Cancer Immune Therapy Research Articles

Mechanisms of Anti-PD Therapy Resistance in Digestive System Neoplasms.

Digestive system neoplasms are highly heterogeneous and exhibit complex resistance mechanisms that render anti-programmed cell death protein (PD) therapies poorly effective. The tumor microenvironment (TME) plays a pivotal role in tumor development, apart from supplying energy for tumor proliferation and impeding the body's anti-tumor immune response, the TME actively facilitates tumor progression and immune escape via diverse pathways, which include the modulation of heritable gene expression alterations and the intricate interplay with the gut microbiota. In this review, we aim to elucidate the mechanisms underlying drug resistance in digestive tumors, focusing on immune-mediated resistance, microbial crosstalk, metabolism, and epigenetics. We will highlight the unique characteristics of each digestive tumor and emphasize the significance of the tumor immune microenvironment (TIME). Furthermore, we will discuss the current therapeutic strategies that hold promise for combination with cancer immune normalization therapies. This review aims to provide a thorough understanding of the resistance mechanisms in digestive tumors and offer insights into potential therapeutic interventions.

Current Understanding of PCSK9 and Its Relevance to Cancer Prognosis and Immune Therapy: A Review.

The effectiveness of immunotherapy for most cancer patients remains low, with approximately 10-30% of those treated surviving. Thus, much effort is being put into finding new ways to improve immune checkpoint therapy. Our review concludes that inhibition of proprotein convertase subtilisin/Kexin type 9 (PCSK9), which plays a critical role in regulating cholesterol metabolism, can cause movement of T cells toward tumors, with increased sensitivity to immune checkpoint therapies. We searched PubMed, NCBI, Scopus, and Google Scholar for the published articles without limitations on publication dates. We used the following terms: "PCSK9", "Cancer", "Immune Checkpoint", and "Cancer Prognosis" in the title and/or abstract. Our search initially revealed 600 records on the subject and stored them in the used databases under EndNote X8 management software. A total of 161 articles were selected and through a careful review, 76 were included in our research. We concluded that PCSK9 reduces the number of LDL receptors (LDL-R) on the cell surface, which is linked to its ability to regulate cholesterol levels in the body. Also, we discuss how suppressing PCSK9 leads to the MHC-1 accumulation on the surface of cancer cells, which results in T lymphocyte invasion. Finally, we believe that inhibiting PCSK9 may be an effective strategy for improving cancer immunotherapy.

Investigating the role of CD47 gene regulation in Iraqi cancer patients: a promising prognostic factor

Background: Innate immune checkpoints have been one of the most studied and promising approaches in cancer immune therapy. Therefore, it is considered valuable when investigating these checkpoints at the gene expression and protein levels. However, this approach still has variations from study to study due to various types of cancer. Therefore, it is hypothesized that CD47 could be one of those checkpoints that affect the survival of cancer cells. CD47 is an integrin like marker that has been identified for its role as “do not eat me signal” which enhances the ability of cancer cells to escape from being phagocytized by macrophages. Aim of the study: This study aims to investigate CD47 expression in ten solid tumors and five blood cancers at the gene level and at the protein level.Methods: Clinical specimens were collected from patients with different cancers attending Kirkuk oncology center. The patients were categorized into two main groups: solid tumor group and blood-derived cancer group together with their control counterparts. CD47 gene expression was investigated using q RT-PCR techniques while its protein levels were investigated using ELISA techniques.Results: The results showed a significant up-regulation of CD47 gene expression in gastric cancer, cholangiocarcinoma; CCA (bile duct), chronic myeloid leukemia (CML), and breast cancer, respectively. These results were also confirmed at the protein level, suggesting that those types of malignancy are more susceptible to an increase in CD47 expression than other types of cancer that were involved in this study. Conclusion: CD47 tends to be over-expressed in solid tumors and blood cancers, and could be used as diagnostic and prognostic markers, especially for solid tumors.

