Heart J."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2334","DOI":"10.1161\/01.CIR.98.21.2334","article-title":"Sudden Cardiac Death","volume":"98","author":"Zipes","year":"1998","journal-title":"Circulation"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"euad203","DOI":"10.1093\/europace\/euad203","article-title":"Risk Stratification of Sudden Cardiac Death: A Review","volume":"25","author":"Garcia","year":"2023","journal-title":"Europace"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1818","DOI":"10.1016\/j.jacc.2022.02.041","article-title":"Incidence of Sudden Cardiac Death in the European Union","volume":"79","author":"Empana","year":"2022","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1657","DOI":"10.1016\/j.hrthm.2021.05.005","article-title":"Nationwide Burden of Sudden Cardiac Death: A Study of 54,028 Deaths in Denmark","volume":"18","author":"Lynge","year":"2021","journal-title":"Heart Rhythm"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1007\/s00414-018-1775-y","article-title":"Exome Analysis in 34 Sudden Unexplained Death (SUD) Victims Mainly Identified Variants in Channelopathy-Associated Genes","volume":"132","author":"Neubauer","year":"2018","journal-title":"Int. J. Leg. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1038\/s41431-019-0500-8","article-title":"The Yield of Postmortem Genetic Testing in Sudden Death Cases with Structural Findings at Autopsy","volume":"28","author":"Lahrouchi","year":"2019","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"7143","DOI":"10.1093\/nar\/gkad519","article-title":"A Critical Spotlight on the Paradigms of FFPE-DNA Sequencing","volume":"51","author":"Steiert","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1186\/s40246-024-00657-x","article-title":"Whole-exome sequencing to identify causative variants in juvenile sudden cardiac death","volume":"18","author":"Modena","year":"2024","journal-title":"Hum. Genom."},{"key":"ref_10","first-page":"519","article-title":"MicroRNA Expression Profiling Outperforms MRNA Expression Profiling in Formalin-Fixed Paraffin-Embedded Tissues","volume":"2","author":"Liu","year":"2009","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1038\/cr.2008.282","article-title":"Characterization of MicroRNAs in Serum: A Novel Class of Biomarkers for Diagnosis of Cancer and Other Diseases","volume":"18","author":"Chen","year":"2008","journal-title":"Cell Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"10513","DOI":"10.1073\/pnas.0804549105","article-title":"Circulating MicroRNAs as Stable Blood-Based Markers for Cancer Detection","volume":"105","author":"Mitchell","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Peskoe, S.B., Barber, J.R., Zheng, Q., Meeker, A.K., De Marzo, A.M., Platz, E.A., and Lupold, S.E. (2017). Differential Long-Term Stability of MicroRNAs and RNU6B SnRNA in 12\u201320 Year Old Archived Formalin-Fixed Paraffin-Embedded Specimens. BMC Cancer, 17.","DOI":"10.1186\/s12885-016-3008-4"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1937","DOI":"10.2741\/e514","article-title":"MiRNAs Are Stable in Colorectal Cancer Archival Tissue Blocks","volume":"E4","author":"Bovell","year":"2012","journal-title":"Front. Biosci."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Li, J., Smyth, P., Flavin, R., Cahill, S., Denning, K., Aherne, S., Guenther, S.M., O\u2019Leary, J.J., and Sheils, O. (2007). Comparison of MiRNA Expression Patterns Using Total RNA Extracted from Matched Samples of Formalin-Fixed Paraffin-Embedded (FFPE) Cells and Snap Frozen Cells. BMC Biotechnol., 7.","DOI":"10.1186\/1472-6750-7-36"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1007\/s00428-013-1485-2","article-title":"High Stability of MicroRNAs in Tissue Samples of Compromised Quality","volume":"463","author":"Burgues","year":"2013","journal-title":"Virchows Arch."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Kakimoto, Y., Tanaka, M., Kamiguchi, H., Ochiai, E., and Osawa, M. (2016). MicroRNA Stability in FFPE Tissue Samples: Dependence on GC Content. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0163125"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kakimoto, Y., Kamiguchi, H., Ochiai, E., Satoh, F., and Osawa, M. (2015). MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0129338"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1007\/s00414-014-1066-1","article-title":"Molecular Analysis of Different Classes of RNA Molecules from Formalin-Fixed Paraffin-Embedded Autoptic Tissues: A Pilot Study","volume":"129","author":"Muciaccia","year":"2014","journal-title":"Int. J. Leg. Med."