Project description:Myeloid-derived suppressor cells (MDSCs) are highly immunosuppressive myeloid cells, which increase in cancer patients. The molecular mechanism behind their generation and function is unclear. Whereas granulocytic-MDSCs correlate with poor overall survival in breast cancer, the presence and relevance of monocytic-MDSCs (Mo-MDSCs) is unknown. Here we report for the first time an enrichment of functional blood Mo-MDSCs in breast cancer patients before they acquire a typical Mo-MDSC surface phenotype. A clear population of Mo-MDSCs with the typical cell surface phenotype (CD14+HLA-DRlow/-Co-receptorlow/-) increased significantly first during disease progression and correlated to metastasis to lymph nodes and visceral organs. Furthermore, monocytes, comprising the Mo-MDSC population, from patients with metastatic breast cancer resemble the reprogrammed immunosuppressive monocytes in patients with severe infections, both by their surface and functional phenotype but also at their molecular gene expression profile. Our data suggest that monitoring the Mo-MDSC levels in breast cancer patients may represent a novel and simple biomarker for assessing disease progression. Peripheral blood monocytes were isolated using magnetic cell sorting from 4 patients with metastatic breast cancer, 3 healthy controls, 3 patients with sepsis and 3 patients with active tuberculosis were immediately frozen at -80C in TRIZOL.
Project description:The goal of this study was to determine if blood circulating monocytes of metastatic breast cancer patient would express a different activation profile compared to healthy donors, in order to use this specific changesas biomarkers to monitor then response to therapy
Project description:Myelomonocytic cells (i.e., monocytes or macrophages) play a key role in tumor progression as revealed by numerous mice tumor model studies. However, their contribution in human tumor progression is not well-studied. Using Renal Cell Carcinoma (RCC) as a model for human cancer, we performed a transcriptomal profiling of blood monocytes from RCC patients to investigate the contribution of these cells in cancer progression. As compared to monocytes from healthy donors (Mo), transcriptome analysis of monocytes from RCC patients (RCC-Mo) showed a distinct gene expression signature consisting of cytokines, chemokines and various inflammation-related genes. Validation of these genes by qPCR as well as other functional assays indicated RCC-Mo possess an inflammatory and tumor promoting phenotype. Blood monocytes from RCC patients (with early or metastatic disease) or healthy donors (controls) were isolated, and then differential gene expression was detected with Limma.
Project description:Differences in the activity of monocytes/macrophages, important target cells of Mycobacterium tuberculosis, might influence tuberculosis progression. With the purpose of identifying candidate genes for tuberculosis susceptibility we infected with M. tuberculosis monocytes from both, healthy elders (a tuberculosis susceptibility group) and elderly tuberculosis patients, and performed a microarray experiment. We detected 78 differentially expressed transcripts and confirmed these results by quantitative PCR of selected genes. We found that monocytes from tuberculosis patients showed similar expression patterns of these genes regardless of whether they were obtained from younger or elder patients. Only one of the detected genes corresponded to a cytokine: IL-26, a member of the IL-10 cytokine family that we found downregulated in infected monocytes from tuberculosis patients. We have analyzed total RNA from Mycobacterium tuberculosis infected monocytes. We have isolated CD14+ cells (monocytes) from peripheral blood mononuclear cells by magnetic separation, and infected them for 4 days with 1 bacterium per monocyte. Blood donors were 7 elderly patients with pulmonary tuberculosis (average age: 83 years; sex: 3 men and 4 women) and 8 non-tuberculous volunteers (81 years, 6 men and 2 women).
Project description:Analysis of tumor-educated changes in peripheral blood monocytes at the gene expression level. We analyzed if gene expression in monocytes of patients with colorectal cancer is differential from those of healthy volunteers and found a diagnostic signature that allowed to accurately discriminate patients with colorectal cancer from healthy individuals. Peripheral blood monocytes from 93 distinct individuals are profiled on 8 beadchips. The individuals belong to one of three groups: healthy volunteers (38), patients with non-metastatic colorectal cancer (27) or patients with metastatic colorectal cancer (28).
