bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract B11: Prostate apoptosis response-4 (PAR4) secretion in breast tumor and normal cell lines cultured in hypoxic conditions
Authors: Garcia Simone Aparecida de Bessa , Brobovnitchaia Irina Gueroldovna , Mangone Flavia Rotea , Nagai Maria Aparecida
Published: May 2019

The prostate apoptosis response-4 (PAR4) gene encodes a 38kDa proapoptotic protein with tumor cell selective action and/or cell sensitization to an additional proapoptotic insult. This selective apoptotic ability is associated with PAR4 nuclear translocation and phosphorylation state. PAR-4 acts through intrinsic and extrinsic apoptotic pathways. PAR-4 can also be secreted by normal/immortalized and cancer cells via the classical ER stress pathway mediated by GRP78. The previous study from our group provided evidence that MCF10A cells cultured in three-dimensional conditions display an increased expression of PAR4 in the center of the acini, where the cells are dying to lumen formation and where the O2 levels are expected to be lower. In the present study, we sought to evaluate the effects of hypoxia on PAR4 secretion in mammary epithelial cells and breast cancer cells. MCF10A (immortalized mammary epithelial cells) and the breast cancer cells MCF7, MDA-MB-231, and SKBR3 were cultured in normoxic and hypoxic (1% O2) conditions. The conditioned medium (CM) was concentrated using Amicon Ultra 10kDA filters and resolved by SDS-PAGE gel electrophoresis in parallel to cell lysate analysis. Cells maintained under hypoxic conditions for 6 h, 24 h, and 48 h showed no alteration in intracellular PAR4 protein levels in relation to cells maintained in normoxia. After 72 h we observed a decrease in intracellular PAR4 protein expression. The GRP78 protein expression did not change in the same time periods. However, Caspase-7 activation could be seen in 24 h, 48 h, and 72 h. Furthermore, we observed an increase (2.7 fold) in PAR4 protein expression in the CM after 72 h under hypoxia. Our results demonstrated for the first time that PAR4 secretion is modulated by hypoxia in normal and tumor breast cells. Further experimental and clinical studies are needed to determine the mechanism and role played by PAR4 secretion in cells' sensitivity to standard chemotherapeutics agents.

Supported by FAPESP and CNPq.

Citation Format: Simone Aparecida de Bessa Garcia, Irina Gueroldovna Brobovnitchaia, Flavia Rotea Mangone, Maria Aparecida Nagai. Prostate apoptosis response-4 (PAR4) secretion in breast tumor and normal cell lines cultured in hypoxic conditions [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 B11.
Abstract B45: Rab5C enhances resistance to ionizing radiation in rectal cancer
Authors: Baptistella Antuani Rafael , Dias Marcos Vinicios Salles , Giudice Fernanda , Rodrigues Bruna Roz , Silva Petrus Paulo Combas Eufrazio , Cassinela Edson Kuatelela , Lacerda Tonielli , Marchi Fabio , Leme Adriana Franco Paes , Begnami Maria Dirlei , Junior Samuel Aguiar , Martins Vilma Regina
Published: May 2019

Rectal cancer is currently the second most common cancer in the large intestine, and represents one-third of the colorectal cancers that are diagnosed. Neoadjuvant chemoradiation is a well-established protocol for rectal cancer treatment and it reduces the risk of local recurrence. However, a pathologic complete response is only achieved in 10–40% of cases and the mechanisms associated with chemoradiation resistance are poorly understood. To identify potential targets for preventing therapy resistance, a proteomic analysis of biopsy specimens collected from stage II and III rectal adenocarcinoma patients before neoadjuvant treatment was performed. These results were then compared with a proteomic analysis of residual rectal adenocarcinoma tissues that were removed by surgery after neoadjuvant therapy. Three proteins, Ku70, Ku80, and Rab5C, exhibited a significant increase in expression after chemoradiation treatment. To better understand the possible role of these proteins in therapy resistance, a rectal adenocarcinoma cell line was irradiated to generate a radiotherapy-resistant lineage. These cells overexpressed the same three proteins that were identified in the tissue samples. Furthermore, radiotherapy resistance in this in vitro model was found to involve modulation of epidermal growth factor receptor (EGFR) internalization by Rab5C in response to irradiation, and this affected expression of the DNA repair proteins, Ku70 and Ku80. Taken together, these findings indicate that EGFR and Rab5C are potential targets for the sensitization of rectal cancer cells to neoadjuvant treatment and they should be further investigated.

Supported by FAPESP - Fundação de Amparo à Pesquisa do Estado de São Paulo.

Note: This abstract was not presented at the conference.

