Higher Frequency Of Tumors Research Articles

Prevalence of Viral Neoplastic Diseases with Special Reference to Avian Leukosis and their Infectious Status in Different Pure Line Indigenous Chickens

Viral neoplastic diseases in poultry causes huge economic loss all over the world. The prevalence of avian leukosis in different indigenous breeds of chicken is not known and needs to be addressed for its effective control. A study was designed to record the prevalence of viral neoplasms in poultry, screening of avian leukosis virus/viruses (ALV) antigen in various pure line chicken, and the infectious status of ALV (viremia or shedding) in indigenous birds. Routine necropsy was carried out to identify the birds that were died of neoplasm and the representative samples were collected. Out of 4195 birds necropsied, 90 birds were recorded with neoplastic lesions in various organs with a prevalence of 2.1%. Grossly, tumor lesions were recorded in various organs in which the high frequency of tumors were recorded in liver, spleen, and kidney. Histopathological examination revealed 13.5% of the samples had myeloid leukosis, 25.2% were lymphoid leukosis, 20.3% were histiocytic sarcomas and 40.7% were Marek’s disease. PCR screening of the tumor samples revealed the prevalence rate of 45.5% Marek’s disease virus (MDV), 53.3% ALV, and 1.1% reticuloendotheliosis virus (REV). Co-infections of MDV and ALV were recorded in 37.7% (34/90) samples and MDV, ALV, and REV were recorded in 1.1% (1/90) samples. Among ALV positive cases, 54.1% (26/48) were positive for ALV E and 25% (12/48) were positive for ALV J. Screening of ALV p27 antigen by antigen ELISA in various pure line chickens revealed the prevalence rate of 76.3% in Kadaknath, 73.5% in Ghagus, 76.5% in Aseel, 20.1% in PB-1, 18.8% in PB-2, 19.6% in PD-1, 14.3% in PD-4 and 8.6% in CB. Further molecular detection of ALV was carried out in buffy coat and albumen samples from the birds of Kadaknath, Ghagus, and Aseel that showed high SPR value to determine the infectious status of the ALV. Infectious status of ALV in the birds revealed 40% in Kadaknath, 68.4% in Ghagus and 85.7% in Aseel were viremic shedders (V+S+); 33.3% in Kadaknath, 10.5% in Ghagus and 0% in Aseel were viremic non shedders (V+S-); 16.6% in Kadaknath, 10.5% in Ghagus and 14.2% in Aseel were non viremic shedders (V-S+). This is the first study that documented the prevalence of ALV in different indigenous pure line birds and their infectious status in India.

Association Between Metabolic Syndrome and Immunohistochemical Profile at Breast Cancer Diagnosis in Postmenopausal Women

BackgroundThe aim of this study was to evaluate the association between metabolic syndrome (MetS) and the immunohistochemical profile of breast cancer (BC) in postmenopausal women. MethodsThis cross-sectional cohort study included 189 women, aged 45 to 75years and amenorrhea >12 months, with newly diagnosed BC and no previous cancer treatment. Clinical, anthropometric and biochemical data were collected, as well as data on BC hormone status (estrogen receptor, ER; progesterone receptor, PR; human epidermal growth factor receptor-2, HER-2), and epithelial proliferative activity (Ki-67). Tumors were divided into 5 subtypes:luminal A, luminal B HER-2 negative, luminal B HER-2 positive, non-luminal HER-2, and triple negative. Women with three or more of the following criteria were diagnosed with MetS: waist circumference ≥88cm; triglycerides ≥150mg/dL; HDL-cholesterol <50mg/dL; blood pressure ≥130/85mmHg; glucose ≥100mg/dL. ResultsSixty-three (33.3%) of the 189 patients had MetS at the time of diagnosis. Women with MetS had a higher frequency of tumors ≤ 2cm than women without MetS (49.2% vs. 31.8%) (P = .038). There were no differences in histological grade, staging, or axillary lymph node metastasis (P > .05). The proportion of PR-positive (P = .006), HER-2-negative (P = .034), and luminal B HER-2-negative (P = .038) tumors was higher among patients with MetS compared to women without MetS (79.4% vs. 61.8%, 89.9% vs. 78.6% and 44.5% vs. 27.8%, respectively). Multivariate analysis adjusted for age, time since menopause and BMI showed a higher risk for luminal B HER-2-negative tumors among women with MetS (OR 2.00, 95% CI 1.03-3.89), obese patients (OR 2.03, 95% CI 1.06-3.90), and women with abdominal obesity (OR 1.96, 95% CI 1.01-4.03). ConclusionIn postmenopausal women with newly diagnosed BC, the presence of MetS was associated with smaller tumor size, PR-positive and HER-2-negative status, and the luminal B tumor subtype.

