Immune Cells In Microenvironment Research Articles

Highlights from Recent Cancer Literature

Highlights from Recent Cancer Literature

Atlas of immune cell infiltration in breast cancer—high M2 macrophage and low native B cellproportions are associated with poor survival

Background: Knowledge of the tumor immune microenvironment is critical for developing new treatments targeting the immune system in breast cancer. However, no profile of immune cell infiltration in breast cancer has been provided. The aim of this study is to illustrate an atlas of the immune cell microenvironment of breast cancer.

Insights Into the Prognostic Value and Immunological Role of NAAA in Pan-Cancer.

N-Acylethanolamine Acid Amidase (NAAA) is an N-terminal cysteine hydrolase and plays a vital physiological role in inflammatory response. However, the roles of NAAA in tumor immunity are still unclear. By using a series of bioinformatics approaches, we study combined data from different databases, including the Cancer Genome Atlas, the Cancer Cell Line Encyclopedia, Genotype Tissue-Expression, cBioPortal, Human Protein Atlas, TIMER, and ImmuCellAI to investigate the role of NAAA expression in prognosis and tumor immunity response. We would like to reveal the potential correlations between NAAA expression and gene alterations, tumor mutational burden (TMB), microsatellite instability (MSI), DNA methylation, tumor microenvironment (TME), immune infiltration levels, and various immune-related genes across different cancers. The results show that NAAA displayed abnormal expression within most malignant tumors, and overexpression of NAAA was associated with the poor prognosis of tumor patients. Through gene set enrichment analysis (GSEA), we found that NAAA was significantly associated with cell cycle and immune regulation-related signaling pathways, such as in innate immune system, adaptive immune system, neutrophil degranulation, and Toll-like receptor signaling pathways (TLRs). Further, the expression of NAAA was also confirmed to be correlated with tumor microenvironment and diverse infiltration of immune cells, especially tumor-associated macrophage (TAM). In addition to this, we found that NAAA is co-expressed with genes encoding major histocompatibility complex (MHC), immune activation, immune suppression, chemokine, and chemokine receptors. Meanwhile, we demonstrate that NAAA expression was correlated with TMB in 4 cancers and with MSI in 10 cancers. Our study reveals that NAAA plays an important role in tumorigenesis and cancer immunity, which may be used to function as a prognostic biomarker and potential target for cancer immunotherapy.

Role of EZH2 in bone marrow mesenchymal stem cells and immune-cancer interactions.

In recent years, methylation modification has been determined to be vital for the biological regulation of normal cells, tumor cells, and tumor microenvironment immune cells. Enhancer of zeste homology 2 (EZH2), a component of the Polycomb Repressive Complex 2 (PRC2), catalyzes the trimethylation of the downstream gene in the tri-methylates histone three lysine 27 (H3K27me3) position, which causes chromatin pyknosis, and thus, silences the expression of related genes. In this paper, we reviewed the role of EZH2 in regulating bone marrow mesenchymal stem cell differentiation and the immune cell function in tumor microenvironment, summarized all types of existing EZH2 inhibitors and the main clinical trials, and proposed relevant ideas for potential clinical applications.

Diffuse large B-cell lymphoma-derived exosomes push macrophage polarization toward M2 phenotype via GP130/STAT3 signaling pathway

Growing evidence shows that cancer progression links with both heterogeneity of the tumor microenvironment and dysregulated activity of immune cells. Cancer-secreted exosomes are being recognized as indispensable mediators of the exchange cargo between cancer and immune cells. The M2-phenotype tumor-associated macrophages have the function of promoting tumor progression and drug resistance. Diffuse large B-cell lymphoma(DLBCL) is a highly heterogeneous and very common malignant non-Hodgkin's lymphoma. Here, we demonstrate that different subtype DLBCL cell-derived exosomes are internalized by macrophages, which can affect macrophages polarization.The mechanism of DLBCL-derived exosomes on macrophage polarization remains unclear currently. This study showed that DLBCL-secreted exosomes could induce the transformation of macrophages to a protumor M2-like phenotype, and block the drug-induced apoptosis of DLBCL cells in an indirect co-culture system. Different DLBCL-derived exosomes could change the phenotype of macrophages through the STAT3 signaling, which upregulated the expression of oncogenic genes and classical markers of M2-like phenotype macrophages, such as IL-10, CD206, and CD163. The addition of DLBCL-derived exosomes resulted in the activation of the STAT3 signaling pathway of M0/M2 macrophages in an indirect co-culture system. GP130 was highly enriched in DLBCL-derived exosomes, which triggered the activation of STAT3 of macrophages and subsequently induced the downstream targets such as BCL2, SURVIVIN, and BAX. The parallel changes of STAT3 and GP130 in macrophages confirmed that GP130 of DLBCL-derived exosomes promoted macrophage polarization by activating STAT3 signaling. Furthermore, all of these effects could be reversed by the GP130 inhibitor SC144. The data indicated that DLBCL-derived exosomes could trigger macrophages polarization into a pro-survival M2-like phenotype, which was at least partially through the GP130/STAT3 signaling pathway. Collectively, this study showed that DLBCL-derived exosomes could promote macrophages transformation to protumor M2-like phenotype in the tumor microenvironment.

