Poor Survival In Cancer Patients Research Articles

Triggering Receptor Expression on Myeloid Cells-1 (TREM1) Promoter Hypomethylation and Its Overexpression Associated With Poor Survival of Cancer Patients: A Pan-Cancer Analysis

Triggering Receptor Expression on Myeloid Cells-1 (TREM1) Promoter Hypomethylation and Its Overexpression Associated With Poor Survival of Cancer Patients: A Pan-Cancer Analysis

Increased SEC14L2 expression is associated with clinicopathological features and worse prognosis in oral squamous cell carcinoma.

Abnormal expression of SEC14L2 has been implicated in many human cancers. However, the role of SEC14L2 in oral squamous cell carcinoma (OSCC) remains unclear. Therefore, this study aimed to evaluate the expression and prognostic roles of SEC14L2 in OSCC. OSCC tumors and adjacent non-tumors were collected from OSCC patients and used for SEC14L2 mRNA expression by quantitative reverse transcription PCR (RT-qPCR). Additionally, the expression of SEC14L2 was further analyzed using The Cancer Genome Atlas-Head Neck Squamous Cell Carcinoma (TCGA-HNSCC) dataset to identify its relationship with HNSCC clinical characteristics. The Kaplan-Meier plot was used to assess survival rates, and the Tumor Immune Estimation Resource (TIMER) database was used to examine the correlation between SEC14L2 expression and tumor immune cell infiltration. In silico tools also looked at SEC14L2 involvement in cancer pathways through its protein network. The mRNA and protein levels of SEC14L2 are notably higher in both OSCC and HNSCC tissues compared to adjacent normal tissues. Upregulation of SEC14L2 was associated with advanced tumor stages, grades, metastasis, HPV-negative, and TP53 mutations in cancer patients. In addition, the high expression of SEC14L2 was negatively correlated with the poor survival of cancer patients and the infiltration of diverse immune cells in cancer patients. According to the findings of this investigation, SEC14L2 is significantly elevated in OSCC/HNSCC patients and associated with a worse prognosis. More investigation and clinical studies are required to completely understand the therapeutic potential of SEC14L2 in HNSCC and convert these findings into better patient outcomes.

B7-H3 Expression in Breast Cancer and Brain Metastasis.

Brain metastasis is a significant challenge for some breast cancer patients, marked by its aggressive nature, limited treatment options, and poor clinical outcomes. Immunotherapies have emerged as a promising avenue for brain metastasis treatment. B7-H3 (CD276) is an immune checkpoint molecule involved in T cell suppression, which is associated with poor survival in cancer patients. Given the increasing number of clinical trials using B7-H3 targeting CAR T cell therapies, we examined B7-H3 expression across breast cancer subtypes and in breast cancer brain metastases to assess its potential as an interventional target. B7-H3 expression was investigated using immunohistochemistry on tissue microarrays of three clinical cohorts: (i) unselected primary breast cancers (n = 347); (ii) brain metastatic breast cancers (n = 61) and breast cancer brain metastases (n = 80, including a subset of 53 patient-matched breast and brain metastasis cases); and (iii) mixed brain metastases from a range of primary tumours (n = 137). In primary breast cancers, B7-H3 expression significantly correlated with higher tumour grades and aggressive breast cancer subtypes, as well as poorer 5-year survival outcomes. Subcellular localisation of B7-H3 impacted breast cancer-specific survival, with cytoplasmic staining also correlating with a poorer outcome. Its expression was frequently detected in brain metastases from breast cancers, with up to 90% expressing B7-H3. However, not all brain metastases showed high levels of expression, with those from colorectal and renal tumours showing a low frequency of B7-H3 expression (0/14 and 2/16, respectively). The prevalence of B7-H3 expression in breast cancers and breast cancer brain metastases indicates potential opportunities for B7-H3 targeted therapies in breast cancer management.

Abstract 5517: Chronic stress causes metastasis via neutrophil-mediated changes to the microenvironment

