Aggressive Disease In Patients Research Articles

Digoxin and Standard-of-Care Therapy for Heart Failure Patients with COVID-19: Analysis of Data from the US Military Health System (MHS) Data Repository

Cardiac glycosides such as digoxin, digitoxin and ouabain are still used around the world to treat patients with chronic heart failure with reduced ejection fraction (HFrEF) and/or atrial fibrillation (AF). However, in the US, only digoxin is licensed for treating these illnesses, and the use of digoxin for this group of patients is increasingly being replaced in the US by a new standard of care with groups of more expensive drugs. However, ouabain and digitoxin, and less potently digoxin, have also recently been reported to inhibit SARS-CoV-2 virus penetration into human lung cells, thus blocking COVID-19 infection. COVID-19 is known to be a more aggressive disease in patients with cardiac comorbidities, including heart failure. We therefore considered the possibility that digoxin might provide at least a measure of relief from COVID-19 in digoxin-treated heart failure patients. To this end, we hypothesized that treatment with digoxin rather than standard of care might equivalently protect heart failure patients with regard to diagnosis of COVID-19, hospitalization and death. To test this hypothesis, we conducted a cross-sectional study by using the US Military Health System (MHS) Data Repository to identify all MHS TRICARE Prime and Plus beneficiaries aged 18-64years with a heart failure (HF) diagnosis during the period April 2020 to August 2021. In the MHS, all patients receive equal, optimal care without regard to rank or ethnicity. Analyses included descriptive statistics on patient demographics and clinical characteristics, and logistic regressions to determine likelihood of digoxin use. We identified 14,044 beneficiaries with heart failure in the MHS during the study period. Of these, 496 were treated with digoxin. However, we found that both digoxin-treated and standard-of-care groups were equivalently protected from COVID-19. We also noted that younger active duty service members and their dependents with HF were less likely to receive digoxin compared with older, retired beneficiaries with more comorbidities. The hypothesis of equivalent protection by digoxin treatment of HF patients in terms of susceptibility to COVID-19 infection appears to be supported by the data.

Metabolic Enzyme DLST Promotes Tumor Aggression and Reveals a Vulnerability to OXPHOS Inhibition in High-Risk Neuroblastoma.

High-risk neuroblastoma remains therapeutically challenging to treat, and the mechanisms promoting disease aggression are poorly understood. Here, we show that elevated expression of dihydrolipoamide S-succinyltransferase (DLST) predicts poor treatment outcome and aggressive disease in patients with neuroblastoma. DLST is an E2 component of the α-ketoglutarate (αKG) dehydrogenase complex, which governs the entry of glutamine into the tricarboxylic acid cycle (TCA) for oxidative decarboxylation. During this irreversible step, αKG is converted into succinyl-CoA, producing NADH for oxidative phosphorylation (OXPHOS). Utilizing a zebrafish model of MYCN-driven neuroblastoma, we demonstrate that even modest increases in DLST expression promote tumor aggression, while monoallelic dlst loss impedes disease initiation and progression. DLST depletion in human MYCN-amplified neuroblastoma cells minimally affected glutamine anaplerosis and did not alter TCA cycle metabolites other than αKG. However, DLST loss significantly suppressed NADH production and impaired OXPHOS, leading to growth arrest and apoptosis of neuroblastoma cells. In addition, multiple inhibitors targeting the electron transport chain, including the potent IACS-010759 that is currently in clinical testing for other cancers, efficiently reduced neuroblastoma proliferation in vitro. IACS-010759 also suppressed tumor growth in zebrafish and mouse xenograft models of high-risk neuroblastoma. Together, these results demonstrate that DLST promotes neuroblastoma aggression and unveils OXPHOS as an essential contributor to high-risk neuroblastoma. SIGNIFICANCE: These findings demonstrate a novel role for DLST in neuroblastoma aggression and identify the OXPHOS inhibitor IACS-010759 as a potential therapeutic strategy for this deadly disease.

Abstract 2872: Methods of assessment of polysialyltransferase inhibitors for treatment of tumor cell dissemination