Diversity of draining lymph node-tumor overlapping T cell receptor repertoire reflects the anti-tumor effect of cancer immunotherapies

Abstract CD8+ tumor-infiltrating lymphocytes, the main effector cells of anti-tumor immune responses, are composed of various T cell clones. Numerous studies have demonstrated the presence of tumor-reactive T cell clones with high frequency in tumors. However, it remains unclear as to which of the two mechanisms, the expansion of limited tumor-specific clones or the induction of diverse tumor-reactive ones, is more beneficial for the action of immune checkpoint inhibitors (ICIs). To address this question, we performed a T cell receptor repertoire analysis on B16F10 and on Lewis lung carcinoma tumor-bearing mice treated with immunotherapy including anti-PD-L1 monoclonal antibody (mAb) and anti-CD4 depletion mAb, which enhance the expansion of tumor-reactive CD8+ T cells. We focused on the T cell clones that overlapped between the tumor and draining lymph node (dLN) to analyze the T cell clones that were mobilized into the anti-tumor response after treatment. In anti-PD-L1 and anti-CD4 treated mice, the total frequency of CD8+ dLN-tumor overlapping clones in the tumor was increased. Surprisingly, although the expansion of the top10 overlapping clones did not differ, ICI treatment increased the total frequency of the 11th or lower overlapping clones, indicating the broadening of the tumor-reactive T cell repertoire. Moreover, the administration of FTY720 to tumor-bearing mice, which inhibits egress of T cells from the LNs to circulation, reduced the total frequency of the 11th or lower overlapping clones in the tumor and reduced the long-term anti-tumor effect of anti-PD-L1 treatment. These results suggest that mobilization of diverse tumor-reactive clones is essential for enhancing and persistent anti-tumor effect of ICIs.

B7-H3 and PD-L1 Expression Are Prognostic Biomarkers in a Multi-racial Cohort of Patients with Colorectal Cancer

BackgroundImmunotherapy has emerged as an effective and durable treatment modality for solid cancers. However, its use in colorectal cancer (CRC) is limited to deficient mismatch repair (dMMR) tumors. As such, assessing immune regulatory proteins from the B7-CD28 family, other than PD-1, PD-L1, and CTLA-4, is critical. This study aimed to evaluate the expression of novel protein regulators in a racially diverse population of patients with CRC. MethodsA tumor microarray was created for 214 samples from a multiracial patient population with metastatic CRC, and expression of HHLA2, B7-H3, PD-L1, CK7, CK20, and CDX2 was determined. The expression pattern was scored as 0 to 12, based on tumor tissue prevalence and the intensity. Clinical information was obtained by chart review and vital statistics from the National Death Index. Associations between low and high expression groups for each protein by race/ethnic groups were assessed, and Kaplan–Meier curves were plotted to evaluate association with survival. ResultsThe median age at diagnosis was 61 years, with a female predominance. The majority of the patients were diagnosed with de novo metastatic disease with left-sided, moderately differentiated tumors. There were no racial disparities in the expression of any protein. Overall, a high frequency of tumors had no expression of B7-H3 (62.5%) or PD-L1 (43.5%). Low expression of both PD-L1 and B7-H3 was a significant prognostic biomarker associated with better survival (median overall survival, 43.3 months vs. 24.6 months; P < .01). ConclusionIn this multiracial tumor microarray of CRC samples, low PD-L1 and B7-H3 expression was associated with an improved prognosis. There was no significant variation among races with respect to the relevant CRC protein markers.

Abstract PS7-76: Association between metabolic syndrome and immunohistochemical profile at breast cancer diagnosis in postmenopausal women

Abstract Objective: To evaluate the association between metabolic syndrome (MetS) and the immunohistochemical profile of breast cancer (BC) in postmenopausal women.Methods: This cross-sectional cohort study included 189 women, aged 45-75 years and amenorrhea &amp;gt; 12 months, with newly diagnosed BC and no previous cancer treatment. Clinical, anthropometric and biochemical (total cholesterol, HDL, LDL, triglycerides, and glucose) data were collected, as well as data on BC (histopathology, grade, tumor stage, lymph node metastasis hormone status (estrogen receptor, ER; progesterone receptor, PR; human epidermal growth factor receptor 2, HER-2), and epithelial proliferative activity (Ki-67). Tumors were divided into five subtypes: luminal A, luminal B HER-2 negative, luminal B HER-2 positive, non-luminal HER-2, and triple negative. Women with three or more of the following criteria were diagnosed with MetS: waist circumference ≥ 88 cm; triglycerides ≥ 150 mg/dL; HDL-cholesterol &amp;lt; 50 mg/dL; blood pressure ≥ 130/85 mmHg; glucose ≥ 100 mg/dL. The Student t-test, gamma distribution (asymmetric variables), chi-square test, and logistic regression (odds ratio, OR) were used for statistical analysis.Results: Sixty-three (33.3%) of the 189 patients had MetS at the time of diagnosis. The mean age, time since menopause and BMI were 59.0 ± 10.6 years, 11.4 ± 9.6 years and 28.5 ± 5.5 kg/m2, respectively, without difference between women with and without MetS (control). Women with MetS had a higher frequency of tumors ≤ 2 cm than women without MetS (49.2% vs 31.8%) (p=0.038). There were no differences in histological grade, staging, or axillary lymph node metastasis (p&amp;gt;0.05). The proportion of PR-positive (p=0.006), HER-2-negative (p=0.034), and luminal B HER-2-negative tumors was higher among patients with MetS (p=0.038) compared to women without MetS (79.4% vs 61.8%, 89.9% vs 78.6% and 44.5% vs 27.8%, respectively). Multivariate analysis adjusted for age, time at menopause and BMI showed a higher risk for luminal B HER-2-negative tumors among women with MetS (OR 2.00, 95% CI 1.03-3.89), obese patients (OR 2.03, 95% CI 1.06-3.90), and women with abdominal obesity (OR 1.96, 95% CI 1.01-4.03). The other BC subtypes were not associated with MetS or its components.Conclusion: In postmenopausal women with newly diagnosed BC, the presence of MetS was associated with a more favorable immunohistochemical profile of BC, such as a smaller tumor size, PR-positive and HER-2-negative status, and the luminal B tumor subtype. Citation Format: Andre Hideo Motoki, Daniel de AraujoBrito Buttros, Ana LuisaCamolezi Gaspar, Eduardo Carvalho Pessoa, Benedito de Sousa Almeida Filho, Jorge Nahas Neto, Heloisa de Luca Vespoli, Eliana AguiarPetri Nahas. Association between metabolic syndrome and immunohistochemical profile at breast cancer diagnosis in postmenopausal women [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS7-76.

