bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A06: Association of environmental risk factors, family history, and polygenic risk scores with chronic lymphocytic leukemia
Authors: Kleinstern Geffen , Robinson Dennis , Call Tim G. , Liebow Mark , Sanjosé Silvia de , Benavente Yolanda , Cerhan James R. , Slager Susan L.
Published: May 2019

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR03) of the Conference Proceedings.

Citation Format: Geffen Kleinstern, Dennis Robinson, Tim G. Call, Mark Liebow, Silvia de Sanjosé, Yolanda Benavente, James R. Cerhan, Susan L. Slager. Association of environmental risk factors, family history, and polygenic risk scores with chronic lymphocytic leukemia. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr A06.
Abstract B04: Development of a comprehensive colorectal cancer risk prediction tool (CRiPT) incorporating known and unknown major genes and polygenes
Authors: Win Aung Ko , Jenkins Mark A. , Dowty James G. , Antoniou Antonis C. , Lee Andrew , Zheng Yingye , Lindor Noralane M. , Newcomb Polly A. , Hopper John L. , MacInnis Robert J.
Published: May 2019

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR10) of the Conference Proceedings.

Citation Format: Aung Ko Win, Mark A. Jenkins, James G. Dowty, Antonis C. Antoniou, Andrew Lee, Yingye Zheng, Noralane M. Lindor, Polly A. Newcomb, John L. Hopper, Robert J. MacInnis. Development of a comprehensive colorectal cancer risk prediction tool (CRiPT) incorporating known and unknown major genes and polygenes. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr B04.
Abstract B06: Utah familial colorectal cancer risk model
Authors: MacInnis Robert J. , Jenkins Mark A. , Hopper John L. , Cannon-Albright Lisa A.
Published: May 2019

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR11) of the Conference Proceedings.

Citation Format: Robert J. MacInnis, Mark A. Jenkins, John L. Hopper, Lisa A. Cannon-Albright. Utah familial colorectal cancer risk model. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr B06.
Abstract B16: Development of breast cancer risk prediction for the UK population using the UK Biobank dataset
Authors: Alajmi Kawthar , Lophatananon Artitaya , Muir Kenneth
Published: May 2019

Breast cancer is the most common female cancer and is the second most common cause of cancer death among females. The UK has the highest age standardised incidence and mortality rates in the world, with two in every 1000 women aged 50 and above developing breast cancer annually. Several risk prediction models have been developed to estimate the likelihood for developing breast cancer based on specific risk factors in currently healthy individuals within a specific period of time. The available models are derived principally from either genetic or non-genetic factors. The majority of these models are however not user-friendly, do not focus on modifiable factors entirely and are not specifically designed for the general public.

Our research group is developing an individualised risk prediction model for breast cancer focusing on the modifiable risk factors using the UK Biobank data. A nested case-control study within the 273,467 female participants is being used to develop the model. We have split the data into training and testing sets and will carry out all statistical tests to ensure our model calibrates well. For model validation, we will further seek external validation cohorts. The model will provide risk scores derived from the presence or absence of specific risk factors and will be compared to the general public score. The model will allow people to modify their risk profile with appropriate prevention measures. The main goal of the model is to be used in cancer education and prevention.

The results from exploratory analyses suggested positive associations between breast cancer risk and age, breast cancer family history, menopause age, age at first child, BMI, height, null-parity, smoking, alcohol intake, and family history of other cancer. An algorithmic model will be developed based on these factors. We will also evaluate public perceptions using focus group technique. We will be presenting the results of the model development from the training set and the results of the internal validation from the testing set.

In conclusion, we are developing an individualised breast cancer risk prediction model for the UK population based on the modifiable risk factors. The model will enable us to educate and to design appropriate interventions tailored to the individual with the aim of assisting them to make appropriate changes to modify their cancer risk profile.

Citation Format: Kawthar Alajmi, Artitaya Lophatananon, Kenneth Muir. Development of breast cancer risk prediction for the UK population using the UK Biobank dataset. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr B16.
Abstract B23: Detection of susceptibility to childhood Acute Lymphoblastic Leukemia (ALL)
Authors: Tebbi Cameron K.
Published: May 2019