Abstract Polysialic acid (polySia) decorates the surface of NCAM (neuronal cell adhesion molecule) on neuroendocrine tumors, notably neuroblastoma and small cell lung cancer, and is strongly associated with poor prognosis and aggressive disease in patients in the clinic [1]. PolySia modulates tumor cell-cell and cell-matrix adhesion, migration, invasion and metastasis. SiRNA knockdown of polysialyltransferase (polyST) ST8SiaII, the enzyme primarily responsible for polySia synthesis in tumors, abrogates tumor cell migration and invasion. PolyST is a selective and largely unexplored therapeutic target for neuroblastoma dissemination [1]. While assays are available to assess polyST enzyme activity, there is no methodology available specifically optimized for identification of novel polyST inhibitors. We describe the development of cell-free and cell-based assays that enable assessment of polysialyltransferase inhibition. Development of the HPLC-fluorescence-based enzyme assay includes a comprehensive optimization of assay conditions, including evaluation of metal ion composition, enzyme concentration, substrate and acceptor concentration, temperature, pH and tolerance to DMSO, followed by validation using known polyST inhibitors. Thorough analysis of each of the assay components provided a set of optimized conditions. Under these optimized conditions, the experimentally observed Ki value for CMP, a competitive polyST inhibitor, was strongly correlated with the predicted Ki value, based on the classical Cheng-Prusoff equation [average fold error (AFE) = 1.043]. These results indicate that this assay can provide medium-throughput analysis for enzyme inhibitors with high accuracy. We additionally report optimized HPLC-based and ELISA-based methodologies for assessment of polyST inhibition in neuroblastoma cells, using endoneuraminidase N as control, and assessment of ICT3176 (a polysialylation inhibitor) as a test agent. In conclusion, in vitro cell-free and cell-based assays for accurate measurement of polysialyltransferase (polyST) inhibition are described, specifically designed for routine identification of potential polyST inhibitors, generation of kinetics data and assessment of mode of inhibition, and assessment of effects on cellular polysialylation. Given the considerable and growing interest in the polySTs as important anti-metastatic targets in cancer drug discovery, these are vital tools to enable preclinical identification and evaluation of novel polyST inhibitors for neuroblastoma therapy. [1] Falconer, R.A. et al., Curr. Cancer Drug Targets, 2012, 12, 925-939. Citation Format: Xiaoxiao Guo, Jodie R. Malcolm, Anjana Patel, Marrwa M. Ali, Goreti Ribeiro Morais, Steven D. Shnyder, Paul M. Loadman, Laurence H. Patterson, Robert A. Falconer. Methods of assessment of polysialyltransferase inhibitors for treatment of tumor cell dissemination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2872.

Enhanced triacylglycerol catabolism by carboxylesterase 1 promotes aggressive colorectal carcinoma.

The ability to adapt to low-nutrient microenvironments is essential for tumor cell survival and progression in solid cancers, such as colorectal carcinoma (CRC). Signaling by the NF-κB transcription factor pathway associates with advanced disease stages and shorter survival in patients with CRC. NF-κB has been shown to drive tumor-promoting inflammation, cancer cell survival, and intestinal epithelial cell (IEC) dedifferentiation in mouse models of CRC. However, whether NF-κB affects the metabolic adaptations that fuel aggressive disease in patients with CRC is unknown. Here, we identified carboxylesterase 1 (CES1) as an essential NF-κB-regulated lipase linking obesity-associated inflammation with fat metabolism and adaptation to energy stress in aggressive CRC. CES1 promoted CRC cell survival via cell-autonomous mechanisms that fuel fatty acid oxidation (FAO) and prevent the toxic build-up of triacylglycerols. We found that elevated CES1 expression correlated with worse outcomes in overweight patients with CRC. Accordingly, NF-κB drove CES1 expression in CRC consensus molecular subtype 4 (CMS4), which is associated with obesity, stemness, and inflammation. CES1 was also upregulated by gene amplifications of its transcriptional regulator HNF4A in CMS2 tumors, reinforcing its clinical relevance as a driver of CRC. This subtype-based distribution and unfavorable prognostic correlation distinguished CES1 from other intracellular triacylglycerol lipases and suggest CES1 could provide a route to treat aggressive CRC.

Hydroa vacciniforme‐like lymphoproliferative disorder: A clinicopathological, immunohistochemical, and prognostic study of 24 cases in China

Hydroa vacciniforme‐like lymphoproliferative disorder (HV‐LPD) is a rare cutaneous disease associated with Epstein–Barr virus infection. We retrospectively analyzed the clinical presentation, histopathological characteristics, and prognostic study of HV‐LPD in 24 Chinese patients. All patients presented with recurrent papulovesicular and necrotic eruptions on the face, neck, and extremities, with 11 showing systemic symptoms. Twenty patients were diagnosed with HV‐LPD in childhood (age < 18 years) and four in adulthood (age ≥ 18 years). The median age at diagnosis was 8.5 years old (range, 2–50). Histopathology revealed variably dense lymphocyte infiltration throughout the dermis. All cases were strongly positive for CD3 and Epstein–Barr encoding region based on in situ hybridization. Of 18 cases with a T‐cell phenotype, 15 harbored monoclonal rearrangements in T‐cell receptor (TCR) genes. Four cases with a natural killer cell phenotype carried polyclonal rearrangements in TCR genes. Among 24 patients, eight (33.3%) received chemotherapy, two (8.3%) allogeneic hematopoietic stem cell transplantation, and both are currently alive without disease. The median follow‐up period was 24 months (range, 7–120) and 23 patients were available: 15 (62.5%) were alive, and eight (33.3%) had died. Fourteen cases had a relapse of disease and three developed lymphoma within 24 months of diagnosis. The mean survival time of childhood‐onset patients was longer than that of adult‐onset patients (36.4 vs. 20.8 months). In summary, the wide clinical course and representative presentation of cases in our center reflect the pedigree characteristics of HV‐LPD. Allogeneic hematopoietic stem cell transplantation should be a preferred choice for relapse and refractory patients due to the poor effect of chemotherapy. Adult‐onset and high serum EBV DNA loads may indicate an increased risk of aggressive disease in patients with HV‐LPD.

