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Lookup NU author(s): Dr Ashwin Sachdeva, Dr Christopher CareyORCiD, Dr Amy VincentORCiD, Professor Laura GreavesORCiD, Professor Rakesh Heer, Emeritus Professor Doug Turnbull
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
Advances in multiplex immunofuorescence (mIF) and digital image analysis has enabled simultaneous assessment of protein defects in electron transport chain components. However, current manual methodology is time consuming and labour intensive. Therefore, we developed an automated highthroughput mIF workfow for quantitative single-cell level assessment of formalin fxed parafn embedded tissue (FFPE), leveraging tyramide signal amplifcation on a Ventana Ultra platform coupled with automated multispectral imaging on a Vectra 3 platform. Utilising this protocol, we assessed the mitochondrial oxidative phosphorylation (OXPHOS) protein alterations in a cohort of benign and malignant prostate samples. Mitochondrial OXPHOS plays a critical role in cell metabolism, and OXPHOS perturbation is implicated in carcinogenesis. Marked inter-patient, intra-patient and spatial cellular heterogeneity in OXPHOS protein abundance was observed. We noted frequent Complex IV loss in benign prostate tissue and Complex I loss in age matched prostate cancer tissues. Malignant regions within prostate cancer samples more frequently contained cells with low Complex I & IV and high mitochondrial mass in comparison to benign–adjacent regions. This methodology can now be applied more widely to study the frequency and distribution of OXPHOS alterations in formalin-fxed tissues, and their impact on long-term clinical outcomes.
Author(s): Sachdeva A, Hart CA, Carey CD, Vincent AE, Greaves LC, Heer R, Oliveira P, Brown MD, Clarke NW, Turnbull DM
Publication type: Article
Publication status: Published
Journal: Scientific Reports
Year: 2022
Volume: 12
Pages: 6660
Online publication date: 22/04/2022
Acceptance date: 01/04/2022
Date deposited: 03/05/2022
ISSN (electronic): 2045-2322
Publisher: Nature
URL: https://doi.org/10.1038/s41598-022-10588-z
DOI: 10.1038/s41598-022-10588-z
PubMed id: 35459777
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