The immune microenvironment of the hydatidiform mole

Gestational trophoblastic diseases (GTDs) are a heterogeneous group of lesions, the most frequent being the hydatidiform mole (HM). HMs are usually cured after surgical treatment or after chemotherapy in the case of a persistent trophoblastic activity. Immunotherapy could be an interesting alternative as a first-line or second-line treatment. However, only a few studies have explored the immune microenvironment of HMs. In the present retrospective study including 19 complete and 17 partial moles, we examined the composition of the immune cell microenvironment by immunohistochemistry using the following antibodies: CD4, CD8, CD56, PD-L1, S100, CD83, CD207, CD123, CD1a, CD11c, CD163, PAX5, and MUM1. In the decidual cells compartment, CD11c+cells were the predominant population, followed by CD4+ cells, CD56+ NK cells, CD163+ macrophages, and CD8+ T lymphocytes.In the endometrial glands compartment, CD11c+cells were the predominant population, followed by CD4+ cells, CD56+ NK cells, and CD8+ T lymphocytes. In the villi compartment, the predominant immune cells were CD4+ cells, followed by CD163+ macrophages and CD11c+cells. Statistically significant differences were observed between partial and complete moles in all three compartments. The immune microenvironment of HMs is immunosuppressive, but it differs between complete and partial moles, the latter having a higher infiltrate of cells with phenotypes suggestive of immunosuppressive activities.

Intrahepatic immune infiltrate in chronic hepatitis B and chronic hepatitis C: Similar but not the same.

In chronic hepatitis B (CHB) and C (CHC) infections, the composition of the immune cell microenvironment at the site of infection is poorly understood. Thus, our aim was to characterize and compare liver infiltrates to identify shared and exclusive hepatic immune components. Immunohistochemistry was performed on 26 CHB and 42 CHC liver biopsies to determine Th (CD4+), Th1 (T-bet+), Th17 (IL-17A+), Treg (Foxp3+) and CTL (CD8+) cells frequency in portal/periportal and intralobular areas and relate them to liver damage. CHB and CHC cases shared a portal/periportal CD4+ lymphocyte predominance and a lobular CD8+ lymphocyte majority. However, CHC exhibited a concomitant lobular T-bet+ cell dominance while in CHB FoxP3+ cells prevail. CHC disclosed higher frequencies of P/P FoxP3+, IL-17A+ and T-bet+ cells and intralobular CD4+, IL-17A+ and T-bet+ lymphocytes. HBeAg+ chronic hepatitis and CHC cell frequencies were similar except for lobular T-bet+ that remained higher among CHC cases. Comparison among cases with less severe liver disease revealed lower lymphocyte frequencies in CHB samples, while no differences were observed between patients with more severe stages. Interestingly, in CHB portal/periportal CD4+ and lobular CD4+, CD8+ and IL-17A+ cells were associated with severe hepatitis. Even when all studied populations were identified in both infections preferential lymphocyte frequencies and prevalence at different areas along with their association with liver damage highlighted that CHB and CHC immune responses are not the same.

Analysis of Single Cell Transcriptomics in Paired Pediatric T-ALL Samples Collected at Diagnosis and Following End of Induction Therapy Reveals an MRD-Associated Stem Cell Signature

