Ovarian Cancer Microenvironment Research Articles

The Immune Subtypes and Landscape of Advanced-Stage Ovarian Cancer.

Immunotherapy has played a significant role in the treatment of a variety of hematological and solid tumors, but its application in ovarian cancer (OC) remains unclear. This study aimed to identify immune subtypes of OC and delineate an immune landscape for selecting suitable patients for immunotherapy, thereby providing potent therapeutic targets for immunotherapy drug development. Three immune subtypes (IS1–IS3) with distinctive molecular, cellular, and clinical characteristics were identified from the TCGA and GSE32062 cohorts. Compared to IS1, IS3 has a better prognosis and exhibits an immunological “hot”. IS3, in contrast, exhibits an immunological “cold” and has a worse prognosis in OC patients. Moreover, gene mutations, immune modulators, CA125, CA199, and HE4 expression, along with sensitivity either to immunotherapy or chemotherapy, were significantly different among the three immune subtypes. The OC immune landscape was highly heterogeneous between individual patients. Poor prognosis was correlated with low expression of the hub genes CD2, CD3D, and CD3E, which could act not only as biomarkers for predicting prognosis, but also as potential immunotherapy targets. Our study elucidates the immunotyping and molecular characteristics of the immune microenvironment in OC, which could provide an effective immunotherapy stratification method for optimally selecting patients, and also has clinical significance for the development of new immunotherapy as well as rational combination strategies for the treatment of OC patients.

Effects of Autophagy-Related Genes on the Prognosis and Immune Microenvironment of Ovarian Cancer.

Ovarian cancer (OC) is among the most malignant tumors of the female reproductive system. The role of autophagy in cancer is complex, and the functional relationship between autophagy-related genes and OC remains unclear. Here, the prognostic value of autophagy-related genes in OC and relationships between autophagy and immune function were evaluated. OC data from The Cancer Genome Atlas and the Human Autophagy Database were obtained to identify autophagy-related genes. Univariate and multivariate Cox analyses were used to construct a prognostic model based on autophagy-related genes. Relationships between risk scores and clinical traits were evaluated. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Cytoscape were used to analyze gene functions and their effects on the immune microenvironment. Relationships between autophagy genes and long noncoding RNAs (lncRNAs) were evaluated by Pearson's correlation coefficients, and lncRNAs corresponding to the autophagy-related genes associated with OC prognosis were used to construct a model. Relationships between risk scores and survival and prognosis were evaluated. Finally, a gene set enrichment analysis was performed. Seven autophagy-related genes (CAPN1, CDKN1B, DNAJB1, GNAI3, MTMR14, RHEB, and SIRT2) were identified as independent predictors of prognosis. Three lncRNAs corresponding to autophagy genes independently influenced prognosis. Autophagy genes are closely related to immunity. Fifteen immune cell types showed different levels of infiltration between the high- and low-risk groups. Moreover, immune cell infiltration differed between the high- and low-risk groups based on the model. Our analysis of genes and lncRNAs related to prognosis clarifies the role of autophagy in OC and provides a theoretical basis for further research.

Novel LncRNA ZFHX4-AS1 as a Potential Prognostic Biomarker That Affects the Immune Microenvironment in Ovarian Cancer.

BackgroundOvarian cancer (OvCa) is a malignant disease of the female reproductive system with a high mortality rate. LncRNA has been confirmed to play a crucial role in the development and progression of various cancer types. Novel lncRNA ZFHX4-AS1 has been reported in several cancers, albeit its functional mechanisms in OvCa remain unclear.MethodsWith reference to the public databases and based on integrating bioinformatics analyses, we explored the expression of ZFHX4-AS1 and its roles in the prognosis of OvCa. We employed the Kaplan-Meier curves to investigate the outcome of patients with different ZFHX4-AS1 expressions. Furthermore, its biological function and the related hallmark pathways were assessed through Gene Ontology (GO) annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, and Gene-set enrichment analysis (GSEA). We explored the correlation between lncRNA ZFHX4-AS1 and tumor-infiltrating immune cells through CIBERSORT. The immune checkpoints associated with lncRNA ZFHX4-AS1 and its related genes were investigated. The effect of lncRNA ZFHX4-AS1 on proliferation, invasion and migration of OvCa cells was verified through Cell Counting Kit (CCK)-8, colony formation, wound healing and transwell assays.ResultsThe expression of lncRNA ZFHX4-AS1 was upregulated in OvCa relative to that in normal tissues. Increased lncRNA ZFHX4-AS1 expression was associated with poor overall survival and progression-free survival in OvCa. The GO and KEGG pathway analyses revealed the role of lncRNA ZFHX4-AS1 in cell metabolism, protein synthesis, cell proliferation, and cell cycle. GSEA indicated the hallmark gene sets that were significantly enriched in the high and low expression groups. The CIBERSORT database revealed M2 macrophages, memory B-cells, naïve B cells, and activated NK cells were affected by lncRNA ZFHX4-AS1 expression (all P < 0.05). The expression of lncRNA ZFHX4-AS1 and its related differential genes MRPS11, NSA2, and MRPL13 were significantly correlated with the immune checkpoints. Knockdown of lncRNA ZFHX4-AS1 could inhibit the proliferation, invasion and migration of OvCa cells.ConclusionsThe results suggested that lncRNA ZFHX4-AS1 is a novel prognostic biomarker associated with cell proliferation, metabolism, infiltration, and distribution of tumor-infiltrating immune cells in OvCa, indicating that lncRNA ZFHX4-AS1 can be used as a potential therapeutic target for OvCa in the future.