Somatic and Germline HLA Determinants of Immune Surveillance and Escape in Myelodysplastic Syndromes

While the contribution of canonical myeloid genetic drivers to myelodysplastic syndromes/neoplasms (MDS) pathobiology is well established, other genetic events operative under special circumstances, and by itself not constituting intrinsic drivers, may affect the disease course. These include genetic defects conveying resistance to specific therapies, clonally restricted DNA repair alterations favoring the accumulation of additional mutations, and/or lesions facilitating the resistance to inflammation or immune escape. Here, we focus on this latter aspect as it may shed light on the pressure exerted by immune tumor surveillance or acquired resistance to cancer immune therapies, a field that has lagged in myeloid neoplasia as compared to the dramatic changes obtained in solid tumors. We hypothesize that during MDS ontogeny and leukemia evolution a spectrum of immunological mechanisms may shape the oncogenic landscape and facilitate the emergence of clones with diverse immunogenic potential as targets, leading to selective immunotherapeutic liabilities. We accrued a clinically and molecularly well annotated cohort of 101 MDS patients at various stages of the disease spectrum [N=51 low risk (LR)-MDS, N=32 high risk (HR)-MDS, N=18 secondary AML-sAML)]. This cohort included specific clinical sub-entities wherein the contribution of immune forces appears to be a prominent feature i.e., hypocellular MDS (hMDS, N=9) or MDS with co-occurring T large granular lymphocytosis (LGL) (N=5). By combining human leukocyte antigen (HLA) and T cell receptor (TCR) immunosequencing at disease onset, we leveraged our bioinformatics expertise to impute HLA mutations and losses, HLA evolutionary divergence (HED), and characteristics of T cell repertoires. The median age of the whole cohort was 71.2 years (IQR: 63.4-77.7), and median bone marrow (BM) blast count was 3% (1-11%). Cytogenetics revealed normal karyotype in 41 patients (40%), while complex karyotype was observed in 21 (20%) and partial or complete loss of chromosome 7 in 10 cases (10%). Comparisons with a cohort of 784 ethnicity-matched healthy individuals (HC), showed no differences in terms of locus-specific HED configurations. However, when HED scores were analyzed across the disease spectrum, we observed a heterogeneous distribution in MDS patients for the HLA-C locus, with higher values in hypoplastic and LGL-associated MDS as compared to HR diseases (p= 0.0068) and sAML (p= 0.0409). The screening for immune evasion highlighted the presence of both somatic mutations and losses in the HLA region in 14 patients, with both type of alterations found in 1 case. Of those, 6 patients (20% LGL-MDS, 5% LR-MDS and 17% sAML) harbored HLA mutations (in HLA-A, B, C and DPB1 loci), whereas HLA losses were observed in 9 patients (22% hMDS, 5% LR-MDS, 10% HR MDS, 11% sAML) almost exclusively in class II alleles (chiefly DPB1, DRB1 and DPA1). When exploring the degree of immunocompetence by TCR immunosequencing as a proxy, we found a higher degree of diversity (imputed via inverse Simpson index) as compared to healthy controls (p=1.5e-07), possibly in relationship to diverse T cell responses across the disease spectrum. However, no difference between MDS sub-entities was found. Of interest is that the maximal productive frequency (an indicator of the degree of T cell expansion) was higher in patients with hMDS and LGL-associated MDS as compared to HR-MDS (p=0.0079). Finally, inter-sample comparisons of shared clonotypes showed a high degree of overlap with regards to TCR repertoires among MDS as compared to healthy subjects. Our data emphasize the adaptive immune-molecular network underpinning MDS development. HLA constellations and their interactions with TCR repertoires may create selective immune pressure. Somatic HLA aberrations may serve as a marker of ongoing immune pressure and correspond to oligoclonal T-cell expansions and variations in TCR diversity. While virtually no HLA mutations have been previously found in AML/MDS, we show a relatively higher frequency of HLA aberrations in hMDS and MDS-LGL, immunologically stressed scenarios reminiscent of aplastic anemia and post-allogenic transplant AML/MDS cases, whereby respectively 36% 1 and 38% 2 of patients harbored HLA lesions as a mean of escape from cytotoxic lymphocytes or allogeneic Graft-vs-Leukemia reactions.