Citation Format: Antuani Rafael Baptistella, Marcos Vinicios Salles Dias, Fernanda Giudice, Bruna Roz Rodrigues, Petrus Paulo Combas Eufrazio Silva, Edson Kuatelela Cassinela, Tonielli Lacerda, Fabio Marchi, Adriana Franco Paes Leme, Maria Dirlei Begnami, Samuel Aguiar Junior, Vilma Regina Martins. Rab5C enhances resistance to ionizing radiation in rectal 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 B45.
Abstract B49: Perillyl alcohol: Dynamic interactions with the lipid bilayer and implications for long-term inhalational chemotherapy for gliomas
Authors: Fonseca Clovis
Published: May 2019

Gliomas display a high degree of intratumor heterogeneity, including changes in physiologic parameters and lipid composition of the plasma membrane, which may contribute to the development of drug resistance. Biophysical interactions between therapeutic agents and the lipid components at the outer plasma membrane interface are critical for effective drug uptake. Amphipathic molecules such as perillyl alcohol (POH) have a high partition coefficient and generally lead to altered lipid acyl tail dynamics near the lipid-water interface, impacting the lipid bilayer structure and transport dynamics. We therefore hypothesized that glioma cells may display enhanced sensitivity to POH-induced apoptosis due to plasma membrane alterations, while in nontransformed cells, POH may be expelled through thermal agitation. Interactions between POH and the plasma membrane was studied using molecular dynamics simulations. In this phase I/II trial, we set up to evaluate the clinical effectiveness of long-term (up to 5 years) daily intranasal administration of POH in a cohort of 19 patients with low-grade glioma (LGG). Importantly, in a series of clinical studies previously published by our group, we have successfully established that intranasal delivery of POH to patients with malignant gliomas is a viable and effective therapeutic strategy. POH altered the plasma membrane potential of the lipid bilayer of gliomas and prolonged intranasal administration of POH in a cohort of patients with LGG halted disease progression with virtually no toxicity. Altogether, the results suggest that POH-induced alterations of the plasma membrane might be contributing to its therapeutic efficacy in preventing LGG progression.

Citation Format: Clovis Fonseca. Perillyl alcohol: Dynamic interactions with the lipid bilayer and implications for long-term inhalational chemotherapy for gliomas [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 B49.
Abstract B53: Sonic Hedgehog inhibition in glioblastoma potentializes temozolomide effect?
Authors: Honorato Jessica , Lopes Giselle Pinto de Faria , Basile Gabriela , Moura-Neto Vivaldo , Spohr Tania
Published: May 2019

Gliomas are malignant intracranial tumors and may occur in any region of central nervous system (CNS). Among them, glioblastoma (GBM) is the most aggressive and frequent brain tumors. In GBM treatment, the alkylating agent temozolomide (TMZ) is the first-line chemotherapy used; however, the patients' prognosis remains limited. During tumorigenesis, several signaling pathways vital for embryonic development are activated or inhibited, such as transforming growth factor beta (TGFbeta), Wingless (Wnt), and Sonic hedgehog (Shh). Currently, it is known that Shh pathway is involved in the chemoresistance of GBMs, and most of them have this pathway constitutive activated, which is one of the factors responsible to resistance to TMZ treatment. The limitation of using TMZ treatment can be solved by the identification of cell type with stem cell-like properties, the cancer stem cells (CSCs). To overcome this problem, some selective inhibitory drugs of Shh pathway have been developed, such as GANT-61. In this work, we investigated if Shh inhibition potentiates the effects of TMZ and verified the mechanisms involved in this process. To achieve our goals, GBM11 and T98G cell lines were treated with TMZ (100, 200, and 400 μM) and GANT-61 (20 μM) at different times of incubation. Our results suggest that both cell lines are resistant to TMZ, and the combination of GANT-61 with TMZ during 72 h reduced the cell viability of T98G lineage by MTT assay. Also, our results suggest that among the mechanisms involved in the cell viability reduction observed by combination of TMZ+GANT, cell death occurs due to DNA fragmentation and loss of membrane integrity, as well as formation of acidic vesicles that are indicative of autophagy and reactive oxygen species (ROS) production. In short, our results suggest that selective inhibition of the Shh pathway by GANT-61 is potentiating the effect of TMZ, being a promising strategy for GBMs treatment.

Citation Format: Jessica Honorato, Giselle Pinto de Faria Lopes, Gabriela Basile, Vivaldo Moura-Neto, Tania Spohr. Sonic Hedgehog inhibition in glioblastoma potentializes temozolomide effect? [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 B53.
Abstract B55: Targeting indoleamine 2,3 dioxygenase to improve the efficacy of a therapeutic vaccine against human papillomaviruses-associated tumors
Authors: Moreno Ana Carolina Ramos , Porchia Bruna F. M. M. , Souza Patrícia da Cruz , Pagni Roberta Liberato , Pegoraro Rafael , Ferreira Luís Carlos de Souza
Published: May 2019

The activity of the enzyme indoleamine 2,3 dioxygenase-1 (IDO) has been associated with the maintenance of the immunosuppressive microenvironment in many types of cancers. IDO catalyzes the degradation of tryptophan via the kynurenine pathway, which impacts on the effector activity of immune cells. The role of IDO in immune tolerance observed in human papillomaviruses-associated tumors is poor understood. Here, we targeted IDO to improve the efficacy of a therapeutic protein vaccine against HPV-16-associated tumors. The combination of IDO inhibitors with immunotherapy showed evidence that can support a new antitumor therapeutic approach. In the murine model C57BL/6, different biologic parameters were evaluated in order to determine the activation of the protective immunity obtained by the combined immunotherapy. We observed that isoforms of 1-methyl tryptophan, combined with our therapeutic vaccine, led to an activation of cytotoxic CD8 (+) T cells in mice grafted with tumor cells expressing the HPV-16 oncoproteins. In addition, the combination of melatonin enhanced the efficacy of the immunotherapeutic approach, resulting in the complete eradication of tumor masses. Altogether, our results indicate that blocking the immunosuppressive mechanisms mediated by IDO may improve the antitumor response induced by immunotherapies.

