HiIDDD: a high-throughput imaging pipeline for the quantitative detection of DNA damage in primary human immune cells

DNA damage is a prominent biomarker for numerous diseases, including cancer, as well as for the aging process. Detection of DNA damage routinely relies on traditional microscopy or cytometric methods. However, these techniques are typically of limited throughput and are not ideally suited for large-...

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Veröffentlicht in:Scientific reports Jg. 12; H. 1; S. 6335 - 13
Hauptverfasser: Gallant, Kelsey, Bektas, Arsun, Kaileh, Mary, Lustig, Ana, Ferrucci, Luigi, Pegoraro, Gianluca, Misteli, Tom
Format: Journal Article
Sprache:Englisch
Veröffentlicht: London Nature Publishing Group UK 15.04.2022
Nature Publishing Group
Nature Portfolio
Schlagworte:
631/80/103
631/80/2373
Aging
Batch processing
Biological Assay
CD4 antigen
CD8 antigen
Cryopreservation
Deoxyribonucleic acid
Diagnostic Imaging
DNA
DNA Damage
High-Throughput Screening Assays - methods
Humanities and Social Sciences
Humans
Lymphocytes B
Lymphocytes T
Monocytes
multidisciplinary
Population studies
Science
Science (multidisciplinary)
ISSN:2045-2322, 2045-2322
Online-Zugang:Volltext
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Zusammenfassung:DNA damage is a prominent biomarker for numerous diseases, including cancer, as well as for the aging process. Detection of DNA damage routinely relies on traditional microscopy or cytometric methods. However, these techniques are typically of limited throughput and are not ideally suited for large-scale longitudinal and population studies that require analysis of large sample sets. We have developed HiIDDD (High-throughput Immune cell DNA Damage Detection), a robust, quantitative and single-cell assay that measures DNA damage by high-throughput imaging using the two major DNA damage markers 53BP1 and γ -H2AX. We demonstrate sensitive detection with low inter-assay variability of DNA damage in various types of freshly isolated and cryopreserved primary human immune cells, including CD4 + and CD8 + T cells, B cells and monocytes. As proof of principle, we demonstrate parallel batch processing of several immune cell types from multiple donors. We find common patterns of DNA damage in multiple immune cell types of donors of varying ages, suggesting that immune cell properties are specific to individuals. These results establish a novel high-throughput assay for the evaluation of DNA damage in large-scale studies.
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ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-022-10018-0