Diverse Cancer Types Research Articles

Abstract LB138: DAXX interacts with sterol regulatory element-binding proteins (SREBPs) to promote oncogenic lipogenesis and tumorigenesis in triple-negative breast cancer

Abstract Elevated lipid metabolism including lipogenesis is a major metabolic feature in cancer cells. In breast and other cancer types, genes involved in lipogenesis are highly upregulated, but the mechanisms that control their expression remain poorly understood. DAXX modulates gene expression through binding to numerous transcription factors although the functional impact of these diverse interactions remains to be defined. Our recent analysis indicates that DAXX is overexpressed in diverse cancer types and metastases. However, mechanisms underlying DAXX’s oncogenic function remains elusive. Using global integrated transcriptomic and lipidomic analyses, we show that DAXX plays a key role in lipid metabolism in triple-negative breast cancer (TNBC) cells. DAXX depletion attenuates, while its overexpression enhances, lipogenic gene expression, lipid synthesis and tumor growth. Mechanistically, DAXX interacts with SREBP1 and SREBP2 and activates SREBP-mediated transcription. DAXX associates with lipogenic gene promoters through SREBPs. Underscoring the critical roles for the DAXX-SREBP interaction for lipogenesis, SREBP2 knockdown attenuates tumor growth in cells with DAXX overexpression, and a DAXX mutant unable to bind SREBPs are incapable of promoting lipogenesis and tumor growth. In TNBC patients, DAXX expression levels are increased in breast cancer brain metastasis and correlate with poor patient survival. Our results identify the DAXX-SREBP axis as an important pathway for tumorigenesis in TNBC. (This work is supported by Florida Department of Health Grants.) Citation Format: Iqbal Mahmud, Guimei Tian, Tarun Hutchison, Brandon Kim, Daiqing Liao. DAXX interacts with sterol regulatory element-binding proteins (SREBPs) to promote oncogenic lipogenesis and tumorigenesis in triple-negative breast 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 LB138.

Abstract 3372: Comprehensive analysis of estrogen-regulated testis specific lncRNAs in breast cancer

Abstract Long non-coding RNAs (lncRNAs) have transcripts longer than 200 nucleotides and recent studies have established they are abnormally expressed in cancer and have the potential to function as either oncogenes or tumor suppressors; Therefore, their potential utility as biomarkers and targets for different types cancer. Breast cancer is the most common cancer diagnosed (excluding skin cancers) among women in the United States, accounting for nearly 1 in 3 cancers. Thus, the need for effective targets and biomarkers to prevent/treat this disease. Available datasets to study gene expression in human diseases are a valuable asset to better understand complex disorders and phenotypes. In this study, we are using three publicly available datasets: The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx) and Cancer Cell Line Encyclopedia (CCLE) to study breast cancer and identify coding and lncRNA transcripts differentially expressed in tumors and highly specific to testis. At the end of our analysis pipeline, we identified a total of 1,668 upregulated genes in breast cancer and 1,163 downregulated genes in breast cancer, from which a total of 31 were lncRNAs. Characterization of the lncRNAs show highly specificity in diverse types of breast cancer (Basal, Luminal A, Luminal B and HER2) compared to protein coding genes. Cell line analysis of breast cancer RNA-Seq samples by the CCLE- Broad Institute show clustering of gene expression for the identified lncRNAs according to molecular subtype. Moreover, categorization of the lncRNAs using TCGA is described by ER- and ER+ status, molecular subtype, comparison between breast tumor, breast normal and GTEx breast, and gene ontology analysis of KEGG Pathways using DAVID. Furthermore, Kaplan-Meier analysis is shown to estimate the potential survival outcome of patients with high/low expression of lncRNAs in breast cancer. Lastly, we compared results from qRT-PCR with our in-silico data for the top lncRNAs and findings show a good correlation between both platforms. Overall, we identify a total of 31 lncRNAs that are estrogen-regulated testis specific in breast cancer by differential expression analysis. In addition, we provide a full genomic characterization of the lncRNAs that show expression according to different molecular subtypes using data from TCGA, GTEx and CCLE. Finally, we show potential survival outcome of high/low expression of identified lncRNAs in breast cancer patients. Taken together, we present lncRNAs specific in breast cancer and high expression in testis, along with their comprehensive characterization of expression signatures. Further studies, are needed to assess their efficacy as potential biomarkers and therapeutic targets in breast cancer. S.S.G. is supported by a grant from the Cancer Prevention and Research Institute of Texas (CPRIT; RR170020). S.S.G. is also supported by a grant from Lizanell and Colbert Coldwell foundation. Citation Format: Enrique I. Ramos, Barbara Yang, Ramesh Choudhari, Melina J. Sedano, Shrikanth S. Gadad. Comprehensive analysis of estrogen-regulated testis specific lncRNAs in breast 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 3372.