Citation Format: Ana Carolina Ramos Moreno, Bruna F. M. M. Porchia, Patrícia da Cruz Souza, Roberta Liberato Pagni, Rafael Pegoraro, Luís Carlos de Souza Ferreira. Targeting indoleamine 2,3 dioxygenase to improve the efficacy of a therapeutic vaccine against human papillomaviruses-associated tumors [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 B55.
Abstract B70: Identification of reference genes for normalizing gene expression for RT-qPCR analysis in established cervical cancer cell lines and their derived cancer stem like cells
Authors: Campos Rafael Paschoal de , Schultz Iago Carvalho , Mello Paola de Andrade , Davies Samuel , Gasparin Manuela Sangalli , Bertoni Ana Paula Santin , Azevedo Jéssica Gonçalves , Buffon Andréia , Wink Márcia Rosângela
Published: May 2019

Cervical cancer is the fourth most common cancer affecting women worldwide. Cancer stem cells are a subpopulation of tumor cells related to malignancy, resistance, and poor survival. Considering the importance of gene expression studies to understand cervical cancer stem cells (CCSC) biology and the need of proper validation of reference genes to normalize RT-qPCR data, this work identifies stable reference genes for the cervical cancer cell lines SiHa, HeLa, and ME180 as well as their respective derived cancer stem like cells. A literature review was performed to identify whether validation of reference genes was available for RT-qPCR assays in cervical cancer cell lines. Then cell lines were cultured in regular monolayer or in tumor sphere formation conditions. RT-PCR was carried out to validate cells grown as spheres as CCSC. RT-qPCR was performed using five commonly used reference genes: ACTB, B2M, GAPDH, HPRT1, and TBP. Stability was assessed through analysis of dispersion data and the Normfinder software to validate the selected genes as suitable reference genes. According to the literature review, no validation of reference genes for SiHa, HeLa, or ME180 cells in standard culture conditions or their CCSC subpopulations has been done so far. Our evaluation validated B2M, GAPDH, HPRT1, and TBP in these set conditions. Among them, GAPDH presented the lowest stability value (0.222), being therefore the most adequate gene to normalize the combination of all samples. Moreover, associating GAPDH and TBP led to a more stable value (0.184). These results suggest that B2M, GAPDH, HPRT1, and TBP are suitable reference genes to normalize RT-qPCR data of established cervical cancer cell lines SiHa, HeLa, and ME180 as well as their derived cancer stem like cells. Furthermore, GAPDH seems to be the most convenient choice for studying the combination of these cells.

Citation Format: Rafael Paschoal de Campos, Iago Carvalho Schultz, Paola de Andrade Mello, Samuel Davies, Manuela Sangalli Gasparin, Ana Paula Santin Bertoni, Jéssica Gonçalves Azevedo, Andréia Buffon, Márcia Rosângela Wink. Identification of reference genes for normalizing gene expression for RT-qPCR analysis in established cervical cancer cell lines and their derived cancer stem like cells [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 B70.
Abstract B71: Stochastic model of contact inhibition and the proliferation of melanoma in situ
Authors: Morais Mauro Cesar C. , Stuhl Izabella , Sabino Alan U. , Lautenschlager Willian W. , Queiroga Alexandre S. , Tortelli Tharcisio C. , Chammas Roger , Suhov Yuri , Ramos Alexandre F.
Published: May 2019

Contact inhibition of proliferation can be described as the decrease of proliferation rates when the cell density increases. Its loss is a key feature of tumor development. Cell density of human metastatic melanoma cell line (SK-MEL-147) and immortalized keratinocyte cells (HaCaT) was evaluated daily in culture experiments. Keratinocytes reach lower cell density than melanoma cells at confluence, indicating that the population growth arrest is associated with contact inhibition (KHaCaT = 1779.6 ± 130.5 cells/mm2; KSK-MEL-147 = 5043.5 ± 316.5 cells/mm2). We performed a coculture experiment to assess the proliferation restriction between both cell lines. After 8 days a spatial pattern was detected, characterized by the formation of clusters of melanoma cells surrounded by keratinocytes constraining their proliferation. Despite showing approximately the same proliferation rates when data were fitted with a logistic model (ρHaCaT = 1.1 ± 0.1 days-1; ρSK-MEL-147 = 1.1 ± 0.3 days-1), the proportion of melanoma cells within the total population has increased from 1:10 to 1:4 approximately. To explain our results we propose a spatial stochastic model where we consider cell proliferation, death, migration, and cell-to-cell interaction through contact inhibition. The agreement between experimental results and numerical simulation indicates that loss of contact inhibition is important for the analysis of the development of melanoma in situ, and explains the increase in the proportion of tumor cells and generation of spatial patterns established in conducted experiments. In addition, the distribution of the distances between cells of the same line indicates the higher proximity of melanoma cells and allowed us to identify their clustering pattern. This work provides results of a quantitative study of the impact of contact inhibition upon formation and growth of tumor-like melanoma cell clusters in a coculture with keratinocytes.

