Low-grade Disease Research Articles

225 - Pre-fibrotic Status in the Lung of a Mouse Model of Metabolic Syndrome: Implication of Oxidative Stress and Inflammation

Obesity is a low-grade chronic inflammatory disease resulting from adipose tissue inflammation. Metabolic syndrome (MS) is the most serious consequence of obesity. Although many organs are affected in the MS, its impact on lung biochemistry is less known. Herein we aimed at determining the impact of MS on redox/inflammatory status and potential consequences in the lung using a previously developed mouse model of MS. To achieve this goal, 6 week-old male C57BL / 6J mice were fed for 16 weeks with a low fat- (LFD) or a high fat-diet (HFD, chicken fat) and water supplemented with 10% fructose (F). Thus, the experimental design included 2 groups: LFD (control) HFD+F (MS). Compared to control, the lung parenchyma of MS mice showed increased neutrophils content, MPO activity, oxidative stress (NOX-2, carbonyls, TBARS and nitrotyrosine) and inflammation markers (iNOS, ICAM-1, TNF-α and IL-6). In addition and compared to LFD, the lung parenchyma of HFD+F mice had a lower total antioxidant capacity, GSH/GSSG ratio; and activity of catalase, superoxide dismutase and glutathione peroxidase. Interestingly and compared to LFD, the lung parenchyma of HFD+F mice had an increased pro-fibrotic status as assessed by trichromic stain and TGF-β expression. In this study, we report a mouse model of MS showing lung oxidative stress/inflammation and a profibrotic status.

Reconstruction in the Metastatic Breast Cancer Patient: Results From the National Cancer Database

Modern treatments are prolonging life for metastatic breast cancer patients. Reconstruction in these patients is controversial. The purpose of this study was to characterize de novo metastatic breast cancer patients who undergo mastectomy and reconstruction and to report complication and survival rates. We queried the National Cancer Database for de novo metastatic breast cancer patients, who underwent systemic therapy and mastectomy with reconstruction (R) or without reconstruction (NR) between 2004 and 2013. Patient-tumor characteristics, mortality, and readmissions were compared. Propensity score matched analysis was performed, and survival was calculated using the Kaplan–Meier method. A total of 8554 patients fulfilled study criteria (n = 980/11.5% R vs. n = 7574/88.5% NR). There was a significant increase in reconstruction rates by year: 5.2% in 2004, 14.3% in 2013 (p < 0.0001). Compared with the NR patients, R patients were younger (mean age 49 vs. 58 years, p < 0.0001), more hormone receptor-positive (76.1% vs. 70.5%, p = 0.0004), had lower grade disease (p = 0.0082), and fewer sites of metastases (85.7% had 1 metastasis; 14.3% had ≥ 2 R vs. 79% had 1; 21% had ≥ 2 NR, p = 0.0002). R patients received more hormonal and chemotherapy than NR but equally received radiation. Median overall survival of the total cohort was 45 months, and median overall survivals of R and NR groups by matched analysis were 56.7 and 55.3 months respectively (p = 0.86). Thirty-day mortality (0.2%-R, 0.3%-NR, p = 0.56) and readmissions (5.9%-R, 5.8%-NR, p = 0.81) were similar; 90-day mortality also was similar (1.1%-R vs. 1.6%-NR, p = 0.796). There is an increasing trend to reconstruct metastatic breast cancer patients with low complication rates, without survival compromise. Impact on quality of life warrants further assessment.

Characteristics of Radiation Therapy for Medullary Carcinoma of the Breast in the National Cancer Database

Medullary breast carcinoma (MedBC) is an uncommon histology of invasive breast cancer that has a more favorable prognosis compared to invasive ductal carcinoma (IDC). There are no specific consensus guidelines for radiotherapy (RT) for MedBC. We aim to determine whether there are differences in the utilization of RT using the National Cancer Data Base (NCDB), a national oncology outcomes database. Patients with MedBC and IDC reported to the NCDB from 2004 to 2014 who had undergone either lumpectomy or mastectomy followed by adjuvant radiotherapy were analyzed for patient and treatment characteristics. Characteristics analyzed between the two histologies included age, race, primary insurance payer, Charles-Deyo score, grade, NCDB analytic stage, regional radiation dose, boost radiation dose, and types of surgery performed. Regional and boost dose for grades of MedBC were evaluated. A total of 1862 patients with MedBC and 589,358 patients with IDC were included in the analysis. Median follow-up was 59.6 months. MedBC patients were more likely to be black (26.4% vs. 11.5%, p< 0.001). There was a higher proportion of Stage II patients with MedBC (47% vs. 29.4% p < 0.001) and less Stage III (6.0% vs. 11.8% p< 0.001). MedBC patients were more likely to have triple negative receptor status compared to IDC (60.7% vs. 13.9% p< 0.001). MedBC had lower grade disease, 52.4%, 34.4% and 13.1% (well, moderately, poorly differentiated) compared to IDC - 20.6%, 41.8%, and 37.6 respectively (p<0.001). There was no significant difference in mean regional dose - 4865.6 and 4893 cGy for MedBC and IDC respectively and a small difference in boost dose, 1061.5 cGy for MedBC and 956.5 cGy for IDC (p<0.001). A greater proportion of patients with IDC received no boost dose compared to MedBC – 20.1% vs. 14.2% (p<0.001). There was no significant difference in the mean regional dose for well, moderate and poorly differentiated grade MedBC (4842.3 Gy, 4913.4 cGy, and 4894.9 cGy, p = 0.701) or boost dose (1165.9 cGy, 1036.0 cGy, and 1062.5 cGy, p = 0.592). There was a significant difference in 5 and 10 year survival, 83.7% and 77.1% for IDC compared to 89.1% and 84.4% for MedBC, respectively, (log-rank p = 0.012). On univariate and multivariate analysis, neither regional nor boost dose was associated with increased HR. On UVA, poorly differentiated grade had an associated HR=2.78 (p=0.035), with a trend on MVA (HR=2.62, p=0.071). The use of radiotherapy for MedBC does not appear to differ greatly for IDC with no difference in regional doses. A small difference in boost dose may be explained by a lower proportion of IDC patients receiving boost dose as compared to MedBC. The grade of disease does not appear to effect radiation dosing, and dose of therapy was not associated with survival. The improved outcomes of MedBC compared to IDC is likely related to disease biology and other factors.