Abstract 2878: Mediating NK cell function against solid tumors, via targeting of tumor stroma, using a TEM8-targeting Tri-specific Killer Engager (TriKE®)

Abstract Immunotherapies for solid tumors need to overcome barriers within the solid tumor microenvironment (TME), including stromal cells, cancer-associated fibroblasts, endothelial cells, pericytes, and immune cells. These can comprise a significant portion of the tumor mass in many common carcinomas and contribute to immune cell dysfunction within the tumor. Stromal cells support cancer cells through several mechanisms and can promote metastasis. Tumor vasculature supplies cancer cells with nutrients and oxygen necessary for tumor growth and dissemination. Thus, targeting these components of cancer can enhance tumor rejection directly and increase sensitization of surviving tumors to immune infiltration and chemotherapeutic treatments through weakening of the tumor architecture. While there are current antiangiogenic therapies being tested clinically, they have associated toxicities in normal angiogenic processes, limiting clinical efficacy. Tumor Endothelial Marker 8 (TEM8, encoded by the ANTXR1 gene) is a highly conserved integrin-like adhesion molecule that was discovered on the endothelium of colorectal cancer, and subsequent studies identified its expression on multiple stromal cells—endothelial cells, fibroblasts, and pericytes—in the tumor microenvironment of diverse human cancer types. Little to no expression has been demonstrated in normal human stroma, indicating that this would be a good target for immunotherapy. Using a mammalian expression system, we designed and expressed a TEM8 targeting TriKE (cam1615TEM8) consisting of a humanized anti-CD16 single domain antibody, a wild-type IL-15 moiety, and an anti-TEM8 scFv. cam1615TEM8 induces NK cell degranulation and cytokine production against TEM8+ tumor and stromal cell lines (endothelial cells and fibroblasts). Using TEM8- cancer lines and a TEM8-CRISPR knockout version of A549 lung cancer lines, we show that cam1615TEM8 only activates NK cells in the presence of tumor antigen. cam1615TEM8 also preferentially stimulates TEM8+ tumor spheroid killing. Moreover, the camelid anti-CD16 nanobody selectively enhances signaling by the IL-15 moiety to NK cells, specifically promoting NK cell survival and proliferation in vitro and in vivo. In an in vivo xenogeneic mouse model, containing human NK cells and tumor, cam1615TEM8-treated mice had enhanced NK cell tumor infiltration, significantly decreased tumor growth, and enhanced survival when compared to IL-15 treated mice. Since TEM8 is highly conserved, and the scFv within the TriKE is reactive to mouse TEM8, we were able to show reduced tumor endothelial density in the tumors of mice treated with cam1615TEM8. Given the findings of this study cam1615TEM8 could be leveraged in several clinical settings by inducing NK cell specific targeting of the tumor itself and/or the tumor endothelium and stroma. Citation Format: Martin Felices, Michael Kaminski, Peter Hinderlie, Rachel Hopps, Laura Bendzick, Melissa Geller, Jeffrey S. Miller. Mediating NK cell function against solid tumors, via targeting of tumor stroma, using a TEM8-targeting Tri-specific Killer Engager (TriKE®) [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 2878.

Abstract 3492: Exosome-based plasma RNA biomarkers for high-risk prostate cancer

Abstract Introduction: There is an unmet need for liquid biopsy tests to support the management of patients with intermediate and high-risk prostate cancer (PCa). In treatment naïve patients, risk stratification by standard pathology and tissue based genomic testing can be influenced by sampling errors inherent to PCa biopsy. Exosome-based liquid biopsies are emerging as a clinically effective platform for minimally invasive and highly sensitive diagnostics for diverse types of cancer. In this project we undertake the development of a plasma exosome-based liquid biopsy to stratify patients with clinically significant PCa that may circumvent the sampling challenges associated with tissue-based genomic tests. Exosomes are small double-lipid membrane vesicles that cells actively shed into various biofluids and that provide stable packages for RNA, DNA and protein molecules. Here we take advantage of a well-developed exosomal biomarker analysis platform to profile plasma exosomal RNA markers that differentiate treatment-naïve men diagnosed with NCCN low risk vs high risk PCa as the first step in the development of a plasma exosomal liquid biopsy to support the management of patients with intermediate and high-risk disease. Methods: We compared plasma exosomal RNA profiles from 11 high-risk and 9 low-risk, treatment-naïve, biopsy-confirmed patients together with 4 healthy controls. Exosomes were isolated from 1 ml plasma samples and exosomal RNA prepared using the ExosomeDx ExoLution platform. A hybrid-capture RNA Next Generation Sequencing analysis was performed to enrich for transcripts of exons, 3’ and 5’ untranslated RNA and long non-coding RNA (lncRNA) to identify differentially expressed RNAs. Results: We detected over 10,000 protein coding genes and ~400 lncRNAs in each plasma sample using exosomal RNAseq. 273 genes were differentially expressed between high-risk vs control but not in low-risk vs control. We identified four potential plasma exosomal biomarkers of high-risk PCa. These include three protein coding mRNA transcripts that showed greater than 20-fold lower expression in plasma from high-risk in comparison to low-risk patients; TCGA data show that two of these mRNA markers are also downregulated in PCa tissue in patients with worse survival. In addition, we identified one lncRNA that showed greater than 20-fold higher expression in plasma from high-risk in comparison to low-risk patients. We also detected multiple alternate androgen receptor transcripts in plasma exosomal RNA from the high-risk patients. Together, these represent early data for potentially novel liquid biopsy RNA biomarkers for high-risk PCa. Conclusions: Here we report results of a liquid biopsy biomarker discovery study to develop an exosome-based long RNA test for to support the management of patients with intermediate and high-risk PCa. Plasma exosomal RNA provides a rich reservoir of currently untapped biomarkers for high-risk PCa. Citation Format: Sandra M. Gaston, Johan K. Skog, Benjamin Spieler, Alan Dal Pra, Radka Stoyanova, Christian Fischer, Yevgenia Khodor, Grannum Sant, Seth Yu, Vasisht Tadigotla, Sudipto K. Chakrabortty, Sanoj Punnen, Alan Pollack. Exosome-based plasma RNA biomarkers for high-risk prostate 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 3492.

