bluuebunny/crossref_metadata_2025 huggingface dataset:
Association between Serum 25-Hydroxyvitamin D Level and Rheumatoid Arthritis
Authors: Cen Xiaomin , Liu Yuan , Yin Geng , Yang Min , Xie Qibing
Published: May 2015

The objective of this study is to examine and evaluate whether serum 25(OH)D is associated with disease activity in patients with rheumatoid arthritis (RA). Our results suggested that serum 25(OH)D in RA groups has significant lower level (35.99±12.59 nmol/L) than that in the normal groups (54.35±8.20 nmol/L,P<0.05). Based on the DAS28, patients with RA were divided into four subgroups, and no differences were found in the four groups (P>0.05). The 25(OH)D levels in complete remission, low disease activity, middle disease activity, and high disease activity group were32.86±12.26,33.97±13.28,38.41±10.64, and38.94±13.35 nmol/L, respectively. Based on the serum 25(OH)D levels, patients with RA were divided into inadequate group and normal group, and there were no significant differences in baseline characteristics and disease activity in the two groups. Our results showed that serum 25(OH)D levels in the inadequate group are significantly lower than those in the normal group. However, no correlations were found between 25(OH)D levels and disease activity among 116 patients with RA. The present findings will help to understand the association between 25(OH)D and disease activity of RA.
Hypomagnesemia Induced by Long-Term Treatment with Proton-Pump Inhibitors
Authors: Janett Simone , Camozzi Pietro , Peeters Gabriëlla G. A. M. , Lava Sebastiano A. G. , Simonetti Giacomo D. , Goeggel Simonetti Barbara , Bianchetti Mario G. , Milani Gregorio P.
Published: May 2015

In 2006, hypomagnesemia was first described as a complication of proton-pump inhibitors. To address this issue, we systematically reviewed the literature. Hypomagnesemia, mostly associated with hypocalcemic hypoparathyroidism and hypokalemia, was reported in 64 individuals on long-term proton-pump inhibitors. Hypomagnesemia recurred following replacement of one proton-pump inhibitor with another but not with a histamine type-2 receptor antagonist. The association between proton-pump inhibitors and magnesium metabolism was addressed in 14 case-control, cross-sectional studies. An association was found in 11 of them: 6 reports found that the use of proton-pump inhibitors is associated per se with a tendency towards hypomagnesemia, 2 found that this tendency is more pronounced in patients concurrently treated with diuretics, carboplatin, or cisplatin, and 2 found a relevant tendency to hypomagnesemia in patients with poor renal function. Finally, findings likely reflecting decreased intestinal magnesium uptake were observed on treatment with proton-pump inhibitors. Three studies did not disclose any relationship between magnesium metabolism and treatment with histamine type-2 receptor antagonists. In conclusion, proton-pump inhibitors may cause hypomagnesemia. In these cases, switching to a histamine type-2 receptor antagonist is advised.
Measurement of Coherence Bandwidth in UHF Radio Channels for Narrowband Networks
Authors: Sadowski Jaroslaw
Published: May 2015

This paper presents results of investigation on the coherence bandwidth of narrowband radio channels in 430 MHz band. The coherence bandwidth values were estimated from a power delay profile obtained by recording CDMA2000 forward channel signals during real-field measurements in various environments: medium city, flat terrain, and hilly terrain in northern Poland. The results of measurements are compared with characteristic parameters of UHF radio channel models defined for exemplary narrowband digital system from the TETRA standard. In all of the tested environments, the coherence bandwidth values during most of an observation time were much higher than 25 kHz. Therefore, the fading in tested UHF narrowband channels should be classified as flat fading.
Abstract 1449: p70 S6 kinase signals tristetraprolin/Dicer-mediated maturation of microRNA-145 to regulate tumor metastasis
Authors: Lam Sophia S.N. , Ip Carman K.M. , Wong Alice S.T.
Published: May 2015