Currently, there are no known methods to predict of susceptibility to, and means for prevent Acute Lymphoblastic Leukemia (ALL). We have evaluated and patented a group of proteins dubbed Protein X from a certain strain of Aspergillus Flavus (AF) and developed methods for screening and identifying totally asymptomatic patients in remission of ALL, including long term survivors of this disease, distinguishing them from normal controls. Subject to institutional approved consents/assents, 15-20 ml of blood was obtained from 40 cases of ALL in children and young adults, including long term survivors of ALL. Controls were normal individuals, sickle cell patients undergoing partial exchange transfusion and patients with solid tumors. Mononuclear leukocytes (MNL) of ALL patients in remission and controls were separated using Ficoll Paque Plus (GE Healthcare). Epstein Barr virus (EBV) was obtained commercially. Positive and negative controls for Protein X were aflatoxin and Mycocladus Corymbifera (MC). Avian leukosis virus (ALV) was used as control for EBV. MNL were co-incubated with Protein X ± EBV ± irradiation, for periods of 1-72 hours. Controls were treated identically with appropriate substitutions. Test and control MNLs were examined for genetic markers, NF-κB and cell surface markers (CSM) including CD10/CD19, CD34/CD19, and CD34/CD117. Changes were expressed as percentage of control. Using ELISA, plasmas were tested for antibodies against Protein X ± EBV time experiments reveled 72 hours was optimum for achieving results. Upon 72 hours exposure of MNL from ALL to Protein X ± EBV, cells from ALL patents in remission developed cell surface phenotypes typical of ALL. This was not seen in controls. Addition of EBV ± radiation to Protein X, enhanced these effects in MNL of ALL and not controls. Changes were statistically significant and clearly separated ALL from controls. Evaluation of NF-κB revealed enhancement in ALL and not controls. Aflatoxin indiscriminately induced cell surface marker changes in both, normal and ALL, while ALV and supernatant of MC had no effect. ELISA, using Protein X ± EBV, distinguished ALL from controls. Gene array and biomarkers confirmed transformation to leukemic cell markers upon exposure to Protein X in cells from ALL patients but not controls. These studies reveal, in vitro, upon exposure to Protein X, unlike normal controls, MNL from ALL patients in remission develop cell surface phenotypes and genetic markers typical of ALL. These techniques have potential for screening for ALL and may have implications for etiology of ALL and its prevention.

Citation Format: Cameron K. Tebbi. Detection of susceptibility to childhood Acute Lymphoblastic Leukemia (ALL). [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr B23.
Abstract IA10: Targeted imaging of the serrated pathway for early detection of colorectal cancer
Authors: Wang Thomas D.
Published: May 2019

Background and Significance: Colorectal cancer (CRC) is a leading cause of cancer-related mortality with ~1.4 million new cases diagnosed globally and ~690,000 annual deaths. The serrated pathway is a primary cause of CRC in the proximal colon. Sessile serrated adenomas (SSA) have flat and subtle features that are difficult to detect with conventional white light colonoscopy.

Aims: We aim to identify a peptide imaging agent that binds specifically to SSA to improve visualization of pre-malignant lesions in the proximal colon.

Methods: We used phage display technology with subtractive biopanning against cells with a V600E point mutation in BRAF. We performed a first-in-humans clinical study by topically administering this FITC-labeled peptide in the proximal colon of n = 38 human subjects, and performed in vivo imaging using a wide-field endoscope that is sensitive to fluorescence.

Results: We identified the peptide sequence KCCFPAQ, and measured an apparent dissociation constant of kd = 72 nM and an apparent association time constant of kd = 0.174 min-1 (5.76 min). On in vivo fluorescence images, we measured a 2.43-fold greater target-to-background ratio for SSA than for normal colonic mucosa, and could distinguish SSA with 89% sensitivity and 92% specificity. We found 84% sensitivity and 91% specificity for discriminating SSA from traditional adenomas. No toxicity was attributed to the peptide in either animal or patient studies. On ex vivo images, we found the mean fluorescence intensity for SSA to be significantly greater than that for hyperplastic polyps.

Conclusions: We have identified a fluorescently-labeled peptide that is safe for clinical use, and is specific for detecting SSAs in the proximal colon with wide-field imaging. This targeted imaging methodology may be useful for early detection of pre-malignant serrated lesions on routine colonoscopy.

Citation Format: Thomas D. Wang. Targeted imaging of the serrated pathway for early detection of colorectal cancer. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr IA10.
Abstract IA12: Risk-assessment in HPV-based cervical cancer screening
Authors: Herrero Rolando
Published: May 2019