Abstract 1221: Comprehensive cell-type classification of tumor and normal cells from single cell RNA sequencing in pan cancer settings

Abstract Single-cell RNA sequencing (scRNA-seq) allows for the study of the transcriptome at a cellular level, where populations of cells are annotated based on the expression of marker genes, providing a tool to gain cell-specific, subtle insights on cancer biology. However, precise annotation of cell type remains a challenge, hindering the efficiency of data interpretation. Several existing tools for cell-type annotation have been developed to improve resolution and reproducibility, yet their performance is reduced when the reference dataset contains many cell types, subclasses of similar cell types, or malignant cells. Interpatient malignant cell heterogeneity often leads to reduced accuracy when classifying cancer cells as most methods rely on correlations to a reference from a different source. Given the challenges in the annotation of scRNA-seq data of cancer and its high impact for elucidating mechanisms associated with tumor heterogeneity, pathogenesis, and treatment, we developed a comprehensive, hierarchically organized, multi-layered classifier spanning diverse malignant and normal cells of the tumor microenvironment. We found that performance improves when each layer focuses on a smaller number of classes and each cell sequentially moves down a series of classifiers with increased cell type resolution. When applied to an external validation dataset of over 300 primary solid tumor biopsies spanning diverse cancer types, the classifier accurately annotated the tissue of origin of malignant cells, and relevant subtypes of stromal and blood cells, with average F1 scores of 0.91, 0.95 and 0.99 respectively. Using confidence thresholds at each layer, the classifier abstains from classifying ambiguous cells. We applied 4 existing annotators provided with the same reference to the external test dataset and found that cancer cells are misclassified or unclassified, while the blood and stromal cells are accurately classified, highlighting our tool’s unique ability to classify cancer cells. Moreover, we applied our classifier’s to scRNA-seq data derived from breast cancer metastasis to the liver and were able to uncover the tissue of origin, demonstrating the potential use for determining the source of a metastatic tumour. Finally, given that our classifier is modular, we leveraged two recently published single cell breast cancer atlases to add a breast cancer subtype classification layer, that consistently identified the correct clinical subtype of single breast cancer cells in external data. This study provides a flexible model for the annotation of cells comprising the tumor microenvironment in pan cancer settings, while existing methods require tissue-specific references for every cancer type. Our classifier provides a powerful method for investigating intercellular communication pathways between tumor cells and non-malignant cells of the tumor microenvironment. Citation Format: Ido Nofech-Mozes, Philip Awadalla, Sagi Abelson. Comprehensive cell-type classification of tumor and normal cells from single cell RNA sequencing in pan cancer settings [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 1221.

Identification and Validation of lncRNA-AC087588.2 in Lung Adenocarcinoma: A Novel Prognostic and Diagnostic Indicator.

Lung adenocarcinoma (LUAD) is the most common histological lung cancer, and it is the leading cause of cancer-related deaths worldwide. Long non-coding RNAs (lncRNAs) have been implicated in the initiation and progression of various cancers. LncRNA-AC087588.2 (ENSG00000274976) is a novel lncRNA that is abnormally expressed in diverse cancer types, including LUAD. However, the clinical significance, prognostic value, diagnostic value, immune role, and the potential biological function of AC087588.2 LUAD remain elusive. In this study, we found that AC087588.2 was upregulated and associated with a poor prognosis in LUAD. In addition, univariate and multivariate Cox regression analysis indicated that AC087588.2 could be an independent prognostic factor for LUAD. Functionally, the knockdown of AC087588.2 restrained LUAD cell proliferation and migration in vitro. Finally, we constructed a ceRNA network that included hsa-miR-30a-5p and four mRNAs (ANLN, POLR3G, EHBP1, and ERO1A) specific to AC087588.2 in LUAD. The Kaplan–Meier survival analysis showed that lower expression of hsa-miR-30a-5p and higher expression of ANLN, POLR3G, EHBP1, and ERO1A were associated with adverse clinical outcomes in patients with LUAD. This finding provided a comprehensive view of the AC087588.2-mediated ceRNA network in LUAD, thereby highlighting its potential role in the diagnosis and prognosis of LUAD.