IntroductionDespite recent improvement in outcomes for de novo disease, pediatric T-cell acute lymphoblastic leukemia (T-ALL) remains challenging to treat at relapse. Investigation into genomic markers of treatment response and therapy resistance offers an opportunity to further enhance outcomes for these patients. We previously identified a T-ALL blast-associated gene signature at diagnosis (Dx) and characterized the immune microenvironment in Dx T-ALL marrow samples using single cell transcriptome analysis (Bhasin et al. Blood 2020(ASH)). This approach allowed us to generate a granular expression map of both the T-ALL landscape and the Dx bone marrow (BM) immune microenvironment. Here we expand this work by evaluating samples collected from the same patients Dx and End of Induction (EOI) BM samples from pediatric T-ALL patients. The use of paired samples provides insight into treatment-induced changes in the microenvironment. Further, the inclusion of both minimal residual disease (MRD) positive and MRD negative samples allowed us to compare differences between these groups.MethodsUsing the 10X genomics platform, we profiled the single cell transcriptome of ~18,000 BM and immune microenvironment cells from viably frozen samples collected from T-ALL patients at Dx or EOI. Five paired Dx and EOI samples and one EOI sample from a patient with relapsed T-ALL were evaluated, for a total of 11 samples. Three paired samples were MRD positive at EOI and two were MRD negative; the relapsed sample was MRD negative. Cell clustering was performed using the Seurat package and differential expression analysis was performed using R/Bioconductor packages (Hao et al. Cell 2021). Cell communication analysis was conducted using the CellChat R tool (v 1.0.0) to infer cell-cell communication within the EOI MRD positive and MRD negative subsets and compare their communication networks (Jin et al. Nature Comm 2021).ResultsUsing our previously described blast-associated gene signature (Bhasin et al. ASH 2020) we were able to identify residual blast populations at EOI in MRD-positive samples. Comparative analysis of gene profiles at Dx and EOI showed significant changes in the microenvironment cell populations with highest increase in erythroid cell populations after induction therapy. The gene expression profiles were significantly different for immune cells at Dx and EOI and the relapsed sample had greater similarity to the Dx samples indicating a persistent immunosuppressive environment. Clustering analysis of the EOI samples (3 MRD positive and 2 MRD negative) demonstrated the presence of patient specific blast cells in MRD positive samples that retained patient-specific transcriptomeheterogeneity at EOI (Fig.1A). Analysis of communication networks between different cell types based on receptor and ligand expression levels between different cell types identified a CD34 + cluster of stem cells that had different interactions with other immune populations in the MRD positive and negative subsets. Differential expression analysis between the MRD positive and MRD negative cells in this CD34 + stem cell cluster identified higher expression of myeloid associated genes such as CEBPB, CEBPD, AZU1 in the MRD negative group relative to the MRD positive cells, which showed higher expression of B-cell related genes such as IGHM, VPREB1, CD79A/ B along with upregulation of P13K signaling in B-lymphocytes, B-cell receptor signaling and autophagy pathways. Analysis of upstream regulators based on the differential gene signature between the MRD positive and MRD negative group demonstrated upregulation of MYC and TCF3 activity and inhibition of TGFB1, CSF3 and CEBPA in MRD positive compared to MRD negative samples (Fig.1B).Conclusions: Leukemic blasts exhibit patient-specific gene expression signatures that are present at EOI in MRD positive samples. Exploration of the impact of minimal residual disease at EOI revealed differential gene expression patterns in stem cells from MRD positive samples, characterized by activation of B cell related signaling pathways and regulators such as MYC and TCF3. In contrast, a more myeloid-like expression signature was observed in stem cells from MRD negative samples. These findings open the avenues for exploration of therapeutic targets of T-ALL progression. [Display omitted] DisclosuresDeRyckere: Meryx: Other: Equity ownership. Graham: Meryx: Membership on an entity's Board of Directors or advisory committees, Other: Equity ownership.

Pediatric Single Cell Cancer Atlas: An Integrative Web-Based Resource for Single Cell Transcriptome Data from Pediatric Leukemias

