bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3919: Inhibition of HER2 and IGF2 in trastuzumab-resistant HER2 positive JIMT1 cells decreases cell growth and reduces activation of MAPK and AKT
Authors: Helu Xousaen M. , DeLeon Daisy
Published: August 2015

Breast cancer is the second leading cause of death in women. Overexpression of human epidermal growth factor receptor 2 (HER2) is detected in 20-30% of breast cancers. Increased levels of HER2 are linked to poor prognosis, more aggressive phenotype, and resistance to chemotherapy and hormonal treatment. HER2 is part of a four-member family of cell surface receptors that are implicated in transmission of signals controlling cell growth and differentiation. HER2 has an intracellular domain with tyrosine kinase catalytic activity. Dimerization of HER2 activates the MAPK and AKT pathways. The HER2 receptor also hetero-dimerizes with the IGF1 receptor (IGF1R) in trastuzumab-resistant cells. The insulin-like growth factor 2 (IGF2) has an important role in fetal and cancer development signaling through the IGF1R, insulin receptor (InsR), and a cross-talk with the estrogen receptors alpha and beta (ERα, ERß). Trastuzumab resistance occurs in 70% of patients with HER2 positive breast cancer. Therefore it is important to understand the mechanism of resistance so we can develop new therapies to help patients with trastuzumab-resistant rumors. Previous findings in our lab demonstrate an increased expression of IGF2 in the trastuzumab-resistant cell line JIMT1 when compared to trastuzumab-sensitive cell lines BT474, SKBR3, and ZR75-30. We hypothesize that to overcome trastuzumab-resistance in JIMT1 cells, dual inhibition of the HER2 and IGF2 is necessary to induce cell death. When JIMT1 cells were treated with trastuzumab and an IGF2 blocking antibody (IBA) a decrease in cell viability was observed, but changes in PARP cleavage were not present. We also analyzed the protein and phosphorylation levels of HER2, IGF1R, MAPK, and AKT. Our studies suggest that blocking both HER2 and IGF2 decreases cell viability and reduces phosphorylation of MAPK and AKT.

Citation Format: Xousaen M. Helu, Daisy DeLeon. Inhibition of HER2 and IGF2 in trastuzumab-resistant HER2 positive JIMT1 cells decreases cell growth and reduces activation of MAPK and AKT. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3919. doi:10.1158/1538-7445.AM2015-3919
Abstract 3922: Novel hepatocyte growth factor and macrophage stimulating protein antagonists for the treatment of castration-resistant prostate cancer
Authors: Vanderwerff Brett , Church Kevin , Kawas Leen , Harding Joseph
Published: August 2015

Castration-resistant prostate cancer (CRPC) is an advanced form of prostate cancer that is refractory to androgen deprivation therapy and characterized by aggressive growth, extensive metastasis formation, and poor prognosis. CRPC is temporarily manageable by the first-line chemotherapeutic docetaxel, but will eventually progress as a chemoresistant cancer, prompting a need for additional therapeutics to either replace docetaxel or to be used in combination therapies that increase the effectiveness of chemotherapy. The aberrant activation of the tyrosine kinase receptor c-Met by the active, dimeric form of hepatocyte growth factor (HGF) and the tyrosine kinase receptor RON by macrophage stimulating protein (MSP) results in the overactivation of signaling pathways that encourage cell proliferation, migration, and survival, contributing to the malignant behavior and chemoresistance of many cancers. The onset of metastatic prostate cancer is associated with both increased serum levels of HGF and expression of c-Met and RON in metastatic growths. Our laboratory has developed small molecules that inhibit HGF dimerization, c-Met activation, and malignant cell behaviors in several cancer cell types. Considering the high degree of homology between HGF and MSP, we predict that our strategy for inhibiting the activation of HGF can be extended to develop compounds that dually inhibit HGF and MSP. We hypothesize that inhibiting HGF and MSP with our antagonists will correspondingly reduce activation of c-Met and RON and suppress tumor growth and metastasis in CRPC. We further anticipate a therapy that combines our antagonists and docetaxel will increase the efficacy of chemotherapy by blunting the pro-survival properties of the HGF/c-Met and MSP/RON systems. Preliminary results indicate that our HGF dimerization antagonist Norleual [Nle-Tyr-Ile-ψ-(CH2-NH2)3-4-His-Pro-Phe] significantly inhibits the migration of CRPC cells and increases their susceptibility to docetaxel treatment in vitro. The primary objective of this study is to investigate the feasibility of HGF, MSP, and HGF/MSP dual antagonists as a therapy for CRPC either alone or in combination with docetaxel.

Citation Format: Brett Vanderwerff, Kevin Church, Leen Kawas, Joseph Harding. Novel hepatocyte growth factor and macrophage stimulating protein antagonists for the treatment of castration-resistant prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3922. doi:10.1158/1538-7445.AM2015-3922
Abstract 3923: A role for fibroblast growth factor receptors in growth of atypical teratoid rhabdoid tumor cells and as potential therapeutic targets
Authors: Azorsa David O. , Pepper Oliver B. , Chakravadhanula Madhavi , Lee David W. , Montoya Justin J. , Ozols Victor , Bhardwaj Ratan , Arceci Robert J.
Published: August 2015