Essential Roles of TDO2 in Gastric Cancer: TDO2 Is Associated with Cancer Progression, Patient Survival, PD-L1 Expression, and Cancer Stem Cells

Introduction: Tryptophan metabolism has been shown to be involved in tumor development. Two main tryptophan-degrading enzymes, tryptophan 2,3-dioxygenase (TDO2) and indoleamine 2,3-dioxygenase 1 (IDO1), may potently promote cancer cell survival and distant metastasis in diverse types of cancer, such as lung and breast cancer. IDO1 overexpression is an independent prognosticator in gastric cancer (GC). This work aimed to uncover the expression of TDO2 and its clinicopathologic significance in GC. Methods: TDO2 expression was evaluated in public data of The Cancer Genome Atlas cohort STAD and in two different GC cohorts. Correlation between TDO2 and immune cell infiltrates as well as PD-L1 tumor staining was investigated. The biofunction of TDO2 was examined with MTT, colony formation, and spheroid formation assays by RNA interference. Results: TDO2 expression was correlated with both progressive disease and clinical outcome, and its expression was an independent predictor of prognosis in GC. TDO2 expression was correlated with infiltration of immune cells and tumor expression of PD-L1. Inhibition of TDO2 expression suppressed cell proliferation, colony formation, and cell invasion of GC cells. Additionally, suppression of TDO2 expression inhibited spheroid body-formation and viability of GC organoids. Conclusion: Our data show that TDO2 might be a crucial marker for predicting prognosis and targeted therapy in GC.

Novel Variation in Acyl-CoA Synthetase Long Chain Family Member 6 (ACSL6) Results in Protein Structural Modification and Multiple Non-Related Neoplasia in a 46-Year-Old: Case Report

Multiple non-related neoplasia does not have an established approach or benefits for performing whole-exome sequencing (WES) analysis. We report on a 46-year-old woman who developed astrocytoma, thyroid, and breast cancer within 10 years. The WES analysis found a novel missense variant in the ACSL6 gene, and the protein modeling showed altered secondary and tertiary structures, which modify the binding to cofactors and substrates. ACSL6 is involved in lipid metabolism, expressed in the brain, thyroid, and breast tissues, and is associated with diverse types of cancer. Our study demonstrates the benefit of WES analysis compared with commercial panels in patients with non-related neoplasia.

Rediscovery of Traditional Plant Medicine: An Underestimated Anticancer Drug of Chelerythrine

In many studies, the extensive and significant anticancer activity of chelerythrine (CHE) was identified, which is the primary natural active compound in four traditional botanical drugs and can be applied as a promising treatment in various solid tumors. So this review aimed to summarize the anticancer capacities and the antitumor mechanism of CHE. The literature searches revolving around CHE have been carried out on PubMed, Web of Science, ScienceDirect, and MEDLINE databases. Increasing evidence indicates that CHE, as a benzophenanthridine alkaloid, exhibits its excellent anticancer activity as CHE can intervene in tumor progression and inhibit tumor growth in multiple ways, such as induction of cancer cell apoptosis, cell cycle arrest, prevention of tumor invasion and metastasis, autophagy-mediated cell death, bind selectively to telomeric G-quadruplex and strongly inhibit the telomerase activity through G-quadruplex stabilization, reactive oxygen species (ROS), mitogen-activated protein kinase (MAPK), and PKC. The role of CHE against diverse types of cancers has been investigated in many studies and has been identified as the main antitumor drug candidate in drug discovery programs. The current complex data suggest the potential value in clinical application and the future direction of CHE as a therapeutic drug in cancer. Furthermore, the limitations and the present problems are also highlighted in this review. Despite the unclearly delineated molecular targets of CHE, extensive research in this area provided continuously fresh data exploitable in the clinic while addressing the present requirement for further studies such as toxicological studies, combination medication, and the development of novel chemical methods or biomaterials to extend the effects of CHE or the development of its derivatives and analogs, contributing to the effective transformation of this underestimated anticancer drug into clinical practice. We believe that this review can provide support for the clinical application of a new anticancer drug in the future.

Novel platform for identifying multiple cancer-specific antigens.