Cervical cancer screening is rapidly evolving to incorporate highly accurate molecular methods that are able to categorize women's risk of cervical cancer and allow recommendations for follow-up or treatment. HPV nucleic acid testing provides highly sensitive and reproducible detection of HPV infection, a necessary cause of cervical cancer. Thus, virtually all women who have a cervical cancer precursor or who will develop one in the following years are HPV positive. On the other hand, the vast majority of HPV positive women have transient infections that will disappear in a few months. In this context, further evaluation of HPV positive women using triage methods allows further risk stratification that avoids unnecessary evaluations and overtreatment. Women positive for the triage test are referred for immediate colposcopic evaluation and follow-up (or, in some contexts, immediate treatment) as needed, while those who test negative usually receive another screening test in a year (or more). Triage with cytology or co-testing with HPV and cytology selects a group at clearly higher risk and generally above the recommended threshold for immediate colposcopy referral based on current consensus guidelines. For HPV positive women, genotyping for HPV 16 and 18 allows further risk stratification, but the additional benefit of other HPV genotypes is unclear. Ideally, triage methods are performed on the same specimen where the HPV test was carried out, to avoid additional visits. Risk stratification is different in different populations and age groups as HPV prevalence and type distribution, as well as screening histories are highly variable around the world. The programmatic decisions need to take into account the feasibility and logistics of different approaches and their respective burden on health services. Further research, particularly in low and middle income countries and in vaccinated cohorts is required.

Citation Format: Rolando Herrero. Risk-assessment in HPV-based cervical cancer screening. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr IA12.
Abstract IA21: Translating evidence to action: Yourdiseaserisk.wustl.edu
Authors: Colditz Graham A.
Published: May 2019

Not very long ago scientific publication was viewed as the primary dissemination goal of scientific discovery. This viewpoint, however, has evolved substantially over the past 10 - 20 years. While scientific discovery and publication remain key to dissemination of findings, it is now often viewed as a single stage in the spectrum from discovery to the application of research results. The view of effective dissemination must now also include the practical world of policy makers, clinicians, health care organizations, and the public – groups that need good data and good tools to make informed decisions that drive individual, national, and global health. The development of health risk assessment and prevention tools can play a key role in doing this. And such development moves through three general translation stages – with each subsequent stage marked by greater difficulty to achieve. 1) Creation of accurate risk prediction calculation(s) from current evidence base. 2) Development of a practical, usable tool that incorporates the calculation(s) and provides actionable messages – for clinical, public policy, or public use.  3) Integration of risk prediction tools with the social, structural, and financial support for translating recommended action messages into actual action – whether we're talking about doctors counseling patients, government representatives making health policy, or the public working to improve their own health. This process by necessity requires a multi-disciplinary approach – drawing on expertise from epidemiology, biostatistics, communication theory, coding, and design – among others. With the addition of precision medicine and big data to long-established data analysis techniques, the field of risk prediction is set to expand in coming years. Along with that expansion, it is important to assure that our efforts are valid, useful, reliable, and applicable.

Citation Format: Graham A. Colditz. Translating evidence to action: Yourdiseaserisk.wustl.edu. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr IA21.
Abstract IA22: Using risk assessment tools to motivate behavior change
Authors: Waters Erika A.
Published: May 2019

Epidemiology identifies risk factors for cancer and other diseases based on the idea that conveying such information to healthcare providers, the general public, and policy makers will result in population-wide improvements in healthy behaviors and, consequently, population-wide improvements in health outcomes. These actions assume that the audience understands and uses the information to make health-related decisions. However, the language of epidemiology, which is steeped in probabilistic thinking, is not necessarily the language of the general public. Furthermore, growing evidence suggests that the language of epidemiology is not particularly well-understood by policy makers or even, disconcertingly, by some healthcare providers.

This presentation will accomplish three objectives. First, it will demonstrate how a user-friendly, Internet-based, personalized risk assessment tool that uses established principles of risk communication and is grounded in health behavior change theories can increase motivation to change behavior. It will accomplish this in the context of using Your Disease Risk to inform women about the association between physical activity and breast cancer risk.

Second, it will illustrate the development of a novel personalized risk assessment tool that increases understanding of the link between lifestyle behaviors and overall health and wellness. Specifically, it translates cumulative incidence data about five diseases that cause significant morbidity and mortality (i.e., colon cancer, breast cancer (women), heart disease, diabetes, and stroke) into a tool that conveys personalized risk estimates in a comprehensible and useful way for laypeople from diverse socio-demographic backgrounds. The premise is that illustrating how a single behavior can affect the likelihood of developing several diseases could foster a more coherent and meaningful picture of the behavior's importance in reducing health risks and could increase motivation to engage in the behavior. The behavioral context for this study is also physical activity.

Third, this presentation discuss the limits of tools that convey only risk information and do not help users bridge the gap between wanting to change their behavior and having the knowledge, skills, and confidence to actually initiate and maintain such changes.

Citation Format: Erika A. Waters. Using risk assessment tools to motivate behavior change. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr IA22.
Abstract IA25: Implementation of risk prediction to improve health: The promises and challenges of precision
Authors: Kramer Barnett S.
Published: May 2019

Advances in the understanding of risk factors have transformed oncology, providing leads for prevention and, in particular, screening. Part of this transformation has been fueled by studies of molecular changes and genetic mutations that are inherited or that precede cancer by years, potentially leading the way to a new era of precision screening and prevention. However, these new leads bring important challenges that demand caution. As Sean Carroll has pointed out, it turns out that life from the molecular scale all the way up to the ecological scale is usually governed by longer chains of interactions than we first imagine, with more links in between (Serengeti Rules: The Quest to Discover How Life Works and Why It Matters, 2016).

