Immune Cell Components Research Articles

A disulfidptosis-related glucose metabolism and immune response prognostic model revealing the immune microenvironment in lung adenocarcinoma.

Lung adenocarcinoma accounts for the majority of lung cancer cases and impact survival rate of patients severely. Immunotherapy is an effective treatment for lung adenocarcinoma but is restricted by many factors including immune checkpoint expression and the inhibitory immune microenvironment. This study aimed to explore the immune microenvironment in lung adenocarcinoma via disulfidptosis. Public datasets of lung adenocarcinoma from the TCGA and GEO was adopted as the training and validation cohort. Based on the differences in the expression of disulfidptosis -related genes, a glucose metabolism and immune response prognostic model was constructed. The prognostic value and clinical relationship of the model were further explored. Immune-related analyses were performed according to CIBERSORT, ssGSEA, TIDE, IPS. We verified that the model could accurately predict the survival expectancy of lung adenocarcinoma patients. Patients with lung adenocarcinoma and a low-risk score had better survival outcomes according to the model. Moreover, the high-risk group tended to have an immunosuppressive effect, as reflected by the immune cell components, phenotypes and functions. We also found that the clinically relevant immune checkpoint CTLA-4 was significantly higher in low-risk group (P<0.05), indicating that the high-risk group may suffer worse tumor immunotherapy efficacy. Finally, we found that this model has accurate predictive value for the efficacy of immune checkpoint blockade in non-small cell lung cancer (P<0.05). The prognostic model demonstrated the feasibility of predicting survival and immunotherapy efficacy via disulfidptosis-related genes and will facilitate the development of personalized anticancer therapy.

Identification of shared mechanisms and targets between immune checkpoint inhibitor-associated myocarditis and autoimmune myocarditis

Objective This study aimed to explore the shared mechanisms and targets between immune checkpoint inhibitor-associated myocarditis (ICIM) and autoimmune myocarditis. Methods Relevant data were retrieved from public datasets and Gene Expression Omnibus (GEO) database. Gene set enrichment analysis (GSEA) of differentially expressed genes (DEGs) was used to identify significant shared signaling pathways between ICIM and non-ICI associated autoimmune myocarditis (NICIAM) represented by ICIM model and experimental autoimmune myocarditis (EAM) model, respectively. Cell type enrichment analysis and immune infiltration analysis by clusterProfiler and ImmuCellAI were performed to identify critical immune cell component involved in ICIM and NICIAM. Additionally, core shared genes across ICIM and NICIAM were identified and validated by various models and methods. Results Interferon-γ response, inflammatory response and allograft rejection signaling were identified as the shared signaling pathways associated with ICIM and NICIAM. Enrichment analysis of cell type supported an important role of increased infiltration of T cells and macrophages in both ICIM and NICIAM. However, the predominant increase of infiltrated T cells was CD4+ T cells in NICIAM, while that were CD8+ T cells in ICIM. Core shared genes Lck and Cd3d expression were found increased in both ICIM and NICIAM, and Lck inhibition was further identified and validated as potential therapeutic approach. Conclusions Our study initially established a comorbidity model to identify potential molecular mechanism including interferon-γ response, inflammatory response and allograft rejection signaling accounting for the concerns of myocarditis risk in patients with preexisting autoimmune disease (PAD) receiving ICI treatment, and supported the therapeutic potential of targeting Lck or Cd3d.

Hepatocellular carcinoma cells induce γδ T cells through metabolic reprogramming into tumor-progressive subpopulation.

Tumor immune microenvironment (TIME) is a tiny structure that contains multiple immune cell components around tumor cells, which plays an important role in tumorigenesis, and is also the potential core area of activated immunotherapy. How immune cells with tumor-killing capacity in TIME are hijacked by tumor cells during the progression of tumorigenesis and transformed into subpopulations that facilitate cancer advancement is a question that needs to be urgently addressed nowadays. γδ T cells (their T cell receptors are composed of γ and δ chains), a unique T cell subpopulation distinguished from conventional αβ T cells, are involved in a variety of immune response processes through direct tumor-killing effects and/or indirectly influencing the activity of other immune cells. However, the presence of γδ T cells in the tumor microenvironment (TME) has been reported to be associated with poor prognosis in some tumors, suggesting that certain γδ T cell subsets may also have pro-tumorigenic effects. Recent studies have revealed that metabolic pathways such as activation of glycolysis, increase of lipid metabolism, enhancement of mitochondrial biosynthesis, alterations of fatty acid metabolism reshape the local TME, and immune cells trigger metabolic adaptation through metabolic reprogramming to meet their own needs and play the role of anti-tumor or immunosuppression. Combining previous studies and our bioinformatics results, we hypothesize that γδT cells compete for resources with hepatocellular carcinoma (HCC) cells by means of fatty acid metabolic regulation in the TME, which results in the weakening or loss of their ability to recognize and kill HCC cells through genetic and epigenetic alterations, thus allowing γδT cells to be hijacked by HCC cells as a subpopulation that promotes HCC progression.