Introduction: The emergence and optimization of single cell profiling as a powerful tool to characterize the tumor microenvironment has revealed the heterogeneity of pediatric cancers, particularly different leukemia types/subtypes. Ease of access and analysis of the data from studies on different leukemia types is critical for improving diagnosis as well as therapy.Currently, there are no single cell data based resources available for pediatric leukemias. We have developed a comprehensive resource, Pediatric Single Cell Cancer Atlas (PedScAtlas), with the goal of developing a pan-leukemia genomics signature as well as highlighting the heterogeneity of different types of leukemia. This resource facilitates exploration and visualization of expression signatures in different leukemias without requiring extensive analysis and bioinformatics support.Methods: The PedScAtlas was built based on single cell data from various leukemias and normal bone marrow (BM) cells that have been pre-processed and analyzed using a uniform approach to generate normalized expression data (M. Bhasin et al. Blood 2020 (ASH), S. S. Bhasin et al. Blood 2020 (ASH), Panigraphy et al. JCI 2019, Stroopinsky et al. Haematologica 2021, Thomas et al. Blood 2020 (ASH)). The current version of PedScAtlas contains data from 30 local leukemia samples (Bhasin, et al. Blood 2020 (ASH), Thomas et al. Blood 2020 (ASH)) and those available on public resources such as GEO (Bailur et al. JCI Insight 2020). The PedScAtlas dataset and the Immune cell dataset each underwent quality control, integration, normalization, and dimensionality reduction using the Uniform Manifold Approximation and Projection (UMAP) method. Unsupervised UMAP analysis identified cellular clusters with similar transcriptome profiles that were annotated based on expression of cell-specific markers. Differential expression comparing different types of leukemia including acute myeloid leukemia (AML), B-cell acute lymphoblastic leukemia (B-ALL), T-cell acute lymphoblastic leukemia (T-ALL), and mixed phenotype acute leukemia (MPAL) samples was performed. To further ascertain genes specifically expressed in malignant blasts, the atlas also contains data from normal BM samples (Bailur et al. JCI Insight 2020) and healthy immune cells from the census of Immune Cells by the Human Cell Atlas Project (https://data.humancellatlas.org/). The web resource source code is written in R programming language and the interactive webserver has been implemented using the R Shiny package (Fig 1). The tool has been extensively tested on multiple operating systems (Linux, Mac, Windows) and web-browsers (Chrome, FireFox, and Safari). The tool is currently hosted on a 64bit CentOS 6 backend server running the Shiny Server program designed to host R Shiny applications.Results: The PedScAtlas contains data that facilitate exploration of gene expression profiles across leukemia types/subtypes and tumor microenvironment (TME) cell types. The atlas includes data from 33,930 AML, 25,744 T-ALL, 13,404 MPAL, and 6,252 B-ALL blast cells. It also contains single cell profiles of healthy BM samples from publicly available studies. The user can select data sets from the 5 major types of leukemia and normal BM in any combination of their choice to explore the expression profile of a gene of interest. The data can be visualized as UMAP (Fig. 1), or violin plots with annotations based on cluster ID, cell type, disease type, sample ID, and future continuous remission or relapse outcome. The UMAP with gene expression analyses allows the user to visualize the distribution of cell expressing a given gene on the UMAP plot. The Biomarker tool shows expression of different leukemia biomarker gene sets in the entire leukemia dataset. The Immune Cell section contains BM data from the Human Cell Atlas Project; the purpose of this section is to validate leukemia biomarkers by checking that the gene does not have significant expression in the healthy immune microenvironment.Conclusions: The PedScAtlas resource provides a unique and straightforward tool for biomarker identification, analysis of leukemia subtype heterogeneity, and transcriptome profile of the immune cell microenvironment. The resource is available online at https://bhasinlab.bmi.emory.edu/PediatricSC/. [Display omitted] DisclosuresDeRyckere: Meryx: Other: Equity ownership. Graham: Meryx: Membership on an entity's Board of Directors or advisory committees, Other: Equity ownership.

Low Percentage of Microenvironment Immune Cells in Bone Marrow and Lymph Node Measured By Flow Cytometry Is Associated with Inferior Overall Survival in Diffuse Large B Cell Lymphoma

INTRODUCTIONTumor microenvironment (TME) in diffuse large B-cell lymphoma (DLBCL) has recently been in the spotlight. The predictive role of immune cells (IC) in peripheral blood (PB), bone marrow (BM) and lymph nodes (LN) has been individually assessed, often using techniques not widely available. However, data regarding the relative prognostic impact of TME in each compartment evaluated simultaneously in DLBCL is still lacking.AIMSTo determine the prognostic impact on survival of the monocyte-lymphocyte prognostic score (MLS) in PB and the percentage (%) of IC in the BM and LN measured by flow cytometry (FC) in DLBCL and whether it could improve the conventional risk assessment of the R-IPI.METHODSWe retrospectively collected information of patients (pts) with DLBCL and available BM aspiration and LN FC data at diagnosis, treated at our center between 2012 and 2019. Pts were stratified according to the MLS (Wilcox et al, Leukemia 2011) in two groups: low (PB monocyte counts <630/ml and lymphocyte counts ≥1000/ml) and intermediate-high risk. FC analysis was performed with 8-color panels according to Euroflow protocols. % of BM and LN IC by FC were compared to normal values determined by Matarraz (Cytometry part B 2010) and Battaglia et al (Immunology 2003), and analyzed in 3 categories: low, normal and high. Survival analysis was performed with Kaplan-Meier (variables compared with log-rank) and Cox regression.RESULTS71 pts were included with a median age of 59 years. Table 1 shows frequency analysis of TME variables and R-IPI in our cohort. All pts received immunochemotherapy. Complete remission rate was 82%. Median overall survival (OS) was 120.5 months (m), with a median follow up time of 38.7 m.On univariate analysis, poor R-IPI and TME variables from the 3 compartments: low LN T lymphocyte (TL) %, both high and low BM monocyte (Mo) %, low polyclonal BM B lymphocyte (BL) %, and intermediate-high MLS showed prognostic impact on OS. BM IC levels did not statistically differ in involved vs not involved BM.On multivariate analysis, poor R-IPI, low LN TL %, low BM Mo and polyclonal BL, remained independent predictors of survival (Table 2). Pts with the 4 unfavorable variables showed a median OS of only 4 m and 100% mortality rate. In contrast, in pts with none of these adverse risk variables OS rate was 100% (p<0.001), Figure 1.CONCLUSIONSLow LN TL %, low BM Mo and polyclonal BL measured by FC were associated with inferior OS in our cohort of DLBCL pts. These TME variables combined with R-IPI can identify both a subgroup of pts with high early mortality rate and a group with excellent long-term prognosis. Immunotherapy with CART cells and BiTEs has shown encouraging results in relapsed refractory DLBCL pts. These strategies could help to improve outcomes in this high risk subset of pts while restoring previously deficient antitumoral immunity. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Anti-Tumor Effects of Chinese Medicine Compounds by Regulating Immune Cells in Microenvironment.