Malignant rhabdoid tumors are characterized by the loss of the SWI/SNF complex subunit SMARCB1. Atypical Teratoid Rhabdoid Tumor (ATRT) is a rare form of pediatric MRT that occurs in the CNS. Current treatment modalities for this tumor type continue to be surgical resection, chemotherapy and radiotherapy, but overall survival is still relatively low for younger children with this disease. We initially tested the sensitivity of three ATRT cell lines to 48 chemotherapeutic agents and selective inhibitors. Our results indicated that two cell lines CHLA-04-ATRT and CHLA-05-ATRT were sensitive to FGFR inhibition. Further analysis of a panel of five ATRT cell lines indicated that the cell lines CHLA-04-ATRT, CHLA-05-ATRT, CHLA-266 and BT-12 were sensitive to the FGFR inhibitors PD173074, BGJ398 and AZD4547. Interestingly, the ATRT cell line CHLA-06-ATRT did not show sensitivity to FGFR inhibition. Analysis of FGFR protein expression by immunoblot analysis indicated that CHLA-04-ATRT and CHLA-266 cells expressed both FGFR1 and FGFR2, while the cell lines CHLA-05-ATRT and BT-12 expressed only FGFR1. The CHLA-06-ATRT cell line did not show expression of any FGFRs. We further demonstrated functional FGFR activity in the ATRT cell lines by treating growth factor-deprived ATRT cells with FGF-1 or FGF-2, both of which led to dose dependent cell growth only in the cell lines expressing FGFR1 or FGFR2. We are currently investigating the expression of FGFR isoforms in ATRT patient samples. Taken together, our results demonstrate a potential role for FGFR1 and FGFR2 in the survival and proliferation of ATRT cells and indicate that FGFRs could serve as potential targets for therapy in ATRT.

Citation Format: David O. Azorsa, Oliver B. Pepper, Madhavi Chakravadhanula, David W. Lee, Justin J. Montoya, Victor Ozols, Ratan Bhardwaj, Robert J. Arceci. A role for fibroblast growth factor receptors in growth of atypical teratoid rhabdoid tumor cells and as potential therapeutic targets. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3923. doi:10.1158/1538-7445.AM2015-3923
Abstract 3930: Targeted cancer gene sequencing identifies potential causative novel candidate mutations among Caucasian colon carcinogenesis
Authors: Ashktorab Hassan , Nickerson Michael , Varma Sudhir , Mokarram Pooneh , Naghibalhossaini fakhraddin , Moore Lee , Boland Joseph , Bass Sara , Yeager Meredith , Haung Wen-Yi , Brim Hassan
Published: August 2015

Much of the sporadic colorectal cancer's (CRC) underlying genetic ‘cancer driver’ mutations are unknown in patients from specific ethnic groups. Here, we report the identification of distinct novel variants from adenoma and CRC patients in mismatch repair (MMR) genes MHS3 and MSH6, and in PIK3CA and APC.

We developed a panel of 20 frequently altered colon cancer genes comprised of ACVR2A, APC, ARID1A, BRAF, FAM123B, FBXW7, KRAS, MSH2, MSH3, MSH6, NRAS, PIK3CA, POLE, PTEN, SMAD2, SMAD4, SOX9, TCF7L2, TGFBR2, and TP53 for targeted sequencing in 65 colon tissues, comprised of 1 normal tissue sample, 2 adenomas, and 63 tumors. Multiplex PCR and Ion Torrent sequencing was used to examine 98.8% of the targeted exons and splice junctions at a depth of sequencing that allowed for high confidence variant calling (most bases were covered by ≥ 500 reads). After alignment and variant calling, we annotated the variants with information from the 1000 Genomes Project, COSMIC, Polyphen2, and PFAM domain and transcription factor motifs. Excluding synonymous SNVs, 3 variants in adenoma, and 692 variants in tumors were detected. Two were known pathogenic variants (MSH6 p.R965H and APC p.R1432X). Novel variants (286, 244, and 115) were found in MMR genes (MSH6 and MSH3), APC, and PIK3CA, respectively. Most of the MMR (n = 98), APC (n = 128), and PIK3CA (n = 43) variants are deleterious. Notably, among the 63 CRC cases, [(29/61 = 46%) 46%) for MSH3, MSH6] [(19/63 = 30%) (30%) for PIK3CA] [31/63 = 49%) (49%) for APC)] carried likely deleterious MMR, PIK3CA and APC mutations, respectively, suggesting the value of a broad cancer gene panel. These finding further highlight the relevance of APC, PIK3CA genes in CRC onset but also the potential underestimation of the MSI-H in sporadic CRC as many of the novel mutations in MMR gens detected here were of a deleterious nature.

Citation Format: Hassan Ashktorab, Michael Nickerson, Sudhir Varma, Pooneh Mokarram, fakhraddin Naghibalhossaini, Lee Moore, Joseph Boland, Sara Bass, Meredith Yeager, Wen-Yi Haung, Hassan Brim. Targeted cancer gene sequencing identifies potential causative novel candidate mutations among Caucasian colon carcinogenesis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3930. doi:10.1158/1538-7445.AM2015-3930
Abstract 3938: MafB and ETS2 potentiate Notch signaling in T-ALL
Authors: Pajcini Kostandin , Xu Lanwei , Lee Curtis , Cherry Sara , Pear Warren
Published: August 2015