The apparent diffusion coefficient (ADC) on 3-T MRI differentiates myometrial invasion depth and histological grade in patients with endometrial cancer.

Diffusion-weighted magnetic resonance imaging (DW-MRI) with apparent diffusion coefficient (ADC) measurement provides additional information about tumor microstructure with potential relevance for staging and predicting aggressive disease in patients with endometrial cancer (EC). To determine whether ADC values in EC diverge according to the tumor's histologic grade and myometrial invasion depth. A sample of 48 pathologically confirmed cases of EC were reviewed retrospectively. The sample was distributed as follows: G1 (n = 9); G2 (n = 18); G3 (n = 21); with myometrial invasion <50% (n = 31); and with myometrial invasion ≥50% (n = 17). DW images were performed at 3.0T with b factors of 0-1000/mm2. The region of interest (ROI) was defined within the tumor with T1-weighted and T2-weighted imaging and copied manually to an ADC map. The tumor's grade and myometrial invasion's depth were determined by postoperative histopathological tests. The means of ADCmin and ADCmean values were significantly lower for patients with G2 and G3 endometrial tumors than G1. The same tendency was observed in myometrial invasion, as both ADCmin and ADCmean values were lower for patients with deep than for those with superficial myometrial invasion. The cut-off values of the ADCmin and ADCmean that predicted high-grade tumors were 0.69 × 10-3 mm2/s and 0.82 × 10-3 mm2/s, respectively, while those for myometrial infiltration were 0.70 × 10-3 mm2/s (ADCmin) and 0.88 × 10-3 mm2/s (ADCmean). ADCmin and ADCmean values correlated with histologic tumor grade and myometrial invasion depth; therefore, it is suggested that ADC on MRI may be a useful indicator to predict malignancy of ECs.

Transcription phenotypes of pancreatic cancer are driven by genomic events during tumor evolution.

Pancreatic adenocarcinoma presents as a spectrum of a highly aggressive disease in patients. The basis of this disease heterogeneity has proved difficult to resolve due to poor tumor cellularity and extensive genomic instability. To address this, a dataset of whole genomes and transcriptomes was generated from purified epithelium of primary and metastatic tumors. Transcriptome analysis demonstrated that molecular subtypes are a product of a gene expression continuum driven by a mixture of intratumoral subpopulations, which was confirmed by single-cell analysis. Integrated whole-genome analysis uncovered that molecular subtypes are linked to specific copy number aberrations in genes such as mutant KRAS and GATA6. By mapping tumor genetic histories, tetraploidization emerged as a key mutational process behind these events. Taken together, these data support the premise that the constellation of genomic aberrations in the tumor gives rise to the molecular subtype, and that disease heterogeneity is due to ongoing genomic instability during progression.

Abstract 241: An efficient cell-based assay for quantification of cellular polysialic acid in neuroblastoma

