Biology subjects

Wiesheu, R.

Publications and source records attributed to Wiesheu, R..

8 recordsLinked to original sources

bioRxiv2024

Uridine Phosphorylase-1 supports metastasis of mammary cancer by altering immune and extracellular matrix landscapes of the lung

Understanding the mechanisms that facilitate early events in metastatic seeding is key to developing therapeutic approaches to reduce metastasis - the leading cause of cancer-related death. Using whole animal screens in genetically engineered mouse models of cancer we have identified circulating metabolites associated with metastasis. Specifically, we highlight the pyrimidine uracil as a prominent metastasis-associated metabolite. Uracil is generated by neutrophils expressing the enzyme uridine phosphorylase-1 (UPP1), and neutrophil specific Upp1 expression is increased in cancer. Altered UPP1 activity influences expression of adhesion molecules on the surface of neutrophils, leading to decreased neutrophil motility in the pre-metastatic lung. Furthermore, we find that UPP1-expressing neutrophils suppress T-cell proliferation, and the UPP1 product uracil can increase fibronectin deposition in the extracellular microenvironment. Consistently, knockout or inhibition of UPP1 in mice with mammary tumours increases the number of T-cells and reduces fibronectin content in the lung and decreases the proportion of mice that develop lung metastasis. These data indicate that UPP1 influences neutrophil behaviour and extracellular matrix deposition in the lung and suggest that pharmacological targeting of this pathway could be an effective strategy to reduce metastasis.

cancer biology↗

bioRxiv2024

Integrin inactivation slows down neutrophils congesting the pre-metastatic lung in a model of breast cancer

Neutrophils are thought to be critical to the process whereby breast cancers establish an immunosuppressive and tumour cell nurturing pre-metastatic niche before overt metastasis can be detected. However, the spatial localization of neutrophils and their interaction with other cell types in the lung pre-metastatic niche is not well described. We used a spontaneously metastatic mammary cancer model combined with a multiplexed three- and four-dimensional imaging approach to investigate the behaviour of neutrophils in the pre-metastatic niche. Volume fixed tissue three-dimensional imaging showed that approximately 40% of CD8+ T cells are adjacent to neutrophils at this stage. In live tissue, we found neutrophils with impaired intravascular motility congested the capillaries of pre-metastatic lungs potentially obstructing CD8+ T cells. Slowed neutrophil transit was dependent on the conformation of {beta}2-integrin and could be recapitulated by treating non-tumour bearing mice with G-CSF, a potent systemic mediator of granulopoiesis. We found a decrease in L-selectin (CD62L) on neutrophils in the lungs of both mammary tumour bearing and G-CSF treated mice. Finally, we observed differential accumulation of intravenously injected micro-beads in the lung, suggestive of transient circulatory dead spaces which were also dependent on {beta}2-integrin inactivation. Overall, our study proposes that integrin-mediated neutrophil congestion of the alveolar capillaries could contribute to the generation of the pulmonary pre-metastatic niche.

immunology↗

bioRxiv2023

IL-27 maintains cytotoxic Ly6C+ gamma delta T cells that arise from immature precursors

In mice, {gamma}{delta} T cells that express the co-stimulatory molecule, CD27, are committed to the IFN{gamma}-producing lineage in the thymus, and in the periphery, these cells play a critical role in host defence and anti-tumor immunity. Unlike {beta} T cells that rely on MHC-presented peptides to drive their terminal differentiation, it is unclear whether MHC-unrestricted {gamma}{delta} T cells undergo further functional maturation after exiting the thymus. Here, we provide evidence of phenotypic and functional diversity within peripheral IFN{gamma}-producing {gamma}{delta} T cells. We found that immature CD27+Ly6C-- cells convert into mature CD27+Ly6C+ cells, and these mature cells control cancer progression while the immature cells cannot. The gene signatures of these two subsets were highly analogous to human immature and mature {gamma}{delta} T cells, indicative of conservation across species. We show that IL-27 supports the cytotoxic phenotype and function of mouse CD27+Ly6C+ cells and human V{delta}2+ cells, while IL-27 is dispensable for mouse CD27+Ly6C-- cells and human V{delta}1+ cells. These data reveal increased complexity within IFN{gamma}-producing {gamma}{delta} T cells, comprising of immature and terminally differentiated subsets, that offer new insights into unconventional T cell biology.