Tumor-intrinsic RGS1 potentiates checkpoint blockade response via ATF3-IFNGR1 axis

ABSTRACT Background Non-responsiveness is a major barrier in current cancer immune checkpoint blockade therapies, and the mechanism has not been elucidated yet. Therefore, it is necessary to discover the mechanism and biomarkers of tumor immunotherapeutic resistance. Methods Bioinformatics analysis was performed based on CD8+ T cell infiltration in multiple tumor databases to screen out genes related to anti-tumor immunity. Associations between Regulator of G-protein signaling 1 (RGS1) and IFNγ-STAT1 signaling, and MHCI antigen presentation pathway were examined by RT-qPCR, western blotting, and flow cytometry. The modulatory mechanisms of RGS1 were investigated via CHIP-qPCR and dual-luciferase assay. The clinical and therapeutic implications of RGS1 were comprehensively investigated using tumor cell lines, mouse models, and clinical samples receiving immunotherapy. Results RGS1 was identified as the highest gene positively correlated with immunogenicity among RGS family. Inhibition of RGS1 in neoplastic cells dampened anti-tumor immune response and elicited resistance to immunotherapy in both renal and lung murine subcutaneous tumors. Mechanistically, RGS1 enhanced the binding of activating transcription factor 3 (ATF3) to the promoter of interferon gamma receptor 1 (IFNGR1), activated STAT1 and the subsequent expression of IFNγ-inducible genes, especially CXCL9 and MHC class I (MHCI), thereby influenced CD8+ T cell infiltration and antigen presentation and processing. Clinically, lower expression level of RGS1 was associated with resistance of PD1 inhibition therapy and shortened progression-free survival among 21 NSCLC patients receiving immunotherapy. Conclusions Together, these findings uncover a novel mechanism that elicits immunotherapy resistance and highlight the function of tumor-intrinsic RGS1, which brings new insights for future strategies to sensitize anti-PD1 immunotherapy.

Discrepancies in tumor mutation burden reporting from sequential endobronchial ultrasound transbronchial needle aspiration samples within single lymph node stations - brief report.

Tumour Mutation Burden (TMB) is a potential biomarker for immune cancer therapies. Here we investigated parameters that might affect TMB using duplicate cytology smears obtained from endobronchial ultrasound transbronchial needle aspiration (EBUS TBNA)-sampled malignant lymph nodes. Individual Diff-Quik cytology smears were prepared for each needle pass. DNA extracted from each smear underwent sequencing using large gene panel (TruSight Oncology 500 (TSO500 - Illumina)). TMB was estimated using the TSO500 Local App v. 2.0 (Illumina). Twenty patients had two or more Diff-Quik smears (total 45 smears) which passed sequencing quality control. Average smear TMB was 8.7 ± 5.0 mutations per megabase (Mb). Sixteen of the 20 patients had paired samples with minimal differences in TMB score (average difference 1.3 ± 0.85). Paired samples from 13 patients had concordant TMB (scores below or above a threshold of 10 mutations/Mb). Markedly discrepant TMB was observed in four cases, with an average difference of 11.3 ± 2.7 mutations/Mb. Factors affecting TMB calling included sample tumour content, the amount of DNA used in sequencing, and bone fide heterogeneity of node tumour between paired samples. TMB assessment is feasible from EBUS-TBNA smears from a single needle pass. Repeated samples of a lymph node station have minimal variation in TMB in most cases. However, this novel data shows how tumour content and minor change in site of node sampling can impact TMB. Further study is needed on whether all node aspirates should be combined in 1 sample, or whether testing independent nodes using smears is needed.

Hyperbaric oxygen augments chemodynamic effect induced by probiotic-derived selenium nanoparticles to enhance cancer immune checkpoint blockade therapy

Hyperbaric oxygen augments chemodynamic effect induced by probiotic-derived selenium nanoparticles to enhance cancer immune checkpoint blockade therapy

Copper-Bacteriochlorin Nanosheet as a Specific Pyroptosis Inducer for Robust Tumor Immunotherapy.