Abstract Chronic stress is associated with increased risk of metastasis and poor survival in cancer patients, yet the reasons are unclear. We show that chronic stress increased lung metastasis from disseminated cancer cells 2- to 4-fold in mice. Chronic stress significantly altered the lung microenvironment, with fibronectin accumulation, reduced T cell infiltration, and increased neutrophil infiltration. Depleting neutrophils abolished stress-induced metastasis, indicating their critical role. Chronic stress shifted neutrophils’ normal circadian rhythm, and, via glucocorticoid release, caused increased neutrophil extracellular trap (NET) formation. Importantly, in mice with neutrophil-specific glucocorticoid receptor deletion, chronic stress failed to increase NETs and metastasis. Furthermore, digesting NETs with deoxyribonuclease (DNase) I prevented chronic stress-induced metastasis. Together, our data show that glucocorticoids released during chronic stress cause NET formation and establish a metastasis-promoting microenvironment. Therefore, NETs could be targets for preventing metastatic recurrence in cancer patients, many of whom will experience chronic stress due to their disease. Citation Format: Xue-Yan He, Yuan Gao, David Ng, Evdokia Michalopoulou, Shanu George, Jose M. Adrover, Lijuan Sun, Jean Albrengues, Juliane Daßler-Plenker, Xiao Han, Ledong Wan, Xiaoli S. Wu, Longling S. Shui, Yu-Han Huang, Bodu Liu, Chang Su, David L. Spector, Christopher R. Vakoc, Linda VanAelst, Mikala Egeblad. Chronic stress causes metastasis via neutrophil-mediated changes to the microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5517.

Chronic stress increases metastasis via neutrophil-mediated changes to the microenvironment

Chronic stress is associated with increased risk of metastasis and poor survival in cancer patients, yet the reasons are unclear. We show that chronic stress increases lung metastasis from disseminated cancer cells 2- to 4-fold in mice. Chronic stress significantly alters the lung microenvironment, with fibronectin accumulation, reduced Tcell infiltration, and increased neutrophil infiltration. Depleting neutrophils abolishes stress-induced metastasis. Chronic stress shifts normal circadian rhythm of neutrophils and causes increased neutrophil extracellular trap (NET) formation via glucocorticoid release. In mice with neutrophil-specific glucocorticoid receptor deletion, chronic stress fails to increase NETs and metastasis. Furthermore, digesting NETs with DNase I prevents chronic stress-induced metastasis. Together, our data show that glucocorticoids released during chronic stress cause NET formation and establish a metastasis-promoting microenvironment. Therefore, NETs could be targets for preventing metastatic recurrence in cancer patients, many of whom will experience chronic stress due to their disease.

Abstract C014: Pan-cancer analysis portrays NVL as a diagnostic and prognostic biomarker

Abstract NVL (also known as NVL2), an AAA+ ATPase, has recently been linked with various cancers. By systematic bioinformatics analyses, here we predict the prospect of NVL as a cancer biomarker. Our pan-cancer analyses reveal that NVL is overexpressed in cancers [GEPIA and UALCAN (TCGA) databases] and the top five cancers with elevated expression (more than twofold) of NVL are Breast Invasive Carcinoma, Testicular Germ Cell Tumors, Cholangiocarcinoma, Diffuse large B cell lymphoma, and Thymoma. NVL is also highly expressed in all stages of most cancers. Additionally, this high expression was found to be correlated to the poor survival of cancer patients (Kaplan-Meier Plotter and OncoDB databases). Since, genetic alteration is one of the major cause of cancer, we investigated mutation and copy number amplification status (cBioPortal and COSMIC databases) of NVL across the cancers. Many potential mutation sites were predicted (cBioPortal and COSMIC databases) and most strikingly, four mutation sites at the ATPase domain of NVL. This signifies that these are the mutations that might have role in carcinogenesis. The copy number amplification study also revealed the high amplification percentage of NVL across different cancers. We further focused on the functional characteristics of the protein NVL across cancers. The protein-protein interaction analyses found the proteins like SKIV2L2, WDR12, WDR74, UTP18, NAT10, TERT, DKC1, GTPBP4, BOP1, TEP1 directly interact with NVL. The top canonical pathways of NVL also indicate that NVL has crucial functions in cancer cell proliferation (DAVID). We, therefore, surmise that NVL could be considered as a potential diagnostic and prognostic marker for multiple cancers and a very promising biomarker and target for cancer. Citation Format: Mousumi Datta, Dr. Aditi Nayak. Pan-cancer analysis portrays NVL as a diagnostic and prognostic biomarker [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr C014.