Identification and Verification of Necroptosis-Related Gene Signature With Prognosis and Tumor Immune Microenvironment in Ovarian Cancer.

Ovarian cancer is the most lethal heterogeneous disease among gynecological tumors with a poor prognosis. Necroptosis, the most studied way of death in recent years, is different from apoptosis and pyroptosis. It is a kind of regulated programmed cell death and has been shown to be closely related to a variety of tumors. However, the expression and prognosis of necroptosis-related genes in ovarian cancer are still unclear. Our study therefore firstly identified the expression profiles of necroptosis-related genes in normal and ovarian cancer tissues. Next, based on differentially expressed necroptosis-related genes, we clustered ovarian cancer patients into two subtypes and performed survival analysis. Subsequently, we constructed a risk model consisting of 5 genes by LASSO regression analysis based on the differentially expressed genes in the two subtypes, and confirmed the strong prognostic ability of the model and its potential as an independent risk factor via survival analysis and independent risk factor analysis. Based on this risk model, patients were divided into high and low risk groups. By exploring differentially expressed genes, enrichment functions and tumor immune microenvironment in patients in high and low risk groups, the results showed that patients in the low risk group were significantly enriched in immune signaling pathways. Besides, immune cells content, immune function activity was significantly better than the high-risk group. Eventually, we also investigated the sensitivity of patients with different risk groups to ICB immunotherapy and chemotherapy drugs. In conclusion, the risk model could effectively predict the survival and prognosis of patients, and explore the tumor microenvironment status of ovarian cancer patients to a certain extent, and provide promising and novel molecular markers for clinical diagnosis, individualized treatment and immunotherapy of patients.

Abstract 6106: The effect of neoadjuvant chemotherapy on tumor immune microenvironment in ovarian cancer

Abstract Background: Ovarian cancer (OC) is a highly lethal gynecologic malignancy, neoadjuvant chemotherapy (NACT) with interval de-bulking surgery (IDS) is an established strategy for woman with advanced epithelial ovarian cancer (EOC). Tumor infiltrating lymphocytes (TILs) play an important role in the pathogenesis of OC. However, the effect of NACT on tumor immune microenvironment (TIME) has not been evaluated. Methods: All patients underwent NACT with IDS. Formalin-fixed paraffin-embedded tissue samples were collected from biopsy specimens and the corresponding post-NACT surgical specimens. The densities and percentage of specific TILs [CD8+, CD3+, CD4+, CD20+, tumor associated macrophages (TAMs), dim CD56+NK, bright CD56+NK, PD-1+ etc.] within the TIME were analyzed by multiplex immunohistochemistry (3DMedcines Inc.). Serum CA 125 was measured 3-weekly per protocol and evaluated the response to NACT using GCIG CA125 response criteria. Results: A total of 28 patients were enrolled in this study with planned IDS after NACT. Of 25 patients evaluated by GCIG CA125 response criteria, 76% (19/25) had a CA125 response. In CA125 response patients, we observed significant recruitments of the densities and percentage of CD8+ in tumor, percentage of PD-1+ cell in tumor after-NACT vs before-NACT (34.85/mm3 vs 110.54/mm3, p=0.0258; 0.50% vs 1.73%, p=0.0132; 0.53% vs 2.66%, p=0.0576, respectively), while no significant change was found in TAMs, NK cells and CD20+ cells. However, in CA125 non-response patients, the TILs in tumor did not change significantly. Conclusion: NACT could release the tumor associated antigen by killing tumor cells for uptake by tumor-infiltrating dendritic cells and promote the infiltration of CD8+ and PD-1+ T lymphocyte in CA125 response OC patients. Given the limited clinical prognostic information and the number of patients in this study, the results of this study will need confirmation in larger studies in the future. Citation Format: Guangming Cao, Dingchao Hua, Lina Cui, Xiaochen Zhao, Shuzhen Wang, Lei Zhu. The effect of neoadjuvant chemotherapy on tumor immune microenvironment in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6106.