Outcomes of sinonasal squamous cell carcinoma with and without association of inverted papilloma: A multi-institutional analysis.

Sinonasal squamous cell carcinoma (SCC) accounts for <1% of all malignancies but represents 70% of sinonasal cancer. Up to 10% of SCCs are associated with inverted papilloma (IPSCC). Studies that compare patients, treatment, and outcomes of SCC and IPSCC are absent in the literature. A retrospective review of patients with SCC and those with IPSCC at Cleveland Clinic and Indiana University from 1995 to 2015. The records were analyzed for demographics, tumor characteristics, treatment, and outcomes. The study comprised 117 patients with SCC, of whom, 29 had IPSCC. The mean age at diagnosis was similar: 63 and 64 years for patients with SCC and patients with IPSCC, respectively; with female patients representing 36% and 34%, respectively (p > 0.99).Smokers represented 64% of the patients with SCC and 55% of patients with IPSCC (p = 0.3); excessive alcohol intake was noted in 16% of the patients with SCC and 21% of the patients with IPSCC (p = 0.56).The maxillary sinus was most commonly involved, followed by the nasal cavity (51% versus 35% SCC, 45% versus 38% IPSCC). Frontal ethmoid and sphenoid sinuses contained primary tumors only in patients with SCC. Upfront treatment was surgery in 84% of patients with SCC and 97% of patients with IPSCC (p = 0.18); 68 and 55% received radiation, respectively, and 25 and 21% received chemotherapy, respectively.Overall survival averaged 5.5 and 3.4 years for patients with SCC and patients with IPSCC, respectively (p = 0.12); disease-free survival was 4.8 and 2.9 years, respectively (p = 0.18). Nodal metastasis was more likely in patients with SCC (18 versus 0%; p = 0.02). When divided into high- and low-stage disease: more common nodal metastases were demonstrated in high-stage SCC than in low-stage disease (p = 0.03). Overall survival was decreased between high- and low-grade disease but not when subdivided between SCC and IPSCC. Although SCC with and without IP association are considered different diseases, their demographics and outcomes seem similar. Nodal metastasis was noted to be higher in the SCC cohort, which may indicate different tumor biology. Further study is warranted.

Novel imaging in advanced prostate cancer.

Prostate cancer (PCa) is the most commonly diagnosed noncutaneous cancer and second leading cause of death in men. Imaging evaluation of PCa is challenging because of the prostate's deep pelvic location, its complex zonal anatomy and its multifocal nature. Novel imaging modalities are needed to improve detection, reassessment in biochemical relapse, and disease progression in advanced metastatic stages. Current imaging modalities have distinct strengths. However, all lack the ability to diagnose micrometastases, differentiate high from low-grade disease and diagnose advanced disease, especially at low prostate specific antigen values. There is a need to combine the existing imaging methods with concepts utilizing tumor biology to differentiate biologically aggressive from indolent tumors. PET imaging with novel tracers facilitate improved imaging of PCa, but also usher in new compounds that could be useful for directing treatment as well. Most tracers have limited sensitivity, with the exception of prostate-specific membrane antigen (PSMA)-targeting tracers, that offer relatively higher sensitivity and specificity. PSMA-PET appears promising in improving the imaging yield particularly in recurrent and advanced disease states. Incorporating PSMA-PET in these settings could open or prolong windows along the trajectory of the disease that could allow new treatments or more effective use of currently existing treatments. Prospective studies focusing on novel imaging enhancement and integration with therapeutic applications are needed.

Effects of cholesterol, C-reactive protein, and interleukin-6 on prostate cancer risk in a population of African ancestry.