Overall survival in 92,991 colorectal cancer patients in Germany: differences according to type of comorbidity

Background Poorer survival in cancer patients with vs. without comorbidity has been reported for various cancer sites. For patients with colorectal cancer (CRC), limited data are available so far. Methods Patients with CRC diagnosed between 2010 and 2018 were identified in a health claims database covering 20% of the German population. We assessed the prevalence of comorbidities at cancer diagnosis and categorized the patients into the groups: ‘none’, ‘somatic only’, ‘mental only’ or ‘both’ types of comorbidities. Hazard ratios (HR, with 95% confidence intervals) for five-year overall survival were estimated by Cox proportional hazard models, adjusted for age, sex and stage at diagnosis (advanced vs. non-advanced). Results We included 92,991 patients (females: 49.1%, median age: 72 years) with a median follow-up of 30 months. The proportions assigned to the groups ‘none’, ‘somatic only’, ‘mental only’ or ‘both’ were 24.7%, 65.5%, 1.4% and 8.4%. Overall, 32.8% of the patients died during follow-up. Compared to patients without comorbidities (‘none’), the adjusted HR regarding death from any cause was 1.11 (95% CI: 1.07–1.14) in the group ‘somatic only’, 1.74 (95% CI: 1.58–1.92) in the group ‘mental only’ and 1.92 (95% CI: 1.84–2.00) in the group ‘both’. For patients with ‘mental only’ comorbidities, the adjusted HR was higher in males than in females (HR = 2.19, 95% CI: 1.88–2.55 vs. HR = 1.55, 95% CI: 1.37–1.75). Conclusions Our results suggest that patients with CRC and with mental comorbidities, particularly males, have a markedly lower overall survival compared to those without any or only somatic comorbidities.

Increased incidence of cancer metastasis in young patients with ischaemic heart disease

Abstract Background Heart disease has been linked to an increased risk of developing certain types of cancer. However, while cancer metastasis is the primary determinant of cancer prognosis, the association between heart disease and cancer metastasis has never been studied in humans. Methods We evaluated 22,057 self-referred adults, free of cancer and IHD at baseline, who were screened in preventive healthcare as private individuals. To eliminate the cases of silent cancer, we implicated a one-year blanking period at the start of the follow-up. Cancer and mortality data were available for all subjects from national registries. Then, multivariable Cox regression model with IHD as a time-dependent covariate was used to evaluate the association of IHD with metastatic cancer during follow-up. Finally, we implicated the same models in evaluating the differences in risk between young (age<55) and old adults. Results During a median follow-up of 6.5 years (Interquartile range 3-13 years), 913 subjects developed IHD and 929 subjects developed metastatic cancer. Compared with subjected who did not develop IHD during follow-up, subjects who developed IHD during follow-up were 64% more likely to develop metastatic cancer during follow-up, in a univariate model (95% CI 1.15-2.36, p=0.0006). Multivariable analysis with adjustment for common risk factors yielded consistent results such that IHD was associated with 50% increased risk of metastatic cancer (Fig-A, p=0.005). Moreover, IHD was associated with poor survival of cancer patients with and without metastasis (HR=1.75 and 1.65, p<0.001 for both). Subgroup analysis demonstrated that the association of IHD with the risk of metastatic cancer was age-dependent: the association was more significant among younger subjects below 55 years of age (Fig-B, n=17,433, n(IHD)=433, HR=2.24, p=0.011), compared with older subjects (n=4624, n(IHD)=480, HR=0.91, p=0.711; p for interaction = 0.085). Notably, IHD emerged as a risk factor for metastatic cancer in younger adults (<55) despite lower rate of cardiovascular risk factors. Conclusions We show, for the first time, that ischaemic heart disease is associated with increased and independent risk of metastatic cancer. Furthermore, we demonstrated that the risk is age-dependent and more pronounced among patients younger than 55. Understanding this relationship could lead to advances in cardiovascular and cancer treatment and could ultimately help to improve patient outcomes.

Prognostic Value of Body Composition and Systemic Inflammatory Markers in Patients with Locally Advanced Cervical Cancer Following Chemoradiotherapy.

Abnormal changes in body composition and systemic inflammation response have been associated with poor survival of cancer patients. Our study was to explore the prognostic value of the association between body composition indicators and systemic inflammation markers among patients with locally advanced cervical cancer (LACC) who underwent concurrent chemoradiotherapy (CCRT). We retrospectively reviewed medical records of LACC patients treated between 2016 and 2019. Subcutaneous, visceral and intra-muscular adipose index (SAI, VAI and IMAI) and skeletal muscle index (SMI) were derived from computed tomography (CT). Kaplan-Meier analysis and Univariate and multivariate Cox analyses were used to evaluate the survival. A nomogram was constructed to assess the prognostic value. The study included 196 patients treated with CCRT. According to multivariable Cox analyses, IIIC1r (P = 0.045), high systemic immune-inflammation index (SII) (P = 0.004), sarcopenia (P = 0.008), high SAI (P = 0.016) and high VAI (P = 0.001) were significantly risk factors for overall survival (OS). Kaplan-Meier analysis showed that patients with low lymphocyte-to-monocyte ratio (LMR) and sarcopenia had longer OS than those with high LMR and sarcopenia (P = 0.023). The high neutrophil-to-lymphocyte ratio (NLR) in non-sarcopenic patients showed better survival (P = 0.022). Low VAI (P = 0.019) or low IMAI (P = 0.019) combined with low SII had a favorable OS. Low LMR combined with low SAI was associated with longer OS (P = 0.022). The calibration plots of nomogram predicting the 3-year and 5-year OS rates were close to the ideal models. Inflammation factors were closely associated with abnormal muscle and fat distribution. The combined prognostic value of body composition indicators and systemic inflammation markers was reliable in predicting survival for LACC patients.