Abstract Introduction Polysialic acid (polySia) decorates the surface of NCAM (neuronal cell adhesion molecule) on neuroendocrine tumors, notably neuroblastoma and small cell lung cancer, and is strongly associated with poor prognosis and aggressive disease in patients in the clinic [1]. PolySia modulates tumor cell-cell and cell-matrix adhesion, migration, invasion and metastasis. SiRNA knockdown of polysialyltransferase (polyST) ST8SiaII, the enzyme primarily responsible for polySia synthesis in tumors, abrogates tumor cell migration and invasion. PolyST is a selective and largely unexplored therapeutic target for neuroblastoma dissemination [1]. Methods and Results We describe the development of a cell-based assay that quantifies the formation of polySia on the cell surface of neuroblastoma cells. This is a sensitive assay that detects the polySia that is released after pH adjustment, organic precipitation and acid hydrolysis. PolySia was first separated from cell lysate by mild acid hydrolysis and acetone precipitation. It was then purified by ethanol precipitation. We further analysed the role of lactonisation of polySia during organic precipitation. The purified polySia is further hydrolysed into sialic acid monomers, followed by DMB labelling and RPLC-fluorescence analysis. This method was validated by analysis of polySia in C6-ST8SiaII cells, following treatment with Endo-NF in a various concentrations. This method proved to be useful to assess polySia changes after treatment of cells with novel polyST inhibitors. The effect of lactonisation of polySia during organic precipitation was also investigated. It was found that lactonised polySia was mainly soluble in 90% acetone/90% ethanol, while natural polySia (non-lactonised) can be precipitated by ethanol in the presence salts. Based on the different solubility of polySia, a cost-efficient assay was designed to purify polySia released from cells with a low level of non-polySia-derived sialic acid contamination before further acid hydrolysis. With this method we quantified the different levels of cell-surface polySia-derived sialic acid between polySia-positive and polySia-negative cell lines. Furthermore, we successfully validated this method by Endo-NF treatment in C6-ST8SiaII cells: the EC50 of Endo-NF digestion of polySia in C6-ST8SiaII cells was approximately 5.4 pM. The assay has been successfully utilised to evaluate novel polyST inhibitors in vitro. [1] Falconer, R.A. et al., Curr. Cancer Drug Targets, 2012, 12, 925-939; [2] Al-Saraireh YMJ et al., PLoS ONE, 2013, 8:e73366. Citation Format: Xiaoxiao Guo, Francis Mprah Barnieh, Jodie Malcolm, Amanda D Race, Goreti Ribeiro Morais, Steven D Shnyder, Paul M Loadman, Laurence H Patterson, Robert A. Falconer. An efficient cell-based assay for quantification of cellular polysialic acid in neuroblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 241.

P3.01-110 Defining Aggressive Disease in Patients With Advanced NSCLC Receiving Second-Line Treatment: A Systematic Review

Recent randomized clinical trials (RCTs) have explored survival benefits of second-line treatments (2LTs) in patients who have rapidly progressed and/or are refractory to first-line treatment, and these trials have determined an existing unmet need for these patients with aggressive non-small cell lung cancer (NSCLC). However, specific characterization of aggressive NSCLC is lacking, thus a systematic literature review was conducted to explore the definitions of aggressive NSCLC. We systematically searched Medline, Embase, BioSciences Information Service, the Cochrane Library, and abstracts from scientific meetings (through October 2017) to identify RCTs reporting the efficacy and/or safety of select 2LTs in patients with advanced NSCLC who have characteristics associated with aggressive disease (AD). Six potential overarching categorizations of these characteristics (based on expert clinical opinion) were explored: (1) refractory and/or progressive disease as best response to prior treatment, (2) rapid progression, (3) short duration on previous treatment, (4) high tumor burden or size, (5) short duration since start of last treatment, and (6) high symptom burden. The 14 identified studies had one or more subgroups within five of the six categorizations (11, 2, 1, 2, and 4 studies presented subgroups within categories 1-5, respectively). No RCTs presenting a subgroup of patients for category 6 were identified. Within each category, the identified subgroup definitions varied (15, 4, 3, 2, and 7 different definitions within categories 1-5, respectively). Reporting of whether a subgroup was prespecified or not was limited and often unclear; 6 studies indicated that subgroup analyses of patients with AD characteristics were preplanned. Moreover, baseline characteristics for the subgroup of patients with AD were often not reported. Definitions of AD varied, both across the identified studies of 2LTs and within the predetermined categorizations, with refractory being the most frequent followed by short duration since start of last treatment. With the emerging clinical importance of AD, more standard use of these definitions within RCTs may allow for greater comparison across 2LTs and will enable indirect treatment comparisons of the results. As with any subgroup, clarity on preplanned versus post hoc analysis is important for interpretation and should be specified. Additional studies powered to assess treatment benefits in advanced NSCLC patients with AD are needed.

Clinical implications of PTEN loss in prostate cancer

Genomic aberrations of the PTEN tumour suppressor gene are among the most common in prostate cancer. Inactivation of PTEN by deletion or mutation is identified in ∼20% of primary prostate tumour samples at radical prostatectomy and in as many as 50% of castration-resistant tumours. Loss of phosphatase and tensin homologue (PTEN) function leads to activation of the PI3K-AKT (phosphoinositide 3-kinase-RAC-alpha serine/threonine-protein kinase) pathway and is strongly associated with adverse oncological outcomes, making PTEN a potentially useful genomic marker to distinguish indolent from aggressive disease in patients with clinically localized tumours. At the other end of the disease spectrum, therapeutic compounds targeting nodes in the PI3K-AKT-mTOR (mechanistic target of rapamycin) signalling pathway are being tested in clinical trials for patients with metastatic castration-resistant prostate cancer. Knowledge of PTEN status might be helpful to identify patients who are more likely to benefit from these therapies. To enable the use of PTEN status as a prognostic and predictive biomarker, analytically validated assays have been developed for reliable and reproducible detection of PTEN loss in tumour tissue and in blood liquid biopsies. The use of clinical-grade assays in tumour tissue has shown a robust correlation between loss of PTEN and its protein as well as a strong association between PTEN loss and adverse pathological features and oncological outcomes. In advanced disease, assessing PTEN status in liquid biopsies shows promise in predicting response to targeted therapy. Finally, studies have shown that PTEN might have additional functions that are independent of the PI3K-AKT pathway, including those affecting tumour growth through modulation of the immune response and tumour microenvironment.