Abstract 2527: Ovarian cancer ascites display altered immune environment featured by enhanced soluble factors and suppressive cellular context

Abstract Background: Tumor microenvironment (TME) of epithelial ovarian cancer is unique among solid tumors as tumor cells create their "malignant ascites" TME. These ascites, acting as a complex mixture of soluble factors and cell components, are known to be rich in macrophages which were suggested to skew to a M2-like phenotype involved in resistance and metastases. However, while macrophages can adopt different phenotypes, it's now postulated that tumor-associated macrophages (TAM) from ovarian cancer patient ascites may even acquire mixed M1/M2 properties, providing a particular pro-inflammatory and tumor-promoting microenvironment. Characterization of cell composition along with the growth factors present in the microenvironment is thus crucial to understand ovarian cancer biology and more particularly the immunosuppressive pathways that would underlie the altered immune cell activity. Methods &amp; Results: Through flow cytometry-based marker expression analysis and quantitation of soluble mediators in ovarian cancer ascites, we investigated here the cell component nature as well as the presence of several growth factors. While these ascites were interestingly enriched in major "immunosuppressive" cell subsets including T and myeloid populations, TAM were intriguingly shown to display a mixed phenotype characterized by high expression of CD163, unrelated to M1/M2 categorization since also expressing Arg1, CD80, and iNOS markers. The immunosuppressive phenotype was also linked to IL6, LIF and IL10 among other factors present in the ascitic environment. In order to assess their role in the occurrence of such an immunosuppressive cell status, soluble mediators were then investigated on healthy monocytes undergoing M1 polarization. We demonstrated that, while LIF did not directly affect IL6 and IL10 levels released by the cells, interestingly IL6 alone or in co-presence with LIF decreased IL6 supernatant content, with respect to the control. These changes were concomitant with a decrease and increase in the expression of CD80 and CD163, respectively. Furthermore, CD14 and CD206 were found to increase in the presence of IL6+/-LIF. Conclusion: All these data highlight a switching activity of IL6 skewing cells from M1 polarization and resulting in a proper macrophage phenotype exhibiting a mixed M1/M2 status and would suggest these ascite-containing factors as keys conferring an immunosuppressive cell phenotype. Altogether, these translational findings highlight ovarian patient-derived ascites as a valuable tool to understand the mechanisms of suppression and to allow for the identification of novel markers to develop innovative targeted therapies. Citation Format: Imane Nafia, Assia Chaibi, Christophe Rey, Jean-Philippe Guegan, Alban Bessede, Coriolan Lebreton, Antoine Italiano. Ovarian cancer ascites display altered immune environment featured by enhanced soluble factors and suppressive cellular context [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2527.

The Effect of Bone Marrow Mesenchymal Stem Cells-Exosomes (BMSC-EXO) on Tumor Angiogenesis and Its Mechanism in Ovarian Cancer Microenvironment

In ovarian cancer microenvironment, BMSC cells can differentiate into a variety of stem cells, thereby reducing the damage to tissues, and this effect lies in the exosomal substances secreted by BMSC cells. Then, in ovarian cancer microenvironment, whether BMSC-exo exhibited an effect on angiogenesis at the tumor site, and its possible molecular mechanism remains unclear. BALA nude mice and ovarian cancer tumor tissues were collected to isolate vascular endothelial cells which were then assigned into Control group, 40 μg/ml BMSC-exo group, 80 μg/ml BMSC-exo group, 120 μg/ml BMSC-exo group in the presence of Wnt/β-catenin inhibitor (PNU-74654) followed by analysis of proliferation and migration of ovarian cancer vascular endothelial cells (OCVECs) and the angiogenesis. 40 μg/ml and 80 μg/mlBMSC-exo group showed significantly higher cell proliferation than control group with higher cell number in 80 μg/ml BMSC-exo group than 40 μg/ml BMSC-exo group (P &lt; 0.05). The number of cell migration after BMSC-exo treatment was increased (P &lt; 0.05) and the tumor tissue showed obvious angiogenesis with more CD31-positive cells (P &lt; 0.05). PNU-74654 group showed significantly downregulated Wnt and β-catenin proteins (P &lt; 0.05) and lower cell number and higher migration rate of vascular endothelial cells (P &lt; 0.05). In conclusion, exosomes secreted by BMSC can repair damaged tissues possibly through activation of Wnt/β-catenin signaling pathway.