The efficacy of fat-free mass index and appendicular skeletal muscle mass index in cancer malnutrition: a propensity score match analysis.

Reduced muscle mass (RMM) is a phenotypic criterion for malnutrition; the appendicular skeletal muscle mass index (ASMI) and fat-free mass index (FFMI) are both applicable indicators in the global leadership initiative on malnutrition (GLIM) guideline. However, their sensitivity and prognostic effect remain unclear. Clinical data of 2,477 patients with malignant tumors were collected. Multi-frequency bioelectrical impedance analysis was used to obtain ASMI and FFMI. RMM was confirmed by ASMI (< 7.0 kg/m2 for men and < 5.7 kg/m2 for women) or FFMI (< 17 kg/m2 for men and < 15 kg/m2 for women). Propensity score match analysis and logistic regression analysis were used to evaluate the efficacy of FFMI and ASMI in diagnosing severe malnutrition and multivariate Cox regression analysis to determine the efficacy of RMM in predicting survival. In total, 546 (22.0%) and 659 (26.6%) participants were diagnosed with RMM by ASMI (RMM.ASMI group) and FFMI (RMM.FFMI group); 375 cases overlapped. Body mass index (BMI), midarm circumference, triceps skinfold thickness, and maximum calf circumference were all significantly larger in the RMM.FFMI group for both sexes (P < 0.05). A 1:1 matched dataset constructed by propensity score match contained 810 cases. RMM.FFMI was an influential factor of severe malnutrition with HR = 3.033 (95% CI 2.068-4.449, P < 0.001), and RMM.ASMI was a predictive factor of overall survival (HR = 1.318, 95% CI 1.060-1.639, P = 0.013 in the RMM.ASMI subgroup, HR = 1.315, 95% CI 1.077-1.607, P = 0.007 in the RMM.FFMI subgroup). In general, RMM indicates negative clinical outcomes; when defined by FFMI, it predicts nutritional status, and when defined by ASMI, it is related to poor survival in cancer patients.

PAD4 controls the anti-tumor response by fine-tuning IFNγ signaling in macrophages

Abstract MHC-II–mediated antigen presentation is essential for the function of the immune system. Recent evidence indicates that MHC-II expression shapes tumor immunity and response to immunotherapy. MHC-II expressing antigen presenting cells (APCs) mediate CD4 +T-cell priming and activation and license them to support the CD8 +T-cell response. Citrullination – catalyzed by peptidyl arginine deiminases (PADs) – is a post-translational modification at arginine residues. Here, we show that PAD4 negatively regulates anti-tumor immunity by tuning IFNγ-signaling gene expression, including that of MHC-II, in tumor associated macrophages (TAMs) in tumor bearing mouse models. Mechanistically, PAD4 citrullinates STAT1, thereby promoting the interaction between STAT1 and PIAS1 (protein inhibitor of activated STAT1) and resulting in reduced MHC-II expression in TAMs. Moreover, low levels of MHC-II expression in TAMs correlate with a reduced response to immunotherapy and are associated with poor cancer patient survival. These data suggest that targeting PAD4-mediated citrullination in macrophages may serve as a potential anti-cancer therapeutic approach. Supported by grants from the NCI (R01CA217510, PA-21-071)

Abstract 62: Untangling the connection between stress and metastatic breast cancer