immunology↗

bioRxiv2023

γδ T cells impair airway macrophage differentiation in lung adenocarcinoma

Protecting mucosal barriers, {gamma}{delta} T cells hold promise for the development of new cancer immunotherapies. In mice, {gamma}{delta} T cells can largely be segregated into CD27+ and CD27- cells, and their functions are modulated by interactions with surrounding cells. However, which cells communicate directly with {gamma}{delta} T cells in lung adenocarcinoma remains unknown. To address this, we combined flow cytometry, confocal microscopy, and scRNA-seq, using an autochthonous genetically engineered mouse model and different {gamma}{delta} T cell-deficient settings. We found that {gamma}{delta} T cells were increased in tumour-bearing lungs, with an altered phenotype. CD27- and CD27+ {gamma}{delta} T cells differed in their localisation and interactions including their tropism for macrophages. Overall, we propose a model where CD27+ {gamma}{delta} T cells undermine the differentiation of tumour-associated macrophages into airway macrophages, fostering a negative outcome in lung adenocarcinoma. Determining its translatability to human health may offer new avenues for immunotherapeutic strategies. SummaryGuardians of pulmonary homeostasis, {gamma}{delta} T cells remain enigmatic regarding their role in lung adenocarcinoma. Raffo-Iraolagoitia et al. report that a subset of {gamma}{delta} T cells impairs the differentiation of tumour-associated macrophages into airway macrophages, relevant for the outcome of lung adenocarcinoma. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=111 SRC="FIGDIR/small/557344v1_ufig1.gif" ALT="Figure 1"> View larger version (49K): org.highwire.dtl.DTLVardef@e679f7org.highwire.dtl.DTLVardef@15152dforg.highwire.dtl.DTLVardef@17eaa5org.highwire.dtl.DTLVardef@2e0e47_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2022

β-catenin obstructs γδ T cell immunosurveillance in colon cancer through loss of BTNL expression

WNT/{beta}-catenin signaling endows cancer cells with proliferative capacity and immune-evasive functions that impair anti-cancer immunosurveillance by conventional, cytoxtoic T cells. However, the impact of dysregulated WNT signalling on unconventional, tissue-resident T cells, specifically in colon cancer is unknown. Here, we show that cancer cells in Apc-mutant mouse models escape immunosurveillance from gut-resident intraepithelial lymphocytes (IELs) expressing {gamma}{delta} T cell receptors ({gamma}{delta}TCRs). Analysis of late-stage tumors from mice and humans revealed that {gamma}{delta}IELs are largely absent from the tumor microenvironment, and that butyrophilin-like (BTNL) molecules, which can critically regulate {gamma}{delta}IEL through direct {gamma}{delta}TCR-interactions, are also downregulated. We could attribute this to {beta}-catenin stabilization, which rapidly decreased expression of the transcription factors, HNF4A and HNF4G, that we found to bind promoter regions of Btnl genes, thereby driving their expression in normal gut epithelial cells. Indeed, inhibition of {beta}-catenin signaling restored Btnl1 gene expression and {gamma}{delta} T cell infiltration into tumors. These observations highlight an immune-evasion mechanism specific to WNT-driven colon cancer cells that disrupts {gamma}{delta}IEL immunosurveillance and furthers cancer progression.

cancer biology↗

bioRxiv2021

Single-cell analysis uncovers differential regulation of lung γδ T cell subsets by the co-inhibitory molecules, PD-1 and TIM-3