Ovarian cancer is a highly metastatic tumor characterized by malignant ascites spheroids, which represent one of the most important prognostic factors of poor clinical outcome. Multicellular aggregates known as spheroids (MCS) are important for anchorage-independent growth and metastasis. MCS are also an important phenomenon of cancer stem cells. However, the factors that regulate MCS formation are largely unknown. MicroRNAs (miRNA) are short non-coding RNAs that are critically involved in different oncogenic events by actively modulating mRNA and protein synthesis. Despite their clinical significance, processes regulating miRNA biogenesis remain obscure. Here, we identify N-cadherin as a key regulator of MCS formation activated in response to p70 S6 kinase (p70S6K) signaling, a downstream effector of phosphatidylinositol 3-kinase/Akt which is hyperactive in human ovarian cancer. The results also identify a new mechanism of p70S6K function that mediates microRNA (miR)-145 coregulation of key transcription factors Twist and Sox-9, thereby enhancing N-cadherin and MCS formation. p70S6K regulates miR-145 by deactivating a distinct tristetraprolin (TPP)/Dicer program. The p70S6K phosphorylates TTP, leading to inhibition of interaction between TTP and Dicer and decreased TTP activity in miR-145 processing. Silencing of p70S6K enhances the TTP/Dicer interaction and its activating in regulating miR-145. These results providea novel regulatory mechanism of p70S6K involved in the formation and spread of MCS and insights on the development of new therapeutic targets (This work was supported by RGC grant HKU782111 and CUHK8/CRF/11R).

Citation Format: Sophia S.N. Lam, Carman K.M. Ip, Alice S.T. Wong. p70 S6 kinase signals tristetraprolin/Dicer-mediated maturation of microRNA-145 to regulate tumor metastasis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1449. doi:10.1158/1538-7445.AM2014-1449
Abstract 1455: Therapeutic synergy between novel tumor suppressor miR-520d-3p and EphA2-targeting siRNA in ovarian cancer
Authors: Shah Maitri , Lopez Gabriel Berestein , Sood Anil , Calin George
Published: May 2015

EphA2 has emerged as an important target for ovarian cancer therapy in recent years. In the present study, we identified miR-520d-3p as an independent favorable prognostic marker for epithelial ovarian cancer patients. We demonstrate that dual targeting of EphA2 in vitro using EphA2-siRNA and miR-520d-3p exhibits synergistic downregulation of EphA2 and inhibits cell proliferation, migration and invasion. In vivo treatment using DOPC (1,2-dioleoyl-sn-glycero-3-phosphatidylcholine) nano-liposomes conjugated with miR-520d-3p and EphA2-siRNA showed synergistic therapeutic efficacy and superior inhibition of tumor proliferation and angiogenesis than either monotherapy alone. We also identified that EphA2(low)/EphB2(low)/miR-520d-3p(high) gene signature predicts favorable clinical prognosis for ovarian cancer patients. This study addresses a new concept of RNA inhibition therapy by combining miRNA and siRNA to target oncogenic pathways altered in ovarian cancer.

Citation Format: Maitri Shah, Gabriel Berestein Lopez, Anil Sood, George Calin. Therapeutic synergy between novel tumor suppressor miR-520d-3p and EphA2-targeting siRNA in ovarian cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1455. doi:10.1158/1538-7445.AM2014-1455
Abstract 1456: MicroRNA improves the efficacy of imatinib on the treatment of chronic myeloid leukemia
Authors: Yi Bin , Ma Ruixia , Feng Xiangling , Xing Zhiwei , Zhang Xiaoguo , Piazza Gary , Xi Yaguang
Published: May 2015

Human chronic myelogenous leukemia (CML) is a malignancy of pluripotent hematopoietic cells that is caused by the deregulated activity of the tyrosine kinase that is encoded by the chimeric bcr-abl oncogene. Given that CML cells are usually highly resistant to chemotherapy, novel and more efficacious strategies against this disease are urgently needed. MicroRNA (miRNA) is a set of newly discovered 20-22 nucleotides RNA molecules, and they broadly repress their cognate target genes at post-transcriptional/trans-lational level. To date, there are approximately 2,500 human miRNA transcripts have been characterized, which are estimated to modulate more than 30% human genes by being involved in many essential normal biological and pathological processes including differentiation, proliferation, apoptosis, and tumorigenesis as well. In this study, we studied let-7a, a miRNA with tumor suppressive signature, for its therapeutic potential in CML. When treating CML K562 and KU812 cell by imatinib, let-7a can be significantly induced. Overexpression of let-7a can sensitize K562 and KU812 cells to imatinib, while knock-down of let-7a can increase the resistance of these cells towards this drug. The functional studies found that let-7a was able to promote apoptosis of CML cells, inhibit the tumor cell proliferation, and lead to a cell cycle arrest in the G0/G1 phase. These results demonstrate that let-7a is a potential target for treating CML by imatinib and/or other anti-cancer drugs. The pro-apoptosis and anti-proliferation roles of let-7a in CML cells may provide novel insights into discovery and development of more efficacious drugs for treating this malignant disease.