Rather than screening for existing asymptomatic disease, we have entered the era of screening for risk factors, or even screening for risk factors for risk factors (disease risk predisposition). This is a double-edged endeavor, because genes may influence fate, but not in a linear or straightforward manner. Therefore, prediction of outcome is far less precise than measurement of the predisposing genetic and molecular alterations. Many people may be labeled as carriers of risk factors that will never develop the clinical forms of the disease predicted by the genetic changes, a form of genetic overdiagnosis. So the new era of risk prediction can bring both benefits and harms. A critical tool in determining the balance of benefits and harms of increasingly sensitive omics technologies is the use of a formal analytic framework, to be discussed in the presentation.

Citation Format: Barnett S. Kramer. Implementation of risk prediction to improve health: The promises and challenges of precision. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr IA25.
Abstract PR09: Prostate cancer prognostication based on an actionable metabolic pathway
Authors: Stopsack Konrad H. , Gerke Travis A. , Mucci Lorelei A. , Rider Jennifer R.
Published: May 2019

Background: We recently discovered that mRNA expression of SQLE, coding for squalene monooxygenase, the second rate-limiting enzyme of cholesterol synthesis, is associated with lethality after prostate cancer diagnosis. Here, we investigate how expression of SQLE and other key regulators of cholesterol homeostasis, identified by prior mechanistic studies, aid risk prediction for lethal prostate cancer.

Methods: The Health Professionals Follow-up Study and the Physicians' Health Study prostate cancer tissue cohorts collected tissue from prostatectomy or transurethral resection of the prostate at cancer diagnosis. Whole-transcriptome profiling was performed. The outcome of interest was lethal cancer defined as prostate cancer mortality or development of metastases in contrast to non-lethal cancer without evidence of metastases after at least eight years of follow up. Discrimination for prostate lethal cancer was assessed by comparing c-statistics using bootstrap resampling.

Results: Combining both cohorts, 112 men had lethal prostate cancer; 290 men had non-lethal cancer. A prognostic model for lethal cancer including Gleason grade, pathologic stage, age, and year of diagnosis had a high c = 0.885; adding body mass index, smoking status, family history of prostate cancer, and diabetes diagnosis increased c to 0.889. A model containing only SQLE (linear) achieved c = 0.663. Adding SQLE to the fully adjusted model increased c to 0.903 (p = 0.027). None of the other cholesterol regulators ABCA1, ACAT1, LDLR, and SCARB1 improved discrimination.

Conclusions: SQLE performs well as a single biomarker of prostate cancer lethality after primary therapy, in contrast to other markers of intratumoral cholesterol regulation. Improvements in prognostication are minimal when SQLE is added to a model that contains a centrally re-reviewed Gleason grade. Most importantly, SQLE may be an actionable, predictive biomarker of benefit from statin therapy, which addresses the cholesterol synthesis pathway regulated by SQLE.

Citation Format: Konrad H. Stopsack, Travis A. Gerke, Lorelei A. Mucci, Jennifer R. Rider. Prostate cancer prognostication based on an actionable metabolic pathway. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr PR09.
Abstract A04: Structural and functional caracterization of BARD1/CDK13 interaction
Authors: Fernandes Vanessa C. , Nepomuceno Thales C. , Carvalho Renato S. , Suarez-Kurtz Guilherme , Monteiro Alvaro N. , Carvalho Marcelo A.
Published: May 2019

DNA damage response (DDR) is crucial for cell survivor through genomic integrity maintenance. Dysfunctions in proteins implicated in DDR prompt cell to diseases, such as cancer. BRCA1 is a central protein involved in DDR and, once damaged, plays a pivotal role in cancer development. BRCA1 structure comprises two major domains, a RING finger domain and two tandem BRCT domains (tBRCT). This structure is shared by BRCA1 major interaction partner: BARD1. The tBRCT domain is commonly found in DDR-associated proteins. To study interactions in DDR, our group charted the tBRCT interactome for 7 different proteins, using three different strategies: tandem affinity purification followed by mass spectrometry (TAP-MS), yeast two hybrid (Y2H) screening, and literature curation. CDK13 was identified as a putative BARD1 tBRCT interactor by both TAP-MS and Y2H strategies. CDK13 is described as involved in several processes, such as chromatin remodeling, transcription regulation, and splicing coordination; however, its biological role remains unclear. To confirm BARD1/CDK13 interaction, whole cell extracts obtained from MCF-7 cells were used in coimmunoprecipitation assays using anti-CDK13 and anti-BARD1. Constructs encoding the HA-tagged regions 1-706aa (N-terminal), 706-982aa (kinase domain), and 1006-1452aa (C-terminal) of CDK13 were ectopically coexpressed with Flag-tagged full-length BARD1 or GFP-tagged BARD1 tBRCT in HEK293Tcells; whole cellular extracts were used in coimmunoprecipitation assays. BARD1 and CDK13 interaction was confirmed both in ectopic and constitutive expression approaches. CDK13 silencing was performed using lentiviral constructs encoding five different short hairpin RNAs (shRNA); MCF-7 cells were transduced and submitted to puromycin selection; extracts were used to evaluate CDK13 silencing by immunoblotting. Our data suggest that BARD1 tBRCT and the CDK13 N-terminal region, as well as the kinase domain, are critical for the formation of BARD1/CDK13 complex. MCF-7 cells lacking CDK13 expression will be used to investigate the kinase role during DDR.