Macrophage-Biomimetic Nanoplatform-Based Therapy for Inflammation-Associated Diseases.

Inflammation-associated diseases are very common clinically with a high incidence; however, there is still a lack of effective treatments. Cell-biomimetic nanoplatforms have led to many breakthroughs in the field of biomedicine, significantly improving the efficiency of drug delivery and its therapeutic implications especially for inflammation-associated diseases. Macrophages are an important component of immune cells and play a critical role in the occurrence and progression of inflammation-associated diseases while simultaneously maintaining homeostasis and modulating immune responses. Therefore, macrophage-biomimetic nanoplatforms not only inherit the functions of macrophages including the inflammation tropism effect for targeted delivery of drugs and the neutralization effect of pro-inflammatory cytokines and toxins via membrane surface receptors or proteins, but also maintain the functions of the inner nanoparticles. Macrophage-biomimetic nanoplatforms are shown to have remarkable therapeutic efficacy and excellent application potential in inflammation-associated diseases. In this review, inflammation-associated diseases, the physiological functions of macrophages, and the classification and construction of macrophage-biomimetic nanoplatforms are first introduced. Next, the latest applications of different macrophage-biomimetic nanoplatforms for the treatment of inflammation-associated diseases are summarized. Finally, challenges and opportunities for future biomedical applications are discussed. It is hoped that the review will provide new ideas for the further development of macrophage-biomimetic nanoplatforms.

Ex Vivo Mathematical Myeloma Advisor (EMMA) - a Clinical, Molecular, and Phenotypic Platform to Tailor Personalized Therapeutic Strategies for Multiple Myeloma

Multiple myeloma (MM) is a genetically heterogeneous disease, lacking molecular markers for patient stratification and prediction of clinical response. We have created a unique platform for clinical-informed decisions, research advancement, and preclinical compound testing called the “Ex Vivo Mathematical Myeloma Advisor” (EMMA). In this pipeline, MM patients who undergo bone marrow (BM) biopsy and consent to participate in the Total Cancer Care protocol have their clinical data (demographics, therapies received, lab results, clinical history, etc.) abstracted, along with the molecular characterization (FISH/cytogenetics, RNA-Seq, and WES) of their disease. In parallel, fresh BM-derived MM blasts are combined with stromal cells in a collagen-based matrix containing patient plasma and used in an ex vivo drug screening assay. The cells are seeded into 384 or 1536-well plates and exposed to serial dilutions of up to 127 single agents and combinations. The entire setup is imaged in bright field every 30 minutes for a duration of 6 days. Image analysis is used to convert membrane integrity into survival curves, and AUC and LD50 values are estimated. The pattern of these curves facilitates the identification of resistant subclones. This data is further combined with augmented synergy models that are parameterized with PK/PD data for each drug. This integration permits the simulation of “in silico clinical trials”, which are compiled into reports that assist clinicians in selecting the best treatment for their patients. In addition, the immune population of the BM aspirate is characterized by flow cytometry, and soluble factors in the patient's BM plasma are quantified by ELISA. The integration of clinical, molecular, and phenotypic data with image analysis and machine learning tools presents a unique opportunity for finding molecular signatures of drug response (Fig. 1). For instance, our pipeline can be utilized to identify clinical factors (such as ethnicity, age, etc.) and molecular features (mutations, cytogenetic abnormalities, enriched transcription factors, differentially expressed biological pathways, etc.) associated with drug resistance/sensitivity. Additionally, it enables the determination of the mechanisms of action of new compounds and the prediction of potential synergistic/antagonistic drug combinations. The longitudinal acquisition of data in our non-destructive ex vivo drug screen assay facilitates the exploration of drug interaction dynamics over time, enabling the pinpointing of the moment when synergism occurs at maximum intensity. This, in turn, allows us to propose optimized temporal-based therapeutic regimens for clinical use. Up to this point, our pipeline has molecularly characterized nearly 1,000 MM samples, and EMMA has been applied to over 600 samples, forming one of the world's most extensive and comprehensively characterized MM cohorts. While EMMA has proven effective in quantifying the activity of monoclonal antibodies, ongoing efforts are focusing on incorporating individual immune cell components to assess ex vivo activity in the context of evolving cellular immunotherapies. As a proof of principle, we demonstrate the system's capability to measure the effect of daratumumab (CD38+ antibody) on autologous macrophage-mediated MM cell death, thereby distinguishing it from apoptosis caused by direct antibody binding (Fig. 2). Additionally, we evaluate the influence of teclistamab (T-cell engager CD3/BCMA bispecific antibody) on CD8+ T cell activity against autologous MM cells, as well as the efficacy of CAR-T cells in killing MM cells. Through the testing of standard-of-care drugs alongside immune agents, our pipeline has identified the combination of bortezomib + daratumumab as a synergistic approach with significant translational potential, warranting further investigation. In summary, our state-of-the-art platform facilitate drug discovery and unravel the intricate biological phenomena underlying drug interactions in MM. The advantages offered by our system are threefold: i) it enables pharmaceutical companies to test new compounds in a bona fide, patient-derived model; ii) it assists clinicians in selecting the best therapy for their patients, and iii) it helps prevent cancer patients from receiving non-suitable therapies. This approach can also be promptly applied to other liquid cancers, such as leukemia and lymphoma.