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Sampaio-Silva, F., Magalh\u00e3es, T., Carvalho, F., Dinis-Oliveira, R.J., and Silvestre, R. (2013). Profiling of RNA Degradation for Estimation of Post Morterm Interval. PLoS ONE, 8.","DOI":"10.1371\/annotation\/cb8b37ac-cbe2-45e3-b9fe-d62e7ced4b25"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1097\/00019606-200209000-00008","article-title":"Postmortem RNA and Protein Stability in Perinatal Human Lungs","volume":"11","author":"Mao","year":"2002","journal-title":"Diagn. Mol. Pathol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Boisen, M.K., Dehlendorff, C., Linnemann, D., Schultz, N.A., Jensen, B.V., H\u00f8gdall, E., and Johansen, J.S. (2015). MicroRNA Expression in Formalin-Fixed Paraffin-Embedded Cancer Tissue: Identifying Reference MicroRNAs and Variability. BMC Cancer, 15.","DOI":"10.1186\/s12885-015-2030-2"},{"key":"ref_23","first-page":"119","article-title":"Unlocking Pathology Archives for MicroRNA-Profiling","volume":"28","author":"Hoefig","year":"2008","journal-title":"Anticancer Res."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"de Biase, D., Visani, M., Morandi, L., Marucci, G., Taccioli, C., Cerasoli, S., Baruzzi, A., and Pession, A. (2012). MiRNAs Expression Analysis in Paired Fresh\/Frozen and Dissected Formalin Fixed and Paraffin Embedded Glioblastoma Using Real-Time PCR. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0035596"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1038\/jid.2008.347","article-title":"MicroRNA Expression in Melanocytic Nevi: The Usefulness of Formalin-Fixed, Paraffin-Embedded Material for MiRNA Microarray Profiling","volume":"129","author":"Glud","year":"2009","journal-title":"J. Investig. Dermatol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Meng, W., McElroy, J.P., Volinia, S., Palatini, J., Warner, S., Ayers, L.W., Palanichamy, K., Chakravarti, A., and Lautenschlaeger, T. (2013). Comparison of MicroRNA Deep Sequencing of Matched Formalin-Fixed Paraffin-Embedded and Fresh Frozen Cancer Tissues. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0064393"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"513","DOI":"10.2353\/jmoldx.2008.080077","article-title":"An Array-Based Analysis of MicroRNA Expression Comparing Matched Frozen and Formalin-Fixed Paraffin-Embedded Human Tissue Samples","volume":"10","author":"Zhang","year":"2008","journal-title":"J. Mol. Diagn."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1038\/nrg3198","article-title":"MicroRNA Profiling: Approaches and Considerations","volume":"13","author":"Pritchard","year":"2012","journal-title":"Nat. Rev. Genet."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"998","DOI":"10.1373\/clinchem.2009.141580","article-title":"Robust MicroRNA Stability in Degraded RNA Preparations from Human Tissue and Cell Samples","volume":"56","author":"Jung","year":"2010","journal-title":"Clin. Chem."},{"key":"ref_30","first-page":"61","article-title":"MiRNA Analysis of Formalin-Fixed Squamous Cell Carcinomas of the Tongue Is Affected by Age of the Samples","volume":"38","author":"Rentoft","year":"2011","journal-title":"Int. J. Oncol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Bo\u0161tjan\u010di\u010d, E., Zidar, N., and Glava\u010d, D. (2012). MicroRNAs and Cardiac Sarcoplasmic Reticulum Calcium ATPase-2 in Human Myocardial Infarction: Expression and Bioinformatic Analysis. BMC Genom., 13.","DOI":"10.1186\/1471-2164-13-552"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.forsciint.2013.02.032","article-title":"Dysregulation of Heart and Brain Specific Micro-RNA in Sudden Infant Death Syndrome","volume":"228","author":"Courts","year":"2013","journal-title":"Forensic Sci. Int."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1016\/j.healun.2018.06.010","article-title":"MicroRNA Signatures in Cardiac Biopsies and Detection of Allograft Rejection","volume":"37","author":"Fedrigo","year":"2018","journal-title":"J. Heart Lung Transplant."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1982","DOI":"10.1002\/ppul.26422","article-title":"MicroRNA Dysregulation in the Heart and Lung of Infants with Bronchopulmonary Dysplasia","volume":"58","author":"Koussa","year":"2023","journal-title":"Pediatr. Pulmonol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Li, L., He, X., Liu, M., Yun, L., and Cong, B. (2022). Diagnostic Value of Cardiac MiR-126-5p, MiR-134-5p, and MiR-499a-5p in Coronary Artery Disease-Induced Sudden Cardiac Death. Front. Cardiovasc. Med., 9.","DOI":"10.3389\/fcvm.2022.944317"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"102867","DOI":"10.1016\/j.fsigen.2023.102867","article-title":"Suitable Biomarkers for Post-Mortem Differentiation of Cardiac Death Causes: Quantitative Analysis of MiR-1, MiR-133a and MiR-26a in Heart Tissue and Whole Blood","volume":"65","author":"Mildeberger","year":"2023","journal-title":"Forensic Sci. Int. Genet."