Abstract There is emerging evidence that exposure to chronic stress is associated with increased risk of metastasis and poor survival in cancer patients. However, the underlying molecular and cellular mechanisms responsible for the effects of stress on tumor progression remain unclear. To determine how stress promotes metastasis, we established new mouse models in which multiple chronic stress exposure promoted breast cancer metastasis to the lungs. We found that chronic stress generates a pro-metastatic lung microenvironment with e.g., reduced T cell infiltration and increased extracellular matrix (ECM) deposition and neutrophil infiltration. Among them, neutrophils were required for these changes as their depletion abolished the pro-metastatic effects of stress and normalized the lung tumor microenvironment. Mechanistically, we analyzed neutrophils from chronically stressed mice and identified dramatically altered transcriptomes, including increased expression of several glucocorticoid receptor regulated genes. Phenotypically, neutrophils from stress exposed mice exhibited increased ROS production, and an increased ability to inhibit T cell activation and to form spontaneous neutrophil extracellular traps (NETs). The stress-associated changes to neutrophils were dependent on glucocorticoid receptor (GR) signaling as neutrophil-specific GR knockout mice showed no increase in NET-formation, a normalized lung microenvironment and no increase in lung metastasis upon stress exposure. Among the changes in the neutrophils caused by stress exposure, we found that the formation of pro-metastatic NETs was critical as directly digesting NETs with DNase I prevented stress-induced metastasis and normalized the lung microenvironment. In sum, this work shows that chronic stress exposure causes a glucocorticoid-mediated increase in NET formation and that NETs are critical drivers in the establishment of a stress-induced, metastasis-promoting microenvironment. Targeting NETs could be an important strategy to prevent metastatic recurrence in cancer patients, many of whom will experience stress due to their cancer diagnosis and subsequent treatments. Citation Format: Xue-Yan He, David Ng, Yuan Gao, Evdokia Michalopoulou, Shanu George, Jose M. Adrover, Lijuan Sun, Jean Albrengues, Ledong Wan, Xiao Han, Christopher Vakoc, Linda Van Aelst, Mikala Egeblad. Untangling the connection between stress and metastatic breast cancer [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 62.

Abstract 1263: Hypoxia induces down-regulation of stimulator of interferon genes (STING) that is synergized with oncometabolites

Abstract The cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway, a component of the innate immune system, functions to detect the presence of cytosolic DNA, and STING signaling is necessary for the anti-cancer immune response. However, STING expression is suppressed or lost in the majority of cancers, which correlates with poor survival in cancer patients. Hypoxia is a key feature of solid tumors that adversely affects prognosis by altering gene expression, creating an immunosuppressive tumor microenvironment, promoting genome instability, and hampering therapy responses. In this study, we found that that prolonged moderate hypoxia induces downregulation of STING expression in multiple cancer cell lines, causing decreased STING pathway activation after treatment with a STING agonist, or with DNA damaging chemotherapy. Mechanically, we found that hypoxia suppressed histone 3 lysine 4 (H3K4) methylation at the STING promoter based on chromatin immunoprecipitation (ChIP) assays in H1975 cells, a non-small cell lung cancer cell line, and knockdown of the H3K4 demethylases, KDM5A or KDM1A, prior to hypoxic exposure prevented hypoxia-induced down-regulation of STING expression. Moreover, decreased STING expression in hypoxic cells was reversed by treatment with the DNA methyltransferase inhibitor, 5-aza-2′-deoxycytidine. Next, we found that prolonged hypoxia increased both D-2-hydroxyglutarate (D-2-HG) and L-2-HG levels in H1975 cells, and that 2-HG treatment, by itself, decreased STING expression under normoxic conditions in H1975 cells. These findings are consistent with analysis of TCGA data showing that STING expression at the mRNA level is reduced in malignancies with mutations in isocitrate dehydrogenase 1 (IDH1) which causes elevated D-2-HG. Furthermore, we found that hypoxia increased Branched Chain Amino Acid Transaminase 1 (BCAT1) expression, and overexpression of BCAT1 accelerated STING downregulation in hypoxic condition. Functionally, expression of interferon (IFN)-stimulated genes (ISGs), CCL5 and interferon-β, was low in hypoxic H1975 cells compared to normoxic cells after treatment with poly(dA:dT), a small molecule STING agonist, and doxorubicin. Immunogenic cell death (ICD) was also reduced in hypoxic H1975 cells. In summary, the results suggest that hypoxia suppresses STING expression through epigenetic regulation, leading to STING silencing, and that this process synergizes with oncometabolites. The decreased STING caused by hypoxia further causes reduction of innate immune responses in cancer cells. Our study provides new understanding of the underlying mechanisms for STING silencing, which will help not only to study cancer immunity but also to develop novel therapeutic strategies. Citation Format: Yuhong Lu, Annali Yurkevicz, Yanfeng Liu, Jonathan Dow, Peter M. Glazer. Hypoxia induces down-regulation of stimulator of interferon genes (STING) that is synergized with oncometabolites [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 1263.