Proteogenomics and de novo sequencing based approach for neoantigen discovery from the immunopeptidomes of patient CRC liver metastases using Mass Spectrometry

Abstract Introduction Using High-resolution Mass Spectrometry (MS), we developed a novel immunopeptidomics workflow using de novo sequencing. This method does not rely on a reference database to deduce full-length peptide sequences. As a consequence, peptide discovery is not limited to the reference database and allows for the discovery of novel peptides directly from MS. Goal Identify HLA-associated peptides that appear in high frequency in tumors through de novo sequencing and missed by reference database searches. Method We employed a highly selective anti-HLA Class I monoclonal antibody, W6/32, to enrich HLA-complexes from 21 CRC liver metastases and identified HLA associated peptides using a Thermo Orbitrap Fusion Lumos on-line with a nanoLC 1200. RNA-Seq was also performed on each tumor. MS raw spectra were analyzed by de novo assisted PEAKS-X (BSI) Results Approximately 1600 unique de novo sequenced HLA-associated peptides were identified in a single LC-MS analysis with high confidence (95% ALC cut-off) from each tumor. Approximately 45% of the 1600 unique de novo sequenced peptides matched to reference databases (e.g Uniprot.), leaving a large percentage of peptides with no match. Interestingly, some of these unmatched peptides have been validated as neoantigens that arise from gene mutation, spliced peptides generated by the proteasomes, and non-coding region derived peptides. Conclusion These de novo sequenced peptides without matches in reference databases may provide a unique source of disease-specific HLA class I-presented peptides and trigger specific immune responses. Some of the higher frequency unmatched de novo sequenced peptides may thus serve as tumor-specific targets for next-generation immunotherapies.

Abstract 3150: Impact of interleukin-1 alpha and EGFR expression on recurrence and survival outcomes in head and neck squamous cell carcinomas

Abstract Interleukin-1 alpha (IL-1α) is a pleiotropic cytokine involved in inflammation and immune response and is upregulated in many solid tumors including head and neck squamous cell carcinomas (HNSCCs). Although IL-1α expression is generally associated with poor prognosis, the implications of the subcellular localization of IL-1α expression in patient outcomes are poorly understood. This study is aimed at investigating the clinical relevance of immunohistochemical IL-1α expression in HNSCCs. Tissue microarrays (TMAs) containing human papillomavirus (HPV)-positive and HPV-negative HNSCCs were analyzed for IL-1α and epidermal growth factor receptor (EGFR) expression by immunohistochemistry. Nuclear and cytoplasmic IL-1α and EGFR expression scores were correlated with clinicopathological parameters and patient outcomes. IL-1α expression was observed in the nuclear and/or cytoplasm compartments in 98% of evaluable tumors and 78% of tumors expressed IL-1α in both compartments. A higher percentage of tumors with combined high nuclear and moderate cytoplasmic IL-1α expression were observed in HPV-negative tumors compared to HPV-positive tumors. In HPV-negative tumors a combined EGFR-negative and high nuclear IL-1α expression profile was associated with a low risk of tumor recurrence and favorable overall survival compared to all other EGFR/IL-1α expression profiles. Lastly, a higher frequency of tumors with combined high nuclear and moderate cytoplasmic IL-1α expression was observed in cetuximab monotherapy responders compared to non-responders. Altogether, IL-1α in combination with EGFR expression may be a strong indicator of risk of recurrence in HPV-negative HNSCCs and warrants further study as a tissue biomarker for predicting HNSCC patient outcomes. Citation Format: Andrean L. Simons, Anand Rajan, Katherine Gibson-Corley, Georgina Ofori-Amanfo, Patrick Ten Eyck, Madelyn Espinosa-Cotton, Sandra Schmitz, Joseph Coppock, Steven Sperry. Impact of interleukin-1 alpha and EGFR expression on recurrence and survival outcomes in head and neck squamous cell carcinomas [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 3150.