Pyroptosis is increasingly considered a new weathervane in cancer immune therapy. However, triggering specific pyroptotic tumor cell death while preserving normal cells still remains a major challenge. Herein, a brand-new pyroptosis inducer, copper-bacteriochlorin nanosheet (Cu-TBB), is designed. The synthesized Cu-TBB canbe activated to an "on" state in the tumor microenvironment with glutathione (GSH) overexpression, leading to the release of Cu+ and TBB, respectively. Intriguingly, the released Cu+ candrive cascade reactions to produce O2 -• and highly toxic ·OH in cells. Additionally, the released TBB canalso generate O2 -• and 1 O2 upon 750nm laser irradiation. Encouragingly, both Cu+ -driven cascade reactions and photodynamic therapy pathways result in potent pyroptosis along with dendritic cell maturation and T cell priming, thus simultaneously eliminating the primary tumors and inhibiting the distant tumor growth and metastases. Conclusively, the well-designed Cu-TBB nanosheet is shown to trigger specific pyroptosis in vitroandin vivo, leading to enhanced tumor immunogenicity and antitumor efficacy while minimizing systemic side effects.

CD300f signalling induces inhibitory human monocytes/macrophages

The CD300 glycoproteins are a family of related leucocyte surface molecules that regulate the immune response via their paired triggering and inhibitory receptors. Here we studied CD300f, an apoptotic cell receptor, and how it modulates the function of human monocytes and macrophages. We showed that CD300f signalling by crosslinking with anti-CD300f mAb (DCR-2) suppressed monocytes causing upregulation of the inhibitory molecule, CD274 (PD-L1) and their inhibition of T cell proliferation. Furthermore, CD300f signalling drove macrophages preferentially towards M2-type with upregulation of CD274, which was further enhanced by IL-4. CD300f signalling activates the PI3K/Akt pathway in monocytes. Inhibition of PI3K/Akt signalling resulting from CD300f crosslinking leads to downregulation of CD274 expression on monocytes. These findings highlight the potential use of CD300f blockade in cancer immune therapy to target immune suppressive macrophages in the tumour microenvironment, a known resistance mechanism to PD-1/PD-L1 checkpoint inhibitors.

Uliledlimab and toripalimab combination therapy in treatment naive advanced NSCLC: Phase 1b/2 clinical trial results using CD73 as a potential predictive biomarker.

2570 Background: CD73 is a rate-limiting enzyme in adenosine pathway promoting immune suppression in tumor microenvironment. The role of CD73 as a novel target in cancer immune therapy is further supported by correlation between its high expression in tumors and poor prognosis in cancer pts. Here we evaluate safety and efficacy of uliledlimab, a differentiated CD73 antibody, in combination with toripalimab, a PD-1 antibody, in treatment naive NSCLC pts. In particular, we investigate the correlation between clinical response and tumor CD73 expression to assess its value as a potential predictive biomarker, following a previous study where a correlation was indicated (Robert F, et al. ASCO 2021). Methods: Pts with treatment naïve advanced NSCLC (EGFR/ALK wild type) were enrolled in a dose-expansion cohort and received uliledlimab (20mg/kg or 30mg/kg Q3W) in combination with toripalimab 240mg Q3W. Safety, the primary endpoint, and clinical efficacy including ORR, determined by iRECIST, were evaluated. Baseline CD73 and PD-L1 expression in tumor specimens were measured by IHC. The cut-off value for CD73High expression was assessed by ROC analysis for correlation with ORR. Results: As of 12/2/22, 66 pts were enrolled (median age 62.5 years, ECOG PS 0 40.6%, male 84.8% and never smokers 22.7%) and received at least one dose. Treatment-related adverse events (TRAEs) occurred in 86.4% of pts (Gr≥3, 15.2%) with no grade 5 TRAE. One patient was discontinued due to TRAE (1.5%). Efficacy was analyzed in 48 pts who were enrolled at least 90 days prior to data cut-off, allowing for two assessments. Baseline PD-L1 and CD73 expression was available in 45 pts. 40% of tumor or immune cells with ≥1+ staining intensity by IHC was determined as a preliminary cut-off for CD73High (the area under ROC curve =0.72, p = 0.02). While ORR and DCR were 31.3% (15/48) and 79.2% (38/48) respectively in all pts, ORR appeared higher in CD73High pts (50%, 9/18) and much lower in CD73Low pts (14.8%, 4/27). Highest ORR was found in a cohort with CD73High and PD-L1 TPS≥1% (57.1%, 8/14). Conclusions: Uliledlimab and toripalimab combination therapy was well-tolerated in treatment naïve advanced NSCLC pts. Clinical response of the therapy seems to correlate with high tumor CD73 expression, indicating a potential value of CD73 as a predictive biomarker. The study warrants a biomarker-guided randomized clinical trial that is currently in preparation. Clinical trial information: NCT04322006 .