Despite the high frequency of activating Notch mutations in human T-cell acute lymphoblastic leukemia (T-ALL), these naturally occurring Notch1 mutant alleles fail to efficiently drive leukemia onset, indicating a requirement for collaborating events that potentiate the Notch oncogenic program. We performed an in vitro gain-of-function screen to identify gene products that enhanced the signaling strength of naturally occurring Notch mutants. Both MafB and ETS2 emerged as candidates that individually potentiated Notch signaling, and when co-expressed, they dramatically enhanced signaling strength comparable to the levels induced by MAML1, a core component of the Notch transcriptional complex. Knockdown of MafB in mouse T-ALL cell lines suppressed the expression of common Notch target genes, including cMyc and Hes1, and sustained knockdown of MafB expression impaired T-ALL cell growth in cell competition assays. Notably, MafB is sufficient to enhance the oncogenic potential of naturally occurring Notch mutants in vivo, by significantly accelerating T-ALL onset. Moreover, this enhanced Notch pathway activation required MafB-ETS2 interaction, and facilitated the recruitment of PCAF and p300. Although all primary human T-ALL samples that express MafB display an elevated Notch pathway signature, the T-ALL samples positive for high levels of MafB expression, exhibit low levels of Notch1 expression, further suggesting a role for MafB as an enhancer. These data present a novel mechanism for the enhancement of the oncogenic potential of weak Notch mutants in leukemia and introduce MafB as a potential therapeutic target in T-ALL.

Note: This abstract was not presented at the meeting.

Citation Format: Kostandin Pajcini, Lanwei Xu, Curtis Lee, Sara Cherry, Warren Pear. MafB and ETS2 potentiate Notch signaling in T-ALL. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3938. doi:10.1158/1538-7445.AM2015-3938
Abstract 3942: Oncogenic BRAF fusion induces MAPK-pathway activation targeted by MEK inhibitor and phosphatidylinositol 3-kinase inhibitor combination treatment in mucosal melanoma
Authors: Kim Han Sang , Jang Kang Won , Jung Minkyu , Kim Soo Hee , Kim Tae-Min , Cho Byoung Chul
Published: August 2015

Background: Although remarkable progress has been made in the genomic characterization of cutaneous melanoma, the genetic alterations of primary mucosal melanoma (PMM) remain largely unknown. This study aims to investigate the recurrent mutations in PMM and to characterize the oncogenic potential of the newly identified ZNF767-BRAF fusion as well as its therapeutic implications in PMM.

Methods: A total of 46 PMMs underwent exon-capture sequencing of 111 cancer-associated genes to identify somatic non-synonymous mutations, copy number alterations, and structural variations. A cancer cell with a novel ZNF767-BRAF fusion was established from a patient with PMM of the respiratory tract, who was refractory to vemurafenib. To characterize the oncogenic functions of the novel ZNF767-BRAF fusion, we performed western blot, soft agar growth assay and in vivo tumor. The inhibitory effects of vemurafenib, trametinib, buparlisib and LEE011 monotherapy and their combination treatment were measured in vitro and in vivo.

Results: Targeted exon-capture sequencing of 46 PMMs revealed a total of 48 somatic non-synonymous mutations in 26 genes. Eight genes harbored recurrent mutations on NRAS (n = 6; 13%) and TP53 (n = 4; 9%) along with BRAF, KIT, MAP3K9, C8A, KRAS, FGFR3 (all observed in two cases; 4%). Of note, a novel ZNF767-BRAF fusion was identified in a PMM of the respiratory tract refractory to vemurafenib treatment. ZNF767-BRAF fusion was validated by Sanger sequencing. We show that ZNF767-BRAF encoded protein promotes the dimerization of RAF and potently activates mitogen-activated protein kinase pathway. The ectopic expression of ZNF767-BRAF fusion induced tumor formation as shown in colony forming assay and tumor xenograft model. The treatment of trametinib inhibited the growth of PMM cells harboring the ZNF767-BRAF fusion but vemurafenib showed no inhibitory effects. Compared with either agent alone, the combination of trametinib and buparlisib showed synergistic antitumor effects in vitro and in vivo.

Conclusions: The genomic makeup of PMM is remarkably diverse and distinct from that of cutaneous melanoma. BRAF fusion defines a new molecular subset of PMM and can be therapeutically targetable by the combination of MEK inhibitor and PI3K Inhibitor.

Citation Format: Han Sang Kim, Kang Won Jang, Minkyu Jung, Soo Hee Kim, Tae-Min Kim, Byoung Chul Cho. Oncogenic BRAF fusion induces MAPK-pathway activation targeted by MEK inhibitor and phosphatidylinositol 3-kinase inhibitor combination treatment in mucosal melanoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3942. doi:10.1158/1538-7445.AM2015-3942
Abstract 3955: A primary cell-based assay to identify initiating oncogenic events in neuroblastoma
Authors: Olsen Rachelle , Otero Joel , Yin Zhirong , Garcia-Lopez Jesus , Freeman Kevin
Published: August 2015

The pediatric cancer neuroblastoma (NBL) forms from immature cells of the sympathetic nervous system (SNS). A prominent category of high-risk NBL is MYCN amplification with loss of heterozygosity (LOH) at 1p36. This LOH includes the chromatin remodeling factor, CHD5, a proposed tumor suppressor of NBL and can include ARID1A, a component of the SWI/SNF chromatin remodeling complex.

To identify genetic changes that cooperate with MYCN amplification in initiating NBL we developed an assay based on oncogenic transformation of neural crest cells (NCCs), the progenitors of the SNS. Isolation of NCCs from E9.5 mouse embryos resulted in a population that was ∼90% positive for the NCC markers p75, Sox10, and Ascl1, suggesting a highly enriched population.