2530 Background: Virtually all cancer treatments that kill cancer cells also adversely affect normal cells and often have debilitating side effects. A key challenge has been to identify antigens that are suitable for antibody-based targeted therapeutics. We have developed a novel platform to detect and identify multiple cell surface antigens that are unique to human cancers. Methods: Breast Cancer cells (Hs578T) or Melanoma cells (Hs895.T) were injected into rabbits to generate polyclonal xeno-antibodies. The resulting serum against each cancer was then incubated with normal cells of the same tissue type derived from the same patients; Breast Epithelium (matched-pair Hs578Bst) or Skin Fibroblasts (matched-pair Hs895.Sk) in order to absorb antibodies that cross-react with antigens on normal tissue (U.S. Patent Application No. 16/243,161). Flow cytometry was used for measuring the binding capacity of the unfiltered serum and filtered serum (lacking cross-reacting antibodies) on both the cancer cells and normal cells. Cytotoxicity assays were performed using a secondary ADC (anti-rabbit IgG specific antibody conjugated to MMAF), to determine the cytotoxicity of the unfiltered antibodies and filtered antibodies on normal and cancer cells. Results: Flow cytometry revealed, for antibodies against breast cancer, there was 90.3% and 88.6% binding to breast cancer cells and normal breast epithelial cells, respectively, pre-filtration; and 78.9% and 23% binding, respectively, post-filtration. For antibodies against melanoma, there was 89.2% and 86.6% binding to melanoma cells and normal skin fibroblasts, respectively, pre-filtration; and 55.1% and 0% binding, respectively, post-filtration. The Table summarizes the cytotoxicity of unfiltered and filtered antibodies on cancer cells and normal cells. Remarkably, there was preferential killing of all the cancer cells tested by both unfiltered and filtered antibodies, while there was no cytotoxicity in the normal cells using filtered antibodies. Cytotoxicity was also observed when antibodies generated against one type of cancer were incubated with other cancer types. Conclusions: This data suggests that our xeno-antibody based method can detect cancer-specific sites, not found on normal cells, and diverse cancer types may share a common antigenic signature. These findings are significant for identifying unique antigens on cancer cells and designing targeted therapies against cancer. [Table: see text]

Examination of Provider and Patient Knowledge, Beliefs, and Preferences in Integrative Oncology at a National Cancer Institute-Designated Comprehensive Cancer Center.

Purpose:The use of integrative approaches for symptom management is highly prevalent among patients undergoing cancer treatment and among cancer survivors and is increasingly endorsed by clinical practice guidelines. However, access to and implementation of integrative oncology (IO) approaches are hindered by barriers at multiple levels, including logistic, geographic, financial, organizational, and cultural barriers. The goal of this mixed-method study was to examine oncology provider and patient knowledge, beliefs, and preferences in IO to identify facilitators, barriers, and recommendations for implementation of IO modalities.Materials and Methods:Data sources included patient surveys and provider semistructured interviews. Patients were in active treatment (n = 100) and survivors (n = 100) of heterogeneous cancer types. Patient and survivor surveys interrogated: (1) interest in types of IO approaches; and (2) preferences for delivery modality, frequency, and location. Providers (n = 18) were oncologists and nurse navigators working with diverse cancer types. Interviews queried their knowledge of and attitudes about IO, about their patients' needs for symptom management, and for recommendations for implementation of IO approaches in their clinic. We used the Consolidated Framework for Implementation Research framework to systematically analyze provider interviews.Results:The primary interests reported among actively treated patients and survivors were massage therapy, acupuncture, and wellness/exercise. Most patients expressed interest in both group and individual sessions and in telehealth or virtual reality options. Emergent themes from provider interviews identified barriers and facilitators to implementing IO approaches in both the internal and external settings, as well as for the implementation process.Conclusion:The emphasis on mind–body interventions as integrative rather than alternative highlights the importance of interventions as evidence-based, comprehensive, and integrated into health care. Gaining simultaneous perspectives from both patients and physicians generated insights for the implementation of IO care into complex clinical systems within a comprehensive cancer center.

Using PD-L1 full-length structure, enhanced induced fit docking and molecular dynamics simulations for structural insights into inhibition of PD-1/PD-L1 interaction by small-molecule ligands

ABSTRACT T-cell activation through the blockade of PD-1/PD-L1 interaction by monoclonal antibodies has demonstrated the clinical benefit for patients with diverse types of metastatic cancers. However, a number of limitations when producing and using antibodies hamper their broader clinical application. This awakened a significant interest in design and discovery of small-molecule inhibitors (SMIs) of PD-1/PD-L1 interaction, and several such inhibitors have been identified. However, up to now, many mechanistic details of their inhibitory action remain not fully understood. In this work, a new protocol of enhanced protein conformational sampling combining RosettaLigand and Schrodinger Induced Fit Docking approaches with Molecular Dynamics simulations, preceded by structural modelling of the full-length PD-L1 – dimer in the heterogeneous environment including the membrane and extracellular solution, are used for the prediction of atomistic structures of the complexes of the PD-L1 dimers with BMS-1016, BMS-2007, BMS-4121, BMS-40210, four SMIs with a high inhibitory activity in vitro against PD-1/PD-L1 interaction. Critical protein–ligand interactions responsible for the stabilisation of the complexes of the PD-L1 dimer with SMIs and high inhibitory activities of these SMIs against the PD-1/PD-L1 interaction are revealed. The roles of the membrane and of interaction of PD-L1 with its cis and trans protein partners are also addressed.

Diet and exercise advice and referrals for cancer survivors: an integrative review of medical and nursing perspectives.