Reduced Retinoblastoma Protein Expression Is Associated with Decreased Patient Survival in Medullary Thyroid Cancer.

The retinoblastoma (RB) transcriptional corepressor 1 protein functions to slow cell-cycle progression. Inactivation of RB by reduced expression and/or hyperphosphorylation allow for enhanced progression through the cell cycle. Murine models develop medullary thyroid carcinoma (MTC) after generalized loss of RB. However, RB expression in MTC has only been evaluated in a small number of tumors, with differing results. The objective of this study was to determine whether reduced expression of RB and/or overexpression of hyperphosphorylated RB predict MTC aggressive behavior. Formalin-fixed, paraffin-embedded primary thyroid tumors and lymph node metastases from MTC patients were evaluated for calcitonin, RB, and phosphorylated RB (pRB) expression by immunohistochemistry. Two expert pathologists evaluated the slides in a blinded manner, and the immunohistochemistry results were compared to disease-specific survival as a primary endpoint. Seventy-four MTC samples from 56 patients were analyzed in this study, including 51 primary tumors and 23 lymph node metastases. The median follow-up time was 6.75 years after surgery (range 0.64-24.30 years), and the median primary tumor size was 30 mm (range 6-96 mm). Sixty-six percent of cases were classified as stage IV. RB nuclear expression was diffusely present in 88% of primary tumors and 78% of lymph node metastases. Nuclear pRB expression was present in 22% of primary tumors and 22% of lymph node metastases. On univariate analysis, reduced RB (<75% tumor cell staining) trended with lower MTC-specific survival for primary tumor and metastatic nodes (primary tumor hazard ratio = 3.54 [confidence interval 0.81-15.47], p = 0.08; and lymph node hazard ratio = 4.35 [confidence interval 0.87-21.83], p = 0.05). For primary tumors, multivariable analysis showed that low nuclear RB expression was independently associated with worse disease-specific (p = 0.01) and overall (p = 0.02) survival. pRB levels were not associated with survival for either primary tumor or lymph node metastases. Reduced RB expression is associated with decreased patient survival in univariate and multivariable analyses, independent from patient age at surgery or advanced TNM stage. Future studies involving larger MTC patient populations are warranted to determine if lower RB expression levels may serve as a biomarker for aggressive disease in patients with MTC.

A definition for aggressive disease in patients with HER-2 negative metastatic breast cancer: an expert consensus of the Spanish Society of Medical Oncology (SEOM).

To converge on an expert opinion to define aggressive disease in patients with HER2-negative mBC using a modified Delphi methodology. A panel of 21 breast cancer experts from the Spanish Society of Medical Oncology agreed upon a survey which comprised 47 questions that were grouped into three sections: relevance for defining aggressive disease, aggressive disease criteria and therapeutic goals. Answers were rated using a 9-point Likert scale of relevance or agreement. Among the 88 oncologists that were invited to participate, 81 answered the first round (92%), 70 answered the second round (80%), and 67 answered the third round (76%) of the survey. There was strong agreement regarding the fact that identifying patients with aggressive disease needs to be adequately addressed to help practitioners to decide the best treatment options for patients with HER2-negative mBC. The factors that were considered to be strongly relevant to classifying patients with aggressive HER2-negative mBC were a high tumor burden, a disease-free interval of less than 12-24months after surgery, the presence of progressive disease during adjuvant or neoadjuvant chemotherapy and having a triple-negative phenotype. The main therapeutic goals were controlling symptoms, improving quality of life and increasing the time to progression and overall survival. High tumor burden, time to recurrence after prior therapy and having a triple-negative phenotype were the prognostic factors for which the greatest consensus was found for identifying patients with aggressive HER2-negative mBC. Identifying patients with aggressive disease leads to different therapeutic approaches.