Evaluation of PD-1/PD-L1 and CTLA-4 immune checkpoints and angiogenic activity in ovarian cancer after oncotherad immunotherapy associated to erythropoietin.

e17560 Background: Ovarian cancer (OC) ranks fifth among cancer deaths in women and remains the deadliest of all gynecological cancers. OncoTherad is a nanostructured complex developed by University of Campinas/Brazil, which exhibits immunomodulatory and antitumor properties. Erythropoietin (EPO) can exert non-hematopoietic functions and the association of immunotherapy and EPO is a reality in anemic patients due to cancer or chemotherapy who may use immunotherapies. We evaluated the effects of OncoTherad and EPO, alone or in combination, on PD-1/PDL-1 and CTLA-4 checkpoints, pro-angiogenic (VEGF) and anti-angiogenic (Endostatin) markers, and microvessel density (CD31-labeled capillaries) in chemically induced ovarian cancer in rats. Methods: Thirty-five Fischer rats were distributed into groups: Control (Sham surgery); Cancer (7,12-dimethylbenzoanthracene – DMBA injection in the ovarian bursa, 1.25 mg/kg); OncoTherad (20mg/kg); EPO (8.4 µg/kg); and OncoTherad+EPO (same doses of the isolated treatments). Intraperitoneal doses were administered twice a week for 4 weeks. Immunohistochemistry was analyzed as total immunoreactivity and intensity of immunoreaction. Results: The Cancer group showed an increase (p &lt; 0.05) in microvascular density and histopathological lesions, predominantly high-grade dysplasias with non-metastatic phenotype, which was related to the little involvement of PD-1/PD-L1 and CTLA-4 checkpoints in this carcinogenesis stage. The treatments altered the frequency of ovarian lesions, especially OncoTherad promoted a significant reduction of cystic lesions, while in the EPO group there was marked hyperplasia of the interstitial tissue and the associated treatment led to intermediate results. OncoTherad immunotherapy was effective in reducing angiogenesis, promoting a decrease (p &lt; 0.05) in VEGF immunostaining and microvessel count, and an increase (p &lt; 0.05) in endostatin immunoreactivity. EPO treatment decreased (p &lt; 0.05) endostatin levels in comparison with OncoTherad group and led to a weaker (p &lt; 0.05) immunoreaction intensity of PD-1/PD-L1 and CTLA-4 compared to the Cancer group. These effects might be related to the modification of the tumor microenvironment modulated by EPO with a decrease in the population of immune cells with immunosuppressive phenotype as well as histopathological alterations including reduction of folliculogenesis and luteogenesis. Conclusions: OncoTherad was able to prevent histopathological lesions and reduce the angiogenesis involved in the ovarian carcinogenesis. EPO treatment decreased endostatin, which possibly had effects on angiogenic activity. However, EPO also modulated the OC microenvironment by reducing PD-1/PD-L1 and CTLA-4. The association of OncoTherad+EPO might have triggered a compensatory effect between the isolated treatments.

A methylation-driven genes prognostic signature and the immune microenvironment in epithelial ovarian cancer.

Aberrant gene methylation has been implicated in the development and progression of tumors. In this study, we aimed to identity methylation-driven genes involved in epithelial ovarian cancer (EOC) to establish a prognostic signature for patients with EOC. We identified and verified 6 MDGs that are closely related to the prognosis of ovarian cancer. A prognostic risk score model and nomogram for predicting the prognosis of ovarian cancer were constructed based on the six MDGs. It can also effectively reflect the immune environment and immunotherapy response of ovarian cancer. These MDGs have great significance to the implementation of individualized treatment and disease monitoring of ovarian cancer patients.

A Potential Indicator ARRDC2 Has Feasibility to Evaluate Prognosis and Immune Microenvironment in Ovarian Cancer.