As the main cause of death in the world, cancer is one of the major health threats for humans. In recent years, traditional Chinese medicine has gained great attention in oncology due to the features of multi-targets, multi-pathways, and slight side effects. Moreover, lots of traditional Chinese medicine can exert immunomodulatory effects in vivo. In the tumor microenvironment, tumor cells, immune cells as well as other stromal cells often coexist. With the development of cancer, tumor cells proliferate uncontrollably, metastasize aggressively, and modulate the proportion and status of immune cells to debilitate the antitumor immunity. Reversal of immunosuppressive tumor microenvironment plays an essential role in cancer prevention and therapy. Immunotherapy has become the most promising strategy for cancer therapy. Chinese medicine compounds can stimulate the activation and function of immune cells, such as promoting the maturation of dendritic cells and inducing the differentiation of myeloid-derived suppressor cells to dendritic cells and macrophages. In the present review, we summarize and discuss the effects of Chinese medicine compounds on immune cells in the tumor microenvironment, including innate immune cells (dendritic cells, natural killer cells, macrophages, and myeloid-derived suppressor cells) and adaptive immune cells (CD4+/CD8+ T lymphocytes and regulatory T cells), and the various immunomodulatory roles of Chinese medicine compounds in cancer therapy such as improving tumor-derived inflammation, enhancing the immunity after surgery or chemotherapy, blocking the immune checkpoints, et al., aiming to provide more thoughts for the anti-tumor mechanisms and applications of Chinese medicine compounds in terms of tumor immunity.

TUBA1C is a Prognostic Marker in Low-grade Glioma and Correlates with Immune Cell Infiltration in the Tumor Microenvironment.

TUBA1C, a microtubule component, contributes to the development of several cancers. Our purpose was to study the expression of TUBA1C, its potential prognostic value, and its effects on the infiltration of immune cells of low-grade glioma (LGG). Through applying multiple bioinformatics analyses, we extracted and analyzed datasets from TCGA, TIMER, GTEx, GEPIA, and HPA to investigate the potential oncogenic mechanisms of TUBA1C, including the correlation between TUBA1C and prognosis, immune-checkpoints, tumor microenvironment (TME), and infiltration of immune cells in LGG. GO functional annotations and KEGG pathway analyses were further applied to investigate the potential action of TUBA1C in LGG. We revealed that the mRNA levels of TUBA1C were increased in LGG tumor tissues than in normal tissues. Additionally, TUBA1C was up-regulated in the grade III of LGG than in grade II. Moreover, we found that TUBA1C may be an independent prognostic factor of LGG, and high TUBA1C expression correlated to a poor prognosis of LGG. TUBA1C expression was positively associated with the infiltration of B cells, CD8 T+ cells, CD4+ T cells, macrophages, dendritic cells, and neutrophils. TUBA1C was also verified to be co-expressed with immune-related genes and immune-checkpoints. GO and KEGG pathway analyses indicated that TUBA1C may potentially regulate the pathogenesis of LGG through immune-related pathways, including chemokine pathway; JAK-STAT pathway; natural killer cell mediated cytotoxicity; T cell receptor pathway; leukocyte migration; negative regulation of immune system process; regulation of lymphocyte activation; T cell activation and other pathways. In conclusion, TUBA1C expression is increased in LGG and high TUAB1C expression is related to a poor prognosis. TUBA1C may influence tumor development by regulating the tumor-infiltrating cells in the TME. TUBA1C may be a potential target for immunotherapy.