Citation Format: Vanessa C. Fernandes, Thales C. Nepomuceno, Renato S. Carvalho, Guilherme Suarez-Kurtz, Alvaro N. Monteiro, Marcelo A. Carvalho. Structural and functional caracterization of BARD1/CDK13 interaction [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A04.
Abstract A06: MicroRNA-195 acts as a tumor suppressor miRNA in human melanoma cells by targeting Prohibitin 1
Authors: Cirilo Priscila D. R. , Côrrea Bruna R. S. , Qiao Mei , Andrade Luciana N. S. , Chammas Roger , Penalva Luiz O. F.
Published: May 2019

Melanoma is the most lethal type of skin cancer. Since chemo-resistance is a significant barrier, identification of regulators affecting chemo-sensitivity is necessary in order to create new forms of intervention. Here, we investigate the role of miR-195, its impact on Prohibitin 1 (PHB1) expression, and on chemo-sensitivity in melanoma cells. TCGA-RNAseq data obtained from 341 melanoma patient samples as well as a panel of melanoma lines were used in an expression correlation analysis between PHB1 and predicted miRNAs. miR-195 impact on PHB1 mRNA and protein levels and relevance of this regulation were investigated in UACC-62 and SK-MEL-5 melanoma lines by qRT-PCR and Western blot, luciferase reporter, and rescue experiments. Cell proliferation, hypodiploid, and caspase 3/7 assays were performed to investigate the potential action of miR-195 as chemo-sensitizer in melanoma cells treated with cisplatin and temozolomide. Analysis of the TCGA-RNAseq revealed a significant negative correlation (Pearson) between miR-195 and PHB1 expression. Moreover, qRT-PCR data showed that miR-195 is downregulated while PHB1 is upregulated in a collection of melanoma lines. We demonstrated that miR-195 regulates PHB1 directly by qRT-PCR and Western blot in melanoma cells and luciferase assays. To establish PHB1 as a relevant target of miR-195, we conducted rescue experiments in which we showed that PHB1 transgenic expression could antagonize the suppressive effect of miR-195 on the proliferation of melanoma cells. Finally, transfection experiments combined with drug treatments performed in the UACC-62 and SK-MEL-5 melanoma cells corroborated miR-195 as a tumor suppressor and potential chemo-sensitivity agent.

Citation Format: Priscila D. R. Cirilo, Bruna R. S. Côrrea, Mei Qiao, Luciana N. S. Andrade, Roger Chammas, Luiz O. F. Penalva. MicroRNA-195 acts as a tumor suppressor miRNA in human melanoma cells by targeting Prohibitin 1 [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A06.
Abstract A17: Methylation profile of SEPT9, NDRG4, p16 genes in colorectal cancer
Authors: Sobanski Thais , Arantes Lidia , Santos Wellington dos , Durães Ronilson , Matsushita Marcus , Reis Rui Manuel , Guimaraes Denise Peixoto
Published: May 2019

Background: Colorectal cancer (CRC) is the third most common cancer in Brazil, where it appears as the fifth most deadly cancer. Brazilian National Cancer Institute estimates for the year of 2016 in Brazil 16,660 newly diagnosed cases of CRC in men and 17,620 in women. There is evidence that the incidence and mortality rates can be reduced through the detection of precursor lesions and early cancer in the context of CRC screening. Aberrant hypermethylation of cancer-related genes has emerged as a promising strategy for the molecular CRC screening. However, the methylation profile of colorectal cancer of the Brazilian population has limited information about its characterization and frequency.

Objective: To investigate the frequency of DNA methylation in the promoter region of tumor suppressor genes SEPT9, NDRG4, and CDKN2A (p16) in sporadic CRCs of Brazilian patients.