Citation Format: Mauro Cesar C. Morais, Izabella Stuhl, Alan U. Sabino, Willian W. Lautenschlager, Alexandre S. Queiroga, Tharcisio C. Tortelli, Jr., Roger Chammas, Yuri Suhov, Alexandre F. Ramos. Stochastic model of contact inhibition and the proliferation of melanoma in situ [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 B71.
Abstract IA17: Liquid biopsies: Concepts and applications
Authors: Camargo Anamaria A.
Published: May 2019

The presence of cell-free DNA (cfDNA) in human blood was first described in 1948 by Mandel and Métais, but mutated KRAS DNA sequences derived from tumor cells were first reported in the plasma of patients with pancreatic cancer almost 50 years later. Since then, our ability to analyze cfDNA has significantly improved with the development of molecular technologies, such as digital PCR and next-generation sequencing, which enable the detection of circulating tumor DNA molecules (ctDNA) with high specificity and sensitivity. ctDNA is shed into the bloodstream by cells undergoing apoptosis or necrosis and genomic profiles of ctDNA have been shown to closely match those of the corresponding tumors. Recent studies have demonstrated the translational potential of ctDNA detection—commonly referred to as liquid biopsies—for diagnosis and prognosis, for treatment decision, and for monitoring residual disease, treatment response, and disease progression. This is an opportune time to discuss how liquid biopsies can be exploited to guide cancer patient care and ultimately be integrated into clinical practice.

Citation Format: Anamaria A. Camargo. Liquid biopsies: Concepts and applications [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 IA17.
Abstract IA18: Cancer evolution as a therapeutic target
Authors: Bardelli Alberto
Published: May 2019

When metastatic cancers are challenged with targeted agents, almost invariably a subset of cells insensitive to the drug emerges. As a result, in most instances, targeted therapies are only transiently effective in patients. Strategies to prevent or overcome resistance are therefore essential to design the next generation of clinical trials. How can we overcome the near-certainty of disease recurrence following treatment with targeted agents? Addressing this question means considering as a target not only individual oncogenes but also the evolving nature of human tumors.

We used colorectal cancer (CRC) as a model system to test the hypothesis that by understanding tumor's evolution, the emergence of drug resistance can be controlled. We find that clonal dynamics can be monitored in real time in the blood of patients, and liquid biopsies can be used to intercept the emergence of resistant clones before relapses are clinically manifest. We discovered that a multistep clonal evolution process driven by progressive increases in drug fitness underlies the development of resistance in cells and patient avatars.

To have long-term efficacy, the use of targeted therapies must take into account the continuous evolution of cancer cells, that is to say, therapies must adapt to tumor evolution. One possibility is to anticipate the changes the tumors will make. For example, by knowing in advance how CRC cells overcome resistance to EGFR blockade, we devised further rounds of therapy. Another approach is to unleash the ability of the immune system to recognize drug-resistant cells. We tested this possibility in syngeneic mouse models of CRC sensitive to targeted therapies. Our findings indicate that manipulation of the mutational loads can trigger prolonged therapeutic responses that are not observed when cancer cells are challenged with targeted drugs alone. We postulate that rationally combined targeted and immunotherapies can restrain tumor evolution and can limit the emergence of drug resistance, thus leading to long-term responses.

Citation Format: Alberto Bardelli. Cancer evolution as a therapeutic target [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 IA18.
Abstract A12: Vicoryx: A phase I/IIa clinical trial using a p16INK4a derived peptide as vaccine in patients with advanced human papillomavirus-associated cancer
Authors: Reuschenbach Miriam , Karbach Julia , Faulstich Franziska , Sauer Madeleine , Kloor Matthias , Rafiyan Mohammad-Reza , Pauligk Claudia , AlBatran Salah , Jaeger Elke , Doeberitz Magnus von Knebel
Published: May 2019

Objectives: The tumor suppressor p16INK4a is strongly overexpressed in HPV-associated neoplasia, whereas in normal tissues barely any p16INK4a expression is detectable. Targeting this HPV type-independent antigen by vaccination could represent an interesting complementary therapeutic approach to recently developed HPV-E6/E7-based therapeutic or secondary preventive vaccines. We performed a phase I/IIa peptide vaccination trial to monitor toxicity and immunogenicity of p16INK4a vaccination in patients with advanced HPV-associated cancers.