To investigate the association between serum cholesterol and prostate cancer and whether any effect may be mediated through inflammatory markers. Data from a case-control study of 40-80years old Jamaicanmale patients (229 cases; 252 controls) were used. Cases had incident histologically-confirmed prostate cancer and controls were men with normal digital rectal examination and prostate-specific antigen (PSA)<4μg/Lorfree: total PSA>0.15 obtained from the same clinic. Total and HDL cholesterol, interleukin-6 (IL-6), and C-reactive protein (CRP) were measured from a non-fasting sample. Multivariable logistic regression models were used to evaluate the associations between these factors and prostate cancer, adjusting for age, body mass index, waist circumference, family history of prostate cancer, diabetes, hypertension, use of cholesterol-lowering drugs, and smoking. Total cholesterol [Mean (cases, 4.71±1.07; controls, 4.64±1.07mmol/L)], CRP [median (cases, 2.11; controls, 2.09µg/ml)], and IL-6: [median (cases, 3.34; controls, 3.24pg/ml)] did not differ by PCA status. Higher total cholesterol was associated with an increased risk of low-grade disease after adjusting for potential confounders [multivariable-adjusted OR (95% CI): tertile 2: 3.32(1.66, 6.45), tertile 3: 2.14(1.07, 4.32)]. Total cholesterol was unrelated to overall prostate cancer or high-grade disease. There was no significant association between HDL cholesterol or any of the inflammatory markers with prostate cancer. Increasing total cholesterol but not inflammatory markers were associated with low-grade prostate cancer in Caribbean men.

Short- and Long-Term Survival in Metastatic Pancreatic Adenocarcinoma, 1993-2013.

Background: During the past 2 decades, numerous clinical trials have focused on improving outcomes in patients with metastatic pancreatic cancer (mPDAC). The efficacy of new treatments has been demonstrated among highly selected patients in randomized phase III trials; hence, it is not clear to what extent these advances are reflected within the broader mPDAC population. Materials and Methods: Survival statistics were extracted from the SEER database for patients diagnosed with mPDAC between 1993 and 2013. Survival was analyzed using the Kaplan-Meier method and proportional hazard models. Results: The study population consisted of 57,263 patients diagnosed with mPDAC between 1993 and 2013; 52% were male, with a median age of 69 years (range, 15-104). Superior prognosis correlated with younger age, being married, tumor located within the head of the pancreas, lower grade disease, and more recent year of diagnosis. Median overall survival (OS) remained stable at 2 months between 1993 and 2013. Improvements in OS were seen for younger patients (age <50 years) and those with a more recent year of diagnosis (2009-2013). The percentage of patients who died within 2 months of initial diagnosis decreased between 1993 and 2013 (from 63.5% to 50.6%; P<.0001). The percentage of patients surviving ≥12 months improved from 4.9% in 1993 to 12.7% in 2013 (P<.0001). Conclusions: In recent years a modest improvement in OS has been seen among younger patients with mPDAC. The percentage of patients living beyond 1 year has significantly increased over time; however, the percentage of those dying within 2 months remains substantial.

Epigenetic risk score improves prostate cancer risk assessment.

Early detection of aggressive prostate cancer (PCa) remains crucial for effective treatment of patients. However, PCa screening remains controversial due to a high rate of overdiagnosis and overtreatment. To better reconcile both objectives, more effective methods for assessing disease severity at the time of diagnosis are needed. The relationship between DNA-methylation and high-grade PCa was examined in a cohort of 102 prospectively enrolled men who received standard 12-core prostate biopsies. EpiScore, an algorithm that quantifies the relative DNA methylation intensities of GSTP1, RASSF1, and APC in prostate biopsy tissue, was evaluated as a method to compensate for biopsy under-sampling and improve risk stratification at the time of diagnosis. DNA-methylation intensities of GSTP1, RASSF1, and APC were higher in biopsy cores from men diagnosed with GS ≥ 7 cancer compared to men with diagnosed GS 6 disease. This was confirmed by EpiScore, which was significantly higher for subjects with high-grade biopsies and higher NCCN risk categories (both P < 0.001). In patients diagnosed with GS ≥ 7, increased levels of DNA-methylation were present, not only in the high-grade biopsy cores, but also in other cores with no or low-grade disease (P < 0.001). By combining EpiScore with traditional clinical risk factors into a logistic regression model, the prediction of high GS reached an AUC of 0.82 (95%CI: 0.73-0.91) with EpiScore, DRE, and atypical histological findings as most important contributors. In men diagnosed with PCa, DNA-methylation profiling can detect under-sampled high-risk PCa in prostate biopsy specimens through a field effect. Predictive accuracy increased when EpiScore was combined with other clinical risk factors. These results suggest that EpiScore could aid in the detection of occult high-grade disease at the time of diagnosis, thereby improving the selection of candidates for Active Surveillance.

Preoperative High Neutrophil-to-Lymphocyte Ratio Is Associated with High-grade Bladder Cancer.