Improving Access to Therapy for Patients With Inborn Errors of Immunity: A Call to Action.

Breakthroughs in sequencing technology, targeted immunotherapy, and immune reconstituting treatment have increased the pool of patients with inborn errors of immunity, requiring expertise from clinical immunologists. A growing category of immunodeficiency, presenting as primary immune regulatory disorder and secondary immunodeficiency due to targeted immune therapy for cancer and autoimmunity, has added to the growing burden of patients needing access to immune-supportive therapy. The confluence of a growing population of patients needing a clinical immunologist, complex payer structures, and inadequate health care representation will exacerbate current problems with access to therapy. Patients, health care providers, researchers, public and private payers, and industry must come together to find solutions to improve access to therapy. In this article, we reviewed the major topics regarding access to therapy for patients with immunodeficiency.

Prediction of cancer treatment response from histopathology images through imputed transcriptomics.

1551 Background: In recent years, the use of tumour molecular profiling within the clinic has allowed for more accurate cancer diagnostics, as well as the delivery of precision oncology. Rapid advances in digital histopathology have allowed the extraction of clinically relevant information embedded in tumor slides by applying machine learning methods, capitalizing on recent advancements in image analysis via deep learning. However, as in many supervised learning approaches, predicting response to therapy using whole slide images (WSI) of tissue stained with hematoxylin and eosin (H&E) requires large datasets comprising matched imaging and response data, severely restricting the applicability of this approach. Methods: To overcome this critical challenge, we introduce here for the first time a generic methodology for generating WSI-based predictors of patients’ response for a broad range of cancer types and therapies, which does not require matched WSI and response data for training. The approach, termed ENLIGHT-DeepPT, consists of: (1) DeepPT, a new deep-learning framework that predicts exome-wide tumor mRNA expression from slides, and (2) ENLIGHT, a recently published response prediction algorithm, applied here to predict response based on the DeepPT predicted expression values, instead of measured values. Results: First, we study the ability to predict tumor expression, showing that DeepPT successfully predicts transcriptomics in all 16 TCGA cohorts tested and generalizes well to two independent datasets. Second, aligned with our key aim, we analyze six independent clinical trial datasets of patients with different cancer types that were treated with various targeted and immune therapies, on which DeepPT was never trained. We show that ENLIGHT, without any adaptation, can successfully predict the true responders from the expression values imputed by DeepPT, using only H&E images: ENLIGHT-DeepPT successfully predicts true responders in these cohorts with an overall odds ratio of 2.19, increasing the baseline response rate by 38.8% on average among predicted responders. Conclusions: ENLIGHT-DeepPT is the first approach to successfully predict response to multiple targeted and immune cancer therapies directly from H&E slides, without requiring any drug-specific training data. ENLIGHT-DeepPT can provide rapid treatment recommendations without requiring tumor sequencing, bringing precision oncology to low-income countries and other situations where NGS is prohibitive.

Altering Landscape of Cancer Vaccines: Unique Platforms, Research on Therapeutic Applications and Recent Patents.