Experimental verification and identifying biomarkers related to insomnia

IntroductionInsomnia is the most common form of sleep deprivation (SD) observed in clinics. Although there are differences between insomnia and SD, they have similar symptoms and the same animal model. Currently, there is a lack of microarray data on insomnia. Therefore, for now, we are going to apply the SD data to insomnia. Although many studies have explained the possible mechanisms associated with insomnia, no previous studies have considered the key genes associated with insomnia or the relationship between insomnia and immune cells. In this study, we analyzed the relationship between key genes and immune cells by identifying biomarkers for the diagnosis of insomnia. Next, we verified the efficacy of these biomarkers experimentally.MethodsFirst, we downloaded four microarrays (GSE11755, GSE12624, GSE28750, and GSE48080) from the Gene Expression Omnibus (GEO) database, which included data from 239 normal human blood samples and 365 blood specimens from patients with SD. Then, we analyzed two groups of differentially expressed genes (DEGs) and used Support Vector Machine Recursive Feature Elimination (SVM-RFE) analysis and the Least Absolute Shrinkage and Selection Operator (LASSO) regression model to investigate these key genes. Next, we used CIBERSORT to investigate the composition of 22 immune cell components of key genes in SD patients. Finally, the expression levels of key biomarkers in sleep-deprived patients were examined by quantitative real-time polymerase chain reaction (qRT-PCR).ResultsA total of 50 DEGs were identified: six genes were significantly upregulated, and 44 genes were significantly downregulated. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that Salmonella infection, NOD-like receptor (NLR) signaling pathway, Kaposi sarcoma-associated herpesvirus infection, and Th17 cell differentiation were significant. Based on machine learning, we identified C2CD2L, SPINT2, APOL3, PKNOX1, and A2M as key genes for SD; these were confirmed by receiver operating characteristic (ROC) analysis. Immune cell infiltration analysis showed that C2CD2L, SPINT2, APOL3, PKNOX1, and A2M were related in different degrees to regulatory T cells (Tregs), follicular T helper cells, CD8 cells, and other immune cells. The qRT-PCR experiments confirmed that the expression levels of C2CD2L concurred with the results derived from machine learning, but PKNOX1 and APOL3 did not.DiscussionIn summary, we identified a key gene (C2CD2L) that may facilitate the development of biomarkers for insomnia.

Dual effects of radiotherapy on tumor microenvironment and its contribution towards the development of resistance to immunotherapy in gastrointestinal and thoracic cancers.

Successful clinical methods for tumor elimination include a combination of surgical resection, radiotherapy, and chemotherapy. Radiotherapy is one of the crucial components of the cancer treatment regimens which allow to extend patient life expectancy. Current cutting-edge radiotherapy research is focused on the identification of methods that should increase cancer cell sensitivity to radiation and activate anti-cancer immunity mechanisms. Radiation treatment activates various cells of the tumor microenvironment (TME) and impacts tumor growth, angiogenesis, and anti-cancer immunity. Radiotherapy was shown to regulate signaling and anti-cancer functions of various TME immune and vasculature cell components, including tumor-associated macrophages, dendritic cells, endothelial cells, cancer-associated fibroblasts (CAFs), natural killers, and other T cell subsets. Dual effects of radiation, including metastasis-promoting effects and activation of oxidative stress, have been detected, suggesting that radiotherapy triggers heterogeneous targets. In this review, we critically discuss the activation of TME and angiogenesis during radiotherapy which is used to strengthen the effects of novel immunotherapy. Intracellular, genetic, and epigenetic mechanisms of signaling and clinical manipulations of immune responses and oxidative stress by radiotherapy are accented. Current findings indicate that radiotherapy should be considered as a supporting instrument for immunotherapy to limit the cancer-promoting effects of TME. To increase cancer-free survival rates, it is recommended to combine personalized radiation therapy methods with TME-targeting drugs, including immune checkpoint inhibitors.

Investigating the prognostic value of constructing disulfidptosis-related gene models for lung adenocarcinoma patients.

