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      High hypoxia status in pancreatic cancer is associated with multiple hallmarks of an immunosuppressive tumor microenvironment

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          Abstract

          Introduction

          Pancreatic ductal adenocarcinoma (PDAC), the most common form of pancreatic cancer, is a particularly lethal disease that is often diagnosed late and is refractory to most forms of treatment. Tumour hypoxia is a key hallmark of PDAC and is purported to contribute to multiple facets of disease progression such as treatment resistance, increased invasiveness, metabolic reprogramming, and immunosuppression.

          Methods

          We used the Buffa gene signature as a hypoxia score to profile transcriptomics datasets from PDAC cases. We performed cell-type deconvolution and gene expression profiling approaches to compare the immunological phenotypes of cases with low and high hypoxia scores. We further supported our findings by qPCR analyses in PDAC cell lines cultured in hypoxic conditions.

          Results

          First, we demonstrated that this hypoxia score is associated with increased tumour grade and reduced survival suggesting that this score is correlated to disease progression. Subsequently, we compared the immune phenotypes of cases with high versus low hypoxia score expression (Hypoxia HI vs. Hypoxia LOW) to show that high hypoxia is associated with reduced levels of T cells, NK cells and dendritic cells (DC), including the crucial cDC1 subset. Concomitantly, immune-related gene expression profiling revealed that compared to Hypoxia LOW tumours, mRNA levels for multiple immunosuppressive molecules were notably elevated in Hypoxia HI cases. Using a Random Forest machine learning approach for variable selection, we identified LGALS3 (Galectin-3) as the top gene associated with high hypoxia status and confirmed its expression in hypoxic PDAC cell lines.

          Discussion

          In summary, we demonstrated novel associations between hypoxia and multiple immunosuppressive mediators in PDAC, highlighting avenues for improving PDAC immunotherapy by targeting these immune molecules in combination with hypoxia-targeted drugs.

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          Most cited references92

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          Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

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            edgeR: a Bioconductor package for differential expression analysis of digital gene expression data

            Summary: It is expected that emerging digital gene expression (DGE) technologies will overtake microarray technologies in the near future for many functional genomics applications. One of the fundamental data analysis tasks, especially for gene expression studies, involves determining whether there is evidence that counts for a transcript or exon are significantly different across experimental conditions. edgeR is a Bioconductor software package for examining differential expression of replicated count data. An overdispersed Poisson model is used to account for both biological and technical variability. Empirical Bayes methods are used to moderate the degree of overdispersion across transcripts, improving the reliability of inference. The methodology can be used even with the most minimal levels of replication, provided at least one phenotype or experimental condition is replicated. The software may have other applications beyond sequencing data, such as proteome peptide count data. Availability: The package is freely available under the LGPL licence from the Bioconductor web site (http://bioconductor.org). Contact: mrobinson@wehi.edu.au
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              Robust enumeration of cell subsets from tissue expression profiles

              We introduce CIBERSORT, a method for characterizing cell composition of complex tissues from their gene expression profiles. When applied to enumeration of hematopoietic subsets in RNA mixtures from fresh, frozen, and fixed tissues, including solid tumors, CIBERSORT outperformed other methods with respect to noise, unknown mixture content, and closely related cell types. CIBERSORT should enable large-scale analysis of RNA mixtures for cellular biomarkers and therapeutic targets (http://cibersort.stanford.edu).
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                Author and article information

                Contributors
                URI : https://loop.frontiersin.org/people/486152Role: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/1673738Role: Role: Role: Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/2613419Role: Role: Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/350487Role: Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/56767Role: Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/2650647Role: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role:
                Journal
                Front Immunol
                Front Immunol
                Front. Immunol.
                Frontiers in Immunology
                Frontiers Media S.A.
                1664-3224
                06 March 2024
                2024
                : 15
                : 1360629
                Affiliations
                [1] 1 Centre for Health and Life Sciences, Coventry University , Coventry, United Kingdom
                [2] 2 Institute of Cancer and Genomic Sciences, University of Birmingham , Birmingham, United Kingdom
                [3] 3 Independent Scholar, National Coalition of Independent Scholars , Visp, Switzerland
                [4] 4 Institute of Medical Sciences, University of Toronto , Toronto, ON, Canada
                Author notes

                Edited by: Dmitry Aleksandrovich Zinovkin, Gomel State Medical University, Belarus

                Reviewed by: Eldar Nadyrov, Gomel State Medical University, Belarus

                Jale Yuzugulen, Eastern Mediterranean University, Türkiye

                *Correspondence: Hassan Sadozai, sadozaih@ 123456uni.coventry.ac.uk ; Animesh Acharjee, a.acharjee@ 123456bham.ac.uk ; Bernard Burke, ac2561@ 123456coventry.ac.uk

                †These authors have contributed equally to this work

                Article
                10.3389/fimmu.2024.1360629
                10951397
                38510243
                b359fc5e-e423-4eb8-b6eb-2b6c2ba4ccd1
                Copyright © 2024 Sadozai, Acharjee, Kayani, Gruber, Gorczynski and Burke

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 23 December 2023
                : 12 February 2024
                Page count
                Figures: 8, Tables: 0, Equations: 0, References: 92, Pages: 14, Words: 7400
                Funding
                The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. HS and BB have received funding from the Research Excellence Development Fund at Coventry University. AA acknowledges support from the, HYPERMARKER (Grant agreement ID 101095480), and the MRC Health Data Research UK (HDRUK/CFC/01) and HDRUK midlands regional community project [QQ2], initiatives funded by UK Research and Innovation, Department of Health and Social Care (England) and the devolved administrations, and leading medical research charities. The views expressed in this publication are those of the authors and not necessarily those of the NHS, the National Institute for Health Research, the Medical Research Council or the Department of Health.
                Categories
                Immunology
                Original Research
                Custom metadata
                Cancer Immunity and Immunotherapy

                Immunology
                hypoxia,tumor microenvironment (tme),pancreatic ductal adenocarcinoma (pdac),immune checkpoint,galectins

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