Abstract 2890: Paternal pre-conception stress as a potential risk factor for neuroblastoma

Abstract Neuroblastomas (NBs) are pediatric tumors that lack adequate treatment for their aggressive form, despite many efforts focusing on novel therapies. Although the disease is known to have a genetic background, its etiology cannot be explained solely by genetics. Yet, the role of environment in NB development is understudied. Thus, there are no established risk factors for NB aside from rare genetic predispositions and therefore no preventative measures. NBs arise due to defects in sympathetic neuron (SN) differentiation, which occurs in the fetus. Thus, we hypothesize that prenatal or pre-conception exposures may interfere with this process and promote NB. Maternal stress during pregnancy and paternal pre-conception stress induce neurodevelopmental defects in offspring by direct effects on fetal development or epigenetic changes, respectively. Indeed, we have previously shown that in a mouse model of NB prenatal stress interferes with SN differentiation in offspring, accelerating NB development and leading to its increased frequency and malignancy. The goal of the current study was to determine if a similar increase in risk of NB development can result from paternal pre-conception stress. To test the effect of paternal stress on NB, we subjected TH-MYCN males to 6 weeks of chronic unpredictable stress (CUS) prior to mating with wild type females and assessed the impact of stress on 1) miRNA content in the germline (RNAseq); 2) fertility and fetal survival; 3) NB frequency, latency, growth and metastasis in their hemizygous TH-MYCN offspring (periodic MRI, histological analyses). CUS resulted in significant changes in non-coding RNA content of the germline, including miRNAs and tsRNAs. Of particular interest was a stress-induced down-regulation of sperm miRNAs, including miR-200b and miR-128, which are involved in neuroblast differentiation, inhibit NB invasiveness and are inactivated in NB tumors. Paternal stress decreased the rate of successful pregnancies from 75% in controls to 37.5% and increased frequency of death in utero from 5.5% to 29.2% (p=0.02). Moreover, offspring of stressed fathers exhibited accelerated NB development (60% vs 0% of mice with tumors at 6 weeks of age for CUS and control groups, respectively; p=0.03) and growth (p=0.3), higher tumor frequency (60% vs 25% at 14 weeks of age, p=0.03), as well as a trend toward a more metastatic phenotype. However, these effects on NB formation and phenotype were observed solely in male offspring. In summary, our data provide the first evidence for the role of paternal preconception stress in NB development. If confirmed, our findings may open new avenues for NB prevention and identifying biomarkers of increased NB risk. Citation Format: Sung-Hyeok Hong, Susana Galli, Raquel Santana da Cruz, Lu Jin, Larissa Wetlisbach, Shiya Zhu, Abir Malik, Jason Tilan, Sonia de Assis, Joanna B. Kitlinska. Paternal pre-conception stress as a potential risk factor for 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 2890.

Epigenomics of Pancreatic Cancer: A Critical Role for Epigenome-Wide Studies.

Several challenges present themselves when discussing current approaches to the prevention or treatment of pancreatic cancer. Up to 45% of the risk of pancreatic cancer is attributed to unknown causes, making effective prevention programs difficult to design. The most common type of pancreatic cancer, pancreatic ductal adenocarcinoma (PDAC), is generally diagnosed at a late stage, leading to a poor prognosis and 5-year survival estimate. PDAC tumors are heterogeneous, leading to many identified cell subtypes within one patient’s primary tumor. This explains why there is a high frequency of tumors that are resistant to standard treatments, leading to high relapse rates. This review will discuss how epigenetic technologies and epigenome-wide association studies have been used to address some of these challenges and the future promises these approaches hold.

APOBEC-induced mutations and their cancer effect size in head and neck squamous cell carcinoma

Recent studies have revealed the mutational signatures underlying the somatic evolution of cancer, and the prevalences of associated somatic genetic variants. Here we estimate the intensity of positive selection that drives mutations to high frequency in tumors, yielding higher prevalences than expected on the basis of mutation and neutral drift alone. We apply this approach to a sample of 525 head and neck squamous cell carcinoma exomes, producing a rank-ordered list of gene variants by selection intensity. Our results illustrate the complementarity of calculating the intensity of selection on mutations along with tallying the prevalence of individual substitutions in cancer: while many of the most prevalently-altered genes were heavily selected, their relative importance to the cancer phenotype differs from their prevalence and from their P value, with some infrequent variants exhibiting evidence of strong positive selection. Furthermore, we extend our analysis of effect size by quantifying the degree to which mutational processes (such as APOBEC mutagenesis) contributes mutations that are highly selected, driving head and neck squamous cell carcinoma. We calculate the substitutions caused by APOBEC mutagenesis that make the greatest contribution to cancer phenotype among patients. Lastly, we demonstrate via in vitro biochemical experiments that the APOBEC3B protein can deaminate the cytosine bases at two sites whose mutant states are subject to high net realized selection intensities—PIK3CA E545K and E542K. By quantifying the effects of mutations, we deepen the molecular understanding of carcinogenesis in head and neck squamous cell carcinoma.