Disulfidptosis is a novel mode of cell death, a programmed mode of intracellular disulfide accumulation due to solute carrier family 7 member 11 (SLC7A11)-mediated abnormalities in the cell membrane cystine transport system. Numerous studies have confirmed the prominent role played by SLC7A11 in tumors, but the involvement of SLC7A11 as an important mediator of disulfidptosis in the death process of lung adenocarcinoma cells remains unclear. We obtained 4,107 SLC7A11-related genes and analyzed them using a total of 1,040 lung adenocarcinoma transcriptome sequencing data from The Cancer Genome Atlas (TCGA) cohort and GEO (Gene Expression Omnibus) cohort and 991 relevant clinical data. First, we screened for differential genes and identified molecular subtypes for assessing characteristic differences between lung adenocarcinoma subtypes under the influence of SLC7A11-associated genes. Then, risk score models were constructed to assess the prognosis, immune infiltration, tumor microenvironment, and drug treatment effects in lung adenocarcinoma patients. Finally, we also analyzed the distribution of cell types and expression of characteristic genes within the tumor using a single-cell database. In addition, relevant drug sensitivities were predicted. We screened 956 genes with significant differences and identified 2 molecular subtypes and found significant differences in their prognosis and that subtype B had a significantly better survival prognosis than subtype A. In addition, we found that pathways associated with cell proliferation division and DNA repair were enriched in the high-risk type A samples. Finally, we constructed a robust risk-scoring system, and our risk analysis revealed a general reduction of various immune cell components and tumor stromal components in the immune microenvironment of high-risk lung adenocarcinoma and a distinct immune infiltration pattern of immune cells, which was associated with a lower survival rate. Our comprehensive analysis of SLC7A11-related genes suggests that disulfidptosis has a potential value in the tumor microenvironment, immunity, clinical outcome, and prognosis of lung adenocarcinoma. These findings may increase our understanding of disulfidptosis as a novel cell death paradigm and provide ideas for assessing the prognosis of lung adenocarcinoma and developing new diagnostic and therapeutic strategies.

Identification of Cell-Cell Communications by Single-Cell RNA Sequencing in End Stage Renal Disease Provides New Insights into Immune Cell Heterogeneity.

Impaired immune system characterized by low-grade inflammation is closely associated with kidney chronic kidney disease (CKD) progression. To reveal the alterations of the function, component, and intercellular communication of immune cells during the progression of CKD. We conducted a case-control study enrolling regular hemodialysis patients and healthy controls. Clinical data, serum and peripheral blood mononuclear cell (PBMC) samples were collected. Flow cytometry and single-cell RNA sequencing were performed to quantitatively analyze the immune cell subsets and T-cell subsets of PBMCs. scRNA data of GSE140023 containing mouse unilateral ureteral obstruction (UUO) models were analyzed the heterogeneity of immune cells. Overall reduction in peripheral blood lymphocyte subsets in patients with end-stage renal disease (ESRD) was observed. A higher ratio of Th17/Treg, Th1/Treg, and b-cell/Treg in the ESRD group was associated with a decrease in eGFR, PTH, and ferritin. Among T cell subsets identified by scRNA analysis, Th17 cells were significantly increased in the ESRD and UU0 group. TFH, Th1, and Th2 cells are located at the final stage in the developmental tree, while Treg and memory CD8+ T cells are at the beginning site. Early developmental differentiation of Th17, Th1, and Tfh cells was observed in the ESRD and UUO group. Analysis of intercellular communication between t-cell subpopulations identified two major input and output signaling pathways: the CD40 and macrophage inhibitory factor (MIF) pathways. The MIF signaling pathway primarily mediates intercellular communication among th17 effects, CD8+ t-cell, and Th17-Treg in the ESRD group, the serum level of MIF showed significant upregulation, which was closely related to Th17/Treg cells. A global immune imbalance was closely associated with the deterioration in renal function and complication development. The MIF signaling pathway mediates Th17/Treg communication and promotes the trans-differentiation of Treg cells to Th17 cells in CKD progression.