Next, we tested the ability of N-Myc infected NCCs to form tumors in vivo. In pilot experiments using NCCs with compromised p53, we obtained subcutaneous tumors following injection of NCCs in nude mice. Resulting tumors were diffusely positive for neuronal markers such as synaptophysin, MAP2, and Tyrosine hydroxylase (TH). Tumors also contained focal areas of divergent differentiation characteristic of melanoma, Schwannoma, and osteosarcoma, all cancers that can arise from NCC lineages. To our knowledge, this is the first report of cancer from transformed primary neural crest cells.

To more closely model NBL genetics, we replaced loss of p53 with shRNA to knockdown CHD5 and ARID1A prior to N-Myc infection. Injection of the resulting NCCs also resulted in tumor formation in nude mice. Tumors were uniformly positive for neuronal markers including TH and Ascl1, as well as Phox2B, a marker of undifferentiated NBL, and with no evidence of other cancers. Future studies in understanding the interactions of N-Myc overexpression with loss of these chromatin remodeling factors should further increase our understanding of NBL development.

Citation Format: Rachelle Olsen, Joel Otero, Zhirong Yin, Jesus Garcia-Lopez, Kevin Freeman. A primary cell-based assay to identify initiating oncogenic events in neuroblastoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3955. doi:10.1158/1538-7445.AM2015-3955
Abstract 396: Ibrutinib exerts potent antifibrotic activity in a mouse model of pancreatic adenocarcinoma
Authors: Massó-Vallés Daniel , Jauset Toni , Serrano Erika , Sodir Nicole M. , Pedersen Kim , Affara Nesrine I. , Whitfield Jonathan R. , Beaulieu Marie-Eve , Evan Gerard I. , Elias Laurence , Arribas Joaquín , Soucek Laura
Published: August 2015

Pancreatic ductal adenocarcinoma (PDAC) is a disease with a poor prognosis characterized by a dense stromal fibroinflammatory reaction that is a significant obstacle to effective therapy. The desmoplastic stroma comprises many inflammatory cells, in particular mast cells that are a key component of the PDAC microenvironment and whose infiltration often correlates with poor patient outcome. Reducing the stroma is of key importance for improving the standard of care. We have previously successfully utilized ibrutinib, a clinically-approved Bruton's Tyrosine Kinase inhibitor, as a mast cell blocker in a mouse model of β cell tumorigenesis. Here, we made use of a transgenic mouse model of PDAC and patient derived xenografts (PDXs) to assess ibrutinib's impact on the tumor microenvironment and animal survival. Across these various experimental pre-clinical models, we show that ibrutinib significantly diminishes tissue fibrosis, increases survival and improves the outcome of standard care. Our results suggest that ibrutinib could be deployed to one more type of cancer and open the way to new clinical trials assessing its therapeutic impact in PDAC patients.

Citation Format: Daniel Massó-Vallés, Toni Jauset, Erika Serrano, Nicole M. Sodir, Kim Pedersen, Nesrine I. Affara, Jonathan R. Whitfield, Marie-Eve Beaulieu, Gerard I. Evan, Laurence Elias, Joaquín Arribas, Laura Soucek. Ibrutinib exerts potent antifibrotic activity in a mouse model of pancreatic adenocarcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 396. doi:10.1158/1538-7445.AM2015-396
Abstract 3968: Identification of microRNAs markers in patients with oral squamous cell carcinomas for the presence of lymph nodes metastasis
Authors: Zamunér Fernando Tadeu , Carvalho André Lopes , Kowalski Luiz Paulo , Vettore André Luiz
Published: August 2015

Introduction: Oral cancer is the most common subset of head and neck squamous cell carcinomas (OSCC). These tumors often have an aggressive clinical outcome hallmarked by a propensity for local invasion and regional nodal metastasis. The presence of metastatic disease in cervical lymph nodes is an important prognostic factor and it influences in the choice of the therapeutic strategy to be adopted. Thus, the identification of molecular markers able to select patients who are at high risk for lymph node metastasis and/or locoregional recurrence could be of great value. Recent studies have shown that microRNAs can be regulators of the tumor metastasis process by the activation of various signaling pathways involved in this process being useful in diagnosis, determination of disease prognosis and prediction of therapeutic response. PURPOSE: to identify microRNAs expressed in OSCC associated with the presence of lymph node metastasis. METHODS: Tumor samples were obtained from patients diagnosed with OSCC at the Head and Neck Surgery Department, Cancer Hospital of Barretos. For microRNA microarrays experiments, total RNA from 32 tumor samples (12 without lymph node metastasis and 12 with metastasis) was extracted labelled and hybridizated Analysis of microarray data was performed using GeneSpring GX 12.0 and R program. RESULTS: This analysis allowed the identification of 10 microRNAs differentially expressed in the comparison between metastatic and non-metastatic OCCS cases. These microRNAs will be validate and selected by qRT-PCR in an independent cohort of OSCC patients and correlate the molecular data with the clinical features of the patients.

Citation Format: Fernando Tadeu Zamunér, André Lopes Carvalho, Luiz Paulo Kowalski, André Luiz Vettore. Identification of microRNAs markers in patients with oral squamous cell carcinomas for the presence of lymph nodes metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3968. doi:10.1158/1538-7445.AM2015-3968
Abstract 3971: Genome-wide tissue-based microRNA signature in healthy women predicting breast cancer risk
Authors: Taslim Cenny , Weng Daniel Y. , Brasky Theodore M. , Dumitrescu Ramona G. , Huang Kun , Kallakury Bhaskar V. s. , Krishnan Shiva , Llanos Adana A. , Marian Catalin , Schneider Sallie S. , Spear Scott L. , Troester Melissa A. , Freudenheim Jo L. , Geyer Susan , Shields Peter G.
Published: August 2015

Introduction: Small non-coding microRNAs (miRNAs) play important roles in both normal breast development and breast carcinogenesis. The goal of this study is to identify miRNAs in normal breast tissues which are related to breast cancer risk.