Immunohistochemical analysis of cyclin A expression in Wilms tumor.

BackgroundCyclin A overexpression is found in a variety of human tumors and correlates with unfavorable outcome. We analyzed immunohistochemical expression of cyclin A in Wilms tumor (WT) in relation to clinicopathological characteristics, preoperative chemotherapy (PrOpChTh), and overall survival (OS).MethodsThis retrospective study involved 43 patients who underwent nephrectomy from January 1996 to October 2010. Tumor stage and histological subtype were determined by revised Societé International d’Oncologie Pediatrique protocol, based on histological components/alterations caused by PrOpChTh, within the prognostic group of low, intermediate and high risk, and with criteria for anaplasia. The regressive/necrotic changes in total tumor mass of primary tumor and the proportion of epithelial, blastemal, and stromal components in the remaining viable tumor tissue were also determined. Cyclin A expression was evaluated by immunohistochemistry using a polyclonal rabbit, antihuman antibody (H-432).ResultsCyclin A overexpression was found in 34.3% of WTs, with higher frequency in tumors with epithelial (31.3%) and blastemal (37.1%) components than those with stromal component (17.7%). Regarding histological type, cyclin A overexpression was found most often in focal anaplasia (100%), stromal (60%), and diffuse anaplastic (66.7) WTs. The overexpression was also more frequent in stages 3 and 4 (77.8% and 66.7%, respectively) compared to tumors in stages 1 and 2 (13.3% and 12.5%, respectively; p = 0.004) in all components, as well as in blastemal component in stages 3 and 4 (77.8% and 66.7%, respectively) vs. stages 1 and 2 (13.3% and 25%, respectively, p = 0.009). Cyclin A overexpression in all components was 66.7% in WTs with metastasis and 31.3% in WTs without metastasis (p = 0.265, Fisher test). Log-rank testing revealed differences of OS regarding stage (p = 0.000), prognostic groups (p = 0.001), and cyclin A expression in blastemal component (p = 0.025). After univariate analysis, tumor stage (p = 0.001), prognostic group (p = 0.004), and cyclin A expression in blastemal component (p = 0.042) were significant prognostic factors for OS; however, after multivariate analysis, none of these factors were confirmed as independent predictors of survival.DiscussionThis study showed that cyclin A overexpression might be associated with the development and progression of WT with anaplasia. Also, cyclin A overexpression was more often observed in advanced stages (3 and 4) of WT, in the group of high-risk WTs, and in focal and diffuse anaplasia WTs. There was no relation of cyclin A overexpression and metastatic ability of WT. Although this study has not confirmed the prognostic value of cyclin A overexpression, its association with unfavorable prognosis should be further evaluated.

The Impact of Implant Location on Breast Cancer Characteristics in Previously Augmented Patients: A Systematic Literature Analysis.

BackgroundThere is a paucity of data comparing the oncologic properties of breast cancer among patients previously having undergone breast augmentation in either the subglandular or subpectoral planes. The objective of the present systematic review was to evaluate whether implant location influenced the characteristics of breast tumors in previously augmented women.MethodsA systematic literature search was performed to identify relevant articles reporting tumor characteristics in augmented patients. The search included published articles in three electronic databases; Ovid MEDLINE, EMBASE, and PubMed. Comparative studies (subglandular vs. subpectoral) were included.ResultsAnalysis of data pooled from the included studies showed that subglandular implants had a higher frequency of tumors between 2 to 5 cm (26.5% vs. 9.9%, P = 0.0130). Subglandular implants also had a higher frequency of stage 2 tumors (42.9% vs. 23.7%, P = 0.0308). There was no significant difference in lymphovascular invasion between the 2 groups. These results of this systematic review suggest that the prognosis of patients undergoing augmentation is unaffected by implant location (subpectoral vs. subglandular).ConclusionsWith the absence of large randomized controlled trials, our study provides surgeons with an evidence-based reference to improve informed consent with regards to implant placement.

Precision cancer therapy: profiting from tumor specific defects in the DNA damage tolerance system.

DNA damage tolerance (DDT) enables replication to continue in the presence of a damaged template and constitutes a key step in DNA interstrand crosslink repair. In this way DDT minimizes replication stress inflicted by a wide range of endogenous and exogenous agents, and provides a critical first line defense against alkylating and platinating chemotherapeutics. Effective DDT strongly depends on damage-induced, site-specific PCNA-ubiquitination at Lysine (K) 164 by the E2/E3 complex (RAD6/18). A survey of The Cancer Genome Atlas (TCGA) revealed a high frequency of tumors presents RAD6/RAD18 bi-allelic inactivating deletions. For instance, 11% of renal cell carcinoma and 5% of pancreatic tumors have inactivating RAD18-deletions and 7% of malignant peripheral nerve sheath tumors lack RAD6B. To determine the potential benefit for tumor-specific DDT defects, we followed a genetic approach by establishing unique sets of DDT-proficient PcnaK164 and -defective PcnaK164R lymphoma and breast cancer cell lines. In the absence of exogenous DNA damage, PcnaK164R tumors grew comparably to their PcnaK164 controls in vitro and in vivo. However, DDT-defective lymphomas and breast cancers were compared to their DDT-proficient controls hypersensitive to the chemotherapeutic drug cisplatin (CsPt), both in vitro and in vivo. CsPt strongly inhibited tumor growth and the overall survival of tumor bearing mice greatly improved in the DDT-defective condition. These insights open new therapeutic possibilities for precision cancer medicine with DNA damaging chemotherapeutics and optimize Next-Generation-Sequencing (NGS)-based cancer-diagnostics, -therapeutics, and -prognosis.