Dynamic atlas of immune cells reveals multiple functional features of macrophages associated with progression of pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease with a high mortality rate and unclarified aetiology. Immune response is elaborately regulated during the progression of IPF, but immune cells subsets are complicated which has not been detailed described during IPF progression. Therefore, in the current study, we sought to investigate the role of immune regulation by elaborately characterize the heterogeneous of immune cells during the progression of IPF. To this end, we performed single-cell profiling of lung immune cells isolated from four stages of bleomycin-induced pulmonary fibrosis-a classical mouse model that mimics human IPF. The results revealed distinct components of immune cells in different phases of pulmonary fibrosis and close communication between macrophages and other immune cells along with pulmonary fibrosis progression. Enriched signals of SPP1, CCL5 and CXCL2 were found between macrophages and other immune cells. The more detailed definition of the subpopulations of macrophages defined alveolar macrophages (AMs) and monocyte-derived macrophages (mo-Macs)-the two major types of primary lung macrophages-exhibited the highest heterogeneity and dynamic changes in expression of profibrotic genes during disease progression. Our analysis suggested that Gpnmb and Trem2 were both upregulated in macrophages and may play important roles in pulmonary fibrosis progression. Additionally, the metabolic status of AMs and mo-Macs varied with disease progression. In line with the published data on human IPF, macrophages in the mouse model shared some features regarding gene expression and metabolic status with that of macrophages in IPF patients. Our study provides new insights into the pathological features of profibrotic macrophages in the lung that will facilitate the identification of new targets for disease intervention and treatment of IPF.

Identification of biomarkers related to sepsis diagnosis based on bioinformatics and machine learning and experimental verification.

Sepsis is a systemic inflammatory response syndrome caused by bacteria and other pathogenic microorganisms. Every year, approximately 31.5 million patients are diagnosed with sepsis, and approximately 5.3 million patients succumb to the disease. In this study, we identified biomarkers for diagnosing sepsis analyzed the relationships between genes and Immune cells that were differentially expressed in specimens from patients with sepsis compared to normal controls. Finally, We verified its effectiveness through animal experiments. Specifically, we analyzed datasets from four microarrays(GSE11755、GSE12624、GSE28750、GSE48080) that included 106 blood specimens from patients with sepsis and 69 normal human blood samples. SVM-RFE analysis and LASSO regression model were carried out to screen possible markers. The composition of 22 immune cell components in patients with sepsis were also determined using CIBERSORT. The expression level of the biomarkers in Sepsis was examined by the use of qRT-PCR and Western Blot (WB). We identified 50 differentially expressed genes between the cohorts, including 2 significantly upregulated and 48 significantly downregulated genes, and KEGG pathway analysis identified Salmonella infection, human T cell leukemia virus 1 infection, Epstein-Barr virus infection, hepatitis B, lysosome and other pathways that were significantly enriched in blood from patients with sepsis. Ultimately, we identified COMMD9, CSF3R, and NUB1 as genes that could potentially be used as biomarkers to predict sepsis, which we confirmed by ROC analysis. Further, we identified a correlation between the expression of these three genes and immune infiltrate composition. Immune cell infiltration analysis revealed that COMMD9 was correlated with T cells regulatory (Tregs), T cells follicular helper, T cells CD8, etal. CSF3R was correlated with T cells regulatory (Tregs), T cells follicular helper, T cells CD8, etal. NUB1 was correlated with T cells regulatory (Tregs), T cells gamma delta, T cells follicular helper, etal. Taken together, our findings identify potential new diagnostic markers for sepsis that shed light on novel mechanisms of disease pathogenesis and, therefore, may offer opportunities for therapeutic intervention.

You complete me: tumor cell-myeloid cell nuclear fusion as a facilitator of organ-specific metastasis.

Every cancer genome is unique, resulting in potentially near infinite cancer cell phenotypes and an inability to predict clinical outcomes in most cases. Despite this profound genomic heterogeneity, many cancer types and subtypes display a non-random distribution of metastasis to distant organs, a phenomenon known as organotropism. Proposed factors in metastatic organotropism include hematogenous versus lymphatic dissemination, the circulation pattern of the tissue of origin, tumor-intrinsic factors, compatibility with established organ-specific niches, long-range induction of premetastatic niche formation, and so-called "prometastatic niches" that facilitate successful colonization of the secondary site following extravasation. To successfully complete the steps required for distant metastasis, cancer cells must evade immunosurveillance and survive in multiple new and hostile environments. Despite substantial advances in our understanding of the biology underlying malignancy, many of the mechanisms used by cancer cells to survive the metastatic journey remain a mystery. This review synthesizes the rapidly growing body of literature demonstrating the relevance of an unusual cell type known as "fusion hybrid" cells to many of the hallmarks of cancer, including tumor heterogeneity, metastatic conversion, survival in circulation, and metastatic organotropism. Whereas the concept of fusion between tumor cells and blood cells was initially proposed over a century ago, only recently have technological advancements allowed for detection of cells containing components of both immune and neoplastic cells within primary and metastatic lesions as well as among circulating malignant cells. Specifically, heterotypic fusion of cancer cells with monocytes and macrophages results in a highly heterogeneous population of hybrid daughter cells with enhanced malignant potential. Proposed mechanisms behind these findings include rapid, massive genome rearrangement during nuclear fusion and/or acquisition of monocyte/macrophage features such as migratory and invasive capability, immune privilege, immune cell trafficking and homing, and others. Rapid acquisition of these cellular traits may increase the likelihood of both escape from the primary tumor site and extravasation of hybrid cells at a secondary location that is amenable to colonization by that particular hybrid phenotype, providing a partial explanation for the patterns observed in some cancers with regard to sites of distant metastases.