To evaluate the correlation between the neutrophil-to-lymphocyte ratio (NLR) and histopathological characteristics of bladder cancer. A retrospective analysis was performed on a cohort of patients (n=302) who were diagnosed with bladder cancer and underwent transurethral resection of bladder tumor or cystectomy between 2009 and 2016. The pathological outcomes were compared between patients with low NLR and those with high NLR with a cut-off value of 2.5, and a logistic regression analysis was performed to find potential predictors of pathological tumor outcomes. Patients with high-grade disease had significantly higher NLRs compared to those with low-grade disease (median NLR=4.42 vs. 3.42, p<0.001). Univariate analysis suggested that age, neutrophil count and NLR, as a continuous or binary variable, were significantly associated with high-grade disease. Multivariate analysis suggested that age and NLR, as a continuous variable, were predictors of pathologically high-grade disease. Preoperative NLR was found to be associated with pathological tumor grading, but was not associated with pathological tumor staging in patients with bladder cancer.

Secular Trends in Prostate Biopsy Criteria and Outcomes: The Dartmouth Experience

To evaluate trends in prostate biopsy and cancer diagnosis at a center with conservative screening practices in the pre- and post-2012 era. More restrictive prostate-specific antigen (PSA) screening guidelines have led to lower rates of screening, biopsy, and diagnosis of prostate cancer. It is not clear, however, how regions with low baseline screening rates (the Lebanon, New Hampshire hospital referral region centered on Dartmouth-Hitchcock Medical Center had the lowest rate of screening among Medicare patients in 2012) have responded to these guidelines. We retrospectively analyzed patients who underwent prostate biopsy from January 2011 to March 2016. Demographic and clinical characteristics were analyzed by time. Multivariable analysis assessed for factors associated with higher grade cancer. There were 614 prostate biopsies were performed. PSA at biopsy increased with time (7.2 in 2011 vs 10.1 in 2015, P = .0085); age did not. There was a stable proportion of benign findings; proportions of low-grade disease decreased, whereas intermediate- and high-grade disease increased (2011 vs 2015: 21.1% vs 10.8% Gleason 3 + 3, 32.9% vs 43.3% ≥ Gleason 3 + 4, P = .0454). Factors predictive of higher grade disease included abnormal digital examination (odds ratio [OR] 2.19, P = .0076), higher PSA (OR 1.09, P = .0040), and later biopsy date (OR 1.01, P = .0469). In an environment of conservative baseline screening practices, there has been a shift in prostate biopsy criteria and outcomes, namely a rising PSA threshold for biopsy and a 50% decrease in the diagnosis of low-grade disease. Additional study is needed to ensure these trends are favorably impacting the quality of care.

Impact of Adjuvant Radiotherapy for Malignant Salivary Gland Tumors.

Objective Using the National Cancer Database (NCDB), we investigated the characteristics, outcomes, and benefits of adjuvant therapy for patients diagnosed with malignant salivary gland tumors between 2004 and 2012. Study Design Retrospective analysis. Setting NCDB. Subject and Methods The cases of patients diagnosed with a nonmetastatic major salivary gland tumor who underwent resection between 2004 and 2012 were abstracted from the NCDB. Patients were further included if they had pT1-4NX-1M0 high-grade disease or pT3-4NX-0M0 or pT1-4N1M0 low-grade disease. Patients were identified as having no postoperative radiation therapy or having received postoperative radiation therapy to a dose of 5000 and 7000 cGy to the head and neck region or the parotid region, and their characteristics and outcomes were compared. Results During the study period, 4068 patients met the inclusion criteria for this analysis, of which 2728 (67.1%) received postoperative radiation and 1340 (32.9%) did not. With a median follow-up of 49.1 months, there was a significant improvement in overall survival associated with those receiving postoperative radiation (5 years, 56% vs 50.6%). On multivariable analysis, radiation utilization (hazard ratio, 0.78; 95% CI, 0.71-0.86; P < 0.001) and female sex (hazard ratio, 0.88) were associated with improved survival. When the analysis was limited to patients ≤65 years old, the survival benefit was persistent on multivariable analysis. Conclusion In conclusion, in this large NCDB study of 4068 patients with locally advanced malignant salivary gland carcinoma, administering adjuvant radiotherapy was associated with improved overall survival.

Adipose Tissue Inflammation and Oxidative Stress: the Ameliorative Effects of Vitamin D.