PurposeTo examine the perspectives of medical and nursing health professionals concerning their roles and responsibilities in providing dietary and exercise advice to cancer survivors, and referrals to allied health professionals.MethodsAn integrative review. PubMed, CINAHL, PsycINFO, Embase, Web of Science databases, and bibliographies of relevant studies were searched from December 2011 to June 2021. All studies were eligible for inclusion. The Mixed-Methods Appraisal Tool (MMAT) was used to critically appraise included studies. Data were extracted and synthesised regarding the perspectives of medical and nursing health professionals on their roles, responsibilities, barriers, and facilitators.ResultsTwenty-one studies involving 3401 medical and nursing health professionals and 264 cancer survivors of diverse cancer types were included. Ten quantitative, nine qualitative, and two mixed-methods studies were eligible. All included studies met at least 80% of the quality criteria in the MMAT. Major findings include the following: (1) medical and nursing health professionals were unclear on their roles in providing dietary and exercise advice to cancer survivors but agreed they play a key role in referrals to dietitians and exercise professionals; (2) most cancer survivors valued the involvement of their general practitioner when receiving dietary and exercise advice.ConclusionAlthough medical and nursing health professionals understand that referrals to allied health professionals form part of their role, there is a lack of clarity regarding their roles to provide dietary and exercise advice to cancer survivors. Future studies should address barriers and facilitators of dietary and exercise advice and referral by medical and nursing health professionals.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00520-022-07152-w.

Cannabidiol exerts anti-proliferative activity via a cannabinoid receptor 2-dependent mechanism in human colorectal cancer cells

Colorectal cancer is the third leading cause of cancer incidence and mortality in the United States. Cannabidiol (CBD), the second most abundant phytocannabinoid in Cannabis sativa, has potential use in cancer treatment on the basis of many studies showing its anti-cancer activity in diverse types of cancer, including colon cancer. However, its mechanism of action is not yet fully understood. In the current study, we observed CBD to repress viability of different human colorectal cancer cells in a dose-dependent manner. CBD treatment led to G1-phase cell cycle arrest and an increased sub-G1 population (apoptotic cells); it also downregulated protein expression of cyclin D1, cyclin D3, cyclin-dependent kinase 2 (CDK2), CDK4, and CDK6. CBD further increased caspase 3/7 activity and cleaved poly(ADP-ribose) polymerase, and elevated expression of endoplasmic reticulum (ER) stress proteins including binding immunoglobulin protein (BiP), inositol-requiring enzyme 1α (IRE1α), phosphorylated eukaryotic initiation factor 2α (eIF2α), activating transcription factor 3 (ATF3), and ATF4. We found that CBD repressed cell viability and induced apoptotic cell death through a mechanism dependent on cannabinoid receptor type 2 (CB2), but not on CB1, transient receptor potential vanilloid, or peroxisome proliferator-activated receptor gamma. Anti-proliferative activity was also observed for other non-psychoactive cannabinoid derivatives including cannabidivarin (CBDV), cannabigerol (CBG), cannabicyclol (CBL), and cannabigerovarin (CBGV). Our data indicate that CBD and its derivatives could be promising agents for the prevention of human colorectal cancer.

Characterization of the Estrogen Response Helps to Predict Prognosis and Identify Potential Therapeutic Targets in Cholangiocarcinoma

Cholangiocarcinoma (CCA) is an aggressive malignancy originating from the epithelium of the bile duct. The prognosis of patients is poor regardless of radical resection and chemoradiotherapy. The current classification and prognostic model of CCA are unable to satisfy the requirements for predicting the clinical outcome and exploring therapeutic targets. Estrogen signaling is involved in diverse cancer types, and it has long been established that CCA could be regulated by estrogen. In our study, estrogen response was identified to be significantly and stably correlated with poor prognosis in CCA. Employing several algorithms, CCA was classified into ES cluster A and B. ES cluster B was mainly composed of patients with fluke infection and overlapped with CCA cluster 1/2, and ES cluster A was mainly composed of patients without fluke infection and overlapped with CCA cluster 3/4. COMT and HSD17B1 were identified to be responsible for the differential estrogen response between ES clusters A and B, and the estrogen response may be correlated with the differentiation and cancer stemness of CCA at the single-cell level. Complement activation and the expression of C3 and C5, which are mainly expressed by CCA cells, were significantly downregulated in ES cluster B. An estrogen response risk score (ESRS) model was constructed to predict the prognosis of CCA, followed by a nomogram integrating ESRS and clinical features. Finally, altered pathways, applicable drugs and sensitivity to chemical drugs were analyzed specific to the estrogen response. In summary, our results provide insights into the role of the estrogen response in CCA progression as well as applicable drugs and potential therapeutic targets in estrogen metabolism, the complement system and ESRS-related pathways.