Citation Format: Bin Yi, Ruixia Ma, Xiangling Feng, Zhiwei Xing, Xiaoguo Zhang, Gary Piazza, Yaguang Xi. MicroRNA improves the efficacy of imatinib on the treatment of chronic myeloid leukemia. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1456. doi:10.1158/1538-7445.AM2014-1456
Abstract 1457: Development of miRNA-loaded polymeric nanoformulation for pancreatic cancer therapy
Authors: Arora Sumit , Swaminathan Suresh K. , Srivastava Sanjeev K. , Singh Seema , Panyam Jayanth , Singh Ajay P.
Published: May 2015

MicroRNAs (miRNAs) are small (18-22 nucleotide long) non-coding RNAs that play important roles in biological processes through post-transcriptional regulation of gene expression. Their aberrant expression and functional significance are reported in several human malignancies, including pancreatic cancer (PC). Recently, we identified miR-150 as a novel tumor suppressor microRNA in PC and demonstrated that its overexpression inhibits growth and malignant behavior of PC cells suggesting that restoration of miR-150 could serve as an effective therapeutic approach for pancreatic cancer. In the present study, we have developed a nanoparticle-based miR-150 delivery system and tested its therapeutic efficacy by in vitro assays. Using double emulsion solvent evaporation method, we developed a poly (D, L-lactide-co-glycolide) (PLGA)-based nanformulation (NF) of miR-150 (miR-150-NF). Polyethyleneimine (a cationic polymer) was incorporated in PLGA matrix to increase the encapsulation of miR-150. Physical characterization of miR-150-NF demonstrated that miR-150-loaded nanoparticles were spherical in shape, had high encapsulation efficiency (>75%), were stable at room temperature and exhibited sustained release profile. miR-150-NF efficiently delivered miR-150 mimics to PC cells and led to the downregulation of its target gene (MUC4) expression. Downregulation of MUC4 correlated with a concomitant decrease in the expression of its interacting partner, HER2, and repression of its downstream signaling. Furthermore, treatment of PC cells with miR-NF suppressed their growth, clonogenicity, motility and invasion. Together, these findings suggest that PLGA-based nanovector platform could be a potential candidate for systemic delivery of miR-150 to the pancreatic tumor cells and can be a step forward in miRNA-based cancer therapeutics.

Citation Format: Sumit Arora, Suresh K. Swaminathan, Sanjeev K. Srivastava, Seema Singh, Jayanth Panyam, Ajay P. Singh. Development of miRNA-loaded polymeric nanoformulation for pancreatic cancer therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1457. doi:10.1158/1538-7445.AM2014-1457
Abstract 146: Combining molecular targeted drugs to inhibit both cancer cells and activated stromal cells in colon cancer
Authors: Yuge Ryo , Kitadai Yasuhiko , Shinagawa Kei , Onoyama Mieko , Tanaka Yuichiro , Tanaka Shinji , Yasui Wataru , Chayama Kazuaki
Published: May 2015

Recent studies have shown that tumor growth and metastasis are not determined by cancer cells alone but also by a variety of stromal cells. We have shown previously that platelet-derived growth factor receptors (PDGF-Rs) are overexpressed by various stromal cell populations, but not by cancer cells themselves in human colon cancer tissues. Blockade of PDGF-R signaling pathways in tumor-associated stromal cells has been shown to inhibit the stromal reaction. Activation of phosphoinositide 3-kinase (PI3K)/Akt/ mammalian target of rapamycin (mTOR) signaling is frequently observed in many cancer types and has been implicated in the pathogenesis of various diseases. mTOR is a central regulatory kinase that increases the production of proteins that stimulate key cellular processes such as cell proliferation, metabolism, survival, and angiogenesis. However, the effects of mTOR inhibitors on the stromal component of tumor tissues have not been studied.

In the present study, the effects of nilotinib (PDGF-R tyrosine kinase inhibitor) and everolimus (mTOR inhibitor) on tumor development and metastasis, were examined using orthotopic and experimental liver metastatic models of human colon cancer, whereby KM12SM cells were implanted into the cecum and spleen of nude mice, respectively. Groups of mice received saline (control), nilotinib (100mg/kg/day) or everolimus (1mg/kg/day), or a combination of nilotinib (100mg/kg/day) and everolimus (1mg/kg/day) by daily oral gavage. After 4 weeks of treatment, primary tumors and metastatic lesions were resected, and specimens were analyzed histologically and immunohistochemically for comparison between the treatment group and control group.