Patients and Methods: 21 patients with p16INK4a-overexpressing, HPV DNA-positive advanced cancers (16 cervical, 5 head and neck) were included. The protocol comprised 12 applications of a synthetic 27mer p16INK4a peptide mixed with Montanide® ISA-51 VG over a six months period. Objectives of the study were clinical safety and changes of humoral and cellular immune responses against the p16INK4a peptide. T cell responses were monitored by interferon-gamma ELISpot and antibodies by ELISA.

Conclusions: No vaccine-related toxicity was observed in any of the patients. One patient (head and neck cancer) completed the entire study protocol with stable disease for now 18 months after the last vaccination.11 patients had progressing disease and were excluded from the study after 4 to 12 weeks. 9 patients continue to be vaccinated. While at baseline only one patient had pre-existing T cell responses (CD4) against the p16INK4a peptide, p16INK4a-reactive T cells (CD4) were successfully induced in at least four patients after 4 to 12 vaccine doses. None of the patients had pre-existing p16INK4a antibodies, but 3 patients developed increasing p16INK4a peptide-specific antibody titers after the 5th dose.

This is the first study demonstrating that p16INK4a peptide vaccination is safe and well tolerated. The results show that spontaneous immune responses against p16INK4a are rare, but can be induced by p16INK4a peptide vaccination. Further studies are needed to assess clinical efficacy of the approach. Please refer to the protocol of this trial under http://clinicaltrials.gov/show/NCT01462838.

Citation Format: Miriam Reuschenbach, Julia Karbach, Franziska Faulstich, Madeleine Sauer, Matthias Kloor, Mohammad-Reza Rafiyan, Claudia Pauligk, Salah AlBatran, Elke Jaeger, Magnus von Knebel Doeberitz. Vicoryx: A phase I/IIa clinical trial using a p16INK4a derived peptide as vaccine in patients with advanced human papillomavirus-associated cancer. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr A12.
Abstract A23: Metformin as a potent antiangiogenic factor: From diabetes to cancer angioprevention
Authors: Dallaglio Katiuscia , Bruno Antonino , Cantelmo Anna Rita , Pfeffer Ulrich , Bertolini Francesco , Noonan Douglas M. , Albini Adriana
Published: May 2019

Metformin, an anti-diabetic drug commonly used for type 2 diabetes therapy, is associated with a decreased risk for certain cancer. Due to extensive reports of its in vitro and in vivo anti-tumor activity, we and others have focused on the anti-angiogenic potential as a possible mechanism through which metformin could affect tumor progression, and to propose its use as an angiopreventive drug. Metformin was able to interfere with endothelial cell proliferation, apoptosis and invasion in vitro. Gene arrays analyses were performed to verify the effect of metformin on endothelial gene expression profiles and the matrigel sponge assay to test its effect in vivo.

We show that metformin inhibited endothelial cell proliferation by effecting cell cycle regulation rather than inducing apoptosis. In vitro, metformin reduces endothelial cell invasion and represses their ability to organize into capillary-like networks. These effects are partially reverted by compound C, a specific inhibitor of AMPK, confirming that metformin could interfere with angiogenesis responses by activating AMPK signaling. Gene expression profiles of endothelial cells treated with metformin showed down-regulation of several angiogenesis-related genes but up-regulation of chemokine pathways. In vivo, metformin inhibited angiogenesis induced by VEGF. Considering the strict correlation between type II diabetes and obesity, we evaluated the ability of Metformin to modulate endothelial precursor cells in an obese murine model and we reported that Metformin decreases the number of adipose tissue progenitor endothelial cells in vivo only in obese mice.

These data suggest that metformin has anti-angiogenic effect associated with down-regulation of specific molecule although it also induces some inflammatory mediators, and that it may be most effective in the context of obesity.

Citation Format: Katiuscia Dallaglio, Antonino Bruno, Anna Rita Cantelmo, Ulrich Pfeffer, Francesco Bertolini, Douglas M. Noonan, Adriana Albini. Metformin as a potent antiangiogenic factor: From diabetes to cancer angioprevention. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr A23.
Abstract A51: The heritability of colorectal cancer in the Nordic Twin Study of Cancer (NorTwinCan)
Authors: Graff Rebecca E. , Skytthe Axel , Adami Hans-Olov , Czene Kamila , Harris Jennifer R. , Kaprio Jaakko , Giovannucci Edward , Mucci Lorelei A. , Hjelmborg Jacob B.
Published: May 2019

Background: Family history is a strong risk factor for colorectal cancer, but the extent to which familial risk can be attributed to heritability versus shared environment remains obscure. Whether familial risk is distinct for colon versus rectal cancer also remains unclear and is important to examine given accumulating evidence that their etiologies are distinct.

Methods: We estimated the concordance and heritability of colorectal, colon and rectal cancer using information from 40,634 monozygotic and 111,897 dizygotic twin pairs from the Nordic Twin Study of Cancer. Time-to-event analyses accounted for left-censoring due to variable initiation of cancer registration and right-censoring resulting from end of follow-up or competing risk of death.