Abstract 296: Clinical significance of glycolytic metabolic activity in liver cancer and its implication to immunotherapy

Abstract Background &amp; Aims: High metabolic activity is a hallmark of cancers including hepatocellular carcinoma (HCC), a major form of liver cancer. However, the molecular features of HCC with high metabolic activity contributing to the clinical outcomes and their therapeutic implications are poorly understood. We aimed to uncover the metabolic characteristics of HCC that are associated with clinical relevance such as overall survival and response to immunotherapy. Methods: By applying cross-species comparison of genomic data, we integrated gene expression profile data from well-defined mouse models and human HCC tumor tissues to stratify HCC tumors according to their intrinsic metabolic activity. For stratification of HCC tumors, we developed a robust genomic predictor and validated it in 5 HCC cohorts (n=1038). Statistics and informatics approaches were performed to assess clinical significance such as overall survival and response to immunotherapy in metabolic subtypes. Genomic data from patient derived xenograft (PDX) models were integrated to the analysis. Results: Analysis of systematically integrated genomic data revealed three distinct metabolic subtypes of HCC (high, moderate and low metabolic subtypes). The high metabolic subtype is characterized by poor survival, the strongest stem cell features, high genomic instability, high expression of alpha-fetoprotein and KRT19, and activation of EPCAM and SALL4, and poor response to immunotherapy. Importantly, analysis of immune cell population in HCC tumors showed that immune-suppressive regulatory T-cells (Treg) are highly enriched in high metabolic HCC tumors, suggesting that high metabolic activity of HCC cells may trigger activation or infiltration of Treg cells, leading to evasion of HCC cells from anti-cancer immune cells. Interestingly, recent study showed that Treg cells can effectively utilize lactic acid as metabolic fuel for its proliferation. Because high metabolic activity of HCC tumors eventually leads to accumulation of glycolysis by-product lactic acid in tumor microenvironment, we postulate that accumulated lactic acid in tumor microenvironment might account for aggregation of Treg cells in high metabolic HCC tumors, leading to resistance to immunotherapy. In agreement with this, 18F-FDG PET/CT imaging data showed that high metabolic activity of tumors reflected in SUVmax is significantly associated with poorer survival of cancer patients after immunotherapy. Conclusions: We identified clinically and metabolically distinct subtypes of HCC tumors, potential biomarkers associated with these subtypes, and potential mechanism of metabolism-mediated evasion of HCC cells to anti-cancer immunotherapy. Moreover, the metabolic subtypes are well conserved in PDX models, offering a tool for selecting the best preclinical models for future study. Citation Format: Sowon Grace Park, Joann Jung, Yeonwoo Grace Jang, Sung Hwan Lee, Yun-Seong Jeong, Sun Young Yim, Ju-Seog Lee. Clinical significance of glycolytic metabolic activity in liver cancer and its implication to immunotherapy [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 296.

Perfluorocarbon nanodrug induced oxygen self-enriching sonodynamic therapy improves cancer immunotherapy after insufficient radiofrequency ablation.

Residual lesions and undetectable metastasis after insufficient radiofrequency ablation (iRFA) are associated with earlier new metastases and poor survival in cancer patients, for induced aggressive tumor phenotype and immunosuppression. Programmed cell death protein 1(PD-1) blockade has been reported to enhance the radiofrequency ablation-elicited antitumor immunity, but its ability to eliminate incompletely ablated residual lesions has been questioned. Here, we report a combined treatment modality post iRFA based on integrating an oxygen self-enriching nanodrug PFH-Ce6 liposome@O2 nanodroplets (PCL@O2)-augmented noninvasive sonodynamic therapy (SDT) with PD-1 blockade. PCL@O2 containing Ce6 as the sonosensitizer and PFH as O2 reservoir, was synthesized as an augmented SDT nanoplatform and showed increased ROS generation to raise effective apoptosis of tumor cells, which also exposed more calreticulin to induce stronger immunogenic cell death (ICD). Combining with PD-1 blockade post iRFA, this optimized SDT induced a better anti-tumor response in MC38 tumor bearing mouse model, which not only arrested residual primary tumor progression, but also inhibited the growth of distant tumor, therefore prolonging the survival. Profiling of immune populations within the tumor draining lymph nodes and tumors further revealed that combination therapy effectively induced ICD, and promoted the maturation of dendritic cells, tumor infiltration of T cells, as well as activation of cytotoxic T lymphocytes. While iRFA alone could result in an increase of regulatory T cells (Tregs) in the residual tumors, SDT plus PD-1 blockade post iRFA reduced the number of Tregs in both primary and distant tumors. Moreover, the combined treatment could significantly initiate long-term immune memory, manifesting as elevated levels of CD8+ and CD4+ central memory cells. Therefore, this study establishes the preclinical proof of concept to apply oxygen self-enriching SDT to augment cancer immunotherapy after iRFA.