Materials and Methods: We used a high-throughput digital counting of miRNAs without amplification (Nanostring®) to examine miRNA expression in 161 reduction mammoplasty (RM) tissues from two independent studies. A multivariate model was used to identify miRNAs associated with breast cancer risk (based upon Gail risk scores) in a training study (n = 90) then the model was validated in a replication study (n = 71). Risk-related microRNAs were then evaluated in serum for associations with real breast cancer cases using publically available prospective cohort (Sister Study, n = 410).

Results: We identified a 41-miRNA signature in healthy women distinguishing high risk from low risk women with a prediction accuracy of 82% (95% CI = 80% to 87%) in the training study. Predictive accuracy was 69% (95% CI = 65% to 73%) in the replication study. 34 of 41 serum miRNAs that mapped to public data predicted women who developed breast cancer within 18 months after blood draw from those who remained cancer free with accuracy of 59% (95% CI = 57% to 61%). We have also shown that these accuracies were significantly higher than random chance (P < 0.0001). IPA canonical pathway analysis revealed that the risk-related microRNAs targets were significantly enriched for HER-2 signaling in breast cancer, and estrogen-dependent breast cancer signaling, and other important cancer pathways such as molecular mechanisms of cancer, PI3K/AKT signaling, PTEN signaling, and TGF-beta signaling.

Conclusion: Our results indicate that miRNA profiling from breast tissue of healthy patients may identify clinically useful predictors of breast cancer risk and these miRNAs may also work as non-invasive biomarker for early breast cancer prediction.

Citation Format: Cenny Taslim, Daniel Y. Weng, Theodore M. Brasky, Ramona G. Dumitrescu, Kun Huang, Bhaskar V. s. Kallakury, Shiva Krishnan, Adana A. Llanos, Catalin Marian, Sallie S. Schneider, Scott L. Spear, Melissa A. Troester, Jo L. Freudenheim, Susan Geyer, Peter G. Shields. Genome-wide tissue-based microRNA signature in healthy women predicting breast cancer risk. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3971. doi:10.1158/1538-7445.AM2015-3971
Abstract 3989: MicroRNA-16 and microRNA-193b as serological predictors for chemoradiation response in esophageal squamous cell carcinoma patients
Authors: Lee Nikki P. , Lai Kenneth K. , Chan Kin Tak , Tong Daniel K. , Law Simon
Published: August 2015

Esophageal cancer is one of the most deadly malignancies along the gastrointestinal tract. Esophageal squamous cell carcinoma (ESCC) is the most common cell type occurring at the upper and middle part of the esophagus. Surgery remains the mainstay treatment for this malignancy, however the long-term prognosis is modest with a 5-year overall survival of approximately 30%. To improve the treatment outcome of this cancer, multimodal treatment strategies including neoadjuvant chemoradiation have been implemented. Such pre-surgery treatment indeed leads to better clinical outcome and increases the chance for cure. Unfortunately patients who do not respond may have to undergo prolonged, unnecessary and potentially toxic treatments with no benefits. Currently, there is no reliable method for chemoradiation response prediction. To address this clinical limitation, we have established a non-invasive assay to stratify chemoradiation responders from non-responders. This study is systematically divided into three phases. In the discovery phase (phase I), we found 23 human microRNAs (out of 742 human microRNAs) with more than 2.5-fold elevation in serum from chemoradiation responders when compared to non-responders. In the selection and small-scale validation phase (phase II), small-scale validation in an independent cohort showed that microRNA-16 and microRNA-193b were two most powerful candidates for indicating chemoradiation responders. In the large-scale validation phase (phase III), we have further validated and confirmed the feasibility of microRNA-16 and miroRNA-193b as discriminators for chemoradiation response in a large cohort of more than a hundred patients. Standard curves were also constructed for these two microRNAs for copy number quantitation. In all, we have newly established a highly sensitive and specific blood-based assay with specific cut-offs for discriminating ESCC patients who are responsive to chemoradiation. This assay works by quantitating the serum level of microRNA-16 and microRNA-193b in ESCC patients, such that patients would be advised to receive pre-surgery chemoradiation if they have elevated levels of circulatory microRNA-16 and microRNA-193b. The development of this assay will benefit patient care by formulating individualized treatment plans.

Note: This abstract was not presented at the meeting.

Citation Format: Nikki P. Lee, Kenneth K. Lai, Kin Tak Chan, Daniel K. Tong, Simon Law. MicroRNA-16 and microRNA-193b as serological predictors for chemoradiation response in esophageal squamous cell carcinoma patients. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3989. doi:10.1158/1538-7445.AM2015-3989
Abstract 3990: Identification of microRNAs from exosomes of bulk and stem cells from prostate cancer by next generation sequencing
Authors: Sanchez Catherine A. , Andahur Eliana , Valenzuela Rodrigo , Castellon Enrique , Ramos Christian , Triviño Juan Carlos
Published: August 2015

Cancer stem cells (CSCs) are related with cancer initiation, metastasis, resistance and recurrence, by regulating their environment in different ways, including the release of exosomes that modify surrounding and distant cells. Exosomes contain, among other molecules, miRNAs. By identifying the entire spectrum of miRNAs included in exosomes from prostate cancer (PCa) cells by next generation sequencing (NGS), is possible to identify miRNAs specifically released by CSCs, and those related with microenvironment regulation.