Results: From earliest cancer registration in 1943 through 2009, 2,286 twins were diagnosed with colon cancer and 1,493 were diagnosed with rectal cancer. The cumulative risk of colorectal cancer overall was 4.5%, while the cumulative risk for twins whose co-twins had colorectal cancer was 15.7% for monozygotic and 11.0% for dizygotic twins. The heritability of colon cancer was 16.2% (95% CI: 2.3%-61.8%), with shared environment accounting for 14.2% (95% CI: 2.7%-49.1%) of the variation in disease liability. For rectal cancer, a model including only environmental effects best fit the data; heritability was not statistically important. Shared environment accounted for 16.6% (95% CI: 8.8%-28.8%) of the variation in liability.

Conclusions: These results suggest a greater genetic contribution to the risk of colon cancer than the risk of rectal cancer. The contribution of shared environment to both cancers may be larger than previously thought.

Citation Format: Rebecca E. Graff, Axel Skytthe, Hans-Olov Adami, Kamila Czene, Jennifer R. Harris, Jaakko Kaprio, Edward Giovannucci, Lorelei A. Mucci, Jacob B. Hjelmborg. The heritability of colorectal cancer in the Nordic Twin Study of Cancer (NorTwinCan). [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr A51.
Abstract A62: Alcohol consumption, genetic variation in alcohol dehydrogenase genes, and risk of clear cell renal cell carcinoma
Authors: Parker Alexander Scott , Diehl Nancy , Arnold Michelle L. , Rivera Shauna , Cannon Ashley , Custer Kaitlynn , Leibovich Bradley
Published: May 2019

Introduction: An inverse association between alcohol consumption and risk of renal cell carcinoma (RCC) is well reported in the literature. Despite this, no investigators have explored the potential for effect modification of this association by genetic variations in enzymes that metabolize alcohol. Motivated by this, we conducted a pilot investigation of the role of genetic variation in the alcohol dehydrogenase genes (ADH) as an effect modifier of the association of alcohol and RCC risk.

Methods: We analyzed data on 121 RCC cases and 121 controls. Cases were prospectively collected as part of our Renal Registry in the Department of Urology. Controls were recruited through the Family Medicine and frequency matched to cases on age, gender and state of residence. We collected detailed risk factor data (self-report questionnaire) as well as a DNA sample on cases and controls. We utilized Sequenome analysis to determine presence of published SNPs in the ADH gene cluster (ADH1-7). We employed logistic regression with interaction terms as well as stratified analyses to evaluate effect modification by genetic variants in the ADH gene cluster on the association of alcohol consumption and RCC risk.

Results: Compared to non-drinkers, those that consumed alcohol above the median experienced a lowering in RCC risk (OR=0.80; 95% CI 0.5 to 1.43). We observed evidence of interactions with 6 SNPs in the ADH gene (p-values from 0.05-0.2). As an example, in stratified analysis, the inverse association strengthened in those without the minor allele present at rs1154454 in ADH7 (OR=0.56; 95% CI 0.3 to 1.0; p=0.08) while evidence of an increased in RCC risk was noted among those with the minor allele (OR=2.14; 95CI 0.8 – 5.7; p=0.1). Adjustment for age and gender did not alter our results.

Conclusion: Our pilot data suggest that specific variants in the ADH gene cluster may modify the reported protective effect of alcohol consumption on RCC risk. Given the possible implications for improving our understanding of the biology of this association and informing prevention, future investigations that analyze a larger sample size and examine additional variants are warranted.

Citation Format: Alexander Scott Parker, Nancy Diehl, Michelle L. Arnold, Shauna Rivera, Ashley Cannon, Kaitlynn Custer, Bradley Leibovich. Alcohol consumption, genetic variation in alcohol dehydrogenase genes, and risk of clear cell renal cell carcinoma. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr A62.
Abstract B45: Obesity, diabetes, other comorbidities and survival in a cohort of women with ovarian cancer
Authors: Protani Melinda M. , Nagle Christina M. , deFazio Anna , Blomfield Penelope , Jobling Thomas , Leung Yee , Nicklin James L. , Perrin Lewis C. , Wain Gerard , Webb Penelope M.
Published: May 2019

Purpose: To determine the relationship between obesity, diabetes, other major comorbidities and survival following a diagnosis of ovarian cancer.

Methods: We examined the association between obesity, as measured by body mass index (BMI), diabetes and other comorbidities and survival from ovarian cancer in a large (n=1404), population-based cohort of women diagnosed with epithelial ovarian cancer between 2002-2006. Information about pre-diagnostic BMI and medical history was obtained via a self-administered questionnaire. BMI at the commencement of primary chemotherapy and clinicopathologic data were abstracted from medical records and deaths were ascertained up to October 2011 via linkage with the Australian National Death Index. We also examined short- (≥5 years) and long-term (>5 years) survival, as well as survival differences by histological subtype. Adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) were obtained using Cox regression.

Results: After adjustment, a survival disadvantage was observed for women who were obese prior to diagnosis compared to women in the normal BMI range (BMI 30-34.9:HR 1.21, 95%CI 0.99-1.48; BMI≥35:HR 1.19, 95%CI 0.92-1.55). The association was similar for BMI at the commencement of primary chemotherapy. There was a suggestion that the adverse association between obesity and survival time was restricted to women with serous or endometrioid histological subtypes. Obesity conferred a survival disadvantage for both short- and long-term survival, with a stronger association in long-term survivors. Diabetes and other comorbidities did not appear to influence survival overall; however appeared to be important in the sub-group of women surviving longer than 5 years (HR1.44 and HR1.64, respectively).