Background: The abnormal expression of α-arrestin protein family plays a key regulatory role in the occurrence and development of many cancers, including colorectal cancer and cervical cancer, and is inseparable from changes in the tumor immune microenvironment. However, the role of ARRDC2, an important member of this family, in the malignant biological process of ovarian cancer (OC) has not been reported, and its role in the change of the immune microenvironment is also unknown. Methods: In this study, HPA, TCGA, GEO and other databases were used to explore the role of ARRDC2 in the prognosis assessment of ovarian cancer. Then, GO, KEGG analysis and GSEA analysis of the biological processes and cell signaling pathways that ARRDC2 may be involved in activated or inhibited. In addition, the TIMER and TISIDB database were used to conduct in-depth research on the role of ARRDC2 in the change of the immune microenvironment of ovarian cancer. The CMap database explored and screened drugs that may be used for treatment. Through cell transfection, CCK-8, Ki-67 immunofluorescence, wound healing, transwell and clone formation assay, the effect of ARRDC2 knockdown on the malignant biological behavior of OC cells were explored. Results: There were significant differences between OC and ARRDC2 mRNA and protein levels. High ARRDC2 expression level is associated with poor overall survival and can be used as an independent prognostic factor. Interestingly, ARRDC2 expression is positively correlated with B cells, Neutrophils, Dendritic cells and CD8+ T cells, signifying that ARRDC2 may be related to infiltration of immune cells. ARRDC2 and its co-expressed genes are enriched in cell signaling pathways related to the immune system. We explored two possible drugs for the treatment of ovarian cancer. Finally, the results of in vitro experiments indicated that knockdown of ARRDC2 may inhibit malignant phenotypes such as proliferation and migration of OC cells. Conclusion: The differentially expressed ARRDC2 may be a potential prognostic indicator and can be used as a novel biomarker for exploring the immune microenvironment of ovarian cancer.

Arachidonic acid, a clinically adverse mediator in the ovarian cancer microenvironment, impairs JAK‐STAT signaling in macrophages by perturbing lipid raft structures

Survival of ovarian carcinoma is associated with the abundance of immunosuppressed CD163highCD206high tumor‐associated macrophages (TAMs) and high levels of arachidonic acid (AA) in the tumor microenvironment. Here, we show that both associations are functionally linked. Transcriptional profiling revealed that high CD163 and CD206/MRC1 expression in TAMs is strongly associated with an inhibition of cytokine‐triggered signaling, mirrored by an impaired transcriptional response to interferons and IL‐6 in monocyte‐derived macrophages by AA. This inhibition of pro‐inflammatory signaling is caused by dysfunctions of the cognate receptors, indicated by the inhibition of JAK1, JAK2, STAT1, and STAT3 phosphorylation, and by the displacement of the interferon receptor IFNAR1, STAT1 and other immune‐regulatory proteins from lipid rafts. AA exposure led to a dramatic accumulation of free AA in lipid rafts, which appears to be mechanistically crucial, as the inhibition of its incorporation into phospholipids did not affect the AA‐mediated interference with STAT1 phosphorylation. Inhibition of interferon‐triggered STAT1 phosphorylation by AA was reversed by water‐soluble cholesterol, known to prevent the perturbation of lipid raft structure by AA. These findings suggest that the pharmacologic restoration of lipid raft functions in TAMs may contribute to the development new therapeutic approaches.

Role of the extracellular ATP/adenosine pathway in neutrophil-mediated T cell suppression in ovarian cancer microenvironment

Abstract The extracellular ATP (eATP)/adenosine (ADO) pathway is a key modulator of innate and adaptive immunity and is a target for immunotherapy for cancer. In the tumor microenvironment (TME) and conditions of chronic injury, ATP is released by dying cells and is hydrolyzed by the ecto-nucleotidase CD39 to AMP, and further to ADO by CD73. We previously observed that normal neutrophils (PMN) acquire a T cell suppressor phenotype mimicking PMN-MDSCs when exposed to the TME. Using a coculture system consisting of ascites supernatants (ASC) from patients with ovarian cancer (OC) as an authentic component of the TME and concentrations of healthy donor PMN and T cells that mimic those in OC ascites, we found that ASC-primed PMNs inhibit stimulated T cell proliferation, activation and metabolic responses. This suppressor activity was dependent on several PMN pathways, including complement signaling and NADPH oxidase. Since PMNs regulate and respond to signaling from the eATP/ADO pathway, we asked whether eATP/ADO mediates PMN suppressor function. PMNs had high constitutive expression of CD39 that was unaffected by ASC exposure, and inhibition of CD39 by POM-1 abrogated PMN suppressor function. CD73 inhibitor Adenosine 5′-(α,β-methylene) diphosphate also abrogated PMN suppressor function while using a different inhibitor (AB-680) had no effect. Neither ADO receptor inhibitors nor the addition of adenosine deaminase to deplete ADO affected PMN suppressor function. Together, these studies point to eATP as a modulator of PMN suppressor function in the TME and the potential to abrogate suppression by targeting CD39. Future studies will address the specific signaling functions of eATP on PMN in the TME that drive suppressor function. Supported by grant from NIH (5R01CA188900)