Homologous recombination deficiency in diverse cancer types and its correlation with platinum chemotherapy efficiency in ovarian cancer

BackgroundHomologous recombination deficiency (HRD) is a molecular biomarker for administrating PARP inhibitor (PARPi) or platinum-based (Pt) chemotherapy. The most well-studied mechanism of causing HRD is pathogenic BRCA1/2 mutations, while HRD phenotype is also present in patients without BRCA1/2 alterations, suggesting other unknown factors.MethodsThe targeted next-generation sequencing (GeneseeqPrime® HRD) was used to evaluate the HRD scores of 199 patients (Cohort I). In Cohort II, a total of 85 Pt-chemotherapy-treated high-grade serous ovarian cancer (HGSOC) patients were included for investigating the role of HRD score in predicting treatment efficacy. The concurrent genomic features analyzed along HRD score evaluation were studied in a third cohort with 416 solid tumor patients (Cohort III).ResultsAn HRD score ≥ 38 was predefined as HRD-positive by analyzing Cohort I (range: 0–107). Over 95% of the BRCA1/2-deficient cases of Cohort I were HRD-positive under this threshold. In Cohort II, Pt-sensitive patients have significantly higher HRD scores than Pt-resistant patients (median: 54 vs. 34, p = 0.031) and a significantly longer PFS was observed in HRD-positive patients (median: 548 vs. 343 days, p = 0.003). Furthermore, TP53, NCOR1, and PTK2 alterations were enriched in HRD-positive patients. In Cohort III, impaired homologous recombination repair pathway was more frequently observed in HRD-positive patients without BRCA1/2 pathogenic mutations. The alteration enrichment of TP53, NCOR1, and PTK2 observed in Cohort II was also validated by the ovarian subgroup in Cohort III.ConclusionsUsing an in-house HRD evaluation method, our findings show that overall HRR gene mutations account for a significant part of HRD in the absence of BRCA1/2 aberrations, and suggest that HRD positive status might be a predictive biomarker of Pt-chemotherapy.

Over-Expression of Long Non-Coding RNA-AC099850.3 Correlates With Tumor Progression and Poor Prognosis in Lung Adenocarcinoma.

Lung adenocarcinoma (LUAD) is the most common histological lung cancer, and it is the leading cause of cancer-related deaths worldwide. Long noncoding RNAs (lncRNAs) have been implicated in the initiation and progression of various cancers. LncRNA-AC099850.3 is a novel lncRNA that is abnormally expressed in diverse cancer types including LUAD. However, the clinical significance, prognostic value, diagnostic value, immune role, and potential biological function of AC099850.3 LUAD remain elusive. In this study, we found that AC099850.3 was highly expressed in LUAD and associated with an advanced tumor stage, poor prognosis, and immune infiltration. Receiver operating curve analysis revealed the significant diagnostic ability of AC099850.3 (AUC=0.888). Functionally, the knockdown of AC099850.3 restrained LUAD cell proliferation and migration in vitro. Finally, we constructed a competitive endogenous RNAs (ceRNA) network that included hsa-miR-101-3p and 4 mRNAs (ESPL1, AURKB, BUB3, and FAM83D) specific to AC099850.3 in LUAD. Kaplan–Meier survival analysis showed that a lower expression of miR-101-3p and a higher expression of ESPL1, AURKB, BUB3, and FAM83D, were associated with adverse clinical outcomes in patients with LUAD. This finding provided a comprehensive view of the AC099850.3-mediated ceRNA network in LUAD, thereby highlighting its potential role in the diagnosis and prognosis of LUAD.

Ribavirin inhibits cell proliferation and metastasis and prolongs survival in soft tissue sarcomas by downregulating both protein arginine methyltransferases 1 and 5.

Protein arginine methyltransferases 1 and 5 (PRMT1 and PRMT5) are frequently overexpressed in diverse types of cancers and correlate with poor prognosis, thus making these enzymes potential therapeutic targets. The aim of this study was to assess and elucidate the anti-tumour effect and epigenetic regulatory mechanism of ribavirin in soft tissue sarcomas (STS). We showed that ribavirin inhibited growth and metastasis and prolonged survival in animals bearing STS cells by downregulating the mRNA and protein levels of PRMT1/PRMT5 and attenuating the accumulation of asymmetric and symmetric di-methylation of arginine (ADMA and SDMA). Furthermore, ribavirin lowered the permeability of the peritoneum in KM mice bearing S180 ascites via decreasing the level of vascular endothelial growth factor (VEGF). Ribavirin was a potent inhibitor of cell proliferation and metastasis in STS cells through downregulation of both type I PRMT1 and type II PRMT5. Ribavirin could be used to enhance the efficacy of doxorubicin in STS allograft tumour models.