Conclusion: This is the first study to show that obesity is important in both short- and long-term survival following a diagnosis of ovarian cancer. More research examining diabetes, comorbidities and ovarian cancer survival should be conducted.

Citation Format: Melinda M. Protani, Christina M. Nagle, Anna deFazio, Penelope Blomfield, Thomas Jobling, Yee Leung, James L. Nicklin, Lewis C. Perrin, Gerard Wain, Penelope M. Webb. Obesity, diabetes, other comorbidities and survival in a cohort of women with ovarian cancer. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr B45.
Abstract C16: Effects of cessation counseling types on quit attempts and intentions
Authors: Cornelius Monica E. , Cummings K. Michael
Published: May 2019

Smoking cessation assistance programs aid smokers in quitting. While cessation programs have shown some effectiveness, many smokers do not utilize them. The combination of nicotine addiction, access to cheaper products and a general lack of access to smoking cessation tools present difficulties to smokers. Direct counseling from a healthcare provider has been shown to aid in smokers thinking about quitting their smoking; however greater assistance is often needed beyond the recommendation to quit. The purpose of this study was to examine the effect different types of cessation counseling on (1) quit attempts and (2) intention to quit within the next month. We hypothesized that advice + quit assistance from a healthcare provider would be associated with making a quit attempt and intending to quit within the next month when compared with advice alone. This paper presents data from the first eight waves of the International Tobacco Control (ITC) US adult smoker survey from 2002-2011 (N=6669). Participants who visited a healthcare provider within the past year of survey were eligible for inclusion. The levels of healthcare provider counseling included no advice, advice only, and advice + quit assistance. Repeated measures logistic regression, using generalized estimating equations (GEE) was used to model (1) whether participants made a quit attempt and (2) intention to quit within the next month. Models were controlled for age, sex, daily smoking, time-in-sample, wave, education, income, and region, as well as cessation counseling. Over 60% of participants indicated having seen a healthcare provider, with over 60% receiving advice to quit. Among those who reported receiving advice to quit, less than one third received a referral or quit prescription and less than one half received a pamphlet from their healthcare provider. Those receiving advice + quit assistance were almost twice as likely to have attempted to quit as those who received advice only. Participants receiving assistance were more likely to intend to quit within the next month. Not all smokers get advice to stop smoking when seen by their doctor and even fewer get offered assistance beyond the recommendation to stop smoking. This study shows that those who received both advice and assistance were more likely to report making a quit attempt or at least report a greater intention to stop smoking in the future.

Citation Format: Monica E. Cornelius, K. Michael Cummings. Effects of cessation counseling types on quit attempts and intentions. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr C16.
Abstract C22: Telomere length variation and frequency of short telomeres in blood lymphocytes: Novel biomarkers for lung cancer risk
Authors: Sun Bing , Wang Ying , Kota Krishna , Shi Yaru , Motlek Salaam , Makambi Kepher , Loffredo Christopher A. , Shields Peter G. , Yang Qin , Harris Curtis C. , Zheng Yun-Ling
Published: May 2019

Background: Although it is widely recognized that telomere dysfunction plays an important role in cancer, the relationship between telomere function in somatic cells and lung cancer risk is not well defined. In a case-control study of non-small cell lung cancer, we examined relationship between several key telomere features and lung cancer risk. Method: Telomere fluorescent in situ hybridization was used to measure telomere features using short-term cultured blood lymphocytes. Logistic regression was used to estimate the strength of association between telomere features and lung cancer risk. Results: Telomere length variation across all chromosomal ends was significantly associated with lung cancer risk, with adjusted odds ratios of 4.67 [95% confidence interval (CI) = 1.46 – 14.9] and 0.46 (95% CI = 0.25 – 0.84) for young (age < 60) and old (age > 60) individuals, respectively. Similar strength and direction of associations between percentage of short telomeres and lung cancer risk were also observed. Telomere length variation and average telomere length jointly affect lung cancer risk. When comparing individuals with short telomere and high telomere length variation to those with long telomere and low telomere length variation, adjusted odd ratios were 8.21 (95% CI = 1.71 – 39.5) and 0.33 (95% CI = 0.15 – 0.72) for young and old individuals, respectively. Conclusion: Telomere length variation and percentage of short telomeres in blood lymphocytes are significantly associated with lung cancer risk and the associations are modulated by age. Telomere length variation in combination with average telomere length may be a promising tool for lung cancer risk assessment.