Methods: Retrospective molecular analysis included 57 adult patients with CRC in different clinical stages attended at the Barretos Cancer Hospital. Demographic, clinical, and tumor-related characteristics were obtained from medical records. DNA was extracted from fresh-frozen tumor tissues. Levels of DNA methylation were determined by quantitative pyrosequencing.

Results: The mean ± SD age of the 57 patients' analysis was 64.40 ±14 years; 56.14% (32/57) were males. Almost all tumors (56/57; 98.24%) were located in the left side of the colon, from descending colon to rectum. Regarding stage, 3/57 (5.26%) were stage 0, 15/57 (26.31 %) stage I, 27/57 (47.36%) stage II, 10/57 (17.54%) stage III, and 2/57 (3.53%) stage IV tumors. Among 57 analyzed, 52/57 (91.22%) were methylated for SEPT9, 42/57 (73.68%) for NDRG4, and 15/57 (26.31%) for CDKN2A. In conclusion, in our Brazilian case series, the gene SEPT9 and NDGR4 showed high frequency of methylation CRC tissues, similar with results found in other populations.

Citation Format: Thais Sobanski, Lidia Arantes, Wellington dos Santos, Ronilson Durães, Marcus Matsushita, Rui Manuel Reis, Denise Peixoto Guimaraes. Methylation profile of SEPT9, NDRG4, p16 genes in colorectal cancer [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A17.
Abstract A20: Identification of new promising germline variants in melanoma-prone patients
Authors: Fidalgo Felipe , Facure Luciana , Barros Bruna , Valieris Renan , Torrezan Giovana , Sá Bianca de , Nóbrega Amanda , Achatz Maria Isabel , Duprat João , Rosenberg Carla , Souza Sandro , Krepischi Ana , Carraro Dirce Maria
Published: May 2019

Background: Melanoma is a highly aggressive skin cancer; approximately 10% of melanomas are caused by germline mutations, mainly affecting the p16 isoform of the CDKN2A gene (responsible for up to 40% of the familial melanoma). Other genes exhibiting moderate- to high-penetrant mutations have been recently associated with melanoma susceptibility but in low frequency. Therefore, the majority of the melanoma-prone patients remain without the identification of genetic etiology.

Objective: In order to find new genes potentially related to melanoma predisposition, we performed whole-exome sequencing of 10 affected individuals from 5 unrelated families. All patients were negative for CDKN2A/CDK4/TERT promoter/MITF (E318K) mutations.

Materials and Methods: Melanoma-prone patients were ascertained based on either the classical criteria for familial melanoma syndrome or based on evidence of a strong genetic predisposition (occurrence of ≥2 primary melanomas). Whole-exome sequencing was performed using the Ion Proton Sequencer (Life Technologies). Our experimental design was performed using at least 20x each base pair sequencing as confidence pattern and the most interesting variants were prioritized, when identified on the two probands from the same family, using the VarSeq software 1.4.2. Once we obtained the most interesting germline rare variants we designed a panel for the validation of these variants through the target sequencing (Ion Proton Thermo Fisher Scientific Inc).

Results: We identified 214 rare nonsynonymous variants. We selected a set with 63 rare germline variants potentially damaging the protein function and associated to important cellular pathways to perform validation (4 LoF variants classified as frameshift). All variants were validated by resequencing using probes specifically designed for the rare selected variants; some of them already associated to melanoma, skin pigmentation, melanoma, and/or cancer susceptibility.

Conclusion: This study provides new knowledge about melanoma predisposition in the Brazilian population, in which the risk is smaller to develop this disease than in other populations, and also highlighted possible new risk factor or predisposition genes for hereditary melanoma syndrome.

Citation Format: Felipe Fidalgo, Luciana Facure, Bruna Barros, Renan Valieris, Giovana Torrezan, Bianca de Sá, Amanda Nóbrega, Maria Isabel Achatz, João Duprat, Carla Rosenberg, Sandro Souza, Ana Krepischi, Dirce Maria Carraro. Identification of new promising germline variants in melanoma-prone patients [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A20.
Abstract A29: Omics profile of two immortalized Brazilian glioblastoma cell cultures
Authors: Cruvinel-Carloni Adriana , Silva-Oliveira Renato José da , Oliveira-Silva Viviane Aline , Rosa Marcela Nunes , Bidinotto Lucas Tadeu , Berdinarelli Gustavo Noriz , Torrieri Raul , Evangelista Adriane Feijó , Martinho Olga Catarina , Gonçalves Vera Mónica , Clara Carlos , Almeida Gisele Caravina de , Reis Rui Manuel
Published: May 2019

Background: Glioblastomas are the most frequent and deadly brain tumors. Genomic alterations, as well as its high heterogeneity, are important features of these tumors. Despite the advances in the biology of these tumors, it is essential to develop models, such as primary tumor cell lines, that better mimic their genomic diversity and therapeutic behavior. We aimed to characterize the molecular profile of two (HCB2 and HCB151) established immortalized glioblastoma cell lines.

