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Stability of exosomes in the postmortem serum and preliminary study on exosomal miRNA expression profiling in serum from myocardial infarction cadavers

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Abstract

Exosome-encapsulated miRNAs could potentially be sensitive biomarkers of human diseases. Since a lipid bilayer membrane surrounds exosomes, the exosomal miRNA may stably exist in body fluids with diseases as well as biological fluids. Therefore, exosomal miRNA may be helpful for autopsy diagnosis. Assuming cadaver blood would be most useful, we initially examined serum exosome stability with regard to storage temperatures and periods. Characteristic analyses of the exosome revealed that exosomes and the content, miRNA, were stably preserved until at least three days when stored at below 20 °C. Subsequently, exosomal miRNA expression profiling was performed on the serum of acute myocardial infarction (AMI, 4 cases) autopsy bodies and on hemorrhagic shock bodies used as the control (CT, 3 cases). Results showed that significant twofold up- and downregulations of expression of 18 and 16 miRNAs were detectable in AMI as compared to the CT, respectively. miR-126-3p, which has been reported to be increased in serum of AMI patients and a mouse model, was one of the significantly upregulated miRNAs. Furthermore, dysregulation of exosomal miRNAs, such as miR-145-5p, miR-143-3p, and miR-222-3p, which are involved in cardioprotection, may be associated with AMI pathogenesis. These findings provide a novel perspective on the potential role of exosomal miRNA in determining the cause of death.

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References

  1. Hessvik NP, Llorente A (2018) Current knowledge on exosome biogenesis and release. Cell Mol Life Sci 75:193–208. https://doi.org/10.1007/s00018-017-2595-9

    Article  CAS  PubMed  Google Scholar 

  2. Akers JC, Ramakrishnan V, Yang I, Hua W, Mao Y, Carter BS, Chen CC (2016) Optimizing preservation of extracellular vesicular miRNAs derived from clinical cerebrospinal fluid. Cancer Biomark 17:125–132. https://doi.org/10.3233/CBM-160609

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Iaconetti C, Sorrentino S, De Rosa S, Indolfi C (2016) Exosomal miRNAs in heart disease. Physiol (Bethesda) 31:16–24. https://doi.org/10.1152/physiol.00029.2015

    Article  CAS  Google Scholar 

  4. Barile L, Moccetti T, Marban E, Vassalli G (2017) Roles of exosomes in cardioprotection. Eur Heart J 38:1372–1379. https://doi.org/10.1093/eurheartj/ehw304

    Article  CAS  PubMed  Google Scholar 

  5. Kinoshita T, Yip KW, Spence T, Liu FF (2017) MicroRNAs in extracellular vesicles: potential cancer biomarkers. J Hum Genet 62:67–74. https://doi.org/10.1038/jhg.2016.87

    Article  CAS  PubMed  Google Scholar 

  6. Jaiswal R, Sedger LM (2019) Intercellular vesicular transfer by exosomes, microparticles and oncosomes - implications for cancer biology and treatments. Front Oncol 9:125. https://doi.org/10.3389/fonc.2019.00125

    Article  PubMed  PubMed Central  Google Scholar 

  7. Russo I, Bubacco L, Greggio E (2012) Exosomes-associated neurodegeneration and progression of Parkinson’s disease. Am J Neurodegener Dis 1:217–225

    PubMed  PubMed Central  Google Scholar 

  8. Ambros V (2004) The functions of animal microRNAs. Nature 431:350–355. https://doi.org/10.1038/nature02871

    Article  CAS  PubMed  Google Scholar 

  9. Mori MA, Ludwig RG, Garcia-Martin R, Brandao BB, Kahn CR (2019) Extracellular miRNAs: from biomarkers to mediators of physiology and disease. Cell Metab 30:656–673. https://doi.org/10.1016/j.cmet.2019.07.011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Liu T, Zhang Q, Zhang J, Li C, Miao YR, Lei Q, Li Q, Guo AY (2019) EVmiRNA: a database of miRNA profiling in extracellular vesicles. Nucleic Acids Res 47:D89–D93. https://doi.org/10.1093/nar/gky985