METTL3-mediated downregulation of splicing factor SRSF11 is associated with carcinogenesis and poor survival of cancer patients.

N6-methyladenosine (m6A) is one of the most abundant post-transcriptional modifications in eukaryotic RNA. As m6A modifications play an important role in RNA processing, abnormal m6A regulation caused by aberrant expression of m6A regulators is closely related to carcinogenesis. In this study, we aimed to determine the role of METTL3 expression in carcinogenesis, regulation of splicing factor expression by METTL3, and their effects in survival period and cancer-related metabolisms. We investigated the correlation between each splicing factor and METTL3 in breast invasive ductal carcinoma (BRCA), colon adenocarcinoma (COAD), lung adenocarcinoma (LUAD) and gastric adenocarcinoma (STAD). Survival analysis was performed based on the expression of each splicing factor. To determine the molecular mechanism of SRSF11 in carcinogenesis, gene set enrichment analysis using RNA sequencing data was performed according to SRSF11 expression. Among the 64 splicing factors used for correlation analysis, 13 splicing factors showed a positive correlation with METTL3 in all four cancer types. We found that when METTL3 expression was decreased, the expression of SRSF11 was also decreased in all four types of cancer tissue when compared to that in normal tissue. Decreased SRSF11 expression was associated with poor survival in patients with BRCA, COAD, LUAD, and STAD. Gene set enrichment analysis according to SRSF11 expression showed that the p53/apoptosis, inflammation/immune response, and ultraviolet/reactive oxygen species stimulus-response pathways were enriched in cancers with decreased SRSF11 expression. These results suggest that METTL3 regulates SRSF11 expression, which could influence mRNA splicing in m6A modified cancer cells. METTL3-mediated downregulation of SRSF11 expression in cancer patients correlates with poor prognosis.

A novel tumor suppressor encoded by a 1p36.3 lncRNA functions as a phosphoinositide-binding protein repressing AKT phosphorylation/activation and promoting autophagy.

Peptides/small proteins, encoded by noncanonical open reading frames (ORF) of previously claimed non-coding RNAs, have recently been recognized possessing important biological functions, but largely uncharacterized. 1p36 is an important tumor suppressor gene (TSG) locus frequently deleted in multiple cancers, with critical TSGs like TP73, PRDM16, and CHD5 already validated. Our CpG methylome analysis identified a silenced 1p36.3 gene KIAA0495, previously thought coding long non-coding RNA. We found that the open reading frame 2 of KIAA0495 is actually protein-coding and translating, encoding a small protein SP0495. KIAA0495 transcript is broadly expressed in multiple normal tissues, but frequently silenced by promoter CpG methylation in multiple tumor cell lines and primary tumors including colorectal, esophageal and breast cancers. Its downregulation/methylation is associated with poor survival of cancer patients. SP0495 induces tumor cell apoptosis, cell cycle arrest, senescence and autophagy, and inhibits tumor cell growth in vitro and in vivo. Mechanistically, SP0495 binds to phosphoinositides (PtdIns(3)P, PtdIns(3,5)P2) as a lipid-binding protein, inhibits AKT phosphorylation and its downstream signaling, and further represses oncogenic AKT/mTOR, NF-κB, and Wnt/β-catenin signaling. SP0495 also regulates the stability of autophagy regulators BECN1 and SQSTM1/p62 through modulating phosphoinositides turnover and autophagic/proteasomal degradation. Thus, we discovered and validated a 1p36.3 small protein SP0495, functioning as a novel tumor suppressor regulating AKT signaling activation and autophagy as a phosphoinositide-binding protein, being frequently inactivated by promoter methylation in multiple tumors as a potential biomarker.