Methods: Immortalized glioblastoma cell lines, HCB2 and HCB151, were established from Barretos Cancer Hospital patients diagnosed with glioblastoma. The cells were cultured with DMEM+10%BFS+1%PS and molecularly characterized, using arrayCGH, next-generation sequencing (Ion Torrent) and validated by Sanger sequencing, miRNA expression and CNV using the NanoString platform, and BD Lyoplate™ Human Cell Surface Marker Screening Panel using Accuri™ C6 flow cytometer. The tumorigenic capacity was further determined in vivo by the chicken chorioallantoic membrane (CAM) assay, and its response to temozolomide-based chemotherapy was done by MTS.

Results: The aCGH analysis showed that the most frequent events was the gain of chromosome 7 and loss of chromosomes 10, 13, and 17p. HCB2 exhibited mutations in the TERT gene, while HCB151 showed mutations in the TP53, PTEN, LZTR1, and TERT genes. The immortalized cultures present an unique miRNA profile. The CAM assay showed that HCB2 and HCB151 were able to form vascularized tumors, with HCB151 more aggressive than HCB2. In addition, both HCB2 (IC50>1000 µM) and HCB151 (IC50=783 µM) showed resistance to TMZ when compared to commercial glioblastomas cell lines (23-441 µM).

Conclusion: The present findings showed that the immortalized glioblastoma cell lines showed classic genomic features of glioblastomas and exhibited a similar biologic behavior, suggesting that these cell lines are a good model for glioblastoma preclinical studies.

Citation Format: Adriana Cruvinel-Carloni, Renato José da Silva-Oliveira, Viviane Aline Oliveira-Silva, Marcela Nunes Rosa, Lucas Tadeu Bidinotto, Gustavo Noriz Berdinarelli, Raul Torrieri, Adriane Feijó Evangelista, Olga Catarina Martinho, Vera Mónica Gonçalves, Carlos Clara, Gisele Caravina de Almeida, Rui Manuel Reis. Omics profile of two immortalized Brazilian glioblastoma cell cultures [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A29.
Abstract A36: Clinical and molecular characterization of oropharynx squamous cell carcinoma samples
Authors: Carvalho Ana Carolina de , Sorroche Bruna Pereira , Melendez Matias Eliseo , Sabato Cristina da Silva , Palmero Edenir Inez , Arantes Lidia Maria Rebolho Batista , Neto Cristovam Scapulatempo , Carvalho André Lopes
Published: May 2019

Background: Oropharynx squamous cell carcinomas (OPSCC) associated with high-risk human papilloma virus (HR-HPV) infection represent a distinct clinical and epidemiologic entity, with differences in clinical presentation and treatment outcome when compared to tobacco-associated tumors.

Objective: The aim of this study was to identify clinical and molecular factors that can differentiate OPSCC patients into subgroups according to their outcome.

Materials and Methods: Twenty-five surgically treated OPSCC samples were included in this study. p16 protein expression was used as a surrogate marker for HR-HPV. The methylation status of 5 genes was evaluated by quantitative methylation-specific PCR (qMSP). Exons 4-8 of the P53 gene were evaluated by Sanger sequencing and p53 protein expression was evaluated by immunohistochemistry (clone DO-7).

Results: The majority of patients (88%) were male, had advanced disease (80%), and were tobacco users (88%). Twenty percent of the cases were HR-HPV positive and 62.5% presented deleterious P53 mutations, which was moderately concordant with strong p53 protein expression (Kappa value = 0.467; p = 0.022). CCNA1, CDH8, DAPK, PCDH10, and TIMP3 promoter hypermethylation was observed in 16-36%, of the OPSCC evaluated. DAPK hypermethylation was associated with HPV status (p = 0.023), while CCNA1 methylation was inversely related to p53 mutations in primary tumors (p = 0.042) and associated with a better disease-free survival (22.3% vs. 100%; p = 0.028) and overall survival (8% vs. 100%; p = 0.012).

Conclusions: Even though all samples analyzed were of the oropharynx, several differences regarding molecular and clinical characteristics were identified. More cases should be evaluated in order to assess whether the differences identified are able to classify these patients according to their outcome and help in a more thorough patient management.