Obesity is a low-grade inflammatory disease and is associated with numerous comorbidities. The current study was aimed to evaluate the effects of vitamin D administrations on markers of inflammation and oxidative stress in adipose tissue of high-fat diet-induced obese rats. In the beginning of the study, 40 rats were divided into two groups: normal diet and high-fat diet (HFD) for 16weeks; then, each group was subdivided into two groups including ND, ND + vitamin D, HFD, and HFD+vitamin D. Vitamin D supplementation was done for 5weeks at 500IU/kg dosage. Tumor necrosis factor (TNF)-α, interleukin (IL)-1β, monocyte chemoattractant protein (MCP)-1, transforming growth factor (TGF)-β and IL-6 concentrations and markers of oxidative stress including glutathione peroxidase (GPx), superoxide dismutase (SOD), malondialdehyde (MDA), and catalase (CAT) concentrations in adipose tissue of rats were determined using ELISA kits and spectrophotometry methods, respectively. Vitamin D treatment led to a significant reduction in adipose tissue TNF-α concentrations in both ND + vitamin D and HFD + vitamin D groups (P<0.05). Adipose tissue MCP-1 concentration also reduced in HFD + vitamin D group compared with HFD group. Among markers of oxidative stress in adipose tissue, SOD and GPx concentrations significantly increased in adipose tissue of HFD + vitamin D treated group compared with other groups (P<0.05). Reduced food intake and weight gain was also occurred after vitamin D treatment. Vitamin D improved adipose tissue oxidative stress and inflammatoryparameters in obese rats. Vitamin D treatment was also associated with decreased food intake and decreased weight gain in animals under a high-fat diet. Further studies are needed to better clarify the underlying mechanisms.

Abstract CT086: Treatment of ductal carcinoma in situ (DCIS) with pasireotide, an IGF-I inhibitor

Abstract DCIS is a non-obligate precursor in which epithelial cells with features of cancer reside in and expand otherwise normal appearing mammary ducts and lobules. DCIS increases the relative risk of developing invasive breast cancer (IBC) by up to 10 fold. Treatment of DCIS includes breast conserving surgery (BCS), tumor margins permitting, followed by radiation, or mastectomy. Recently we found that pasireotide, a somatostatin analog inhibitor of IGF-I, was effective in reducing cell proliferation and increasing apoptosis in patients with atypical hyperplasia (AH) and or proliferative disease of the breast. At present there are no recognized medical treatments for DCIS. Here we raise the possibility that pasireotide might also be an effective therapy for some DCIS. To make this determination, 8 patients previously diagnosed with DCIS were treated with pasireotide, 600µg twice daily subcutaneously for a period of 20 days. Lesion size was assessed on DCE-MRIs in 3 planes before and after treatment. Of the eight, two had lesions characterized as low grade DCIS on core biopsy, 4 had intermediate or intermediate to high grade lesions, and two had high grade lesions. After treatment they all had surgical excision biopsies. We also measured proliferation and apoptosis to assess effects of pasireotide by expression of Ki67 and caspase 3. To confirm that the effect of pasireotide was mediated via the IGF-I pathway, p-AKT and p-ERK 1/2 were evaluated. Core and excision samples from 14 untreated patients where used as control. Overall, treatment with pasireotide reduced DCIS lesion size by ≥30% according the RECIST criteria in 4 of 8 patients, as measured by DCE-MRI. Of those, pathology of the surgical excision confirmed that there was no remaining DCIS tissue in the two patients with low grade disease. The two additional lesions that were reduced in size one was intermediate to high grade and the other a high grade DCIS. The remaining 4 patients had tumors that were stable in size. Pasireotide reduced cell proliferation and significantly increased apoptosis in the DCIS lesions. While DCIS is less responsive to pasireotide than AH and proliferative disease, it may nevertheless provide a window of opportunity for treatment and possibly eradication of some low grade or higher grade DCIS as well. The main side effect included hyperglycemia, intestinal discomfort and rash or redness at the injection area. Our data show that hyperglycemia disappears relatively fast after ending treatment with pasireotide. In conclusion, treatment with pasireotide eliminated 2 low grade DCIS lesions, shrunk two higher grade ones and also decreased proliferation and increased apoptosis. Additional evidence in a larger number of patients will be necessary. Citation Format: Irineu Illa-Bochaca, Linda Moy, Julia Smith, Debora Axelrod, Baljit Singh, David L. Kleinberg. Treatment of ductal carcinoma in situ (DCIS) with pasireotide, an IGF-I inhibitor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr CT086. doi:10.1158/1538-7445.AM2017-CT086