Early postoperative decrease of skeletal muscle mass predicts recurrence and poor survival after surgical resection for perihilar cholangiocarcinoma

BackgroundPreoperative sarcopenia is a predictor of poor survival in cancer patients. We hypothesized that sarcopenia could progress as occult metastasis arose, especially after highly invasive surgery for highly aggressive malignancy. This study aimed to evaluate the associations of postoperative changes in skeletal muscle mass volume with survival outcomes in patients who underwent surgery for perihilar cholangiocarcinoma.MethodsFifty-six patients who underwent major hepatectomy with extrahepatic bile duct resection for perihilar cholangiocarcinoma were studied. The skeletal muscle index (SMI) at the third lumbar vertebra was calculated from axial computed tomography images taken preoperatively and 3–6 months postoperatively (early postoperative period). The associations of clinicopathological variables, including changes of SMI after surgery, with overall survival and recurrence-free survival were evaluated. Moreover, the associations of decreased SMI and elevated serum carbohydrate antigen 19–9 level with early recurrence and poor survival was compared.ResultsAmong 56 patients, 26 (46%) had sarcopenia preoperatively and SMI decreased in 29 (52%) in the early postoperative period. During the median follow-up of 57.9 months, 35 patients (63%) developed recurrence and 29 (50%) died. Decreased SMI in the early postoperative period was independently associated with a shorter overall survival (hazard ratio, 2.39; 95% confidence interval, 1.00–6.18; P = 0.049) and a shorter recurrence-free survival (hazard ratio, 2.14; 95% confidence interval, 1.04–4.57; P = 0.039), whereas elevated carbohydrate antigen 19–9 level was not.ConclusionsDecreased SMI in the early postoperative period may be used as a predictor for recurrence and poor survival in patients undergoing surgery for perihilar cholangiocarcinoma.

MTAP deficiency contributes to immune landscape remodelling and tumour evasion.

Methylthioadenosine phosphorylase (MTAP) deficiency occurs in various malignancies and is associated with poor survival in cancer patients. However, the mechanisms underlying tumour progression due to MTAP loss are yet to be elucidated. Utilizing integrated analyses of the transcriptome, proteome and secretome, we demonstrated that MTAP deficiency alters tumour-intrinsic, immune-related pathways and reprograms cytokine profiles towards a tumour-favourable environment. Additionally, MTAP-knockout cells exhibited a marked increase in the immune checkpoint protein PD-L1. Upon co-culturing primary T cells with cancer cells, MTAP loss-mediated PD-L1 upregulation inhibited T cell-mediated killing activity and induced several T cell exhaustion markers. In two xenograft tumour models, we showed a modest increase in average volume of tumours derived from MTAP-deficient cells than that of MTAP-proficient tumours. Surprisingly, a remarkable increase in tumour size was observed in humanized mice bearing MTAP-deficient tumours, as compared to their MTAP-expressing counterparts. Following immunophenotypic characterization of tumour-infiltrating leukocytes by mass cytometry analysis, MTAP-deficient tumours were found to display decreased immune infiltrates with lower proportions of both T lymphocytes and natural killer cells and higher proportions of immunosuppressive cells as compared to MTAP-expressing tumour xenografts. Taken together, our results suggest that MTAP deficiency restructures the tumour immune microenvironment, promoting tumour progression and immune evasion.

Role of methylation-related genes CRYAB and SLC39A11 in the occurrence and development of lung adenocarcinoma.

Previous studies have shown that human crystallin alpha B (CRYAB) is highly expressed in human cancers and associated with poor survival in cancer patients. Here we investigated whether SLC39A11 and CRYAB genes were related to the proliferation and development of lung adenocarcinoma (LUAD) to explore their potential as therapeutic targets and prognostic markers for LUAD. CRYAB and SLC39A11 genes were identified from The Cancer Genome Atlas (TCGA) database and Gene Expression Omnibus (GEO) database. The human lung cancer cell lines A549 and H1975 were cultured, transfected, and subjected to RNA extraction. After genomic DNA removal, the RNA was reverse-transcribed. Differences between 2 groups were compared using t-test. Knockdown of SLC39A11 inhibited the proliferation of LUAD cells in A549 and H1975. Knockdown of CRYAB promoted the increase of LUAD cell clones, while knockdown of SLC39A11 suppressed LUAD cell clones. In both A549 and H1975 cell lines, knockout of CRYAB inhibited the apoptosis of LUAD cells, whereas knockout of SLC39A11 promoted the apoptosis of LUAD cells. In the H1975 cell line, knockout of CRYAB also lowered the proportion of cells in interphase and increased the proportion of mitotic cells, while knockout of SLC39A11 also slowed down the division cycle of tumor cells. Knockdown of CRYAB promoted the migration of LUAD cells in both the A549 cell line and H1975 cell line. In the H1975 cell line, knockout of SLC39A11 also reduced the invasive ability of LUAD cells. CRYAB and SLC39A11 could be used as prognostic indicators and therapeutic targets for LUAD.