Citation Format: Ana Carolina de Carvalho, Bruna Pereira Sorroche, Matias Eliseo Melendez, Cristina da Silva Sabato, Edenir Inez Palmero, Lidia Maria Rebolho Batista Arantes, Cristovam Scapulatempo Neto, André Lopes Carvalho. Clinical and molecular characterization of oropharynx squamous cell carcinoma samples [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A36.
Abstract A65: In vivo neutralization of IL-10 enhances the antitumor effects of a therapeutic vaccine against HPV-associated tumor
Authors: Silva Jamile Ramos da , Sales Natiely Silva , Silva Mariangela de Oliveira , Aps Luana M. M. , Ferreira Luis Carlos de Souza , Diniz Mariana de Oliveira
Published: May 2019

Although experimental therapeutic vaccines against HPV-induced tumors are capable of activating systemic E6 and E7-specific CD8+ T cells and inducing high levels of antitumor protection in murine model, they do not always correlate with clinical efficacy once these vaccines are tested in clinical settings. This observed discrepancy can be attributed to an immunosuppressive microenvironment elicited by the tumor that limits the efficacy of immunotherapeutic approaches. Solid evidence shows that IL-10 cytokine secretion is a relevant mediator of tumor immune evasion mechanisms, which inhibits the function of CD8+ T cells. Our group has sought alternatives to potentiate the effect of a DNA-based therapeutic vaccine (pgDE7h), which encodes the HPV-16 E7 protein fused to the herpes simplex virus type 1 (HSV-1) glycoprotein D (gD), previously developed in our laboratory. The present study aimed to investigate the potential role of immunossupression restriction using a plasmid encoding the soluble IL-10 receptor (pIL10R) to enhance the pgDE7h vaccine potency in the control of tumors expressing HPV-16 E6 and E7 proteins in a murine model (TC-1 cells). Our results showed a tumor growth delay after combining the vaccine with pIL-10R. Additionally, the use of in vivo electroporation as the plasmids delivery method conferred a therapeutic protection of 90% and 60% when the animals were immunized 5 or 14 days after challenge, respectively. The combination of the plasmids was also capable of increasing substantially the numbers of activated E7-specific CD8+ T lymphocytes both systemically and at the tumor site. Altogether, the results obtained are promising for the development of new strategies to increase the antitumor therapeutic effects of the pgDE7h vaccine and may contribute to its use under clinical conditions.

Note: This abstract was not presented at the conference.

Citation Format: Jamile Ramos da Silva, Natiely Silva Sales, Mariangela de Oliveira Silva, Luana M. M. Aps, Luis Carlos de Souza Ferreira, Mariana de Oliveira Diniz. In vivo neutralization of IL-10 enhances the antitumor effects of a therapeutic vaccine against HPV-associated tumor [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A65.
Abstract A73: GD3 ganglioside-enriched extracellular vesicles stimulate melanocyte migration
Authors: Otake Andreia H. , Duarte Ana Paula M. , Saito Renata de Freitas , Ramos Alexandre F. , Chammas Roger
Published: May 2019

Gangliosides are sialic acid-containing glycosphingolipids located mainly in outlet membrane plasma. Tumors usually accumulate gangliosides as disialoganglioside GD3 which is widely expressed in melanomas and neuroblastomas. GD3 has been associated with tumor progression, conferring tumorigenic characteristics to cells such as increased proliferation and migration. In order to investigate the role of GD3 in cell migration, we have transfected the GD3 synthase gene (ST8Sia I) in a melanocyte cell line. In this work we have demonstrated that overexpression of GD3 upon transfection of GD3 synthase was associated with an increased migratory response towards laminin-1. GD3 and 9-O-acetyl-GD3 gangliosides, which colocalize with integrins in cell lamellipodia, are positive modulators of integrin function in the migratory response while GD3 was left behind as a molecular footprint. Indeed, addition of exogenous GD3 to the cells stimulated cell migration whereas exogenously added GM3 inhibited it. However, depletion of gangliosides using a glucosylceramide synthase inhibitor led cells more migratory. Glycosphingolipids content modulates FGF-2-induced cell migration as we demonstrated that FGF-2 induces cell migration. However, this effect was abolished after using glucosylceramide synthase inhibitor, suggesting that GD3 may modulate the motogenic activity of FGF-2. Finally, we showed that GD3 synthase transfected melanocytes release extracellular vesicles (EVs) enriched in GD3. These EVs are responsible for modulating cell migration response due to increase average velocity of GD3 negative cells as we showed in a real-time wound-healing assay. This study shows that EVs from a subset of GD3 positive cells may contribute to the horizontal signaling propagation to a different subset of GD3 negative cells. Thus, data present here led us to an important insight into how EVs carrying GD3 ganglioside could participate in intercellular communication modulating cellular migration.

Citation Format: Andreia H. Otake, Ana Paula M. Duarte, Renata de Freitas Saito, Alexandre F. Ramos, Roger Chammas. GD3 ganglioside-enriched extracellular vesicles stimulate melanocyte migration [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A73.