We stablished adherent primary cell cultures (bulk cells) from PCa tissue from 5 patients. Later, from these cells we obtained CSCs enriched prostatospheres. From bulk and CSCs cultures we purified exosomes by precipitation (ExoQuick-TC, SBI) for miRNAs extraction (mirVana, Ambion). Quality of miRNAs was assessed by Bioanalizer (Agilent Technologies). NGS was performed in the Illumina platform (MiSeq, Illumina) according to manufacturer instructions. Bioinformatic analyses were carried out by Sistemas Genómicos. Overexpressed miRNAs were detected in plasma exosomes by TaqMan miRNA Assays (Lifetechnologies).

We identified 1839 miRNAs in exosomes, and 223 species were differentially expressed between bulk and CSCs, but only 26 significantly. Nineteen of them were known species including 6 that were overexpressed in CSCs respect to bulk cells. These miRNAs target genes related with cell differentiation, matrix remodeling and genic expression. From all miRNA detected, 3 were highly overexpressed among all, and were detected in exosomes isolated from plasma of men with prostatic disease.

Our study extensively characterizes the miRNA from exosomes of PCa cells. Exosomes contain high diversity of miRNAs, different in CSCs and bulk cells, suggesting differential regulation of their microenvironment. From these, CSCs exosome-related miRNAs could be released to bloodstream and play a role in PCa progression and metastasis (FONDECYT 11121525/1140417).

Citation Format: Catherine A. Sanchez, Eliana Andahur, Rodrigo Valenzuela, Enrique Castellon, Christian Ramos, Juan Carlos Triviño. Identification of microRNAs from exosomes of bulk and stem cells from prostate cancer by next generation sequencing. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3990. doi:10.1158/1538-7445.AM2015-3990
Abstract 4001: Array profiling identified upregulated miRNAs and target genes and pathways in laryngeal squamous cell carcinoma
Authors: Lian Meng , Liu Xiaoqin , Han Demin , Fang Jugao , Chen Zhong
Published: August 2015

Background: Laryngeal squamous cell carcinoma (LSCC) is prevalent in the Chinese population, where smoking is a major carcinogen. MicroRNAs (miRNAs) and their gene targets potentially contribute to the multistep regulatory processes of LSCC development.

Objective: The purpose of this study is to investigate and validate aberrant expressed miRNAs in LSCC, and analyze the putative target genes of the screened miRNAs.

Methods: Six pairs of primary Stage II-IV LSCC and adjacent non-neoplastic tissues were surgically removed for miRNA array analysis using the Affymetrix GeneChip miRNA Array 3.0, containing probes for 5683 miRNAs, and the significantly altered miRNAs were validated by the qRT-PCR in an independent cohort of 36 LSCC patients. MiRNA gene targets were predicted using TargetScan, PITA and microRNAorg in GeneSpring 12.5 software. The functional categories and pathways of the target genes were analyzed by GO category and KEGG pathway database.

Results: MiRNAs arrays identified seven up-regulated miRNAs, including hsa-miR-224, hsa-miR-183, hsa-miR-23a, hsa-miR-675, hsa-miR-27a, hsa-miR-503 and hsa-miR-4800-3p in LSCC by the SAM method, and qRT-PCR was performed to validate the screened miRNAs. 72 putative target genes were identified by GeneSpring 12.5 software. These putative target genes are involved in biological processes, such as metabolic, biological regulation, membrane, protein binding, ion binding by GO category analysis, and are the molecular components in MAPK signaling, adherence, and junction pathways by KEGG pathway database analysis.

Conclusions: Seven upregulated miRNAs were identified in LSCC tissues by microarrays, and validated by qRT-PCR. The putative target genes were also identified. These up-regulated miRNAs could serve as potential biomarkers for LSCC diagnosis and therapy.

Citation Format: Meng Lian, Xiaoqin Liu, Demin Han, Jugao Fang, Zhong Chen. Array profiling identified upregulated miRNAs and target genes and pathways in laryngeal squamous cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4001. doi:10.1158/1538-7445.AM2015-4001
Abstract 4006: MiRNA profiling in esophageal precancerous for malignancy progression risk assessment
Authors: Srovnal Josef , Slaby Ondrej , Ehrmann Jiri , Gregar Jan , Radova Lenka , Staffova Katerina , Hajduch Marian
Published: August 2015

Background: Barrett's esophagus (BE) is a metaplastic disease with risk to develop esophageal adenocarcinoma (EAC). Current screening method for BE patients (endoscopy and biopsy) is discomfort, expensive and relatively risky for patients. Moreover, the procedure is complicated by subjective histopathology examination and only 2% of BE patients annually progress to adenocarcinoma. The aim of our study was to identify miRNA profiles in order to predict cancer progression in high-risk BE patients.

Methods and patients: FFPE samples were obtained from 113 patients (35 EAC, 21 low-grade dysplasia of esophagus, 33 BE and 24 healthy control). There were 74 males and 39 females; mean age was 55.9 years, range 15 - 94 years. Total RNA was purified from FFPE biopsy samples using proteinase K treatment followed by RNeasy kit (Qiagen). Microarray analysis was performed using the GeneChip miRNA 3.0 Array (Affymetrix). The data were analyzed using “R” software and the Bioconductor package.