    Article  CAS  PubMed  Google Scholar 

  11. Nabel EG, Braunwald E (2012) A tale of coronary artery disease and myocardial infarction. N Engl J Med 366:54–63. https://doi.org/10.1056/NEJMra1112570

    Article  CAS  PubMed  Google Scholar 

  12. Markwerth P, Bajanowski T, Tzimas I, Dettmeyer R (2021) Sudden cardiac death-update. Int J Legal Med 135:483–495. https://doi.org/10.1007/s00414-020-02481-z

    Article  CAS  PubMed  Google Scholar 

  13. Zheng D, Huo M, Li B, Wang W, Piao H, Wang Y, Zhu Z, Li D, Wang T, Liu K (2020) The role of exosomes and exosomal microRNA in cardiovascular disease. Front Cell Dev Biol 8:616161. https://doi.org/10.3389/fcell.2020.616161

    Article  PubMed  Google Scholar 

  14. Akbar N, Digby JE, Cahill TJ, Tavare AN, Corbin AL, Saluja S, Dawkins S, Edgar L, Rawlings N, Ziberna K, McNeill E, Oxford Acute Myocardial Infarction S, Johnson E, Aljabali AA, Dragovic RA, Rohling M, Belgard TG, Udalova IA, Greaves DR, Channon KM, Riley PR, Anthony DC, Choudhury RP (2017) Endothelium-derived extracellular vesicles promote splenic monocyte mobilization in myocardial infarction. JCI Insight 2 https://doi.org/10.1172/jci.insight.93344

  15. Kanno S, Hirano S, Sakamoto T, Furuyama A, Takase H, Kato H, Fukuta M, Aoki Y (2020) Scavenger receptor MARCO contributes to cellular internalization of exosomes by dynamin-dependent endocytosis and macropinocytosis. Sci Rep 10:21795. https://doi.org/10.1038/s41598-020-78464-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Kosaka N, Iguchi H, Yoshioka Y, Takeshita F, Matsuki Y, Ochiya T (2010) Secretory mechanisms and intercellular transfer of microRNAs in living cells. J Biol Chem 285:17442–17452. https://doi.org/10.1074/jbc.M110.107821

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Park SJ, Jeon H, Yoo SM, Lee MS (2018) The effect of storage temperature on the biological activity of extracellular vesicles for the complement system. In Vitro Cell Dev Biol Anim 54:423–429. https://doi.org/10.1007/s11626-018-0261-7

    Article  CAS  PubMed  Google Scholar 

  18. Ge Q, Zhou Y, Lu J, Bai Y, Xie X, Lu Z (2014) miRNA in plasma exosome is stable under different storage conditions. Molecules 19:1568–1575. https://doi.org/10.3390/molecules19021568

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Schulz E, Karagianni A, Koch M, Fuhrmann G (2020) Hot EVs - How temperature affects extracellular vesicles. Eur J Pharm Biopharm 146:55–63. https://doi.org/10.1016/j.ejpb.2019.11.010

    Article  CAS  PubMed  Google Scholar 

  20. Duan S, Wang C, Xu X, Zhang X, Su G, Li Y, Fu S, Sun P, Tian J (2022) Peripheral serum exosomes isolated from patients with acute myocardial infarction promote endothelial cell angiogenesis via the miR-126-3p/TSC1/mTORC1/HIF-1α pathway. Int J Nanomed 17:1577–1592. https://doi.org/10.2147/ijn.S338937

    Article  CAS  Google Scholar 

  21. Ling H, Guo Z, Shi Y, Zhang L, Song C (2020) Serum exosomal microRNA-21, microRNA-126, and PTEN are novel biomarkers for diagnosis of acute coronary syndrome. Front Physiol 11:654. https://doi.org/10.3389/fphys.2020.00654

    Article  PubMed  PubMed Central  Google Scholar 

  22. Hergenreider E, Heydt S, Treguer K, Boettger T, Horrevoets AJ, Zeiher AM, Scheffer MP, Frangakis AS, Yin X, Mayr M, Braun T, Urbich C, Boon RA, Dimmeler S (2012) Atheroprotective communication between endothelial cells and smooth muscle cells through miRNAs. Nat Cell Biol 14:249–256. https://doi.org/10.1038/ncb2441