Abstract 2266: A prospective study of serum metabolites and glioma risk

Abstract Malignant glioma is one of the most devastating adult malignancies. Although there exists an urgent need for actionable leads regarding its prevention and early detection, the etiology of glioma remains largely unknown. We conducted a prospective serum metabolomic analysis of glioma based on 64 cases and individually matched controls selected from Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study. Median time from collection of baseline fasting serum to diagnosis of glioma was nine years (interdecile range 3-20 years). We used ultra-high performance LC/MS which identified 730 known metabolites for analysis. Conditional logistic regression models examined odds ratios (ORs) and 95% confidence intervals for one-standard deviation differences in metabolite signals on a log-scale. Forty-three serum metabolites were associated with glioma at P&amp;lt;0.05. 2-Oxoarginine, cysteine, alpha-ketoglutarate, chenodeoxycholate and argininate yielded the strongest metabolite signals and were inversely associated overall risk of glioma (0.0065≤P&amp;lt;0.0083). Three serum acylcarnitines (0.016≤P&amp;lt;0.046), and seven xanthine (i.e., caffeine) metabolites (0.017≤P&amp;lt;0.042) were higher in cases than controls. Findings were mostly similar in high-grade glioma cases, although proportionally more metabolites showed positive risk associations such as N-acetylglutamate, ribonate, and 5-methyluridine (0.04≤P&amp;lt;0.02). Notably, xanthine compounds appeared unrelated to high-grade glioma. Prominent metabolite signals for inverse associations were the bile acids glycocholenate sulfate and 3β-hydroxy-5-cholenoic acid, xenobiotic methyl-4-hydroxybenzoate sulfate, sex steroid 5alpha-pregnan-3beta,20beta-diol monosulfate, and cofactor/vitamin oxalate. We found weaker risk associations for lower-grade glioma, of which none achieved significance at P&amp;lt;0.05. The metabolic sub-pathway analysis revealed associations for primary bile acid, urea cycle/arginine and proline, tocopherol, glycolysis/gluconeogenesis/pyruvate metabolites with overall glioma risk (0.005≤P&amp;lt;0.048); ascorbate and aldarate pathways were associated with high-grade glioma (P=0.02), whereas glycolysis/gluconeogenesis/pyruvate, eicosanoid and glutamate pathways were found to be associated with lower-grade disease (0.01≤P&amp;lt;0.04). These associations did not, however, reach the stringent Bonferroni significance threshold for correction of multiple comparisons. In conclusion, we identified a serum metabolomic profile of glioma, years in advance of clinical diagnoses, characterized by altered molecular signals in the arginine/proline, antioxidant, bile acid, and glycolytic pathways. The observed patterns provide potential new leads regarding the molecular basis relevant to etiologic or sub-clinical biomarkers for glioma. The findings require reexamination in larger and more diverse populations, including through glioma consortia. Citation Format: Jiaqi Huang, Stephanie J. Weinstein, Cari M. Kitahara, Edward Karoly, Joshua N. Sampson, Demetrius Albanes. A prospective study of serum metabolites and glioma risk [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2266. doi:10.1158/1538-7445.AM2017-2266

Abstract 2523: Key cluster for prostate cancer diagnosis and therapy; miR-888 cluster

Abstract Prostate cancer (PCa) is the 2nd leading cause of cancer death among men in the U.S. No accurate biomarkers exist and therapeutic options are limited for advanced PCa. MicroRNAs (miRNAs), small non-coding RNAs, act as negative regulators of gene expression. MiRNAs have recently emerged as promising therapeutic tools for cancers. It is poorly understood how they function to promote cancer progression. We found miR-888 was involved with PCa progression. miR-888 belongs to a genomic cluster on human chromosome Xq27.3, linked to hereditary PCa. I hypothesized additional members of this cluster (miR-892c, -890, -892a, -892b, -891b, -891a) function to promote PCa progression. Our lab profiled miR-888 cluster expression in paired syngenic human PCa cell lines (RWPE1 &amp; WPE1-NB26, LNCaP &amp; C4-2, PC3-N &amp; PC3-ML) differing in metastatic status and androgen response. We noted that the miR-888 cluster was preferentially elevated in metastatic PC3-ML and under-expressed in non-aggressive PC3-N cells. I examined miR-888 cluster function in vitro and found that forced expression (via miRNA mimics) of certain miR-888 cluster members, notably miR-888 and miR-891a, promoted cell proliferation, colony formation, migration and invasion in PC3-N cells. Conversely, other cluster members inhibited these activities. Experiments with hormone sensitive LNCaP cells showed similar results in these assays. PC3-ML transfected with miRNA inhibitors exhibited reciprocal effects. In mouse xenograft studies, I used lentiviral vectors to individually overexpress miR-888 and miR-891a in PC3-N cells and injected them subcutaneously into flanks of NOD/SCID mice. The miRNAs accelerated tumor formation and increased tumor size compared to controls. miR-888 may promote tumorigenesis by suppressing the tumor suppressors TIMP2 and SMAD4, which our lab validated as miR-888 targets using luciferase reporter assays. I confirmed by western blot that xenograft tumors overexpressing miR-888 had decreased protein levels for these targets. Studies reveal miRNAs exist in exosomes, small membrane bound vesicles involved in cell-to-cell communication. I postulated that aggressive tumors secrete exosomes with the miR-888 cluster as cargo and promote PCa progression. Indeed, I noted that miR-888 cluster expression is elevated in exosomes isolated from aggressive PC3-ML compared to PC3-N cells as well as in EPS urine exosomes from high-grade PCa patients versus those with low-grade disease. My work indicates miR-888 and miR-891a are novel oncogenic factors in the prostate and may use exosomes to exert their effects. This is the first report to explore the functional role of the miR-888 cluster in any tissue and may lead to promising clinical strategies for PCa. Citation Format: Tsuyoshi Hasegawa, Aurora Kerscher. Key cluster for prostate cancer diagnosis and therapy; miR-888 cluster [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2523. doi:10.1158/1538-7445.AM2017-2523