TREM2 Agonism with a Monoclonal Antibody Attenuates Tau Pathology and Neurodegeneration.

TREM2 is a membrane receptor expressed on microglia that plays a pivotal role in the organization and function of these innate immune cell components within the neurodegenerated brain. Whereas TREM2 deletion has been studied extensively in experimental beta-amyloid and Tau-based models of Alzheimer's disease, its engagement, and subsequent agonism have not been tested in the context of Tau pathology. Herein, we explored the effects of Ab-T1, an agonistic TREM2 monoclonal antibody on Tau uptake, phosphorylation, seeding, and spreading as well as its therapeutic efficacy in a Tauopathy model. Ab-T1 enhanced the uptake of misfolded Tau to microglia and induced a non-cell autonomous attenuation of spontaneous Tau seeding and phosphorylation in primary neurons from human Tau transgenic mice. Ex vivo, incubation with Ab-T1 led to a significant reduction in the seeding of Tau pathology in the hTau murine organoid brain system. Systemic administration of Ab-T1 resulted in reduced Tau pathology and propagation when hTau was stereotactically injected into the hemispheres of hTau mice. Intraperitoneal treatment with Ab-T1 lead to attenuation of cognitive decline in the hTau mice that was associated with reduced neurodegeneration and synaptic preservation with amelioration of the global neuroinflammatory program. Collectively, these observations show that TREM2 engagement with an agonistic antibody result in reduced Tau burden concomitant with attenuated neurodegeneration ascribed to the education of resident microglia. These results may suggest that despite the opposing results with regard to the effect of TREM2 knockout in experimental Tau-based model systems, receptor engagement and activation by Ab-T1 appears to possess beneficial effects with respect to the various mechanisms mediating Tau-driven neurodegeneration.

Targeting molecular cross-talk between tumor cells and tumor associated macrophage as therapeutic strategy in triple negative breast cancer

Triple-negative Breast cancer (TNBC) is a subtype of breast cancer (BC) that lacks expression for ER/PR/Her2 receptors and is associated with aggressive disease pathogenesis and the worst prognosis among other subtypes of BC. Accumulating evidence-based studies indicate the high immunogenic ability of TNBC tumors and the applicability of immunotherapeutic strategies to overcome therapy resistance and tumor recurrence in TNBC patients. However, not all TNBC patients respond equally well to current immunotherapies that mainly target the adaptive immune system for tumor rejection. Recent studies are contemplating the efficacy of tumor-associated macrophage (TAM) targeted therapies since these subpopulations of cells comprise one of the major components of tumor-infiltrating immune cells (TIIs) in the TNBC tumor microenvironment (TME) and play an essential role in priming the adaptive immune response mediators towards both antitumorigenic and pro-tumorigenic response facilitated by intercellular cross-talk between tumor cells and TAM populations present within TNBC-TME. The present review discusses these molecular mechanisms and their consequence on the progression of TNBC tumors. Also, the therapeutic strategies targeting candidate genes/pathways involved in molecular cross-talk between TAM-TNBC cells and their impact on the development and progression of TNBC tumors are also discussed.

EGFR Inhibition by Cetuximab Modulates Hypoxia and IFN Response Genes in Head and Neck Squamous Cell Carcinoma.

While the hypoxic and immunosuppressive TME of HNSCC has been well described, comprehensive evaluation of the immune cell components and signaling pathways contributing to immunotherapy resistance has been poorly characterized. We further identified additional molecular determinants and potential therapeutic targets of the hypoxic TME to fully leverage currently available targeted therapies that can be administered with immunotherapy.

Decoding the structural and cellular composition of the tumor microenvironment in multiple human cancers using Imaging Mass Cytometry

Abstract High-plex imaging techniques such as Imaging Mass Cytometry™ (IMC™) have become key tools in understanding and decoding the spatial complexity of the tumor microenvironment (TME). The TME contains tumor-infiltrating lymphocytes that have been associated with positive therapeutic outcomes. IMC enables detailed assessment of cell phenotype and function using 40-plus markers simultaneously at subcellular resolution on a single slide without spectral overlap or background autofluorescence. We customized the Maxpar® Human Immuno-Oncology IMC Panel Kit using antibodies from the Standard BioTools™ catalog to create panels for tissue-based immuno-oncology research. Data acquisition was performed using a Hyperion™ Imaging System. Cell segmentation was facilitated using an IMC Cell Segmentation Kit. A pixel classification approach and CellProfiler™ were applied for single-cell segmentation. HistoCAT™ was used for single-cell analysis to visualize protein expression in various cancer types via PhenoGraph clustering and t-SNE maps. Our custom panels were applied to normal and cancer human tissue microarrays to phenotype and analyze cell populations in these tissues. We classified the activation state of immune cells, epithelial-to-mesenchymal transition (EMT) progression, and composition of the extracellular matrix. In-depth single-cell analysis quantitatively evaluated the cellular makeup and immune cell component in the TME of cancer tissues. This work demonstrates the capability of IMC to identify subcellular localization of cellular and structural markers, including quantitative and spatial identification of multiple immune parameters in the TME, in tumor microarrays of cancer subject samples.