Results: In total, 113 GeneChip miRNA 3.0 Arrays were performed in 113 patients. Expression profiles of 1733 human miRNAs were analyzed. In supervised clustering analysis we found 8 differentially expressed miRNAs (p<0.001 or logFC 2) in BE patients compared to EAC patients and 10 differentially expressed miRNAs (p<0.001 or logFC 2) in LGD patients compared to EAC patients. One of the dominant findings was decreased expression of miR-223 in EAC and LGD samples, which is known to function as a tumor suppressor blocking E2F1. Using LOOCV classifier with 13 miRNAs signature we were able to correctly classify 86 from 113 samples (AUC = 0.840). Prediction model fails mostly in LGD prediction (18/21), probably due to incorrect pathological classification.

Conclusion: We have identified miRNA profiles and signature allowing for better diagnostics of cancer progression in BE and LGD patients, which, however, will require further validation.

Acknowledgment: This work was financially supported by IGA UP LF 2014_019, NPU LO1304, CZ.1.05/2.1.00/01.0030 and CZ.1.07/2.3.00/30.0004.

Citation Format: Josef Srovnal, Ondrej Slaby, Jiri Ehrmann, Jan Gregar, Lenka Radova, Katerina Staffova, Marian Hajduch. MiRNA profiling in esophageal precancerous for malignancy progression risk assessment. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4006. doi:10.1158/1538-7445.AM2015-4006
Abstract 4014: Whole-genome microRNA profiling identifies prognostic serum biomarkers of esophageal adenocarcinoma
Authors: Wei Yongyue , Su Li , Christiani David C.
Published: August 2015

Background: Cell free circulating microRNAs (cfmiRNAs) have been recognized as robust and stable biomarkers of cancers. However, little is known about the prognostic significance of cfmiRNAs in esophageal adenocarcinoma (EA). In this study, we explored whether specific cfmiRNA profiles could predict EA prognosis and whether H. pylori (HP) infection status could influence the association between cfmiRNAs and EA survival outcomes.

Methods: At discovery stage, we profiled 1075 cfmiRNAs in 6 pooled serum samples from 30 EA patients and 30 cancer-free control subjects. After filtration, a total of 589 cfmiRNAs were stably detectable in the serum samples. The most significantly differential expressed cfmiRNAs were then validated the association with overall survival (OS) in an independent cohort of 82 patients, using Log-Rank test and multivariate Cox regression models in overall samples or stratified by HP infection status. qRT-PCR was used for cfmiRNA profiling in validation. HP infection status was determined by immunoblotting assay.

Results: We identified a 21-cfmiRNA panel that could distinguish EA patients from cancer-free subjects (multiple response analysis by Hotelling T test, P = 3.0E-12). In validation, the OS analysis among HP-positive subtype patients showed that no cfmiRNA was significantly associated with EA prognosis. In HP-negative patients, however, 9 cfmiRNAs were significantly associated with OS (minimal P = 1.9E-4, all FDR adjusted q-value < 0.05). A risk-score analysis by combining the 9 cfmiRNA biomarkers showed a worse outcome in high-risk group (HR = 12.82, P = 0.0075) that showed better prediction of prognosis than individual cfmiRNA alone.

Conclusions: Our findings suggest that deregulated cfmiRNAs may contribute to EA prognosis and HP infection status may modify the association between cfmiRNAs and EA survival.

Citation Format: Yongyue Wei, Li Su, David C. Christiani. Whole-genome microRNA profiling identifies prognostic serum biomarkers of esophageal adenocarcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4014. doi:10.1158/1538-7445.AM2015-4014
Abstract 4046: Interleukin-10 and programmed death-1 cooperate to regulate tumor antigen-specific CD8+ T cells in melanoma patients
Authors: Sun Zhaojun , Fourcade Julien , Pagliano Ornella , Chauvin Joe-Marc , Sander Cindy , Kirkwood John M. , Zarour Hassane M.
Published: August 2015

With the recent success of immune checkpoint blockade in the clinic, it is expected that strategies aimed at counteracting the negative immunoregulatory networks in the tumor microenvironment will develop into potent therapy for melanoma. Here, we report that programmed death-1high (PD-1high) tumor antigen (TA)-specific CD8+ T cells present at periphery and at tumor sites in patients with advanced melanoma upregulate IL-10 receptor (IL-10R) expression. IL-10 is produced by multiple PBMC subsets in melanoma patients and acts directly on IL-10R+ TA-specific CD8+ T cells to impede proliferation and increase apoptosis. PD-1 blockade augments IL-10R expression by TA-specific CD8+ T cells, suggesting that PD-1 blockade renders PD-1+ TA-specific CD8+ T cells more sensitive to the immunosuppressive effects of endogenous IL-10. As a result, we show that IL-10 blockade adds to PD-1 blockade to increase the expansion and functions of TA-specific CD8+ T cells. Collectively, our findings support dual IL-10 and PD-1 blockade to counteract melanoma-induced T cell dysfunction and enhance TA-specific CD8+ T cell responses in patients with advanced melanoma.