Recent developments in several areas are rekindling interest and empowering progress in improving therapeutic cancer vaccines. These advances have been made in target selection, vaccine technology, and approaches for reversing the immunosuppressive mechanisms exploited by cancers. Studies on diverse tumor antigens have revealed target properties, including high cell specificity and adequate immunogenicity, to affect clinical efficiency. Therefore, one of the principal goals of cancer vaccinology is the development of efficient therapeutic cancer vaccines that are capable of eliciting an effector as well as memory T cell response specific to tumor antigens. Neoantigens, which arise from mutated proteins in cancer cells, are cancer-specific and may be highly immunogenic. However, the vast majority of these are distinctive to each patient's cancer and hence require the development of personalised therapies. Novel immunotherapeutic strategies are focused on breaking immune tolerance to tumor antigens, improving the immunogenicity of tumor vaccines, in addition to overcoming mechanisms of tumor escape. However, current developments and patents in cancer immune therapies, together with associated technologies, are significant. Supreme achievements in immune inhibitor-centered therapies and neo-antigen identification tools envisage probable improvements in cancer vaccines with respect to treatments of malignancies. Antibody modulation of T cell function through checkpoint blockade or co-stimulatory activation may restore survival, proliferation, and effector function. Thus, these tumor-infiltrating T cells with genetically engineered therapeutic vaccines have the potential to act as curative cancer immune therapeutics. In this review, target tumor antigens employed in recent years in the development of therapeutic cancer vaccine approaches are described, and the patents involved in the process are highlighted.

Applications of intelligent biomaterials in cancer immunotherapy: A review

Intelligent biomaterials can adapt to the changingphysiological parameters and external stressors, and they tend to provide an influence on many facets of medical technology. They have the ability to accelerate promising medicines and enhance the management of chronic ailments. They find utilization in the domains of regenerative medicine, drug delivery systems, medical gadgets, and immunotherapy. Cancer Immunotherapies, which uses a recipient’s immune-system to combat malignancy, is currently reinventing the domain of cancer treatment. Even though aiming T-cells has demonstrated to be a technologically effective restorative strategic approach for the majority of cancer immunotherapies, just a small number of people benefit from such initiatives. Recently, there has been a growing interest in studying the largely untapped innate immunity as a potential treatmentintervention tool. Via gene editing, metabolic labelling, synthetic complexation, lipophilic incorporation, and some other methods, sensibly proposed practical biomaterials have been employed to fabricateartificial functionalities on the cellular surface to provide an increased immunity against cancer and reduced toxicological effects. The biomaterials can be utilized in providing cell and/or tissue explicit better-refined modulation in immunity of a cancer patient. Thus, in the current review are discussing the advances in immunoengineering employing bio-materials and their implications in supporting specific procedures of cancer immune cell therapy, paying special emphasis on antibody-, cell-, immunomodulator-, and gene-based immunity concerned treatment strategies and their conjunction with pharmacological treatments. The limitations and prospects of using biomaterials design technologies in the creation of potential cancer immunotherapies are highlighted.

Liposomal ATM siRNA delivery for enhancing triple-negaitive breast cancer immune checkpoint blockade therapy.

Triple-negative breast cancer (TNBC), which accounts for 10%-20% of breast cancer cases, is characterized by a higher metastasis rate, higher recurrence risk, and worse prognosis. Traditional treatments such as chemotherapy, surgery, and radiotherapy have limited therapeutic effects. Although immune checkpoint blockade (ICB) therapy represented by anti-programmed death 1 (aPD-1) antibody has made further progress in treating TNBC, its therapeutic effect is still not optimistic. Ataxia telangiectasia mutated (ATM) is a critical factor in the DNA damage response (DDR) pathway, which is associated with the development of tumors. Recent studies have found that it can regulate the tumor immune microenvironment, affecting ICB responsiveness. Inhibition of ATM could enhance ICB therapy by promoting mitochondrial DNA cytoplasmic leakage and activating the innate immune signaling pathway. To explore the effect of ATM siRNA(siATM) on the ICB responsiveness of TNBC, we designed and synthesized nanoparticles using 1,2-dioleoyl-glycero-3-phosphatidylcholine (DOPC) liposomes to deliver siATM. In vitro and in vivo experiments demonstrated that DOPC/siATM could enhance the ability of siRNA to enter tumor cells and effectively inhibit the expression of ATM protein. Our study found that nanoparticles carrying siATM could activate cytotoxic T lymphocytes and regulate the immunosuppressive tumor microenvironment (ITM) by activating the cGAS-STING pathway. Its combination with aPD-1 may be a potential way to improve the efficacy of TNBC.