Abstract 2217: Detection of circulating immune biomarkers of cervical disease using proteome arrays

Abstract Introduction: More than 260,000 women die of cervical cancer every year. Screening methods have reduced the incidence of cervical cancer in high-income countries, but detection continues to lag in low and middle income countries (LMICs). IgG antibody (Ab) immunity to early (E) HPV antigens (Ags) are potential biomarkers of disease progression. Since HPV16 accounts for only ~50% of invasive cervical cancers, we developed protein microarrays expressing the proteomes of 12 HPV types to detect host IgG Abs to a broad spectrum of viral Ags to detect pre-invasive and invasive cervical disease. Methods: We developed custom HPV protein microarrays displaying the proteomes of two low-risk HPV types (HPV6 and 11) and ten oncogenic high-risk HPV types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52 and 58). Arrays were probed with serum samples obtained from women with invasive cervical cancer (ICC; n=80), no or low-grade cervical disease (CIN 0/I; n=60), and high-grade cervical dysplasia (CIN II/III; n=60). To identify positive serologic responses, arrays were scanned and the signal intensity of each protein spot was quantified and normalized. Visual examination of diffused signal (ring) around each spot was performed and the Ab response to each protein was scored on a scale from 0 to 5. Results: To verify array quality and reproducibility, we confirmed high protein expression levels for 98% (96/98) of the antigens printed and high correlation (R ≥ 0.90) of protein expression signals between different randomly selected arrays. Epitope expression was confirmed using four commercial monoclonal Abs raised against HPV16 Ags. Host Abs to at least one early antigen (E1, E2, E4, E6, or E7) were detected in the sera of 31.2% and 43.3% of ICC and CINII/III patients, respectively, compared with 11.7% of women with CIN 0/I. 73.1% of CINII/III cases detected had Abs only to non-HPV16 Ags. Abs to E1, E2, E4, E6, and E7 Ags were detected in 3.8%, 7.7%, 54%, 19%, and 31% of CINII/III versus 16%, 16%, 32%, 32%, and 40% of ICC cases that were positive. The immunodominant Ags in women with CINII/III and ICC were E4 (54%) and E7 (40%), respectively. These results are consistent with the difference in tissue expression levels of these proteins in these two disease stages. This emphasizes the importance of broadening the scope of serological detection to include non-HPV16 proteomes. Conclusions: This study demonstrates that Abs against high risk HPV16 and non-HPV16 serotypes are potential biomarkers for both the diagnosis of cervical cancer and the differentiation between high grade and low-grade pre-invasive cervical lesions. These results suggest that serology is a potential minimally invasive tool for early detection of cervical cancer. High-throughput immunoprofiling of HPV-associated cancers and pre-invasive lesions may be informative for understanding the diversity of host immunity and viral progression. Citation Format: Radwa Ewaisha, Ian Meshay, Jack Resnik, Tirinder Bharaj, Karen S. Anderson. Detection of circulating immune biomarkers of cervical disease using proteome arrays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2217. doi:10.1158/1538-7445.AM2017-2217

Primary endometrioid stromal sarcomas of the ovary: a clinicopathological study of 14 cases with a review of the literature.

PurposePrimary endometrioid stromal sarcomas (ESS) of the ovary are rare mesenchymal tumors with scarce data on their behavior and optimal treatment. We aimed to describe the clinicopathologic features and outcome among patients with primary ovarian ESS.ResultsThe age of the patients ranged from 34 to 61 years (mean: 49.1 years, median: 51.5 years). The most common symptoms were abdominal distention or pain or both. Nine (64.3%) and five patients (35.7%) had low-grade and high-grade disease, respectively. The median duration of follow-up was 65 months (range, 8–311 months). All 9 patients with low-grade ESS were alive, of these, 3 (33.3%) of them developed recurrence after surgery. Only 1 patient (20%) with high-grade ESS was alive with no evidence of disease in a short-term follow-up visit; the remaining 4 (80%) developed recurrence after surgery, and 2 (40%) died of progressive disease.MethodsMedical records of 14 patients with primary ovarian ESS in our institution were collected and analyzed.ConclusionsThe behavior of primary ovarian ESS is similar to that of their uterine counterparts. Low-grade ESS is an indolent tumor with a propensity for late recurrences. The prognosis of high-grade ESS is poor.