    Article  CAS  PubMed  Google Scholar 

  23. Chistiakov DA, Sobenin IA, Orekhov AN, Bobryshev YV (2015) Human miR-221/222 in physiological and atherosclerotic vascular remodeling. Biomed Res Int 2015:354517. https://doi.org/10.1155/2015/354517

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Lai TC, Lee TL, Chang YC, Chen YC, Lin SR, Lin SW, Pu CM, Tsai JS, Chen YL (2020) MicroRNA-221/222 mediates ADSC-exosome-induced cardioprotection against ischemia/reperfusion by targeting PUMA and ETS-1. Front Cell Dev Biol 8:569150. https://doi.org/10.3389/fcell.2020.569150

    Article  PubMed  PubMed Central  Google Scholar 

  25. Pang J, Ye L, Chen Q, Wang J, Yang X, He W, Hao L (2020) The effect of microRNA-101 on angiogenesis of human umbilical vein endothelial cells during hypoxia and in mice with myocardial infarction. Biomed Res Int 2020:5426971. https://doi.org/10.1155/2020/5426971

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Higashi K, Yamada Y, Minatoguchi S, Baba S, Iwasa M, Kanamori H, Kawasaki M, Nishigaki K, Takemura G, Kumazaki M, Akao Y, Minatoguchi S (2015) MicroRNA-145 repairs infarcted myocardium by accelerating cardiomyocyte autophagy. Am J Physiol Heart Circ Physiol 309:H1813–H1826. https://doi.org/10.1152/ajpheart.00709.2014

    Article  CAS  PubMed  Google Scholar 

  27. Ragusa M, Barbagallo C, Statello L, Caltabiano R, Russo A, Puzzo L, Avitabile T, Longo A, Toro MD, Barbagallo D, Valadi H, Di Pietro C, Purrello M, Reibaldi M (2015) miRNA profiling in vitreous humor, vitreal exosomes and serum from uveal melanoma patients: Pathological and diagnostic implications. Cancer Biol Ther 16:1387–1396. https://doi.org/10.1080/15384047.2015.1046021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Zhao Z, Liu G, Zhang H, Ruan P, Ge J, Liu Q (2021) BIRC5, GAJ5, and lncRNA NPHP3-AS1 are correlated with the development of atrial fibrillation-valvular heart disease. Int Heart J 62:153–161. https://doi.org/10.1536/ihj.20-238

    Article  CAS  PubMed  Google Scholar 

  29. Hou J, Wang J, Lin C, Fu J, Ren J, Li L, Guo H, Han X, Liu J (2014) Circulating microRNA profiles differ between Qi-stagnation and Qi-deficiency in coronary heart disease patients with blood stasis syndrome. Evid Based Complement Alternat Med 2014:926962. https://doi.org/10.1155/2014/926962

    Article  PubMed  PubMed Central  Google Scholar 

  30. Gryshkova V, Lushbough I, Palmer J, Burrier R, Delaunois A, Donley E, Valentin JP (2022) MicroRNAs signatures as potential biomarkers of structural cardiotoxicity in human-induced pluripotent stem-cell derived cardiomyocytes. Arch Toxicol https://doi.org/10.1007/s00204-022-03280-8

  31. Jordan NP, Tingle SJ, Shuttleworth VG, Cooke K, Redgrave RE, Singh E, Glover EK, Ahmad Tajuddin HB, Kirby JA, Arthur HM, Ward C, Sheerin NS, Ali S (2021) MiR-126-3p Is dynamically regulated in endothelial-to-mesenchymal transition during fibrosis. Int J Mol Sci 22 https://doi.org/10.3390/ijms22168629

  32. Xue S, Liu D, Zhu W, Su Z, Zhang L, Zhou C, Li P (2019) Circulating MiR-17-5p, MiR-126-5p and MiR-145-3p are novel biomarkers for diagnosis of acute myocardial infarction. Front Physiol 10:123. https://doi.org/10.3389/fphys.2019.00123

    Article  PubMed  PubMed Central  Google Scholar 

  33. Li M, Ren C, Wu C, Li X, Li X, Mao W (2020) Bioinformatics analysis reveals diagnostic markers and vital pathways involved in acute coronary syndrome. Cardiol Res Pract 2020:3162581. https://doi.org/10.1155/2020/3162581