Abstract 3301: Breast cancer subtypes have distinct microbial and immune cytolytic transcriptomic signatures

Abstract Background: The role of genetic mutations in breast cancer is well documented and has formed the basis for novel therapies. However, the impact of viral and bacterial infection in the pathogenesis of breast cancer is not well understood. Methods: We analyzed RNA sequencing (RNASeq) data generated by the Cancer Genome Atlas (TCGA) from over 900 pairs of breast tumor and adjacent normal tissues, using our pathogen discovery tool, PathSeq. We developed a novel metric for quantifying and normalizing microbial sequence abundance from RNASeq data by correcting for both the sequencing coverage and relative genome sizes of bacteria in the dataset. We also performed linear discriminant analyses (LDA) to test the hypothesis that the differences between the canonical breast cancer subtypes can be modeled on microbial abundance data. In addition, we interrogated the human gene expression data derived from the RNASeq dataset to determine the relationships between the breast cancer subtypes and immune cytolytic activity (CYT index; defined as the geometric mean of the expression levels of perforin and granzyme). Results: Viral sequences were rare but we identified bacterial sequences to varying degrees of abundance. Intriguingly, we demonstrated that microbial abundance data can be used to recapitulate the well-established PAM50 gene expression-based subsets. Comparison of tumors and adjacent normal tissues also revealed subtype-associated microbial profiles. In addition, we observed that the basal and Her2 subtypes of breast cancer had a higher frequency of tumors with high CYT index compared with luminal A and B subtypes. Conclusion: This study has revealed that the various breast cancer subtypes have unique microbial transcriptomic signatures as well as differential immune cytolytic activity profiles. These data suggest the potential for differential responses of breast cancer subsets to immunotherapy. Citation Format: Chandra Sekhar Pedamallu, Matthew Meyerson, Akinyemi Ojesina. Breast cancer subtypes have distinct microbial and immune cytolytic transcriptomic signatures. [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 3301.

Abstract 116: Study of TERT promoter and FGFR3 mutations in upper-tumor urothelial carcinomas

Abstract Background: Mutations in the promoter of the telomerase reverse transcriptase (TERT) and fibroblast growth factor receptor 3 (FGFR3) genes represent the most recurrent somatic alterations in urothelial carcinomas of the bladder. However, data regarding frequency and prognostic values of these alterations in upper-tumor urothelial carcinoma (UTUC) remain limited. Methods: DNA was extracted from 31 UTUC including 12 cases arising in the ureter and 19 in renal pelvis. Somatic mutations of TERT promoter and FGFR3 gene were assessed by Sanger sequencing. The generated sequences were analyzed by seqscape and mutational status of each of those genes were correlated with clinico-pathological tumor features. Disease-free survival and overall survival were estimated using the Kaplan-Meier method. Results: Median patient's age was 72 years (range: 41-83) with a male predominance (90.3%). Mutation status of TERT and FGFR3 were obtained in all tumors. Out of 31 UTUC, 81% were classified as high-grade including 56% which were muscle-invasive (≥pT2). TERT and FGFR3 mutations were detected in 19 (61%) and 14 (46%) tumors, respectively. Of note, 8 (26%) out of them harbored both mutations. In the TERT-mutated subgroup, we observed higher incidence of men as compared to TERT non-mutated subgroup (p = 0.04); in addition, we did not identify any other significant differences regarding other clinico-pathological features. This was also the case for FGFR3 mutations, although we observed a higher frequency in tumors arising in the ureter (66.6%) as compared to those located in renal pelvis (28.5%) (p = 0.11). Neither mutations of FGFR3 nor TERT were associated with progression-free survival and patient's overall survival. Conclusion: Our study identified a high rate of somatic mutations in FGFR3 and TERT which were similar between muscle-invasive and non-muscle invasive tumors as well as between tumors arising in the ureter or renal pelvis. As targeted inhibitors are being investigated in this setting, these results might have implications for patient's management. Citation Format: Roger Mouawad, Souheyla Bensalma, Xiaoping Su, Frederick Allanic, Eva Comperat, Morgan Rouprêt, JeanPhillipe Spano, Gabriel Malouf, David Khayat. Study of TERT promoter and FGFR3 mutations in upper-tumor urothelial carcinomas. [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 116.

Checkmate to CHK1 in T-cell ALL?