Citation Format: Zhaojun Sun, Julien Fourcade, Ornella Pagliano, Joe-Marc Chauvin, Cindy Sander, John M. Kirkwood, Hassane M. Zarour. Interleukin-10 and programmed death-1 cooperate to regulate tumor antigen-specific CD8+ T cells in melanoma patients. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4046. doi:10.1158/1538-7445.AM2015-4046
Abstract 405: Kindlin-2 regulates integrin function and sensitivity to docetaxel in prostate cancer cells
Authors: Plow Edward F. , Das Mitali , Hirbawi Jamila , Sossey-Alaoui Khalid
Published: August 2015

Over the past decade, many lines of evidence have emerged to implicate the three members of the kindlin family of FERM domain proteins as important regulators of integrin adhesion receptors. Specifically, kindlins appear to cooperate with talin to be essential regulators of the ligand binding function of integrins and thereby enhance the adhesive and migratory responses of cells. Kindlin-2 (FERMT2) is the most broadly distributed of the kindlins. To explore the relationship between integrin activation and kindlin-2, prostate cancer cell lines were screened and LNCaP prostate cancer cells were found to express relatively low levels of kindlin-2 and endogenous levels of beta-1 integrin on their surface. A portion of this integrin was expressed in an activated state as detected with an activation specific antibody, HUTS-4. These cells were transfected with vectors for kindlin-2, with or without talin-head domain, a combination that leads to activation of β3 integrins in model cells. Transfections with kindlin-2 suppressed β1 integrin activation on the surface of the LNCaP cells. This inhibitory effect was abolished by mutation of kindlin-2 that prevented its binding to integrin cytoplasmic tail. This inhibitory response to kindlin-2 on β1 integrin was in stark contract to the effects of kindlin-2 plus talin-head expression on β3 function, where co-expression of the same constructs enhanced integrin activation. A mutation in kindlin-2, deletion of its last 15 amino acids, that blocks activation of β3 integrins without preventing its binding to the integrin, also prevents its inhibition of β1 function. We next looked at the effects of kindlin-2 on docetaxel induced apoptosis as monitored with Annexin V staining and cell death as monitored by propidium iodide staining (both by flow cytometry). Compared to LNCaP cells expressing EGFP alone, cells overexpressing EGFP-kindlin-2 were protected against apoptosis and cell death. This effect was dependent upon binding of kindlin-2 to the cytoplasmic tail of integrins. Hence, the capacity of kindlin-2 to influence integrin function depends upon the integrin subfamily and can range from activation to suppression of function. Kindlin-2, which is often overexpressed in transformed cells, may desensitize cells towards chemotherapeutic agents.

Citation Format: Edward F. Plow, Mitali Das, Jamila Hirbawi, Khalid Sossey-Alaoui. Kindlin-2 regulates integrin function and sensitivity to docetaxel in prostate cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 405. doi:10.1158/1538-7445.AM2015-405
Abstract 4050: Association of long term NK cell culture and TIMP3 over-expression with NK cell reduced susceptibility to leukemia and epithelial cancer cell induced damage
Authors: Sconocchia Giuseppe , Arriga Roberto , Caratelli Sara , Lanzilli Giulia , Coppola Andrea , Spagnoli Giulio C. , Venditti Adriano , Amadori Sergio , Lauro Davide , Del Principe Maria I. , Maurillo Luca , Buccisano Francesco , Capuani Barbara , Wang Xinhui , Ferrone Soldano
Published: August 2015

Allogeneic leukemia and cancer epithelial (CE) cells have been recently shown to induce,in vitro, NK cell abnormalities (NKCA). The latter include NK cell depletion and apoptosis as well as natural cytotoxic receptor (NCR) CD16 down-regulation. The potential negative impact of leukemia and CE cell induced NKCAs on the in vivo anti-tumor activity of NK cells and the limited information about strategies to protect NK cells from NKCAs have prompted us to characterize NKCAs and to develop strategies to counteract them. NKCA induction by leukemia and CE cells is influenced neither by interleukin-2 (IL-2) treatment nor by HLA class I antigen expression, but is abrogated by a long term culture of peripheral blood mononuclear cells (PBMCs) at 37 ºC. Following a 10 day culture of PBMCs, NK cells become resistant to cancer cell induced NKCA but maintain their cytotoxic activity. Actinomycin D restores the susceptibility of long term NK (LTNK) cells to NKCAs suggesting that the generation of resistance to NKCAs requires RNA transcription. TAPI-0, a functional analogue of the tissue inhibitor of metalloproteinases (TIMP) 3, inhibits cancer cell induced NKCAs indicating a role for a restricted number of metalloproteinases (MMPs) in the generation of this phenomenon. This conclusion is supported by the reduced susceptibility to cancer cell induced NKCAs of LTNK cells in which TIMP3 gene and protein are over-expressed. Finally, subcutaneous infusion of IL-2 stimulated LTNK cells inhibited subcutaneous growth of the monoblastic leukemia cells, ML-2 in CB17scid mice while short term, IL-2 stimulated NK cells did not suggesting that LTNK cells have significant anti-leukemia capacity in vivo. This information may contribute to the rational design of targeted strategies to enhance the efficacy of NK cell-based-immunotherapy for the treatment of myeloid leukemias with haploidentical or allogeneic NK cells.

Citation Format: Giuseppe Sconocchia, Roberto Arriga, Sara Caratelli, Giulia Lanzilli, Andrea Coppola, Giulio C. Spagnoli, Adriano Venditti, Sergio Amadori, Davide Lauro, Maria I. Del Principe, Luca Maurillo, Francesco Buccisano, Barbara Capuani, Xinhui Wang, Soldano Ferrone. Association of long term NK cell culture and TIMP3 over-expression with NK cell reduced susceptibility to leukemia and epithelial cancer cell induced damage. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4050. doi:10.1158/1538-7445.AM2015-4050