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Immunohistochemistry of Human Hsp60 in Health and Disease: Recent Advances in Immunomorphology and Methods for Assessing the Chaperonin in Extracellular Vesicles

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Chaperones

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2693))

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Abstract

The subject matter of this chapter is defined by the title of its two previous editions, “Immunohistochemistry of human Hsp60 in health and disease: From autoimmunity to cancer,” the latest of which appeared in 2018. Since then, considerable advances have been made in the fields of autoimmunity and cancer and some of them are closely linked to progress in the understanding of the chaperone system (CS). This is a physiological system composed of molecular chaperones, co-chaperones, chaperone cofactors, and chaperone interactors and receptors. The molecular chaperones are the chief members of the CS, and here we focus on one of them, Hsp60. Since extracellular vesicles (EVs) have also emerged as key factors in the functioning of the CS and in carcinogenesis, we have incorporated a detailed section about them. This chapter explains how to assess Hsp60 in tissues and in EVs for application in diagnosis, prognostication, and patient monitoring and, eventually, for developing methods using them as therapeutic targets and tools. We describe immunohistochemical techniques, immunofluorescence and double immunofluorescence-confocal microscopy, and methods for collecting and isolating EVs from blood plasma and for assessing their contents in Hsp60 and related microRNAs (miRNAs). All these procedures have proven to be reliable and useful in the study and management of various types of cancer and inflammatory and autoimmune conditions.

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References

  1. Macario AJL, Conway de Macario E, Cappello F (2013) The Chaperonopathies. Diseases with defective molecular chaperones. Springer, Dordrecht/Heidelberg/New York/London

    Book  Google Scholar 

  2. Bross P, Fernandez-Guerra P (2016) Front Mol Biosci 3:49. https://doi.org/10.3389/fmolb.2016.00049

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Gomez-Llorente Y, Jebara F, Patra M et al (2020) Structural basis for active single and double ring complexes in human mitochondrial Hsp60-Hsp10 chaperonin. Nat Commun 11:1916. https://doi.org/10.1038/s41467-020-15698-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Macario AJL, Conway de Macario E (2023) The chaperone system in autoimmunity, inflammation, and virus-induced diseases: role of chaperonins. In: Fink G (ed) Stress: immunology and inflammation. Handbook of stress series, vol 5. Chapter 13. Elsevier/Academic, San Diego, USA

    Google Scholar 

  5. Cappello F, Conway de Macario E, Zummo G, Macario AJL (2011) Immunohistochemistry of human Hsp60 in health and disease: from autoimmunity to cancer. Methods Mol Biol 787:245–254. https://doi.org/10.1007/978-1-61779-295-3_18

    Article  CAS  PubMed  Google Scholar 

  6. Cappello F, Conway de Macario E, Rappa F et al (2018) Immunohistochemistry of human Hsp60 in health and disease: from autoimmunity to cancer. Methods Mol Biol 1709:293–305. https://doi.org/10.1007/978-1-4939-7477-1_21

    Article  CAS  PubMed  Google Scholar 

  7. Caruso Bavisotto C, Alberti G, Vitale AM et al (2020) Hsp60 post-translational modifications: functional and pathological consequences. Front Mol Biosci 7:95. https://doi.org/10.3389/fmolb.2020.00095

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Vitale AM, Conway de Macario E, Alessandro R et al (2020) Missense mutations of human Hsp60: a computational analysis to unveil their pathological significance. Front Genet 11:969. https://doi.org/10.3389/fgene.2020.00969. Section Genetics of Common and Rare Diseases

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Basset CA, Rappa F, Lentini VL et al (2021) Hsp27 and Hsp60 in human submandibular salivary gland: quantitative patterns in healthy and cancerous tissues with potential implications for differential diagnosis and carcinogenesis. Acta Histochem 123:151771. https://doi.org/10.1016/j.acthis.2021.151771

    Article  CAS  PubMed  Google Scholar 

  10. Alberti G, Vergilio G, Paladino L et al (2022) The chaperone system in breast cancer: roles and therapeutic prospects of the molecular chaperones Hsp27, Hsp60, Hsp70, and Hsp90. Int J Mol Sci 23:7792

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Sun B, Li G, Yu Q et al (2021) HSP60 in cancer: a promising biomarker for diagnosis and a potentially useful target for treatment. J Drug Target 30:31–45. https://doi.org/10.1080/1061186X.2021.1920025

    Article  PubMed  Google Scholar 

  12. Tang Y, Zhou Y, Fan S et al (2022) The multiple roles and therapeutic potential of HSP60 in cancer. Biochem Pharmacol 201:115096. https://doi.org/10.1016/j.bcp.2022.115096

    Article  CAS  PubMed  Google Scholar 

  13. Tikhomirova TS, Matyunin MA, Lobanov MY et al (2022) In-depth analysis of amino acid and nucleotide sequences of Hsp60: how conserved is this protein? Proteins 90:1119–1141. https://doi.org/10.1002/prot.26294

    Article  CAS  PubMed  Google Scholar 

  14. Caruso Bavisotto C, Scalia F, Marino Gammazza A et al (2020) Extracellular vesicle-mediated cell-cell communication in the nervous system: focus on neurological diseases. Int J Mol Sci 20:pii: E434. https://doi.org/10.3390/ijms20020434