Immune evasion mechanism and its clinical application value in Hodgkin's lymphoma

Hodgkin's lymphoma (HL) is a unique malignancy in which rare malignant Hodgkin and Reed-Sternberg (HRS) cells are scattered in the inflammatory cell rich microenvironment. This extensive but ineffective inflammatory cell infiltrate indicates that HRS cells have developed mechanisms to evade immune surveillance. The immune escape mechanisms of HL provide prognostic biomarkers and opportunities to develop new drugs. The immune evasion mechanisms in Hodgkin lymphoma include a reduction of human leukocyte antigen (HLA) to affect first signal which is essential for T cell activation; an upregulation of negative co-stimulatory molecules to inhibit T cell activation; resistance to apoptosis or killing by expressing some molecules on HRS cells membrane; an immunosuppressive network formed by HRS cells regulating the microenvironment immune cells. Immune escape mechanisms of HRS cells provide new targets for the development of new drug and the new drug development strategies include drugs on HRS cells and drugs on microenvironment.

Pan-Cancer Analysis of PIMREG as a Biomarker for the Prognostic and Immunological Role.

Phosphatidylinositol binding clathrin assembly protein interacting mitotic regulator (PIMREG) localizes to the nucleus and can significantly elevate the nuclear localization of clathrin assembly lymphomedullary leukocythemia gene. Although there is some evidence to support an important action for PIMREG in the occurrence and development of certain cancers, currently no pan-cancer analysis of PIMREG is available. Therefore, we intended to estimate the prognostic predictive value of PIMREG and to explore its potential immune function in 33 cancer types. By using a series of bioinformatics approaches, we extracted and analyzed datasets from Oncomine, The Cancer Genome Atlas, Cancer Cell Lineage Encyclopedia (CCLE) and the Human Protein Atlas (HPA), to explore the underlying carcinogenesis of PIMREG, including relevance of PIMREG to prognosis, microsatellite instability (MSI), tumor mutation burden (TMB), tumor microenvironment (TME) and infiltration of immune cells in various types of cancer. Our findings indicate that PIMREG is highly expressed in at least 24 types of cancer, and is negatively correlated with prognosis in major cancer types. In addition, PIMREG expression was correlated with TMB in 24 cancers and with MSI in 10 cancers. We revealed that PIMREG is co-expressed with genes encoding major histocompatibility complex, immune activation, immune suppression, chemokine and chemokine receptors. We also found that the different roles of PIMREG in the infiltration of different immune cell types in different tumors. PIMREG can potentially influence the etiology or pathogenesis of cancer by acting on immune-related pathways, chemokine signaling pathway, regulation of autophagy, RIG-I like receptor signaling pathway, antigen processing and presentation, FC epsilon RI pathway, complement and coagulation cascades, T cell receptor pathway, NK cell mediated cytotoxicity and other immune-related pathways. Our study suggests that PIMREG can be applied as a prognostic marker in a variety of malignancies because of its role in tumorigenesis and immune infiltration.

SOX11, CD70, and Treg cells configure the tumor immune microenvironment of aggressive mantle cell lymphoma

AbstractMantle cell lymphoma (MCL) is a mature B-cell neoplasm with a heterogeneous clinical and biological behavior. SOX11 oncogenic expression contributes to the aggressiveness of these tumors by different mechanisms, including tumor and stromal cell interactions. However, the precise composition of the immune cell microenvironment of MCL, its possible relationship to SOX11 expression, and how it may contribute to tumor behavior is not well known. Here, we performed an integrative transcriptome analysis of 730 immune-related genes combined with the immune cell phenotype analysis by immunohistochemistry in SOX11+ and SOX11− primary nodal MCL cases and non-neoplastic reactive lymph nodes. SOX11+ MCL had a significant lower T-cell intratumoral infiltration compared with negative cases. A reduced expression of MHCI/II-like and T-cell costimulation and signaling activation related transcripts was significantly associated with poor clinical outcome. Moreover, we identified CD70 as a SOX11 direct target gene, whose overexpression was induced in SOX11+, but not SOX11− tumor cells by CD40L in vitro. CD70 was overexpressed in primary SOX11+ MCL and it was associated with an immune unbalance of the tumor microenvironment characterized by increased number of effector regulatory T (Treg) cell infiltration, higher proliferation, and aggressive clinical course. CD27 was expressed with moderate to strong intensity in 76% of cases. Overall, our results suggest that SOX11 expression in MCL is associated with an immunosuppressive microenvironment characterized by CD70 overexpression in tumor cells, increased Treg cell infiltration and downmodulation of antigen processing, and presentation and T-cell activation that could promote MCL progression and represent a potential target for tailored therapies.