Urinary DNA Methylation Biomarkers for Noninvasive Prediction of Aggressive Disease in Patients with Prostate Cancer on Active Surveillance

Patients with prostate cancer on active surveillance are monitored by repeat prostate specific antigen measurements, digital rectal examinations and prostate biopsies. A subset of patients on active surveillance will later reclassify with disease progression, prompting definitive treatment. To minimize the risk of under treating such patients on active surveillance minimally invasive tests are urgently needed incorporating biomarkers to identify patients who will reclassify. We assessed post-digital rectal examination urine samples of patients on active surveillance for select DNA methylation biomarkers that were previously investigated in radical prostatectomy specimens and shown to correlate with an increasing risk of prostate cancer. Post-digital rectal examination urine samples were prospectively collected from 153 men on active surveillance who were diagnosed with Gleason score 6 disease. Urinary sediment DNA was analyzed for 8 DNA methylation biomarkers by multiplex MethyLight assay. Correlative analyses were performed on gene methylation and clinicopathological variables to test the ability to predict patient risk reclassification. Using backward logistic regression a 4-gene methylation classifier panel (APC, CRIP3, GSTP1 and HOXD8) was identified. The classifier panel was able to predict patient reclassification (OR 2.559, 95% CI 1.257-5.212). We observed this panel to be an independent and superior predictor compared to current clinical predictors such as prostate specific antigen at diagnosis or the percent of tumor positive cores in the initial biopsy. We report that a urine based classifier panel of 4 methylation biomarkers predicts disease progression in patients on active surveillance. Once validated in independent active surveillance cohorts, these promising biomarkers may help establish a less invasive method to monitor patients on active surveillance programs.

Abstract 4752: Overexpression of enhancer of zeste homolog 2 (EZH2) in endometrial carcinoma: evidence from an NRG Oncology/Gynecologic Oncology Group trial

Abstract Objectives: Enhancer of zeste homolog 2 (EZH2) is a histone methyltransferase that mediates epigenetic silencing of tumor suppressor genes. It is commonly over-expressed in several solid tumors and has been shown to be a prognostic biomarker in breast, renal, and gastric cancers. This study aims to investigate patterns of EZH2 expression in endometrial cancer. Methods: Evaluation of expression of EZH2 was completed on both early and advanced stage endometrioid adenocarcinoma tissues and a subset of matched normal mullerian tissue samples, from participants enrolled in GOG protocol 210, using real time reverse transcription polymerase chain reaction (RT-PCR) and western blot (WB) analysis. Investigators were blinded to clinical data. One-way ANOVA, F test and Fisher's exact test were used to assess differences in log mRNA and protein expression respectively with known clinical and pathologic prognostic factors. Kaplan Meier survival analysis was used to evaluate differences in progression free survival (PFS) based on EZH2 expression. Results: Eight-seven specimens were analyzed including 60 tumors and 27 normal tissue specimens. EZH2 mRNA and protein expression in tumor specimens was significantly higher than in matched controls (paired t test, p &amp;lt; 0.001). EZH2 expression was associated with lympho-vascular space invasion (p = 0.044) but other clinical and pathologic factors including age, stage, grade, nodal involvement or disease status were not associated with significant differences in expression. However, there does appear to be a trend towards decreased progression free survival among patients with increased EZH2 expression levels. Conclusions: Our results confirm that there is differential expression of EZH2 in uterine cancers compared to normal tissues and suggests a potential role of the EZH2 as prognostic marker of aggressive disease in patients with endometrial cancer. In view of the substantial portion of early stage low grade tumors expressing EZH2, it will be interesting to investigate if increased EZH2 protein levels may also have therapeutic implications, since targeted inhibition of EZH2 expression has been shown to repress tumor cell growth in other solid malignancies. Further investigation into this potential prognostic biomarker and therapeutic target is warranted. Citation Format: Lauren Krill, Shamshad Ali, Ramez Eskander, Meenakshi Singh, David Mutch, Susan Zweizig, Michael Birrer, Heather Lankes, Bang Hoang, Leslie Randall. Overexpression of enhancer of zeste homolog 2 (EZH2) in endometrial carcinoma: evidence from an NRG Oncology/Gynecologic Oncology Group trial. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4752.

20 - MYC Gene Rearrangement in Plasma Cell Neoplasms

MYC gene rearrangements and amplification are known to be associated with aggressive disease in patients with acute leukemia and B-cell Non-Hodgkin lymphoma. Up to 15% of Plasma Cell Neoplasm (PCN) patients are reported to have MYC gene rearrangement with evidence indicating an associated poor prognosis and shortened survival ( 1 Walker B.A. Wardell C.P. Brioli A. et al. Translocations at 8q24 juxtapose MYC with genes that harbor superenhancers resulting in overexpression and poor prognosis in myeloma patients. Blood Cancer J. 2014; 4: e191https://doi.org/10.1038/bcj.2014.13 Crossref Scopus (117) Google Scholar ). While it has been historically associated with advanced disease, rearrangement of the gene has also now been identified in patients with “smoldering” myeloma or monoclonal gammopathy ( 2 Affer M. Chesi M. Chen W.D. et al. Promiscuous MYC locus rearrangements hijack enhancers but mostly super-enhancers to dysregulate MYC expression in multiple myeloma. Leukemia. 2014; 28: 1725-1735 Crossref Scopus (174) Google Scholar ).