Project description:Several single-cell studies have reported transcriptomic, genomic and epigenomic dysregulation in breast cancer tissue. However, the peripheral immune landscape of breast cancer patients remains poorly understood. We report the analysis of the single-cell transcriptomes of more than 110,000 peripheral blood mononuclear cells (PBMCs) of healthy and metastatic breast cancer patients. Integrated analysis revealed depleted memory B cells (BMEM) and T cell subpopulations, including transitional CD8 T, CD4 naïve, CD4 ribosome and MAIT cells in the circulating immune landscape of metastatic breast cancer patients. Nevertheless, metastatic breast cancer patients showed marked enrichment of pro-inflammatory NKG7 high monocytes, M1 macrophages, migratory myeloid, T memory stem cells (TSCM), CD4 T effector memory (TEM) and CD4 T central memory (TCM) cells. Further, interrogation of single-cell transcriptomes based on metastatic disease state, i.e. stable vs progressive, revealed distinct compositional and transcriptional changes in PBMCs correlated with the metastatic burden. The antigen inexperienced CD4 naïve T cells and cytotoxic MAIT cells are further depleted in the progressive metastatic stage. Surprisingly, the pro-inflammatory classical monocytes and CD4 T effector memory (TEM) cells are also depleted in the progressive metastatic state, indicating their potential role in stable metastatic disease. Lastly, the receptor-ligand relationships, such as cell-cell contact, ECM-receptor interactions, and cytokine transcription profiles, were tightly associated with metastatic burden. Collectively, our study provides unique molecular insight into the peripheral immune system operating in metastatic breast cancers and identified novel surrogate biomarkers of metastatic disease.
Project description:Cancer dysregulates intratumoral adaptive-innate immune cell crosstalk, however, it remains largely unknown how the systemic immune landscape is modified during breast cancer progression and whether prior chemotherapy treatment exerts persistent changes on circulating immune cells. Here, we comprehensively profiled the systemic immune landscape in patients with triple negative breast cancer (TNBC) at distinct disease stages, to understand how cancer progression and treatment history shape the systemic immune landscape. We performed multi-parameter flow cytometry analysis to assess the global systemic immune landscape, including often overlooked granulocytes. We showed that patients with metastatic TNBC (mTNBC) exhibited increased classical monocytes and neutrophils compared to healthy donors (HDs) irrespective of prior therapy. Conversely, circulating T cells, dendritic cell subsets and differentiated B cells were reduced in patients with mTNBC compared to HDs, which could partially be attributed to prior chemotherapy treatment. Phenotypic and functional characterization of the T cell compartment revealed increased IL17 production by vδ1 γδ T cells, were increased. Transcriptional and proteomic analysis, alongside ex vivo functionality assays, revealed increased migratory capacity, increased granule proteins, and elevated ROS production in circulating neutrophils of mTNBC patients. Our data demonstrate that patient neutrophils are not only more abundant, but also display an altered functionality, underscoring the significant impact of TNBC disease stage and treatment history on the systemic immune composition and function.
Project description:Differences in the activity of monocytes/macrophages, important target cells of Mycobacterium tuberculosis, might influence tuberculosis progression. With the purpose of identifying candidate genes for tuberculosis susceptibility we infected with M. tuberculosis monocytes from both, healthy elders (a tuberculosis susceptibility group) and elderly tuberculosis patients, and performed a microarray experiment. We detected 78 differentially expressed transcripts and confirmed these results by quantitative PCR of selected genes. We found that monocytes from tuberculosis patients showed similar expression patterns of these genes regardless of whether they were obtained from younger or elder patients. Only one of the detected genes corresponded to a cytokine: IL-26, a member of the IL-10 cytokine family that we found downregulated in infected monocytes from tuberculosis patients.
Project description:Metastatic behaviour varies significantly among breast cancers. Mechanisms explaining why the majority of breast cancer patients never develop metastatic outgrowth are largely lacking but could underlie the development of novel immunotherapeutic target molecules. Here we show interplay between non-metastatic primary breast cancer and innate immune response, acting together to control metastatic progression. The primary tumor systemically recruits IFNγ-producing immune effector monocytes to the lung. IFNγ upregulates Tmem173/STING in neutrophils and enhances their killing capacity. The immune effector monocytes and tumoricidal Tmem173/STINGhigh neutrophils target disseminated tumor cells in the lungs, preventing metastatic outgrowth. Importantly, our findings could underlie the development of immunotherapeutic target molecules that augment the function of immune effector monocytes and Tmem173high neutrophils.
Project description:Human peripheral monocytes have been categorized into three subsets based on differential expression levels of CD14 and CD16. However, the factors that influence the distribution of monocyte subsets and the roles which each subset plays in autoimmunity are not well studied. To compare the gene expression profiling 1) on intermediate monocytes CD14++CD16+ monocytes between healthy donors and autoimmune uveitis patients and 2) among 3 monocyte subsets in health donors, here we purified circulating intermediate CD14++CD16+ monocytes from 5 patients with autoimmune uveitis (labeled as P1-5) and 4 healthy donors (labeled as HD1-4) by flow cytometry and isolated total RNA to proceed microarray assay. In addition, we also purified CD14+CD16++ (non-classical monocytes) and CD14++CD16- (classical monocytes) from 4 healthy donors to do microarray. We demonstrate that CD14++CD16+ monocytes from patients and healthy control donors share a similar gene expression profile. The CD14+CD16++ cells (non-classical monocytes) display the most distinctive gene expression profiling when compared to intermediate CD14++CD16+ monocytes and classical CD14++CD16- monocytes.