DNA replication ensures accurate duplication of the original genetic information present in a cell in order for it to be properly transmitted to daughter cells. However, replication can be perturbed, for instance in rapidly dividing cancer cells, in a process referred to as replication stress (RS). Checkpoint kinase 1 (CHK1) is an essential component of the ATR-dependent DNA damage-response pathway that protect cells from RS by preventing replication fork collapse and activating homologous DNA repair. The ATR-CHK1 pathway is triggered upon exposure of single-stranded DNA that arises with the stalling of replication forks [1], and it is required to reset proper origin firing, and to promote fork stability and checkpoint activation, delaying mitosis until replication is completed and thereby avoiding mitotic catastrophe [2]. Whereas these functions point towards a tumor suppressor role for CHK1, mouse models modulating ATR-CHK1 expression and genetic evidence from human tumors suggest otherwise: Atr and Chk1 knock-out models do not display higher tumor frequency; Chk1 favors oncogene-induced transformation in mice; CHK1 is frequently overexpressed in human cancers, while loss-of-function mutations are rare [2, 3]. Moreover, CHK1 affords protection against DNA damaging agents, a fact that prompted the use of CHK1 inhibitors as chemosensitizers [4]. Similarly, tumors whose oncogenic profile fuels RS were proposed to become addicted to ATR-CHK1 response [1, 2]. In our recent study published in Oncogene [5], we hypothesized that T-cell acute lymphoblastic leukemia (T-ALL), an aggressive hematological cancer arising from T-cell precursor clonal expansion, could be one of such tumors and showed that CHK1 plays a key role in T-ALL cell maintenance. T-ALL cells tend to be highly proliferative due to a myriad of genetic lesions that culminate in cyclin-dependent kinase hyperactivation, and deregulated progression of S-phase that may impact on DNA replication [5, 6]. We found that T-ALL cells overexpressed CHK1 mRNA and protein as compared to normal hematopoietic progenitors. This was accompanied by aberrantly high CHK1 kinase activity, likely triggered by high basal levels of RS [5]. Experimental inactivation of CHK1, by a CHK1 selective inhibitor (PF-00477736) or by gene silencing, demonstrated that CHK1 is essential to control the accumulation of RS and to prevent apoptosis of T-ALL cells that appear to enter mitosis without having concluded DNA replication. Furthermore, accumulation of DNA damage in the context of CHK1 loss induced the activation of the ATM-CHK2 DNA double-strand break (DSB) response pathway, likely due to DSB formation upon the collapse of stalled replication forks. T-ALL apoptosis upon CHK1 inactivation was in the first instance dependent on ATM and caspase-3, since ATM inhibition prevented caspase-3 cleavage and rescued T-ALL cell viability despite sustained elevated amounts of RS markers [5]. Following the demonstration that T-ALL cells were eliminated using a CHK1 small molecule inhibitor as single agent, we showed that this effect was leukemia-specific, since normal T-cell progenitors were not sensitive to the low doses of PF-00477736 that killed primary T-ALL patient cells. Moreover, the in vitro anti-leukemia effect of PF-00477736 was not prevented by microenvironment pro-survival factors, and the potential clinical value of CHK1 inhibition was further demonstrated by the fact that PF-00477736 limited the growth of xenografted T-ALL tumors [5]. Interestingly, our preliminary analyses indicated that T-ALL cells expressing higher CHK1 levels appeared more sensitive to CHK1 pharmacological inhibition, suggesting that CHK1 expression could be a suitable drug response marker in T-ALL patients. As clinical trials against ATR-CHK1 pathway may be envisaged, this issue warrants extended T-ALL patient analysis. T-ALL constitutes only a fraction of all ALL cases, but it associates with high-risk. Therapeutic options with less detrimental side-effects and/or effective upon relapse are most desired. Our findings defining CHK1 as a ‘subverted’ tumor suppressor that stands in T-ALL as a major guardian of leukemia cell survival, thereby formally acting as an oncogene, reinforce a new way of viewing the mechanisms of cancer progression [2] and may set the ground for anti-leukemia breakthrough approaches. In this context, it is important to understand the mechanisms of CHK1 upregulation in T-ALL. We thoroughly documented CHK1 transcript overexpression in primary T-ALL [5]. However, how this occurs remains undetermined. Maybe transcription factors known to activate CHK1, such as E2F (downstream of G1/S-phase CDK activity) or MYC (downstream of NOTCH1), are involved in CHK1 overexpression in T-ALL. Or maybe as yet unidentified CHK1 regulatory elements are mutated or epigenetically altered. Curiously, in contrast to our findings, Chk1 mRNA downregulation was documented in a murine T-ALL model [7]. A more integrative view of the role of CHK1 in T-ALL is therefore required. We believe CHK1 downregulation may occur at T-ALL initiation, driving genomic instability secondary to an increase in RS. As the pro-proliferative oncogenic program establishes and RS rises, leukemic cells are naturally selected for their ability to upregulate CHK1 as a means to maintain RS levels compatible with cell viability.

TRIMming down tumor suppressors in breast cancer

TRIMming down tumor suppressors in breast cancer

Concurrent alterations in TERT, KDM6A, and the BRCA pathway in bladder cancer.