Association of inclusion body myositis with T cell large granular lymphocytic leukaemia.

SEE HOHLFELD AND SCHULZE-KOOPS DOI101093/BRAIN/AWW053 FOR A SCIENTIFIC COMMENTARY ON THIS ARTICLE: Inclusion body myositis and T cell large granular lymphocytic leukaemia are rare diseases involving pathogenic cytotoxic CD8+ T cells. After encountering four patients with both disorders, we prospectively screened 38 patients with inclusion body myositis for the presence of expanded large granular lymphocyte populations by standard clinical laboratory methods (flow cytometry, examination of blood smears, and T cell receptor gene rearrangements), and performed muscle immunohistochemistry for CD8, CD57, and TIA1. Most (22/38; 58%) patients with inclusion body myositis had aberrant populations of large granular lymphocytes in their blood meeting standard diagnostic criteria for T cell large granular lymphocytic leukaemia. These T cell populations were clonal in 20/20 patients and stably present on follow-up testing in 15 patients a median of 350 days later. T cell aberrant loss of CD5 or gain of expression of CD16 and CD94 were common (19/42, 45%). In comparison, 2/15 (14%) age-matched patients with dermatomyositis, polymyositis, or necrotizing myopathy, and 0/20 (0%) age-matched healthy subjects had large granular lymphocyte expansions, with none of these patients having T cell aberrant expression of CD5, CD16 or CD94. Reduced blood CD4/CD8 ratio, increased blood CD8 count, and lymphocytosis were additional biomarkers highly correlated with flow cytometry-measured large granular lymphocyte expansions. Cross-sectional data suggested more aggressive disease in patients with such expansions than without. Muscle immunohistochemistry demonstrated invasion of large granular lymphocytes into muscle in 15/15 inclusion body myositis patients but in only 1/28 patients with dermatomyositis or polymyositis. The extent of CD8+ and CD57+ cells in inclusion body myositis muscle correlated with the size of blood large granular lymphocyte populations. Myofibre-invading cells expressed CD57, a marker of persistent T cell exposure to antigen and T cell aggressiveness. In many patients with inclusion body myositis, the autoimmune T cell expansion has evolved into a neoplastic-like or overtly neoplastic disorder, perhaps contributing to its relative refractoriness to immune-directed therapies previously reported.

Association of genetic variations of selenoprotein genes, plasma selenium levels, and prostate cancer aggressiveness at diagnosis.

Genetic variations in some of the selenoprotein genes, alone or together with an individual's selenium status, may influence risk or progression of prostate cancer. We investigated the impact of genetic variants of selenoproteins on plasma selenium levels and cancer aggressiveness at diagnosis in men with localized prostate cancer (PCa). The study cohort comprised 722 patients seen at Dana-Farber Cancer Institute who had localized/locally advanced PCa (i.e., stage T3 or less, N0, and M0) from 1994 to 2001. Fifty-five tagging single nucleotide polymorphisms (SNPs) from six selenoprotein genes (TXNRD1, TXNRD2, SEP15, GPX3, SELENBP1, and SEPP1) were analyzed. Logistic regression is used to examine associations of genotypes and plasma selenium levels with risk of aggressive disease, defined as D'Amico intermediate/high risk categories. Step down permutation was applied to adjust for multiple comparisons. Three hundred and forty-eight patients (48%) had aggressive disease at diagnosis. Two SNPs were associated with cancer aggressiveness at diagnosis (unadjusted P = 0.017 and 0.018, respectively). The odds ratio for aggressive disease in patients carrying TXNRD2 rs1005873-AG/GG genotypes or SELENBP1 rs10788804-AG/AA genotypes was 1.54 (95% CI = 1.08, 2.20) and 1.45 (95% CI = 1.07, 1.98), respectively, compared to TXNRD2 rs1005873-AA or SELENBP1 rs10788804-GG carriers. Four SNPs in TXNRD2 (rs1005873, rs13054371, rs3788310, and rs9606174) and the rs230820 in SEPP1 were associated with plasma selenium levels (unadjusted P < 0.05). Permutation adjusted P-values were not statistically significant for all these comparisons at the cut-off point of 0.05. We identified polymorphisms in selenoproteins that may influence the plasma selenium levels and may be associated with the risk of presenting with aggressive PCa in men with localized or locally advanced PCa. These results should be validated in other independent datasets.