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"29476","DOI":"10.1038\/s41598-024-81114-6","article-title":"Postmortem Analyses of Myocardial MicroRNA Expression in Sepsis","volume":"14","author":"Lehto","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1520","DOI":"10.5858\/arpa.2013-0691-RA","article-title":"A Review of Preanalytical Factors Affecting Molecular, Protein, and Morphological Analysis of Formalin-Fixed, Paraffin-Embedded (FFPE) Tissue: How Well Do You Know Your FFPE Specimen?","volume":"138","author":"Bass","year":"2014","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Petersen, P.H.D., James, J.P., Riis, L.B., H\u00f8gdall, C.K., and H\u00f8gdall, E.V. (2024). Proof of Concept Study: Comparison of Semi-Automated RNA Isolation Methods from Archived Formalin-Fixed, Paraffin-Embedded Tissues with Clinical Routine RNA Isolation Methods. Methods Protoc., 7.","DOI":"10.3390\/mps7060101"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Bernini Di Michele, A., Onofri, V., Melchionda, F., Fiordelmondo, L., Ciarimboli, E., Palpacelli, M., Sablone, S., Turchi, C., and Pesaresi, M. (2025). Cardiac Genetic Variants in Sudden, Unexpected Death in Epilepsy: From Challenging DNA Extraction Methods to Updated NGS Panels for Improved Genetic Analysis. Genes, 16.","DOI":"10.3390\/genes16111272"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1016\/j.jmoldx.2011.06.007","article-title":"Paraffin Embedding Contributes to RNA Aggregation, Reduced RNA Yield, and Low RNA Quality","volume":"13","author":"Evers","year":"2011","journal-title":"J. Mol. Diagn."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1038\/nature09783","article-title":"Pervasive Roles of microRNAs in Cardiovascular Biology","volume":"469","author":"Small","year":"2011","journal-title":"Nature"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2177","DOI":"10.1016\/j.jacc.2014.01.050","article-title":"microRNAs in Cardiovascular Diseases","volume":"63","author":"Condorelli","year":"2014","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1161\/CIRCRESAHA.109.215566","article-title":"Circulating MicroRNAs in Patients With Coronary Artery Disease","volume":"107","author":"Fichtlscherer","year":"2010","journal-title":"Circ. Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2765","DOI":"10.1093\/eurheartj\/ehq167","article-title":"Circulating microRNAs Are New and Sensitive Biomarkers of Myocardial Infarction","volume":"31","author":"Devanna","year":"2010","journal-title":"Eur. Heart J."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Lazzari, J., Scatena, A., Di Paolo, M., and Rocchi, A. (2026). Selection and Validation of Endogenous Reference microRNAs for Post-Mortem Interval Estimation in Vitreous Humor: A Preliminary Study. Int. J. Mol. Sci., 27.","DOI":"10.3390\/ijms27052102"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1439","DOI":"10.1007\/s00414-025-03480-8","article-title":"Are Pre-Analytical Factors Fully Considered in Forensic FFPE Molecular Analyses? A Systematic Review Reveals the Need for Standardised Procedures","volume":"139","author":"Bonin","year":"2025","journal-title":"Int. J. Leg. Med."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Sacco, M.A., Gualtieri, S., Verrina, M.C., Cordasco, F., Monterossi, M.D., Grimaldi, G., Mastrangelo, H., Mazza, G., and Aquila, I. (2026). MicroRNAs in Cardiovascular Diseases and Forensic Applications: A Systematic Review of Diagnostic and Post-Mortem Implications. Int. J. Mol. Sci., 27.","DOI":"10.3390\/ijms27020825"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1735","DOI":"10.1007\/s00414-025-03702-z","article-title":"Molecular Forensics: RNA Degradation as a Marker for Postmortem Interval Determination","volume":"140","author":"Gupta","year":"2026","journal-title":"Int. J. Leg. Med."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Maiese, A., Scatena, A., Costantino, A., Di Paolo, M., La Russa, R., Turillazzi, E., Frati, P., and Fineschi, V. (2021). MicroRNAs as Useful Tools to Estimate Time Since Death. A Systematic Review of Current Literature. Diagnostics, 11.","DOI":"10.3390\/diagnostics11010064"}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/17\/7\/819\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T04:16:11Z","timestamp":1784607371000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/17\/7\/819"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,17]]},"references-count":50,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["genes17070819"],"URL":"https:\/\/doi.org\/10.3390\/genes17070819","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,17]]}}}