Group 1 Innate Lymphocyte-Derived IFN-γ Regulates Macrophage Alternative Activation In Colon Cancer

Background: Tumor-associated macrophage (TAM) is an important innate immune cell-subset in tumor microenvironment, and that is also a pivotal orchestrator of tumor-promoting inflammation and tumor progression. Evidence proved that TAMs are up-regulated in a great number of cancers, and most of them are alternative activated M2 phenotype, which greatly promote the progress of cancer diseases. Group 1 innate lymphocytes including conventional NK cells and type 1 innate lymphocytes (ILC1s), are abundant in intestinal tissue, and characterized by expressing transcription factor T-bet and secreting interferon (IFN)-γ, which can promote the macrophage to classically activated anti-tumor M1 phenotype. However, the relationship between these two cell subsets remains unclear in colon cancer. Methods: Flow cytometry was used to detect the percentage of M1 phenotype macrophage, M2 phenotype macrophage and group 1 innate lymphocytes in colon cancer tissue and paracancer healthy colon tissue of AOM/DSS-induced colon cancer mice model. In vitro isolating group 1 innate lymphocytes and inducing bone marrow-derived macrophage to detect the cross-talk when co-cultured. Adoptively transfer or blocking group 1 innate lymphocytes in vivo to investigate the role of group 1 innate lymphocytes on tumor-infiltrating macrophage and the tumor growth. Results: We found that M1 phenotype macrophage and group 1 innate lymphocytes were down-regulated in colon cancer tissue, and they were positively correlated. Group 1 innate lymphocytes promoted macrophage to classically activated M1 phenotype in vitro, and that could be blocked by anti-IFN-γ. In vivo results showed that the administration of group 1 innate lymphocytes-blocking antibody anti-NK1.1 could decrease the number of M1 phenotype macrophage in tumor tissue of MC38 tumor-bearing mice and promote the tumor growth, while adoptively transferring group 1 innate lymphocytes led to tumor-inhibiting and level of M1 phenotype macrophage up-regulating in MC38 tumor-bearing mice. Conclusions: Our studies preliminarily prove that group 1 innate lymphocytes promote the alternative activation of M1 macrophage by secreting IFN-γ to inhibit the progress of colon cancer for the first time, which may provide an insight in the immunotherapy of colon cancer.

The Role of EREG/EGFR Pathway in Tumor Progression

Aberrant activation of the epidermal growth factor receptor (EGFR/ERBB1) by erythroblastic leukemia viral oncogene homolog (ERBB) ligands contributes to various tumor malignancies, including lung cancer and colorectal cancer (CRC). Epiregulin (EREG) is one of the EGFR ligands and is low expressed in most normal tissues. Elevated EREG in various cancers mainly activates EGFR signaling pathways and promotes cancer progression. Notably, a higher EREG expression level in CRC with wild-type Kirsten rat sarcoma viral oncogene homolog (KRAS) is related to better efficacy of therapeutic treatment. By contrast, the resistance of anti-EGFR therapy in CRC was driven by low EREG expression, aberrant genetic mutation and signal pathway alterations. Additionally, EREG overexpression in non-small cell lung cancer (NSCLC) is anticipated to be a therapeutic target for EGFR-tyrosine kinase inhibitor (EGFR-TKI). However, recent findings indicate that EREG derived from macrophages promotes NSCLC cell resistance to EGFR-TKI treatment. The emerging events of EREG-mediated tumor promotion signals are generated by autocrine and paracrine loops that arise from tumor epithelial cells, fibroblasts, and macrophages in the tumor microenvironment (TME). The TME is a crucial element for the development of various cancer types and drug resistance. The regulation of EREG/EGFR pathways depends on distinct oncogenic driver mutations and cell contexts that allows specific pharmacological targeting alone or combinational treatment for tailored therapy. Novel strategies targeting EREG/EGFR, tumor-associated macrophages, and alternative activation oncoproteins are under development or undergoing clinical trials. In this review, we summarize the clinical outcomes of EREG expression and the interaction of this ligand in the TME. The EREG/EGFR pathway may be a potential target and may be combined with other driver mutation targets to combat specific cancers.