CD301b +DCs orchestrate type 2 immune responses to helminth infection in the lung

Abstract Helminth parasites induce potent host type 2 immune responses (T2I) that promote parasite clearance and prevent reinfection. The parasite-induced T2I involve both innate and adaptive immune cell components, but how the activation of different cell types is coordinated during the initiation of T2I is incompletely understood. Dendritic cells (DCs) are best known for their role in bridging innate and adaptive immunity by sensing immunological insults and presenting antigens to T cells. We previously found that CD301b +DCs, a major subset of migratory type 2 conventional DCs in peripheral organs, are specifically required for the differentiation of T helper type 2 (Th2) cells in the skin- and lung-draining lymph nodes following infection with Nippostrongylus brasiliensis (Nb) in mice, but their overall role in host immune responses against Nb infection remains unclear. Here, we show that the depletion of CD301b +DCs results in reduced T2I in the lung and impaired worm clearance. The reduction in type 2 cytokine production and eosinophil influx was observed as early as 2 days post-infection before the onset of adaptive immunity, as well as in CD4T cell-depleted mice, suggesting that CD301b +DCs play a critical role in the induction of innate T2I independently of their role in Th2 cell priming. Moreover, CD301b +DC-depleted mice exhibited ameliorated lung hemorrhage with reduced neutrophil infiltration and IL-17 production early after infection. These data suggest the role of CD301b +DCs in co-regulating innate type 2 and type 3 inflammation mechanisms during the early phase of Nb infection in the lung. This work was supported by NIH grant R01AI132576.

Abstract 2250: Immuno-oncology study to profile the tumor microenvironment in multiple human cancers using high-plex imaging mass cytometry

Abstract Immune profiling of tumor tissues has become a key tool in understanding the complexity of the tumor microenvironment (TME), for predictive biomarker discovery, and in cancer treatment. The presence of tumor-infiltrating lymphocytes has been associated with benefit from therapy. Furthermore, the TME also contains immunosuppressive elements that can impede immune response. Imaging Mass Cytometry™ (IMC™) enables detailed assessment of cell phenotype and function using 40-plus markers simultaneously at subcellular resolution on a single slide without spectral overlap or background autofluorescence. High-plex IMC has enabled us to evaluate the TME in different cancer histologies including in highly autofluorescent tissue types like lung, hepatocellular carcinoma, and skin melanoma. The Maxpar® Human Immuno-Oncology IMC Panel Kit (201508) was customized using antibodies from the Standard BioTools™ catalog to create panels for tissue-based immuno-oncology research. Data acquisition was performed using a Hyperion™ Imaging System. To facilitate cell segmentation, an IMC Cell Segmentation Kit (TIS-00001) was applied to enhance cell membrane boundaries. We applied a pixel classification approach and CellProfiler™ for single-cell segmentation. We used histoCAT™ for single-cell analysis to visualize protein expression in various cancer types via PhenoGraph clustering and t-SNE maps. Our panels were applied to normal and cancer human tissue microarrays (TMAs) to phenotype and analyze cell populations in these tissues. We provide detailed analysis of the TME by classifying activation state of immune cell populations, epithelial-to-mesenchymal transition (EMT) progression, and composition of the extracellular matrix. In-depth single-cell analysis quantitatively evaluated the cellular makeup and immune cell component in the TME of cancer tissues and identified major tumor, immune, and stromal cell phenotypes. This work demonstrates the capability of IMC for quantitative and spatial identification of multiple immune parameters in the TME on a single slide of cancer patient samples (e.g. tumor microarray). For Research Use Only. Not for use in diagnostic procedures. Citation Format: Thomas D. Pfister, Liang Lim, Shaida Ouladan, Nick Zabinyakov, Qanber Raza, Christina Loh. Immuno-oncology study to profile the tumor microenvironment in multiple human cancers using high-plex imaging mass cytometry [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2250.