    Article  PubMed  PubMed Central  Google Scholar 

  34. Xu C, Jia Z, Cao X, Wang S, Wang J, An L (2022) Hsa_circ_0007059 promotes apoptosis and inflammation in cardiomyocytes during ischemia by targeting microRNA-378 and microRNA-383. Cell Cycle 21:1003–1019. https://doi.org/10.1080/15384101.2022.2040122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Eyyupkoca F, Ercan K, Kiziltunc E, Ugurlu IB, Kocak A, Eyerci N (2022) Determination of microRNAs associated with adverse left ventricular remodeling after myocardial infarction. Mol Cell Biochem 477:781–791. https://doi.org/10.1007/s11010-021-04330-y

    Article  CAS  PubMed  Google Scholar 

  36. Su Y, Sun Y, Tang Y, Li H, Wang X, Pan X, Liu W, Zhang X, Zhang F, Xu Y, Yan C, Ong SB, Xu D (2021) Circulating miR-19b-3p as a novel prognostic biomarker for acute heart failure. J Am Heart Assoc 10:e022304. https://doi.org/10.1161/jaha.121.022304

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Corsten MF, Heggermont W, Papageorgiou AP, Deckx S, Tijsma A, Verhesen W, van Leeuwen R, Carai P, Thibaut HJ, Custers K, Summer G, Hazebroek M, Verheyen F, Neyts J, Schroen B, Heymans S (2015) The microRNA-221/-222 cluster balances the antiviral and inflammatory response in viral myocarditis. Eur Heart J 36:2909–2919. https://doi.org/10.1093/eurheartj/ehv321

    Article  CAS  PubMed  Google Scholar 

  38. Zhang Q, Kandic I, Kutryk MJ (2011) Dysregulation of angiogenesis-related microRNAs in endothelial progenitor cells from patients with coronary artery disease. Biochem Biophys Res Commun 405:42–46. https://doi.org/10.1016/j.bbrc.2010.12.119

    Article  CAS  PubMed  Google Scholar 

  39. Verjans R, Peters T, Beaumont FJ, van Leeuwen R, van Herwaarden T, Verhesen W, Munts C, Bijnen M, Henkens M, Diez J, de Windt LJ, van Nieuwenhoven FA, van Bilsen M, Goumans MJ, Heymans S, González A, Schroen B (2018) MicroRNA-221/222 family counteracts myocardial fibrosis in pressure overload-induced heart failure. Hypertension 71:280–288. https://doi.org/10.1161/hypertensionaha.117.10094

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We acknowledge the assistance of the Research Equipment Sharing Center at Nagoya City University. We would like to thank Mr. Hiroshi Takase of the Research Equipment Sharing Center of Nagoya City University for his assistance with the transmission electron microscopic observations, and Dr. Seishiro Hirano of the National Institute for Environmental Studies for his help with the dynamic light scattering analysis and suggestions.

Funding

JSPS KAKENHI grant (number 21K10526).

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Author notes

  1. Sanae Kanno and Tsubasa Sakamoto contributed equally to this work as co-first authors.

Authors and Affiliations

  1. Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-Cho, Mizuho-Ku, Nagoya, 467-8601, Japan

    Sanae Kanno, Tsubasa Sakamoto, Mamiko Fukuta, Hideaki Kato & Yasuhiro Aoki

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  1. Sanae Kanno
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  2. Tsubasa Sakamoto
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Contributions

S. K. and T. S.: conceived and designed the experiments; S. K. and T. S.: performed the experiments and data analyses; M. F., H. K., and Y. A.: performed the autopsies and examinations; S. K. wrote the manuscript with contributions from co-authors; Y. A.: project administration.

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Correspondence to Sanae Kanno.

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This study in use of human serum and autopsy samples was approved by the Institutional Review Board for the Protection of Human Subjects in Research at Nagoya City University (approval number: 60–18-0151 and 60–21-0144), respectively.

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Kanno, S., Sakamoto, T., Fukuta, M. et al. Stability of exosomes in the postmortem serum and preliminary study on exosomal miRNA expression profiling in serum from myocardial infarction cadavers. Int J Legal Med 137, 825–834 (2023). https://doi.org/10.1007/s00414-022-02913-y

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