Mechanisms Involved in Type II Macrophage Activation and Effector Functions

<p>Autoimmunities are extremely difficult to treat and involved in their pathogenesis are pro-inflammatory immune responses redirected against one's own tissues. Studies in our lab have shown macrophages that are induced to become type II macrophages protect against an animal model of MS, experimental autoimmune encephalomyelitis (EAE), with protection due to immune deviation. Another way to deviate immune responses away from inflammation is by infection with the parasitic helminth Schistosoma mansoni, which also protects against EAE. The contribution of type II macrophages in this protection is unknown, as are the mechanisms involved in promoting the phenotype induced by type II activation. This project investigates key mechanisms involved in type II activation, while also elucidating the possible effect of schistosome exposure on the induction of this activation state. Using a validated model of type II activation in vitro, we compared the effects of schistosome immune complexes on various macrophage properties such as cytokine, surface marker and enzymatic profiles. This thesis identified that exposure to schistosome complexes induces a macrophage state with characteristics of two distinct activation states (type II and alternative activation), as well as completely novel characteristics. This activation state shows many phenotypic properties associated with immune regulation, and may have important consequences for understanding mechanisms involved in protection against inflammatory illnesses. We also investigated key mechanisms involved in the anti-inflammatory responses induced by type II activation. Cytokine, chemokine and surface marker profiles of macrophages were assessed in response to type II activation in vitro, with the main emphasis on determining the effects of IL-10 and CD40 on the type II activation phenotype and function. This investigation found that type II activated macrophages depend on low levels of CD40/CD40L signalling to polarise Th2 development, as the expression of receptors for Th2-inducing cytokines are significantly impaired in the absence of this interaction. This suggests an important role for the low but maintained levels of CD40 on type II activated macrophages, in aiding the deviation of immune responses, while maintaining Th2 polarization. We also suggest a suppressive role of CD40/CD40L in IL-10 production, which is a novel find. The requirement of new treatments for MS is escalating as more people are affected each year. The impact of MS on the quality of life is severe and long lasting. Having a greater understanding of the mechanisms involved in deviating pro-inflammatory or anti-inflammatory responses will enable the development of much more effective treatments and therapies in the future.</p>

Mechanisms Involved in Type II Macrophage Activation and Effector Functions

<p>Autoimmunities are extremely difficult to treat and involved in their pathogenesis are pro-inflammatory immune responses redirected against one's own tissues. Studies in our lab have shown macrophages that are induced to become type II macrophages protect against an animal model of MS, experimental autoimmune encephalomyelitis (EAE), with protection due to immune deviation. Another way to deviate immune responses away from inflammation is by infection with the parasitic helminth Schistosoma mansoni, which also protects against EAE. The contribution of type II macrophages in this protection is unknown, as are the mechanisms involved in promoting the phenotype induced by type II activation. This project investigates key mechanisms involved in type II activation, while also elucidating the possible effect of schistosome exposure on the induction of this activation state. Using a validated model of type II activation in vitro, we compared the effects of schistosome immune complexes on various macrophage properties such as cytokine, surface marker and enzymatic profiles. This thesis identified that exposure to schistosome complexes induces a macrophage state with characteristics of two distinct activation states (type II and alternative activation), as well as completely novel characteristics. This activation state shows many phenotypic properties associated with immune regulation, and may have important consequences for understanding mechanisms involved in protection against inflammatory illnesses. We also investigated key mechanisms involved in the anti-inflammatory responses induced by type II activation. Cytokine, chemokine and surface marker profiles of macrophages were assessed in response to type II activation in vitro, with the main emphasis on determining the effects of IL-10 and CD40 on the type II activation phenotype and function. This investigation found that type II activated macrophages depend on low levels of CD40/CD40L signalling to polarise Th2 development, as the expression of receptors for Th2-inducing cytokines are significantly impaired in the absence of this interaction. This suggests an important role for the low but maintained levels of CD40 on type II activated macrophages, in aiding the deviation of immune responses, while maintaining Th2 polarization. We also suggest a suppressive role of CD40/CD40L in IL-10 production, which is a novel find. The requirement of new treatments for MS is escalating as more people are affected each year. The impact of MS on the quality of life is severe and long lasting. Having a greater understanding of the mechanisms involved in deviating pro-inflammatory or anti-inflammatory responses will enable the development of much more effective treatments and therapies in the future.</p>