Regulatory Role of the Adipose Microenvironment on Ovarian Cancer Progression

Simple SummaryAdipocytes or fat cells are integral part of the ovarian tumor microenvironment. Secreted factors from adipocytes, as well as direct cell-to-cell interaction with ovarian cancer cells have been shown to directly support ovarian tumor progression. Elucidating the molecular pathways involved is crucial in the identification of relevant targets.The tumor microenvironment of ovarian cancer is the peritoneal cavity wherein adipose tissue is a major component. The role of the adipose tissue in support of ovarian cancer progression has been elucidated in several studies from the past decades. The adipocytes, in particular, are a major source of factors, which regulate all facets of ovarian cancer progression such as acquisition of chemoresistance, enhanced metastatic potential, and metabolic reprogramming. In this review, we summarize the relevant studies, which highlight the role of adipocytes in ovarian cancer progression and offer insights into unanswered questions and possible future directions of research.

Construction of a New Dual-Responsive Nano-Drug Delivery System for Matrix Metalloproteinases and Adenosine Triphosphate in Ovarian Cancer Using Nanomicelles.

In this manuscript we constructed a dual-responsive nano-drug delivery system for matrix metalloproteinases and ATP in ovarian cancer microenvironment. The nanomicelle PCL-DNA/DOX-Peptide-PEG was prepared by intercalating doxorubicin hydrochloride between C and G base pairs of DNA double helix structure. Another ATP-responsive nanomicelle PCL-DNA/DOX-PEG was prepared. Then we analyzed the characterization of nanomicelles (particle size, potential, surface morphology, etc.) and drug loading binding and drug release behavior. In addition, the effect of nanomicelles on the viability of mouse ovarian epithelial tumor cell ID-8 was detected by CCK-8 method. CCK-8 assay detected that different concentrations of carrier had no difference on the proliferation of ID-8 cells, and the survival rate of ID-8 cells by different concentrations of DOX preparations was statistically significant and the same results were observed in cytotoxicity comparison. Confocal microscopy showed that DOX in the drug-loaded micelle group was concentrated in the nucleus, while free DOX was concentrated in the cytoplasm. ID-8 cells took up the drug-loaded micelles faster. The semi-quantitative analysis of the DOX uptake of ID-8 cells with different treatments showed extremely significant statistical differences. In conclusion, the prepared self-assembled dual-responsive nanomicelle PCL-DNA/DOX-Peptide-PEG is novel anti-tumor agent, and is expected to have good tumor tissue penetration ability with a low toxicity.

Genomic Instability Is Defined by Specific Tumor Microenvironment in Ovarian Cancer: A Subgroup Analysis of AGO OVAR 12 Trial.

Simple SummaryGiven the importance of genomic instability signatures in the management of ovarian cancer and the difficulties in defining the role of immunotherapy, our objective was to describe the tumor immune microenvironment in the light of genomic instability signatures. Intratumoral CD3+ T lymphocytes confirmed its prognostic value. HLA-E appears to be a robust prognostic biomarker and preferentially overexpressed in homologous recombination deficiency (HRD) ovarian cancers. Our data provide a rationale for future immunotherapy strategies targeting the inhibitory CD94/NKG2A receptor of HLA-E in HRD tumors.Background: Following disappointing results with PD-1/PD-L1 inhibitors in ovarian cancer, it is essential to explore other immune targets. The aim of this study is to describe the tumor immune microenvironment (TME) according to genomic instability in high grade serous ovarian carcinoma (HGSOC) patients receiving primary debulking surgery followed by carboplatin-paclitaxel chemotherapy +/− nintedanib. Methods: 103 HGSOC patients’ tumor samples from phase III AGO-OVAR-12 were analyzed. A comprehensive analysis of the TME was performed by immunohistochemistry on tissue microarray. Comparative genomic hybridization was carried out to evaluate genomic instability signatures through homologous recombination deficiency (HRD) score, genomic index, and somatic copy number alterations. The relationship between genomic instability and TME was explored. Results: Patients with high intratumoral CD3+ T lymphocytes had longer progression-free survival (32 vs. 19.6 months, p = 0.009) and overall survival (OS) (median not reached). High HLA-E expression on tumor cells was associated with a longer OS (median OS not reached vs. 52.9 months, p = 0.002). HRD profile was associated with high HLA-E expression on tumor cells and an improved OS. In the multivariate analysis, residual tumor, intratumoral CD3, and HLA-E on tumor cells were more predictive than other parameters. Conclusions: Our results suggest HLA-E/CD94-NKG2A/2C is a potential immune target particularly in the HRD positive ovarian carcinoma subgroup.

Overexpression of CAPG Is Associated with Poor Prognosis and Immunosuppressive Cell Infiltration in Ovarian Cancer.