    Article  CAS  Google Scholar 

  15. David S, Vitale AM, Fucarino F et al (2021) The challenging riddle about the Janus-type role of Hsp60 and related extracellular vesicles and miRNAs in carcinogenesis and the promises of its solution. Appl Sci 11:1175. https://doi.org/10.3390/app11031175

    Article  CAS  Google Scholar 

  16. Graziano F, Iacopino DG, Cammarata G, Scalia G, Campanella C, Giannone AG et al (2021) The triad Hsp60-miRNAs-extracellular vesicles in brain tumors: assessing its components for understanding tumorigenesis and monitoring patients. Appl Sci 11:2867. https://doi.org/10.3390/app11062867

    Article  CAS  Google Scholar 

  17. Rappa F, Pitruzzella A, Marino Gammazza A et al (2016) Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer. Cell Stress Chaperones 21:927–933. https://doi.org/10.1007/s12192-016-0721-5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. McKee C, Sigala B, Soeda J et al (2015) Amphiregulin activates human hepatic stellate cells and is upregulated in non-alcoholic steatohepatitis. Sci Rep 5:8812. https://doi.org/10.1038/srep08812

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Rappa F, Sciume C, Bello M et al (2014) Comparative analysis of hsp10 and hsp90 expression in healthy mucosa and adenocarcinoma of the large bowel. Anticancer Res 34:4153–4159

    CAS  PubMed  Google Scholar 

  20. Alberti G, Campanella C, Paladino L et al (2022) The chaperone system in glioblastoma multiforme and derived cell lines: diagnostic and mechanistic implications. Front Biosci (Landmark Ed) 27. https://doi.org/10.31083/j.fbl2703097

  21. Mishra A, Mirzaei H, Guidi N et al (2021) Fasting-mimicking diet prevents high-fat diet effect on cardiometabolic risk and lifespan. Nat Metab 3:1342–1356. https://doi.org/10.1038/s42255-021-00469-6

    Article  CAS  PubMed  Google Scholar 

  22. Barone R, Marino Gammazza A, Paladino L et al (2021) Morphological alterations and stress protein variations in lung biopsies obtained from autopsies of COVID-19 subjects. Cell 10:3136. https://doi.org/10.3390/cells10113136

    Article  CAS  Google Scholar 

  23. Théry C, Witwer KW, Aikawa E et al (2018) Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles 7:1535750. https://doi.org/10.1080/20013078.2018.1535750

    Article  PubMed  PubMed Central  Google Scholar 

  24. Théry C, Clayton A, Amigorena S et al (2006) Isolation and characterization of exosomes from cell culture supernatants. Chapter 3. Unit 3.22. https://doi.org/10.1002/0471143030.cb0322s30

  25. Campanella C, Rappa F, Sciumè C et al (2015) Heat shock protein 60 levels in tissue and circulating exosomes in human large bowel cancer before and after ablative surgery. Cancer 121:3230–3239. https://doi.org/10.1002/cncr.29499

    Article  CAS  PubMed  Google Scholar 

  26. Caruso Bavisotto C, Cipolla C, Graceffa G et al (2019) Immunomorphological pattern of molecular chaperones in normal and pathological thyroid tissues and circulating exosomes: potential use in clinics. Int J Mol Sci 20:4496. https://doi.org/10.3390/ijms20184496

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

A.J.L.M. and E.C. de M. were partially supported by IMET and IEMEST.

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Authors and Affiliations

  1. Department of Biomedicine, Neurosciences and Advanced Diagnostic (BiND), Human Anatomy Section, University of Palermo, Palermo, Italy

    Celeste Caruso Bavisotto, Francesco Cappello & Francesca Rappa

  2. Department of Microbiology and Immunology, School of Medicine, University of Maryland at Baltimore-Institute of Marine and Environmental Technology (IMET) – Rita Rossi Colwell Center, Baltimore, MD, USA

    Everly Conway de Macario & Alberto J. L. Macario

Authors
  1. Celeste Caruso Bavisotto
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  2. Francesco Cappello
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  3. Everly Conway de Macario
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  4. Alberto J. L. Macario
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  5. Francesca Rappa
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Corresponding author

Correspondence to Celeste Caruso Bavisotto .

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Editors and Affiliations

  1. Harvard Medical School, Beth Israel Deaconess Medical Center, Boston, MA, USA

    Stuart K. Calderwood

  2. Danville, PA, USA

    Thomas L. Prince

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Caruso Bavisotto, C., Cappello, F., Conway de Macario, E., Macario, A.J.L., Rappa, F. (2023). Immunohistochemistry of Human Hsp60 in Health and Disease: Recent Advances in Immunomorphology and Methods for Assessing the Chaperonin in Extracellular Vesicles. In: Calderwood, S.K., Prince, T.L. (eds) Chaperones. Methods in Molecular Biology, vol 2693. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3342-7_20

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  • DOI: https://doi.org/10.1007/978-1-0716-3342-7_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3341-0

  • Online ISBN: 978-1-0716-3342-7

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