Operative strategies to reduce cerebral embolic events during on- and off-pump coronary artery bypass surgery: A stratified, prospective randomized trial

ObjectiveTo determine the impact of different aortic clamping strategies on the incidence of cerebral embolic events during coronary artery bypass grafting (CABG). MethodsBetween 2012 and 2015, 142 patients with low-grade aortic disease (epiaortic ultrasound grade I/II) undergoing primary isolated CABG were studied. Those undergoing off-pump CABG were randomized to a partial clamp (n = 36) or clampless facilitating device (CFD; n = 36) strategy. Those undergoing on-pump CABG were randomized to a single-clamp (n = 34) or double-clamp (n = 36) strategy. Transcranial Doppler ultrasonography (TCD) was performed to identify high-intensity transient signals (HITS) in the middle cerebral arteries during periods of aortic manipulation. Neurocognitive testing was performed at baseline and 30-days postoperatively. The primary endpoint was total number of HITS detected by TCD. Groups were compared using the Mann–Whitney U test. ResultsIn the off-pump group, the median number of total HITS were higher in the CFD subgroup (30.0; interquartile range [IQR], 22-43) compared with the partial clamp subgroup (7.0; IQR, 0-16; P < .0001). In the CFD subgroup, the median number of total HITS was significantly lower for patients with 1 CFD compared with patients with >1 CFD (12.5 [IQR, 4-19] vs 36.0 [IQR, 25-47]; P = .001). In the on-pump group, the median number of total HITS was 10.0 (IQR, 3-17) in the single-clamp group, compared with 16.0 (IQR, 4-49) in the double-clamp group (P = .10). There were no differences in neurocognitive outcomes across the groups. ConclusionsFor patients with low-grade aortic disease, the use of CFDs was associated with an increased rate of cerebral embolic events compared with partial clamping during off-pump CABG. A single-clamp strategy during on-pump CABG did not significantly reduce embolic events compared with a double-clamp strategy.

A DNA hypermethylation profile reveals new potential biomarkers for the evaluation of prognosis in urothelial bladder cancer.

DNA hypermethylation has emerged as a molecular biomarker for the evaluation of cancer diagnosis and prognosis. We define a methylation signature of bladder cancer and evaluate whether this profile assesses prognosis of patients. Genome-wide methylation analysis was performed on 70 tumor and 10 normal bladder samples. Hypermethylation status of 1505 CpGs present in the promoter region of 807 genes was studied. Thirty-three genes were significantly hypermethylated in ≥10% of the tumors. Three clusters of patients were characterized by their DNA methylation profile, one at higher risk of dead of disease (p = 0.0012). Association between cluster distribution and stage (p = 0.02) or grade (p = 0.02) was demonstrated. Hypermethylation of JAK3 and absence of hypermethylation of EYA4, GAT6, and SOX1 were associated with low-grade non-invasive disease. On the other hand, in high-grade invasive disease hypermethylation of CSPG2, HOXA11, HOXA9, HS3ST2, SOX1, and TWIST1 was associated with muscle invasiveness. A panel of hypermethylated genes including APC, CSPG2, EPHA5, EYA4, HOXA9, IPF1, ISL1, JAK3, PITX2, SOX1, and TWIST1 predicted cancer-specific survival and SOX1 (HR = 3.46), PITX2 (HR = 4.17), CSPG2 (HR = 5.35), and JAK3 hypermethylation (HR = 0.19) did so independently. Silencing of genes by hypermethylation is a common event in bladder cancer and could be used to develop diagnostic and prognostic markers. Combined hypermethylation of SOX1, PITX2, or CSPG2 signals patients at higher risk of death from bladder cancer.

Epigenetic Signature: A New Player as Predictor of Clinically Significant Prostate Cancer (PCa) in Patients on Active Surveillance (AS).

Widespread prostate-specific antigen (PSA) testing notably increased the number of prostate cancer (PCa) diagnoses. However, about 30% of these patients have low-risk tumors that are not lethal and remain asymptomatic during their lifetime. Overtreatment of such patients may reduce quality of life and increase healthcare costs. Active surveillance (AS) has become an accepted alternative to immediate treatment in selected men with low-risk PCa. Despite much progress in recent years toward identifying the best candidates for AS in recent years, the greatest risk remains the possibility of misclassification of the cancer or missing a high-risk cancer. This is particularly worrisome in men with a life expectancy of greater than 10–15 years. The Prostate Cancer Research International Active Surveillance (PRIAS) study showed that, in addition to age and PSA at diagnosis, both PSA density (PSA-D) and the number of positive cores at diagnosis (two compared with one) are the strongest predictors for reclassification biopsy or switching to deferred treatment. However, there is still no consensus upon guidelines for placing patients on AS. Each institution has its own protocol for AS that is based on PRIAS criteria. Many different variables have been proposed as tools to enrol patients in AS: PSA-D, the percentage of freePSA, and the extent of cancer on biopsy (number of positive cores or percentage of core involvement). More recently, the Prostate Health Index (PHI), the 4 Kallikrein (4K) score, and other patient factors, such as age, race, and family history, have been investigated as tools able to predict clinically significant PCa. Recently, some reports suggested that epigenetic mapping differs significantly between cancer patients and healthy subjects. These findings indicated as future prospect the use of epigenetic markers to identify PCa patients with low-grade disease, who are likely candidates for AS. This review explores literature data about the potential of epigenetic markers as predictors of clinically significant disease.