Historically, immunotherapies have only resulted in a partial response from patients with advanced ovarian cancer, resulting in poor clinical efficacy. A full understanding of immune-related gene expression and immunocyte infiltration in ovarian cancer would be instrumental for the improved implementation of immunotherapy. The Capping Actin Protein, Gelsolin-Like (CAPG) gene encodes an actin-regulatory protein, which plays important roles in tumor progression and immune regulation. This study is aimed at identifying the potential therapeutic and prognostic roles of CAPG in ovarian cancer. CAPG expression and clinical information were investigated in the data collected from TCGA, Oncomine, GEPIA, UALCAN, and Kaplan-Meier plotter. CAPG coexpression networks were evaluated by LinkedOmics, GeneMANIA, and NetworkAnalyst. The correlation of CAPG with immune infiltrates was analyzed via TIMER, ImmuCellAI, and GEPIA. Our result showed that patients with high tumoral CAPG expression had significantly shorter 5-year overall survival. Functional enrichment analysis indicated that CAPG-related phenotypes were largely involved in inflammatory response, chemokine and cytokine signaling, cell adhesion, and Toll-like receptor signaling pathways. CAPG expression was positively correlated with infiltrating levels of regulatory T cells (Tregs), tumor-associated macrophages (TAMs), and exhausted T cells (Texs) while being negatively correlated with infiltrating levels of natural killer T cells (NKTs) and neutrophils in ovarian cancer. Moreover, the expression of FOXP3, CD25, CD127, CCR8, and TGFβ in respect to Tregs; CCL2 and CD68 in respect to TAM; CD163, VSIG4, and MS4A4A in respect to M2 macrophages; CD33 and CD11b in respect to myeloid-derived suppressor cells (MDSCs); and PD1, CTLA4, LAG3, TIM3, GZMB, 2B4, and TIGIT in respect to Texs was significantly correlated with CAPG expression in ovarian cancer. These findings suggest that CAPG may contribute to the immunosuppressive tumor microenvironment in ovarian cancer, leading to an exhausted T cell phenotype and tumor progression. Therefore, CAPG can be used as a potential biomarker for determining prognosis and immunotherapy effectiveness in ovarian cancer.

Integrated analysis of prognostic immune-related genes in the tumor microenvironment of ovarian cancer.

BackgroundOvarian cancer (OC) is a major cause of most gynecological cancer deaths, and the rates of incidence and mortality are increasing worldwide. However, factors in the tumor microenvironment (TME) related to OC and certain prognostic markers of OC are still unknown. We aimed to identify biomarkers connected to prognostic immunity based on clinical patients’ data from The Cancer Genome Atlas (TCGA).MethodsWe used the ESTIMATE algorithm to compute the immune and matrix scores of OC patients from TCGA. Next, differentially expressed genes (DEGs) according to the immune and matrix scores were obtained. Subsequently, genes (GZMB, C2orf37, CXCL13, and UBD) connected with prognostic immunity were determined. Moreover, functional enrichment analysis and the protein-protein interaction network showed that these genes were enriched in many biological processes related to immune function. The Tumor Immune Estimation Resource (TIMER) algorithm was also used to analyze the immune prognostic genes according to six immuno-infiltrating cells.ResultsAccording to high/low immune-scores and matrix-score groups, 682 common genes were identified, within 420 upregulated genes and 262 downregulated genes. Gene ontology (GO) analysis of biological process primarily enriched in T cell activation, regulation of lymphocyte activation and lymphocyte differentiation. OS analysis showed 45 genes (6.6%) were relevant in the final results. The Kaplan-Meier plotter database verified the top 10 genes related to prognosis, but only GZMB, C2orf37, CXCL13 and UBD were related to overall survival (OS).ConclusionsGZMB, CXCL13, and UBD may influence prognosis via their effects on the infiltration of immune cells and therefore represent potential targets for OC immunotherapy.

Identification of Hypoxia Signature to Assess the Tumor Immune Microenvironment and Predict Prognosis in Patients with Ovarian Cancer.