Dual Target of EGFR and mTOR Suppress Triple Negative Breast Cancer Cell Growth Via Regulation The Phosphorylation of mTOR Downstream Proteins

Background：Triple-negative breast cancers (TNBC) are the most aggressive subtype of breast cancer, accounting for 15% - 20% of all cases, and have no response to available hormonal therapies and anti-HER2-targeted therapies due to the absence of corresponding targets. Over half of TNBC patients have overexpressed EGFR, but they are insensitive to EGFR inhibitors from monotherapy. Mammalian target of rapamycin (mTOR) connected with EGFR in the downstream signaling and involved in the progress of TNBC. The purpose of this study is to determine the combined effect of everolimus and geftinib in a TNBC cell model and investigate the possible mechanism. Results: This work showed the expression EGFR and p-mTOR protein in TNBC tissues were significantly higher than that in non-TNBC(p<0.05), while the expression of mTOR, S6K1, pEGFR and p-S6K1 were significantly higher in the EGF stimulation. EGFR and p-mTOR protein are related to poor prognosis. EGFR inhibitor gefitinib and mTOR inhibitor everolimus significantly inhibited the proliferation of human triple-negative breast cancer MDA-MB-468 cells and arrested cells in G0/G1 phase when applied separately and in combination in a dose-dependent manner (P<0.05). Meanwhile, the rate of apoptosis of MDA-MB-468 cells was significantly incresased separately by two drugs (P<0.01). Furthermore, the combination of everolimus and geftinib reduced the phosphorylation of mTOR downstream proteins. Instead, the phosphorylation of 4E-BP1 was enhanced after the everolimus and geftinib treatment, indicated an alternative activation pattern. Conclusions: These results suggested that dual inhibition of mTOR and EGFR could be a promising approach to treat TNBC.

Classical Dichotomy of Macrophages and Alternative Activation Models Proposed with Technological Progress

Macrophages are important immune cells that participate in the regulation of inflammation in implant dentistry, and their activation/polarization state is considered to be the basis for their functions. The classic dichotomy activation model is commonly accepted, however, due to the discovery of macrophage heterogeneity and more functional and iconic exploration at different technologies; some studies have discovered the shortcomings of the dichotomy model and have put forward the concept of alternative activation models through the application of advanced technologies such as cytometry by time-of-flight (CyTOF), single-cell RNA-seq (scRNA-seq), and hyperspectral image (HSI). These alternative models have great potential to help macrophages divide phenotypes and functional genes.

CNN Variants for Computer Vision: History, Architecture, Application, Challenges and Future Scope

Computer vision is becoming an increasingly trendy word in the area of image processing. With the emergence of computer vision applications, there is a significant demand to recognize objects automatically. Deep CNN (convolution neural network) has benefited the computer vision community by producing excellent results in video processing, object recognition, picture classification and segmentation, natural language processing, speech recognition, and many other fields. Furthermore, the introduction of large amounts of data and readily available hardware has opened new avenues for CNN study. Several inspirational concepts for the progress of CNN have been investigated, including alternative activation functions, regularization, parameter optimization, and architectural advances. Furthermore, achieving innovations in architecture results in a tremendous enhancement in the capacity of the deep CNN. Significant emphasis has been given to leveraging channel and spatial information, with a depth of architecture and information processing via multi-path. This survey paper focuses mainly on the primary taxonomy and newly released deep CNN architectures, and it divides numerous recent developments in CNN architectures into eight groups. Spatial exploitation, multi-path, depth, breadth, dimension, channel boosting, feature-map exploitation, and attention-based CNN are the eight categories. The main contribution of this manuscript is in comparing various architectural evolutions in CNN by its architectural change, strengths, and weaknesses. Besides, it also includes an explanation of the CNN’s components, the strengths and weaknesses of various CNN variants, research gap or open challenges, CNN applications, and the future research direction.