Less aggressive disease in patients with primary squamous cell carcinomas of the thyroid gland and coexisting lymphocytic thyroiditis.

Aim of the studyPrimary squamous cell carcinoma (SCC) of the thyroid gland is extremely rare. Infrequently, primary SCC of the thyroid gland is accompanied by other thyroid diseases such as Hashimoto's thyroiditis (HT). Recently, studies have demonstrated that differentiated thyroid cancer with coexisting HT has a better prognosis. However, the prognosis of patients with primary SCC of the thyroid gland and coexistent HT has not been clearly identified. We compared the clinical characteristics and disease stages of patients with primary SCC with and without lymphocytic thyroiditis (LT).Material and methodsWe reviewed reports of primary SCC of the thyroid gland published in the English literature.Results and conclusionsWe identified 46 papers that included 17 cases of primary SCC of the thyroid gland with LT and 77 cases of primary SCC of the thyroid gland without LT. Lymph node metastasis and local invasion rates did not differ between these two groups. Distant metastases were absent in patients with LT, and were observed in 13 (16.9%) patients without LT. A greater proportion of patients without LT had advanced stage disease (stage IV A-B-C) than patients with LT (p < 0.05). Patients with primary SCC of the thyroid gland and coexisting LT had lower tumour-node-metastasis stage and frequency of distant metastasis than those without LT. Lymphocytic infiltration in patients with SCC appears to limit tumour growth and distant metastases.

Impact of the Site of Metastases on Survival in Patients with Metastatic Prostate Cancer

BackgroundLimited data exist on the impact of the site of metastases on survival in patients with stage IV prostate cancer (PCa). ObjectiveTo investigate the role of metastatic phenotype at presentation on mortality in stage IV PCa. Design, setting, and participantsOverall, 3857 patients presenting with metastatic PCa between 1991 and 2009, included in the Surveillance Epidemiology and End Results–Medicare database were evaluated. Outcome measurements and statistic analysesOverall and cancer-specific survival rates were estimated in the overall population and after stratifying patients according to the metastatic site (lymph node [LN] alone, bone, visceral, or bone plus visceral). Multivariable Cox regression analyses tested the relationship between the site of metastases and survival. All analyses were repeated in a subcohort of patients with a single metastatic site involved. Results and limitationsRespectively, 2.8%, 80.2%, 6.1%, and 10.9% of patients presented with LN, bone, visceral, and bone plus visceral metastases at diagnosis. Respective median overall survival and cancer-specific survival were 43 mo and 61 mo for LN metastases, 24 mo and 32 mo for bone metastases, 16 mo and 26 mo for visceral metastases, and 14 mo and 19 mo for bone plus visceral metastases (p<0.001). In multivariable analyses, patients with visceral metastases had a significantly higher risk of overall and cancer-specific mortality versus those with exclusively LN metastases (p<0.001). The unfavorable impact of visceral metastases persisted in the oligometastatic subgroup. Our study is limited by its retrospective design. ConclusionsVisceral involvement represents a negative prognostic factor and should be considered as a proxy of more aggressive disease in patients presenting with metastatic PCa. This parameter might indicate the need for additional systemic therapies in these individuals. Patient summaryPatients with visceral metastases should be considered as affected by more aggressive disease and might benefit from the inclusion in clinical trials evaluating novel molecules.

MicroRNA-155 influences B-cell receptor signaling and associates with aggressive disease in chronic lymphocytic leukemia

AbstractHigh-level leukemia cell expression of micro-RNA 155 (miR-155) is associated with more aggressive disease in patients with chronic lymphocytic leukemia (CLL), including those cases with a low-level expression of ζ-chain–associated protein of 70 kD. CLL with high-level miR-155 expressed lower levels of Src homology-2 domain-containing inositol 5-phosphatase 1 and were more responsive to B-cell receptor (BCR) ligation than CLL with low-level miR-155. Transfection with miR-155 enhanced responsiveness to BCR ligation, whereas transfection with a miR-155 inhibitor had the opposite effect. CLL in lymphoid tissue expressed higher levels of miR155HG than CLL in the blood of the same patient. Also, isolated CD5brightCXCR4dim cells, representing CLL that had been newly released from the microenvironment, expressed higher levels of miR-155 and were more responsive to BCR ligation than isolated CD5dimCXCR4bright cells of the same patient. Treatment of CLL or normal B cells with CD40-ligand or B-cell–activating factor upregulated miR-155 and enhanced sensitivity to BCR ligation, effects that could be blocked by inhibitors to miR-155. This study demonstrates that the sensitivity to BCR ligation can be enhanced by high-level expression of miR-155, which in turn can be induced by crosstalk within the tissue microenvironment, potentially contributing to its association with adverse clinical outcome in patients with CLL.