Background The 5-year overall survival rate of ovarian cancer (OC) patients is less than 40%. Hypoxia promotes the proliferation of OC cells and leads to the decline of cell immunity. It is crucial to find potential predictors or risk model related to OC prognosis. This study aimed at establishing the hypoxia-associated gene signature to assess tumor immune microenvironment and predicting the prognosis of OC. Methods The gene expression data of 378 OC patients and 370 OC patients were downloaded from datasets. The hypoxia risk model was constructed to reflect the immune microenvironment in OC and predict prognosis. Results 8 genes (AKAP12, ALDOC, ANGPTL4, CITED2, ISG20, PPP1R15A, PRDX5, and TGFBI) were included in the hypoxic gene signature. Patients in the high hypoxia risk group showed worse survival. Hypoxia signature significantly related to clinical features and may serve as an independent prognostic factor for OC patients. 2 types of immune cells, plasmacytoid dendritic cell and regulatory T cell, showed a significant infiltration in the tissues of the high hypoxia risk group patients. Most of the immunosuppressive genes (such as ARG1, CD160, CD244, CXCL12, DNMT1, and HAVCR1) and immune checkpoints (such as CD80, CTLA4, and CD274) were upregulated in the high hypoxia risk group. Gene sets related to the high hypoxia risk group were associated with signaling pathways of cell cycle, MAPK, mTOR, PI3K-Akt, VEGF, and AMPK. Conclusion The hypoxia risk model could serve as an independent prognostic indicator and reflect overall immune response intensity in the OC microenvironment.

Immunotherapy in Ovarian Cancer: Thinking Beyond PD-1/PD-L1.

Ovarian cancer (OC) is the most lethal gynecologic malignancy, affecting approximately 1 in 70 women with only 45% surviving 5 years after diagnosis. This disease typically presents at an advanced stage, and optimal debulking with platinum-based chemotherapy remains the cornerstone of management. Although most ovarian cancer patients will respond effectively to current management, 70% of them will eventually develop recurrence and novel therapeutic strategies are needed. There is a rationale for immune-oncological treatments (IO) in the managements of patients with OC. Many OC tumors demonstrate tumor infiltrating lymphocytes (TILs) and the degree of TIL infiltration is strongly and reproducibly correlated with survival. Unfortunately, results to date have been disappointing in relapsed OC. Trials have reported very modest single activity with various antibodies targeting PD-1 or PD-L1 resulting in response rate ranging from 4% to 15%. This may be due to the highly immunosuppressive TME of the disease, a low tumor mutational burden and low PD-L1 expression. There is an urgent need to improve our understanding of the immune microenvironment in OC in order to develop effective therapies. This review will discuss immune subpopulations in OC microenvironment, current immunotherapy modalities targeting these immune subsets and data from clinical trials testing IO treatments in OC and its combination with other therapeutic agents.

Significance of CD47 and Its Association With Tumor Immune Microenvironment Heterogeneity in Ovarian Cancer.

BackgroundIt was reported that tumor heterogeneity and the surrounding tumor microenvironment (TME) in ovarian cancer affects immunotherapy efficacy and patient outcomes. And the TME of ovarian cancer is intrinsically heterogeneous. CD47 plays vital roles in cell functional behavior and immune homeostasis relating to cancer prognosis. But how it affects TME and its contribution to heterogeneity in ovarian cancer has not been fully illustrated. Therefore, we aimed to identify a prognostic biomarker which may help explain tumor immune microenvironment heterogeneity of ovarian cancer.MethodsCancer single-cell state atlas (CancerSEA) was used to evaluate functional role of CD47. Several bioinformatics database including Oncomine, Gene Expression Profiling Interaction Analysis (GEPIA), Tumor Immune Estimation Resource (TIMER), The Human Protein Atlas (HPA), Ualcan and Kaplan-Meier plotter (KM plotter) were applied to illustrate correlation of CD47 with ovarian cancer prognosis and immune infiltration. Tumor Immune Single-cell Hub (TISCH) single cell database was employed to evaluate correlation of CD47 with tumor microenvironment. GeneMANIA was implemented to identify regulation networks of CD47. Differentially expressed genes (DEGs) between CD47 high and low expression groups were analyzed with R package DESeq2. Kyoto encyclopedia of genes and genomes (KEGG) and Gene Set Enrichment Analysis (GSEA) were utilized to explore how CD47 affect the immune related cell signaling pathway.ResultsCD47 expression was upregulated and connected to worse OS and PFS in ovarian cancer. Close relation was found between CD47 expression level and immune infiltration in ovarian cancer, especially with Treg cells, Monocytes, Macrophages and T cell exhaustion (P<0.05). The CD47 expression level was relatively low in plasma cells, dendritic cells and Mono/Macro cells of OV_GSE115007, in myofibroblasts, fibroblasts and endothelial cells of OV_GSE118828, compared to malignant cells of OV_GSE118828 dataset. The cell components and distribution in primary and metastatic ovarian cancer are quite distinct, which may lead to TME heterogeneity of ovarian cancer.ConclusionOur results indicated that CD47 is closely correlated to ovarian cancer immune microenvironment and might induce ovarian cancer heterogeneity. Therefore, CD47 may be used as a candidate prognostic biomarker and provide us with new insights into potential immunotherapy in ovarian cancer patients.