IL-17A-producing {gamma}{delta} T cells within the lung consist of both V{gamma}6+ tissue-resident cells and V{gamma}4+ circulating cells that play important roles in homeostasis, inflammation, infection, tumor progression and metastasis. How these {gamma}{delta} T cell subsets are regulated in the lung environment during homeostasis and cancer remains poorly understood. Using single-cell RNA sequencing and flow cytometry, we show that lung V{gamma}6+ cells express a repertoire of cell surface molecules distinctive from V{gamma}4+ cells, including PD-1 and ICOS. We found that PD-1 functions as a co-inhibitory molecule on V{gamma}6+ cells to reduce IL-17A production, whereas manipulation of ICOS signaling fails to affect IL-17A in V{gamma}6+ cells. In a mammary tumor model, ICOS and PD-1 expression on lung V{gamma}6+ cells remained stable. However, V{gamma}6+ and V{gamma}4+ cells within the lung pre-metastatic niche increased expression of IL-17A, IL-17F, amphiregulin (AREG) and TIM-3 in response to tumor-derived IL-1{beta} and IL-23, where the upregulation of TIM-3 was specific to V{gamma}4+ cells. Inhibition of either PD-1 or TIM-3 in mammary tumor-bearing mice further increased IL-17A by V{gamma}6+ and V{gamma}4+ cells, indicating that both PD-1 and TIM-3 function as negative regulators of IL-17A-producing {gamma}{delta} T cell subsets. Together, these data demonstrate how lung {gamma}{delta} T cell subsets are differentially controlled by co-inhibitory molecules in steady-state and cancer.

immunology↗

bioRxiv2020

Ly6C defines a subset of memory-like CD27+ γδ T cells with inducible cancer-killing function

In mice, IFN{gamma}-producing {gamma}{delta} T cells that express the co-stimulatory molecule, CD27, play a critical role in host defence and anti-tumour immunity. However, their phenotypic diversity, composition in peripheral and secondary lymphoid organs, similarity to {beta} T cells as well as homology with human {gamma}{delta} T cells is poorly understood. Here, using single cell RNA sequencing, we show that CD27+ {gamma}{delta} T cells consist of two major clusters, which are distinguished by expression of Ly6C. We demonstrate that CD27+Ly6C-- {gamma}{delta} T cells exhibit a naive T cell-like phenotype, whereas CD27+Ly6C+ {gamma}{delta} T cells display a memory-like phenotype, produce several NK cell-related and cytotoxic molecules and are highly similar to both mouse CD8+ T cells and mature human {gamma}{delta} T cells. In a breast cancer mouse model, depletion of CD27+ {gamma}{delta} T cells failed to affect tumour growth, but these cells could be coerced into killing cancer cells after expansion ex vivo. These results identify novel subsets of {gamma}{delta} T cells in mice that are comparable to human {gamma}{delta} T cells, opening new opportunities for {gamma}{delta} T cell-based cancer immunotherapy research.

immunology↗

bioRxiv2020

Impact of formate supplementation on body weight and plasma amino acids

Current nutritional recommendations are focused on energy, fat, carbohydrate, protein and vitamins. Less attention has been paid to the nutritional demand of one-carbon units for nucleotide and methionine synthesis. Here we investigate the impact of sodium formate supplementation as a nutritional intervention to increase the dietary intake of one-carbon units. A cohort of six female and six male mice received 125 mM sodium formate in the drinking water for three months. A control group of another six female and six male mice was also followed up for the same period of time. Tail vein blood samples were collected once a month and profiled with a haematology Analyser. At the end of the study blood and tissues were collected for metabolomics analysis and immune cell sorting. Formate supplementation has no significant physiological effect on male mice. Formate supplementation has no significant effect on the immune cell counts during the intervention or at the end of the study in either gender. In female mice however, the body weight and spleen wet-weight were significantly increased by formate supplementation, while the blood plasma levels of amino acids were decreased. Formate supplementation also increased the frequency of probiotic bacteria in the stools of female mice. We conclude that formate supplementation induces physiological changes in female mice.

biochemistry↗