Novel Subtypes Based on Microenvironment Immune Signature Provides Prognostic Stratification and Therapeutic Strategy in Peripheral T-Cell Lymphoma

Peripheral T-cell Lymphoma Not Otherwise Specified (PTCL-NOS) is the largest group of peripheral T-cell lymphomas (PTCL) with dismal prognosis. PTCL-NOS does not correspond to any of the specifically defined entities of T-cell lymphoma in the World Health Organization classification and is referred to as the “waste-basket” category that contains heterogeneous disease entities. Its heterogeneity represents a major obstacle to development of standardized therapeutic strategies, and therefore classification of this “waste-basket” diagnosis is necessary. Although previous studies tried to classify PTCL-NOS from the viewpoint of Cell-of-Origin (COO) of tumor cells by utilizing histopathologic methods or gene expression profiling, the prognostic impact of COO-based stratification still remains controversial. In addition, recent studies revealed the prognostic importance and the therapeutic potential of tumor-microenvironment in various types of cancers. These situations prompted us to investigate microenvironment profile of PTCL-NOS.The aim of this study is to elucidate the microenvironment signatures of PTCL-NOS by gene expression profiling, and to establish a new stratification model. For this purpose, the expression levels of 770 genes related to phenotype and function of tumor T-cells and microenvironment immune cells (B-cells, macrophages (Mø), dendritic cells and granulocytes) were analyzed by employing nCounter system, digital RNA counting technology with high reproducibility for the measurement of even lowly-expressed genes from rare microenvironment components. Sixty-eight patients who were diagnosed as PTCL-NOS between 1993 and 2011 were included in this study. These patients were equally treated by anthracycline-based conventional chemotherapies,and the median survival period was 2.8 years (range, 0.2-11.0 years), which was comparable to historical controls.To explore the classifier genes for PTCL-NOS, principal component analysis was performed and 2 distinct clusters of microenvironment-derived genes, B-cell (CD19, CD20, PAX5) and Mø (CD68, CD163) related genes, were identified (Figure 1). By hierarchical clustering analysis based on the expression levels of these genes, 68 PTCL-NOS patients were classified into three groups as B-cell-rich, Mø-rich and unclassified type (Figure 2). We visually confirmed that CD20+ B-cells and CD163+ Mø were enriched in the tumor tissues of B-cell-rich and Mø-rich type, respectively utilizing multiplexed fluorescent immunostaining (Perkin-Elmer, Mantra quantitative pathology workstation). More importantly, this microenvironment-based classification was significantly associated with prognosis: B-cell-rich type patients had better overall survival compared with Mø-rich patients (2-year overall survival: B-cell-rich type, 77.7%; Mø-rich type, 39.3%; others, 55.5%; p = 0.02) (Figure 3). We also validated the impact of these microenvironment signatures on the prognosis by analyzing published microarray datasets of 123 PTCL-NOS patients (GSE58445). Thus, B-cell-rich type patients were expected to respond to conventional chemotherapies and classified as favorable subtype.We further investigated potential therapeutic targets for Mø-rich patients who were classified as unfavorable subtype, and found that gene expression levels of immune checkpoint molecules, such as PD-L1, PD-L2, TIM-3, LAG-3, IDO-1, were up-regulated in the Mø-rich type compared with the others (PD-L1, p < 0.01; PD-L2, p < 0.01; TIM-3, p < 0.01; LAG-3, p = 0.01; IDO-1, p < 0.01). Imaging analysis of Mø-rich type tissues with Mantra system revealed that PD-L1 was highly and exclusively expressed on the CD163+ Mø, suggesting that microenvironment Mø might negatively regulate tumor immunity via expression of immune-checkpoint molecules and that Mø-rich type patients were expected to respond to immunotherapeutic agents including immune checkpoint inhibitors.In conclusion, our findings collectively suggest that tumor microenvironment signature has significant prognostic impact, and novel microenvironment-based stratification model will contribute to the development of optimal therapeutic strategies for PTCL-NOS patients. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Unraveling Immune-Related lncRNAs in Breast Cancer Molecular Subtypes.

Breast cancer (BRCA) is the most leading cause of cancer worldwide. It is a heterogeneous disease with at least five molecular subtypes including luminal A, luminal B, basal-like, HER2-enriched, and normal-like. These five molecular subtypes are usually stratified according to their mRNA profile patterns; however, ncRNAs are increasingly being used for this purpose. Among the ncRNAs class, the long non-coding RNAs (lncRNAs) are molecules with more than 200 nucleotides with versatile regulatory roles; and high tissue-specific expression profiles. The heterogeneity of BRCA can also be reflected regarding tumor microenvironment immune cells composition, which can directly impact a patient’s prognosis and therapy response. Using BRCA immunogenomics data from a previous study, we propose here a bioinformatics approach to include lncRNAs complexity in BRCA molecular and immune subtype. RNA-seq data from The Cancer Genome Atlas (TCGA) BRCA cohort was analyzed, and signal-to-noise ratio metrics were applied to create these subtype-specific signatures. Five immune-related signatures were generated with approximately ten specific lncRNAs, which were then functionally analyzed using GSEA enrichment and survival analysis. We highlighted here some lncRNAs in each subtype. LINC01871 is related to immune response activation and favorable overall survival in basal-like samples; EBLN3P is related to immune response suppression and progression in luminal B, MEG3, XXYLT1-AS2, and LINC02613 were related with immune response activation in luminal A, HER2-enriched and normal-like subtypes, respectively. In this way, we emphasize the need to know better the role of lncRNAs as regulators of immune response to provide new perspectives regarding diagnosis, prognosis and therapeutical targets in BRCA molecular subtypes.