Abstract 301: CENP-E inhibitors activate cGAS-STING pathway by inducing chromosome misalignment and micronucleation after mitotic slippage

Abstract Background: cGAS-STING, which monitors cytosolic DNA and triggers innate immune response, has recently attracted attention as a molecular target to sensitize cancer immune therapies. Since DNAs enclosed in micronucleus frequently leak into cytoplasm, micronucleus generated by centromere-associated protein-E (CENP-E) inhibition is expected to activate cGAS-STING pathways. We validated that small-molecule CENP-E inhibitors profoundly generate micronucleus through misaligned chromosomes. The aim of this study is to elucidate the possibility that micronucleus formation by CENP-E inhibitors activates the cGAS-STING pathway in cancer cells and induces immunological conversion of the tumor microenvironment from cold to hot. Material and Methods: GSK923295 and Cmpd-A were used as CENP-E inhibitors. HeLa and A549 cells were treated with the CENP-E inhibitors at the indicated concentrations. RNA sequencing data were subjected to gene set enrichment analysis (GSEA) to determine the signaling pathways modulated by Cmpd-A treatment compared to DMSO (control). Reporter assays for IRF and NF-kB were performed in A549 dual-reporter cells. High-throughput screening on IRF3 reporter activity was performed in A549 dual reporter cells in combination with -1,400 kinase inhibitor library and GSK923295. Results: The CENP-E inhibitors, GSK-923295 and Cmpd-A, induced chromosome misalignment, resulting in micronucleus formation after mitotic slippage. Phosphorylations of TBK1 and IRF3, cGAS-STING pathway markers, were remarkably upregulated in Cmpd-A-treated cells concordantly with micronucleus formation. The reporter assays also demonstrated that both IRF and NF-kB pathways were significantly activated in Cmpd-A-treated cells in a time-dependent manner. Transcriptome analysis of GSEA demonstrated that the gene ontologies (GO) for innate immune pathways, such as cytosolic DNA sensing pathway and cytokine-cytokine receptor interaction pathway, were significantly enriched in Cmpd-A-treated cells. The kinase library screening revealed that a series of inhibitors targeting PI3K-AKT-mTOR signaling pathways intensively suppressed CENP-E inhibitor-induced IRF activation, which occupied ~40% of hit compounds. Conclusions: The CENP-E inhibitors generated micronucleus activating the cGAS-STING pathway in multiple cancer cell lines. PI3K-AKT-mTOR signaling pathway may play important roles in the CENP-E inhibitor-induced cGAS-STING pathway activation. We are also initiating AI-based drug discovery, aiming to generate novel CENP-E inhibitors by performing virtual screening and generative molecular design. Citation Format: Ryo Kamata, Hitoshi Saito, Yumi Hakozaki, Yukie Kashima, Tomoko Yamamori. Morita, Pinyi Lu, Akihiro Ohashi. CENP-E inhibitors activate cGAS-STING pathway by inducing chromosome misalignment and micronucleation after mitotic slippage [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 301.

Inhibition of PD-L1 and tumor growth in triple-negative breast cancer using a magnetic nanovector with microRNA34a

BackgroundClinical applications of RNA interference for cancer treatment and immune therapy require the development of simultaneous therapy and imaging systems for microRNA. This research was performed to fabricate the miRNA34a-loaded magnetic nanoparticles and investigate its anticancer effects against triple-negative breast cancer (TNBC) in mice model.ResultsUsing two types of polymers to improve their water dispersibility and gene delivery, iron oxide magnetic nanoparticles were prepared for delivery of miRNA34a. The iron oxide magnetic nanoparticles were delivered to TNBC cells, and their efficacy was evaluated in vitro and in vivo. Delivery of miRNA34a reduced TNBC cell migration and decreased the expression of PD-L1 at the mRNA and protein levels. In animal experiments, delivery of miRNA34a reduced tumor growth, and immunostaining and algorithmic analysis confirmed the decrease in PD-L1 expression.ConclusionThis study is the first to modulate PD-L1 by delivering miRNA34a with magnetic nanoparticles, and the results suggest that miRNA34a can be delivered effectively using magnetic nanoparticles and has potential as a molecular imaging contrast agent.

The regulation of N6-methyladenosine modification in PD-L1-induced anti-tumor immunity.