Abstract 4705: Profiling of immune cell components and soluble factors in ovarian cancer ascites highlights impaired immune environment

Abstract Background: It’s now increasingly recognized that ovarian cancer (OC) ascites play a significant role in OC progression - behavior of tumor cells is influenced by the nature of their surrounding microenvironment. Thus, characterization of ascites composition is essential to understand how this milieu affects tumor progression and particularly the immunosuppressive pathways that would underlie immune response dysfunctions and, in turn, how biological ascites effects can be influenced by that composition. Methods: Ascites samples were collected from advanced OC patients. Through respective multiplexed approaches of flow cytometry-based marker expression analysis and quantitation of mediators, immune context was profiled by investigating immune cell composition and levels of a plethora of soluble factors, including cytokines/chemokines, and metabolic pathways of some amino acids known to be involved in immunosuppression. Furthermore, ascites fluids were functionally screened for their biological effects on healthy monocytes, either undifferentiated or undergoing M1 polarization. Results: Unlike healthy PBMCs, OC ascites were mostly “enriched” in regulatory/immunosuppressive immune cell subsets including T and myeloid populations. Also, T cells were shown to highly express immune checkpoints such as PD1 in T cells and TIGIT in Tregs. Intriguingly, a high CD4/CD8 ratio was seen. Also, CD163+ tumor-associated macrophages were shown to express CSF1R, CCR8 and CCR2, and to even display a mixed phenotype since also expressing Arg1, CD80, and iNOS. On acellular fractions, most ascites demonstrated elevated CCL18, IL6, LIF, VEGF, and CCL2 levels, and low IL2, IL4, and IL17 levels. Interestingly, unlike healthy plasmas, these ascites appeared to harbor a metabolically-immunosuppressive profile characterized by high glutaminolysis and tryptophan (Trp) degradation in kynurenine (Kyn). Functionally, we demonstrated that not only ascites basically polarized monocytes into M2 macrophages, but even antagonized with their M1 polarization to ultimately tilt to M2 status. Conclusions: Taken together, our data show that ascites fluids most favorable to the M2 phenotype were associated with high LIF, VEGF, IL6, CCL2, and CCL18 levels, and with elevated Kyn to Trp ratios - Kyn levels being strongly higher than in healthy plasmas. Our results thus highlight a peculiar altered environment of OC ascites where a mixture of suppressive cells and signaling factors mediate extracellular cues leading to immune cell activity dysfunction. Altogether, these translational findings highlight OC ascites as a valuable tool to understand the mechanisms of suppression and develop predictive profiles, and to provide new insights for the identification of new targets and development of targeted-therapies. Citation Format: Assia Chaibi, Coriolan Lebreton, Dominique Bodet, Jean-Philippe Guegan, Guillaume Babin, Antoine Italiano, Alban Bessede, Imane Nafia. Profiling of immune cell components and soluble factors in ovarian cancer ascites highlights impaired immune environment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4705.

The Relationship between Occurrence of aGVHD in Patients with Acute Myeloid Leukemia after Allogeneic Hematopoietic Stem Cell Transplantation and Immune Cell Components in Graft

To explore the relationship between occurrence of acute graft-versus-host disease (aGVHD) and various immune cell composition in patients with acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The clinical data of 104 patients with AML undergoing allo-HSCT in our hospital were retrospectively analyzed, and the hematopoietic reconstitution and occurrence of GVHD were analyzed. Flow cytometry was used to detect the proportion of various types of immune cells in the grafts, the number of graft composition in patients with different degrees of aGVHD was calculated and compared, and to analyze the correlation between the severity of aGVHD in AML patients after allo-HSCT and the immune cell components in the graft. There was no significant difference in the time of hematopoietic reconstitution between the high number group of total number of nucleated cells (TNC) and the low number group, while the time of neutrophil and platelet reconstruction in the high number of CD34 group was significantly faster than that in the low number of CD34 group (P＜0.05), and the total hospital stay also tends to be shorten. Compared with patients in 0-Ι aGVHD group, both HLA-matched and HLA-haploidentical transplantation, the infusion amounts of CD3+ cells, CD3+CD4+ cells, CD3+CD8+ cells, NK cells and CD14+ monocytes were higher in patients of Ⅱ-Ⅳ aGVHD group， but the difference was not statistically significant (P>0.05); In addition, in patients with HLA-haploidentical transplantation, the number of CD4+CD25+ cells in Ⅱ-Ⅳ aGVHD group was significantly lower than that in 0-Ι aGVHD group (P<0.05), and the same trend was also observed in HLA-matched transplanted patients, but the difference was not significant (P=0.078). High number of CD34+ cells in the graft is beneficial to hematopoietic reconstitution in AML patients. To a certain degree, high number of CD3+ cells, CD3+CD4+ cells, CD3+CD8+ cells, NK cells and CD14+ cells tend to increase the occurrence of aGVHD, but high number of CD4+CD25+ regulatory T cells is beneficial to reduce the incidence of aGVHD in AML patients.