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Preanalytical Variables in the Analysis of Mitochondrial DNA in Whole Blood and Plasma from Pancreatic Cancer Patients

Given the crucial role of mitochondria as the main cellular energy provider and its contribution towards tumor growth, chemoresistance, and cancer cell plasticity, mitochondrial DNA (mtDNA) could serve as a relevant biomarker. Thus, the profiling of mtDNA mutations and copy number variations is rece...

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Autores principales: Randeu, Hannah, Bronkhorst, Abel J., Mayer, Zsuzsanna, Oberhofer, Angela, Polatoglou, Eleni, Heinemann, Volker, Haas, Michael, Boeck, Stefan, Holdenrieder, Stefan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9406772/
https://www.ncbi.nlm.nih.gov/pubmed/36010255
http://dx.doi.org/10.3390/diagnostics12081905
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author Randeu, Hannah
Bronkhorst, Abel J.
Mayer, Zsuzsanna
Oberhofer, Angela
Polatoglou, Eleni
Heinemann, Volker
Haas, Michael
Boeck, Stefan
Holdenrieder, Stefan
author_facet Randeu, Hannah
Bronkhorst, Abel J.
Mayer, Zsuzsanna
Oberhofer, Angela
Polatoglou, Eleni
Heinemann, Volker
Haas, Michael
Boeck, Stefan
Holdenrieder, Stefan
author_sort Randeu, Hannah
collection PubMed
description Given the crucial role of mitochondria as the main cellular energy provider and its contribution towards tumor growth, chemoresistance, and cancer cell plasticity, mitochondrial DNA (mtDNA) could serve as a relevant biomarker. Thus, the profiling of mtDNA mutations and copy number variations is receiving increasing attention for its possible role in the early diagnosis and monitoring therapies of human cancers. This applies particularly to highly aggressive pancreatic cancer, which is often diagnosed late and is associated with poor prognosis. As current diagnostic procedures are based on imaging, tissue histology, and protein biomarkers with rather low specificity, tumor-derived mtDNA mutations detected from whole blood represents a potential significant leap forward towards early cancer diagnosis. However, for future routine use in clinical settings it is essential that preanalytics related to the characterization of mtDNA in whole blood are thoroughly standardized, controlled, and subject to proper quality assurance, yet this is largely lacking. Therefore, in this study we carried out a comprehensive preanalytical workup comparing different mtDNA extraction methods and testing important preanalytical steps, such as the use of different blood collection tubes, different storage temperatures, length of storage time, and yields in plasma vs. whole blood. To identify analytical and preanalytical differences, all variables were tested in both healthy subjects and pancreatic carcinoma patients. Our results demonstrated a significant difference between cancer patients and healthy subjects for some preanalytical workflows, while other workflows failed to yield statistically significant differences. This underscores the importance of controlling and standardizing preanalytical procedures in the development of clinical assays based on the measurement of mtDNA.
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spelling pubmed-94067722022-08-26 Preanalytical Variables in the Analysis of Mitochondrial DNA in Whole Blood and Plasma from Pancreatic Cancer Patients Randeu, Hannah Bronkhorst, Abel J. Mayer, Zsuzsanna Oberhofer, Angela Polatoglou, Eleni Heinemann, Volker Haas, Michael Boeck, Stefan Holdenrieder, Stefan Diagnostics (Basel) Article Given the crucial role of mitochondria as the main cellular energy provider and its contribution towards tumor growth, chemoresistance, and cancer cell plasticity, mitochondrial DNA (mtDNA) could serve as a relevant biomarker. Thus, the profiling of mtDNA mutations and copy number variations is receiving increasing attention for its possible role in the early diagnosis and monitoring therapies of human cancers. This applies particularly to highly aggressive pancreatic cancer, which is often diagnosed late and is associated with poor prognosis. As current diagnostic procedures are based on imaging, tissue histology, and protein biomarkers with rather low specificity, tumor-derived mtDNA mutations detected from whole blood represents a potential significant leap forward towards early cancer diagnosis. However, for future routine use in clinical settings it is essential that preanalytics related to the characterization of mtDNA in whole blood are thoroughly standardized, controlled, and subject to proper quality assurance, yet this is largely lacking. Therefore, in this study we carried out a comprehensive preanalytical workup comparing different mtDNA extraction methods and testing important preanalytical steps, such as the use of different blood collection tubes, different storage temperatures, length of storage time, and yields in plasma vs. whole blood. To identify analytical and preanalytical differences, all variables were tested in both healthy subjects and pancreatic carcinoma patients. Our results demonstrated a significant difference between cancer patients and healthy subjects for some preanalytical workflows, while other workflows failed to yield statistically significant differences. This underscores the importance of controlling and standardizing preanalytical procedures in the development of clinical assays based on the measurement of mtDNA. MDPI 2022-08-06 /pmc/articles/PMC9406772/ /pubmed/36010255 http://dx.doi.org/10.3390/diagnostics12081905 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Randeu, Hannah
Bronkhorst, Abel J.
Mayer, Zsuzsanna
Oberhofer, Angela
Polatoglou, Eleni
Heinemann, Volker
Haas, Michael
Boeck, Stefan
Holdenrieder, Stefan
Preanalytical Variables in the Analysis of Mitochondrial DNA in Whole Blood and Plasma from Pancreatic Cancer Patients
title Preanalytical Variables in the Analysis of Mitochondrial DNA in Whole Blood and Plasma from Pancreatic Cancer Patients
title_full Preanalytical Variables in the Analysis of Mitochondrial DNA in Whole Blood and Plasma from Pancreatic Cancer Patients
title_fullStr Preanalytical Variables in the Analysis of Mitochondrial DNA in Whole Blood and Plasma from Pancreatic Cancer Patients
title_full_unstemmed Preanalytical Variables in the Analysis of Mitochondrial DNA in Whole Blood and Plasma from Pancreatic Cancer Patients
title_short Preanalytical Variables in the Analysis of Mitochondrial DNA in Whole Blood and Plasma from Pancreatic Cancer Patients
title_sort preanalytical variables in the analysis of mitochondrial dna in whole blood and plasma from pancreatic cancer patients
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9406772/
https://www.ncbi.nlm.nih.gov/pubmed/36010255
http://dx.doi.org/10.3390/diagnostics12081905
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