There is growing evidence that programmed death ligand-1 (PD-L1) has exciting therapeutic efficacy in hematological malignancy and partial solid tumors. However, many patients still face failure with the treatment of immune checkpoint blockade because of PD-L1 expression regulation during transcription and post-transcription processes, including N6-methyladenosine (m6A). Similar to the epigenetic regulation in DNA and histones, recent research has revealed the essential regulation of m6A modification in RNA nuclear export, metabolism and translation. Recent studies have shown that m6A-induced PD-L1 expression emerges as one of the main reasons for the immunological alteration in this process and contributes to the failure of Tcell-induced anti-tumor immunity. The results of preclinical studies demonstrate the potential of m6A-targeted therapy in combination with immune checkpoint blockade. The comprehensive expression of m6A-related genes also provided the possibility to indicate the prognosis and to optimize the treatment for patients of various cancer types. In this review, we focus on the m6A modification in PD-L1 mRNA as well as the regulation of PD-L1 expression in cancer cells and summarize its clinical value in anti-PD-L1 cancer immune therapy.

Biological knowledge graph-guided investigation of immune therapy response in cancer with graph neural network.

The determination of transcriptome profiles that mediate immune therapy in cancer remains a major clinical and biological challenge. Despite responses induced by immune-check points inhibitors (ICIs) in diverse tumor types and all the big breakthroughs in cancer immunotherapy, most patients with solid tumors do not respond to ICI therapies. It still remains a big challenge to predict the ICI treatment response. Here, we propose a framework with multiple prior knowledge networks guided for immune checkpoints inhibitors prediction-DeepOmix-ICI (or ICInet for short). ICInet can predict the immune therapy response by leveraging geometric deep learning and prior biological knowledge graphs of gene-gene interactions. Here, we demonstrate more than 600 ICI-treated patients with ICI response data and gene expression profile to apply on ICInet. ICInet was used for ICI therapy responses prediciton across different cancer types-melanoma, gastric cancer and bladder cancer, which includes 7 cohorts from different data sources. ICInet is able to robustly generalize into multiple cancer types. Moreover, the performance of ICInet in those cancer types can outperform other ICI biomarkers in the clinic. Our model [area under the curve (AUC = 0.85)] generally outperformed other measures, including tumor mutational burden (AUC = 0.62) and programmed cell death ligand-1 score (AUC = 0.74). Therefore, our study presents a prior-knowledge guided deep learning method to effectively select immunotherapy-response-associated biomarkers, thereby improving the prediction of immunotherapy response for precision oncology.

Cytotoxicity of Human Hepatic Intrasinusoidal Gamma/Delta T Cells Depends on Phospho-antigen and NK Receptor Signaling.

We previously showed that human hepatic intrasinusoidal (HI) natural killer (NK) T cells selectively eliminate hepatocellular carcinoma (HCC) cell lines. In this study, we investigated the underlying mechanisms on how HI γδ T cells, expanded with zoledronate, exhibit a superior cytotoxic effect on HI NK-resistant Huh7 HCC cells. γδ T cells were obtained from living liver transplant donors or from peripheral blood mononuclear cells (PBMC) of healthy volunteers and were expanded in the presence of IL-2, IL-15, and zoledronate for 2 weeks. Cytotoxicity was measured using the lactate dehydrogenase (LDH) assay in vitro and by flow cytometry using carboxyfluorescein succinimidyl ester (CFSE) in vivo. The cytotoxicity of expanded HI γδ T cells against Huh7 cells was associated with a higher pyrophosphate expression in Huh7 cells compared to SNU398 cells. In contrast, the cytotoxicity of HI γδ T cells against SNU398 cells depended on NKG2D. HI γδ T cells expressed less PD-1 than PB γδ T cells. The cytotoxicity of HI γδ T cells against Du145 and PC3 prostate cancer cells was also associated with pyrophosphate expression in these cells, as well as NKG2D and DNAM-1. The expression levels of phospho-antigen in tumor cells determined the cytotoxicity of HI γδ T cells, although the NK activating receptors, death ligands, and immune checkpoint molecules also contribute to their cytotoxicity. γδ T cells are attractive candidates for cancer immune cell therapy.