Citation Format: Bing Sun, Ying Wang, Krishna Kota, Yaru Shi, Salaam Motlek, Kepher Makambi, Christopher A. Loffredo, Peter G. Shields, Qin Yang, Curtis C. Harris, Yun-Ling Zheng. Telomere length variation and frequency of short telomeres in blood lymphocytes: Novel biomarkers for lung cancer risk. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr C22.
Abstract C54: Mechanism and preclinical prevention of increased breast cancer risk caused by pregnancy
Authors: Li Yi
Published: May 2019

While a first pregnancy before age 22 lowers breast cancer risk, a pregnancy after age 35 significantly increases life-long breast cancer risk. Pregnancy causes several changes in the normal breast that raise barriers to transformation, but how pregnancy can also increase long-term cancer risk remains unclear. Precancerous lesions accumulate in the breast with age. We show in mice that pregnancy has different effects on the few early lesions that have already accumulated in the otherwise normal breast'it causes apoptosis evasion and accelerated progression to cancer. The apoptosis evasion is due to the normally tightly controlled STAT5 signaling going astray' in response to pregnancy/lactation hormones, these precancerous cells can activate STAT5; but during involution, the oncogenic signaling can prevent STAT5 deactivation. The resulting persistent STAT5 signaling prevents the apoptosis normally initiated by oncogene activation and involution. Short-term anti-STAT5 treatment of lactation-completed, early lesions-bearing mice eliminates the increased risk after a pregnancy. This chemoprevention strategy has important implications for preventing increased human breast cancer risk caused by pregnancy as it may lower long-term breast cancer risk in women who have had a late-age pregnancy and thus are more likely to harbor STAT5-activated progressing early lesions.

Citation Format: Yi Li. Mechanism and preclinical prevention of increased breast cancer risk caused by pregnancy. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr C54.
Abstract CN01-03: Frameshift peptide vaccination: Towards cancer prevention in Lynch syndrome
Authors: Kloor Matthias , Reuschenbach Miriam , Karbach Julia , Rafiyan Reza , Al-Batran Salah-Eddin , Pauligk Claudia , Jaeger Elke , Doeberitz Magnus von Knebel
Published: May 2019

Lynch syndrome is one of the most common hereditary cancer syndromes. It is caused by germ line mutations of DNA mismatch repair genes and predisposes affected individuals to the development of colorectal cancer and extracolonic malignancies with a lifetime risk of over 50%. Lynch syndrome-associated cancers show the high-level microsatellite instability (MSI-H) phenotype and are characterized by signs of a pronounced local anti-tumoral immune response, suggesting that the immune system plays an active role in the surveillance and natural history of these cancers. The progression of MSI-H cancers is triggered by mutations in microsatellite sequences within gene-encoding regions, which can cause shifts of the translational reading frame and lead to the generation of immunogenic frameshift peptides (FSP) at the carboxy-terminal end of the respective proteins. FSP-specific immune responses have been observed frequently in MSI-H cancer patients and Lynch syndrome mutation carriers. We have therefore initiated a clinical phase I/IIa vaccination trial that evaluates vaccination with a combination of three FSP antigens in patients with MSI-H colorectal cancer, UICC stage III and IV. Preliminary data demonstrate that no treatment-associated severe adverse events have been observed after FSP vaccination. Moreover, significant T cell and humoral immune responses against FSP antigens were induced in the majority of vaccinated patients. The observation of strong FSP-specific immune responses in vaccinated patients suggests that FSP-based vaccination may represent a novel option for the treatment of MSI-H colorectal cancer patients. Moreover, FSP vaccination may be used for tumor prevention in Lynch syndrome mutation carriers in the future.

Citation Format: Matthias Kloor, Miriam Reuschenbach, Julia Karbach, Reza Rafiyan, Salah-Eddin Al-Batran, Claudia Pauligk, Elke Jaeger, Magnus von Knebel Doeberitz. Frameshift peptide vaccination: Towards cancer prevention in Lynch syndrome. [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr CN01-03.
Abstract CN05-01: Genomics and cancer management: Mind the gap!
Authors: Jankowski Janusz A.
Published: May 2019

All diseases are a combination of nature versus nurture. In the former understanding of the inherited genetics through genomics is vital to understanding the predisposition to diseases and in some cases prognosis and response to therapy. The application of genome wide analyses studies identify several diseases has been particularly helpful especially in Crohn's disease, Diabetes and Psoriasis. Outside autoimmune diseases genomics whilst improving the understanding of cancer and its etiology has not made a difference in the clinical management of diseases. As an exemplar the progression from oesophagitis to metaplasia to esophageal adenocarcinoma presents challenges which are generalizable to other cancers. These include delineating differential genomic drivers for premalignant intermediates from those for the subsequent neoplastic lesions. In addition developments in cancer genomics by Exome sequencing or whole genome sequencing don't readily match with inherited genomic changes from GWAS. Moreover genomics has yet to reveal robust pathways for the design of new targets.

When these issues are coupled with failure in many cases to assess other factors responsible for inherited variation including CNVs and methylation patterns, it seems likely progress will be slow and need very large sample sets.

Citation Format: Janusz A. Jankowski. Genomics and cancer management: Mind the gap! [abstract]. In: Proceedings of the Twelfth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2013 Oct 27-30; National Harbor, MD. Philadelphia (PA): AACR; Can Prev Res 2013;6(11 Suppl): Abstract nr CN05-01.