Genetic analysis of bladder cancer has revealed a number of frequently altered genes, including frequent alterations of the telomerase (TERT) gene promoter, although few altered genes have been functionally evaluated. Our objective is to characterize alterations observed by exome sequencing and sequencing of the TERT promoter, and to examine the functional relevance of histone lysine (K)-specific demethylase 6A (KDM6A/UTX), a frequently mutated histone demethylase, in bladder cancer. We analyzed bladder cancer samples from 54 U.S. patients by exome and targeted sequencing and confirmed somatic variants using normal tissue from the same patient. We examined the biologic function of KDM6A using in vivo and in vitro assays. We observed frequent somatic alterations in BRCA1 associated protein-1 (BAP1) in 15% of tumors, including deleterious alterations to the deubiquitinase active site and the nuclear localization signal. BAP1 mutations contribute to a high frequency of tumors with breast cancer (BRCA) DNA repair pathway alterations and were significantly associated with papillary histologic features in tumors. BAP1 and KDM6A mutations significantly co-occurred in tumors. Somatic variants altering the TERT promoter were found in 69% of tumors but were not correlated with alterations in other bladder cancer genes. We examined the function of KDM6A, altered in 24% of tumors, and show depletion in human bladder cancer cells, enhanced in vitro proliferation, in vivo tumor growth, and cell migration. This study is the first to identify frequent BAP1 and BRCA pathway alterations in bladder cancer, show TERT promoter alterations are independent of other bladder cancer gene alterations, and show KDM6A loss is a driver of the bladder cancer phenotype.

Maternal exercise during pregnancy reduces risk of mammary tumorigenesis in rat offspring (886.1)

Breast cancer is the most common cancer among women. Emerging research indicates that modifying lifestyle factors during pregnancy may convey long‐term health benefits to offspring. This study was designed to determine whether maternal exercise during pregnancy leads to reduced mammary tumorigenesis in female offspring. Pregnant rats were randomly assigned to exercised and sedentary groups, with the exercised group having free access to a running wheel and the sedentary group housed with a locked wheel during pregnancy. Female pups from exercised or sedentary dams were weaned at 21 days of age and fed a high fat diet without access to a running wheel. At 6 weeks, all pups were injected with the carcinogen N‐methyl‐N‐nitrosourea (MNU). Mammary tumor development in all pups was monitored for 15 weeks. Pups from exercised dams had a substantially lower tumor incidence (42.9%) compared to pups from sedentary dams (100%). Of the pups with tumors, the pups from exercised dams had a higher tumor frequency (1.7±0.4 tumors/rat) than pups from sedentary dams (1.2±0.2 tumors/rat). Neither tumor latency nor histological grade differed between the two groups. These data are the first to demonstrate that exercise during pregnancy potentiates reduced tumorigenesis in offspring. This study provides an important foundation towards developing more effective modes of behavior modification for cancer prevention.Grant Funding Source: Supported by NIH R25CA128770 and ICTSI NIH/NCRR TR000006

Perfil imuno-histoquímico e variáveis clinicopatológicas no câncer de mama

OBJETIVO: Descrever as principais caracteristicas em mulheres com câncer de mama, de acordo com o perfil imuno-histoquimico. METODOS: A populacao foi composta a partir de coorte hospitalar formada por mulheres com diagnostico de câncer de mama efetuado entre 2003 e 2005 (n = 601) e atendidas em centro de referencia em assistencia oncologica de Juiz de Fora-MG. Foram selecionadas apenas 397 mulheres que possuiam imunohistoquimica completa. Para definicao dos grupos segundo perfil imuno-histoquimico, optou-se por classificacao baseada na avaliacao dos receptores de estrogeno e de progesterona, indice de proliferacao celular Ki67 e superexpressao de HER2. De acordo com os diferentes fenotipos, foram definidos cinco subtipos: luminal A, luminal B-HER2 negativo, luminal B-HER2 positivo, triplo negativo e HER2 superexpresso. RESULTADOS: A maioria dos pacientes tinha pele branca (80,7%) e era pos-menopausada (64,9%), com idade media de 57,4 anos (±13,5). Ao diagnostico, 57,5% delas tinha tumor de tamanho > 2,0 cm, e 41,7% exibiam comprometimento linfonodal. Os subtipos mais frequentes foram luminal B-HER2 negativo (41,8%) e triplo negativo (24,2%). No subtipo luminal A, 72,1% das pacientes eram pos-menopausadas, enquanto que os maiores percentuais na pre-menopausa foram observados nos subtipos luminal B-HER2 positivo e triplo negativo (45,2% e 44,2%, respectivamente). Verificou-se maior frequencia de tumores > 2,0 cm e com linfonodos comprometidos nos subtipos triplo negativo e HER2 positivo. CONCLUSAO: Esta pesquisa possibilitou avaliar a distribuicao das principais caracteristicas clinicopatologicas e relacionadas aos servicos de saude em coorte de mulheres brasileiras com câncer de mama, segundo os subtipos tumorais imuno-histoquimicos.