bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

Abstract
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

Abstract
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

Abstract
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&gt;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

Abstract
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

Abstract
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

Abstract
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.
Abstract B12: HPV-positive tumor cell lines exhibit major alterations in Toll-like receptor pathways and depend on HMGB1 expression
Authors: Morale Mirian Galliote , Abjaude Walason da Silva , Silva Aline Montenegro da , Villa Luisa Lina , Boccardo Enrique 
Published: May 2019

Abstract
Cervical cancer is one of the leading causes of cancer death in women worldwide. Persistent infection with high-risk human papillomavirus (HPV) types is the main risk factor for the development of cervical cancer precursor lesions and is usually characterized by innate immune system evasion. The innate immune system is the first line of defense against pathogens and homeostatic changes. Toll-like receptors (TLR) are a family of membrane proteins that actively participate in this process by recognizing molecular patterns derived from pathogens and from cellular danger signals. Alterations in TLR expression and activation may be important for the control of HPV infections and could play a role in the progression of HPV-associated lesions and cancers. In the present study, we analyzed the expression of 84 genes involved in TLR signaling pathways in cervical cancer cell lines and primary human keratinocytes. We observed that 80% of the differentially expressed genes were downregulated in cervical cancer cell lines relative to normal keratinocytes. Major alterations were detected in genes coding for several TLR signaling proteins, including adaptor molecules MyD88 and SARM1, NFκB activation complex components such as Ube2N and TRAF6, and effector proteins such as HMGB1. Specifically, HMGB1, a protein related to stimulation of inflammation after TLR activation, showed increased expression levels in the HPV-positive cell lines HeLa and SiHa relative to normal keratinocytes. HMGB1 silencing also reduced cell viability, proliferation, and colony formation capacity of cervical cancer cell lines. In addition, silenced cervical cancer cell lines exhibited increased activated caspase-3 levels and hypodiploid cell population suggestive of apoptosis triggering. HPV infection may interfere with TLR pathways at different levels, resulting in the impairment of their activation and function and potentially leading to innate immune system evasion. Our results suggest that TLR pathway proteins such as TLR4, SARM1, and HMGB1 may play other roles related to tumor development. In conclusion, HMGB1 is a key factor in the survival of HPV-positive tumor cells, which warrants further studies to evaluate its potential as a therapeutic target or tumor biomarker.
Citation Format: Mirian Galliote Morale, Walason da Silva Abjaude, Aline Montenegro da Silva, Luisa Lina Villa, Enrique Boccardo. HPV-positive tumor cell lines exhibit major alterations in Toll-like receptor pathways and depend on HMGB1 expression [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 B12.
Abstract B34: Pyrimidobenzimidazoles as a potential new drug to treat human melanoma
Authors: Azevedo Jéssica Gonçalves , Schultz Iago Carvalho , Santos Jacqueline Fraga de Souza , Maria-Engler Silvya Stuchi , Lenz Guido , Wink Marcia Rosângela 
Published: May 2019

Abstract
Melanoma is the most aggressive skin tumor due to its high capacity to metastasize and resistance to therapy, leading to poor prognosis and high mortality rates. Thus, the aim of this study was to investigate a possible antitumoral action of pyrimidobenzimidazole compounds in human melanoma lineages, SK-Mel-103, SK-Mel-28, and SK-Mel-28 resistant to vemurafenib. Using acute treatment (72 hours) it was possible to select two, among the seven compounds tested, by their effectiveness in decrease the cell viability, by MTT assay, in low concentrations. Further, using chronic assay (120 hours) it was possible to find the IC50 of the best compounds, PBZs 1 and 4. For cell line SK-Mel-103 the IC50 value was 3.2 µM and 4.0 µM, for the two more potent compounds respectively, and for cell line SK-Mel-28 IC50 of 9.7 µM and 9.6 µM was obtained for PBZ01 and PBZ04, respectively. These compounds, alone or in combination with DTIC, were used to analyze the cumulative population doubling, in an attempt to simulate the treatment cycle used for patients. The results show that PBZs 1 and 4, especially when in combination with dacarbazine, were able to cause a significant decrease in the population growth rate and in the number of colonies in clonogenic assay, of the SK-Mel-28, better than dacarbazine alone and in lower concentration. Also, it is important to highlight that the effect of PBZs was selective to melanoma, considering that it did not cause relevant effect in melanocyte and in the proliferative nontumoral cell line, MRC5. The cell cycle analysis showed that the treatment alters the cell complexity, increasing the size (FSC) and granularity (SSC). Furthermore, when vemurafenib-resistant cells were treated with PBZ (cotreatment with vemurafenib), there were a population decrease in the population doubling assay more than vemurafenib alone. These results are important and encourage the study of the mechanism by which these compounds affect the melanomas' viability as well as the development of a preclinical model to confirm their effectiveness in vivo.
Citation Format: Jéssica Gonçalves Azevedo, Iago Carvalho Schultz, Jacqueline Fraga de Souza Santos, Silvya Stuchi Maria-Engler, Guido Lenz, Marcia Rosângela Wink. Pyrimidobenzimidazoles as a potential new drug to treat human melanoma [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 B34.