Innate Immune Cells in the Esophageal Tumor Microenvironment

Esophageal cancer (EC) is one of the most common mucosa-associated tumors, and is characterized by aggressiveness, poor prognosis, and unfavorable patient survival rates. As an organ directly exposed to the risk of foodborne infection, the esophageal mucosa harbors distinct populations of innate immune cells, which play vital roles in both maintenance of esophageal homeostasis and immune defense and surveillance during mucosal anti-infection and anti-tumor responses. In this review, we highlight recent progress in research into innate immune cells in the microenvironment of EC, including lymphatic lineages, such as natural killer and γδT cells, and myeloid lineages, including macrophages, dendritic cells, neutrophils, myeloid-derived suppressor cells, mast cells and eosinophils. Further, putative innate immune cellular and molecular mechanisms involved in tumor occurrence and progression are discussed, to highlight potential directions for the development of new biomarkers and effective intervention targets, which can hopefully be applied in long-term multilevel clinical EC treatment. Fully understanding the innate immunological mechanisms involved in esophageal mucosa carcinogenesis is of great significance for clinical immunotherapy and prognosis prediction for patients with EC.

Autophagy-Mediated Exosomes as Immunomodulators of Natural Killer Cells in Pancreatic Cancer Microenvironment.

Pancreas ductal adenocarcinoma is a highly aggressive cancer with an incredible poor lifespan. Different chemotherapeutic agents’ schemes have been tested along the years without significant success. Furthermore, immunotherapy also fails to cope with the disease, even in combination with other standard approaches. Autophagy stands out as a chemoresistance mechanism and is also becoming relevant as responsible for the inefficacy of immunotherapy. In this complex scenario, exosomes have emerged as a new key player in tumor environment. Exosomes act as messengers among tumor cells, including tumor microenvironment immune cells. For instance, tumor-derived exosomes are capable of generating a tolerogenic microenvironment, which in turns conditions the immune system behavior. But also, immune cells-derived exosomes, under non-tolerogenic conditions, induce tumor suppression, although they are able to promote chemoresistance. In that way, NK cells are well known key regulators of carcinogenesis and the inhibition of their function is detrimental for tumor suppression. Additionally, increasing evidence suggests a crosstalk between exosome biogenesis and the autophagy pathway. This mini review has the intention to summarize the available data in the complex relationships between the autophagy pathway and the broad spectrum of exosomes subpopulations in pancreatic cancer, with focus on the NK cells response.

Susceptibility for Some Infectious Diseases in Patients With Diabetes: The Key Role of Glycemia.

Uncontrolled diabetes results in several metabolic alterations including hyperglycemia. Indeed, several preclinical and clinical studies have suggested that this condition may induce susceptibility and the development of more aggressive infectious diseases, especially those caused by some bacteria (including Chlamydophila pneumoniae, Haemophilus influenzae, and Streptococcus pneumoniae, among others) and viruses [such as coronavirus 2 (CoV2), Influenza A virus, Hepatitis B, etc.]. Although the precise mechanisms that link glycemia to the exacerbated infections remain elusive, hyperglycemia is known to induce a wide array of changes in the immune system activity, including alterations in: (i) the microenvironment of immune cells (e.g., pH, blood viscosity and other biochemical parameters); (ii) the supply of energy to infectious bacteria; (iii) the inflammatory response; and (iv) oxidative stress as a result of bacterial proliferative metabolism. Consistent with this evidence, some bacterial infections are typical (and/or have a worse prognosis) in patients with hypercaloric diets and a stressful lifestyle (conditions that promote hyperglycemic episodes). On this basis, the present review is particularly focused on: (i) the role of diabetes in the development of some bacterial and viral infections by analyzing preclinical and clinical findings; (ii) discussing the possible mechanisms by which hyperglycemia may increase the susceptibility for developing infections; and (iii) further understanding the impact of hyperglycemia on the immune system.