MP27-12 DEVELOPMENT OF EXOSOME-BASED PLASMA RNA BIOMARKERS FOR HIGH-RISK PROSTATE CANCER

You have accessJournal of UrologyCME1 May 2022MP27-12 DEVELOPMENT OF EXOSOME-BASED PLASMA RNA BIOMARKERS FOR HIGH-RISK PROSTATE CANCER Sandra Gaston, Benjamin Spieler, Alan Dal Pra, Radka Stoyanova, Yevgenia Khodor, Sudipto Chakrabortty, Vasisht Tadigotla, Seth Yu, Grannum Sant, Christian Fischer, Sanoj Punnen, Johan Skog, and Alan Pollack Sandra GastonSandra Gaston More articles by this author , Benjamin SpielerBenjamin Spieler More articles by this author , Alan Dal PraAlan Dal Pra More articles by this author , Radka StoyanovaRadka Stoyanova More articles by this author , Yevgenia KhodorYevgenia Khodor More articles by this author , Sudipto ChakraborttySudipto Chakrabortty More articles by this author , Vasisht TadigotlaVasisht Tadigotla More articles by this author , Seth YuSeth Yu More articles by this author , Grannum SantGrannum Sant More articles by this author , Christian FischerChristian Fischer More articles by this author , Sanoj PunnenSanoj Punnen More articles by this author , Johan SkogJohan Skog More articles by this author , and Alan PollackAlan Pollack More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002570.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: There is an unmet need for liquid biopsy tests to support the management of patients with intermediate and high-risk prostate cancer (PCa). In treatment naïve patients, risk stratification by standard pathology and tissue based genomic testing can be influenced by sampling errors inherent to PCa biopsy. Exosome-based liquid biopsies are emerging as a clinically effective platform for minimally invasive and highly sensitive diagnostics for diverse types of cancer. Exosomes are small double-lipid membrane vesicles that cells shed into various biofluids and that provide packages for RNA, DNA and protein molecules. In this project we undertake the development of a plasma exosome-based liquid biopsy to stratify patients with clinically significant PCa that may circumvent the sampling challenges associated with tissue-based genomic tests. METHODS: Our pilot study included plasma samples from 11 NCCN high-risk and 9 NCCN low-risk, treatment-naïve, biopsy-confirmed PCa patients together with 4 healthy controls. Exosomes were isolated from 1 ml plasma using the ExosomeDx ExoLution platform. A hybrid-capture RNA Next Generation Sequencing analysis was performed to enrich for transcripts of exons, 3’ and 5’ untranslated RNA and long non-coding RNA (lncRNA). Data analysis included identification of differentially expressed RNAs in high-risk vs low-risk vs control samples. RESULTS: In each plasma exosome sample we detected transcripts of over 10,000 protein coding genes and ∼400 lncRNAs using exosomal RNAseq. 273 genes were differentially expressed between high-risk vs control but not between low-risk vs control. We identified four potential plasma exosomal biomarkers of high-risk PCa. These include three protein coding mRNA transcripts that showed > 20-fold lower expression in plasma from high-risk compared to low-risk patients; TCGA data show that two of these transcripts are also downregulated in PCa tissue in patients with worse survival. In addition, we identified one lncRNA that showed > 20-fold higher expression in plasma from high-risk compared to low-risk patients. We also detected multiple alternate androgen receptor transcripts in plasma exosomal RNA from high-risk patients. Together, these represent early data for potentially novel liquid biopsy RNA biomarkers for high-risk PCa. CONCLUSIONS: Here we report results of a liquid biopsy biomarker discovery study to develop an exosome-based RNA test to support the management of patients with intermediate and high-risk PCa. Plasma exosomal RNA provides a rich reservoir of currently untapped biomarkers for high-risk PCa. Source of Funding: NIH Grants R01CA189295, U01CA239141 and P30CA240139 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e454 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Sandra Gaston More articles by this author Benjamin Spieler More articles by this author Alan Dal Pra More articles by this author Radka Stoyanova More articles by this author Yevgenia Khodor More articles by this author Sudipto Chakrabortty More articles by this author Vasisht Tadigotla More articles by this author Seth Yu More articles by this author Grannum Sant More articles by this author Christian Fischer More articles by this author Sanoj Punnen More articles by this author Johan Skog More articles by this author Alan Pollack More articles by this author Expand All Advertisement PDF DownloadLoading ...

A critical ETV4/Twist1/Vimentin axis in Ha-RAS-induced aggressive breast cancer.

RAS oncogenes are major drivers of diverse types of cancer. However, they are largely not druggable, and therefore targeting critical downstream pathways and dependencies is an attractive approach. We have isolated a tumorigenic cell line (FE1.2), which exhibits mesenchymal characteristics, after inoculating Ha-Ras-expressing retrovirus into mammary glands of rats, and subsequently isolated a non-aggressive revertant cell line (FC5). This revertant has lost the rat Ha-Ras driver and showed a more epithelial morphology, slower proliferation in culture, and reduced tumorigenicity in vivo. Re-expression of human Ha-RAS in these cells (FC5-RAS) reinduced mesenchymal morphology, higher proliferation rate, and tumorigenicity that was still significantly milder than parental FE1.2 cells. RNA-seq analysis of FC5-RAS vs FC5-Vector cells identified multiple genes whose expressions were regulated by Ha-RAS. This analysis also identified many genes including those controlling cell growth whose expression was altered by loss of HA-Ras in FC5 cells but remained unchanged upon reintroduction of Ha-RAS. These results suggest that targeting the Ha-Ras driver oncogene induces partial tumor regression, but it still denotes strong efficacy for cancer therapy. Among the RAS-responsive genes, we identified Twist1 as a critical mediator of epithelial-to-mesenchymal transition through the direct transcriptional regulation of vimentin. Mechanistically, we show that Twist1 is induced by the ETS gene, ETV4, downstream of Ha-RAS, and that inhibition of ETV4 suppressed the growth of breast cancer cells driven by the Ha-RAS pathway. Targeting the ETV4/Twist1/Vimentin axis may therefore offer a therapeutic modality for breast tumors driven by the Ha-RAS pathway.