After treatment with nilotinib, the stromal reaction was significantly decreased. After treatment with everolimus, the stromal reaction was not decreased, but tumor cell proliferation and microvessel density were decreased. With the combination therapy, both tumor cell proliferation and the stromal reaction were decreased and apoptosis of tumor cells was increased, resulting in a remarkable decrease in tumor size and the number of metastatic liver lesions.

Therefore, concurrent inhibition of tumor cells and activated stromal cells by combination therapy may represent a novel molecular-targeted approach for patients with colon cancer.

Citation Format: Ryo Yuge, Yasuhiko Kitadai, Kei Shinagawa, Mieko Onoyama, Yuichiro Tanaka, Shinji Tanaka, Wataru Yasui, Kazuaki Chayama. Combining molecular targeted drugs to inhibit both cancer cells and activated stromal cells in colon cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 146. doi:10.1158/1538-7445.AM2014-146
Abstract 1461: A miR-221 multigene pharmacodynamic signature for assessing miR-221 inhibition
Authors: Pavlicek Adam , Vincent Thomas , Guicherit Oivin , Chau Nelson , Zabludoff Sonya , Gibson Neil , Lee Edmund C.
Published: May 2015

miR-221 targets key cancer pathways, inhibits translation of important tumor suppressors, and is overexpressed in hepatocellular carcinoma (HCC). Aberrant miR-221 expression has been linked to HCC progression, while inhibition of miR-221 has been shown to reduce tumor growth. In preclinical models, miR-221 inhibition can be assessed by measuring de-repression of known miR-221 target genes such as p27. However, changes in individual microRNA target genes are often small and model-dependent; and comprehensive assessment of pharmacodynamic (PD) effects following microRNA inhibition is often challenging. Herein, we present a miR-221 multigene PD signature that can assess miR-221 target engagement with a large dynamic window and work across different human HCC models following miR-221 inhibition. In the first stage, we transfected five different human HCC lines with two chemically-modified oligonucleotides that inhibit miR-221 functions (miR-221 anti-miRs) and a microRNA-duplex that mimics miR-221/222 functions (mimic). The resulting global mRNA expression changes were compared to mock treated cells using GeneChip Human Genome U133 Plus 2.0 Array (Affymetrix). Ninety unique human genes with transcripts containing conserved and/or 8-mer target seeds (TargetScan), that significantly went up and down after anti-miR-221s and mimic treatment respectively, were further profiled with qPCR in the second stage. Out of the 90 tested transcripts, 62 of them were significantly up-regulated by anti-miR-221s but not by control oligonucleotides (either mismatch or targeting unrelated microRNAs). From the 62 genes, an optimal multigene signature consisted of 13 unique human genes normalized by 6 reference housekeeping genes was identified. We also devised a “miR-221 signature score” and demonstrated that our multigene signature can separate miR-221 anti-miRs treatment from control oligonucleotides with a high degree of sensitivity and specificity. We further validated this multigene signature by demonstrating that it correctly distinguished miR-221 anti-miRs treatment from controls in three additional HCC cell lines in vitro and one HCC model in vivo. In summary, our miR-221 multigene PD signature analysis provided a comprehensive assessment of miR-221 inhibition, with significantly larger dynamic window compare to individual gene analysis, that may be used to discover, rank order and guide the development of novel miR-221 anti-miRs for the treatment of human cancers.

Citation Format: Adam Pavlicek, Thomas Vincent, Oivin Guicherit, Nelson Chau, Sonya Zabludoff, Neil Gibson, Edmund C. Lee. A miR-221 multigene pharmacodynamic signature for assessing miR-221 inhibition. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1461. doi:10.1158/1538-7445.AM2014-1461
Abstract 1463: Long non-coding RNA RoR and microRNA-145 regulate tumor cell invasion in triple-negative breast cancer via targeting of ADP-ribosylation factor 6
Authors: Eades Gabriel L. , Zhou Qun
Published: May 2015

Triple-negative (ER-, HER2-, PR-) breast cancer (TNBC) is an aggressive disease with a poor prognosis. Currently, there is no available molecularly targeted therapy for TNBC. MicroRNA-145 (miR-145) silencing was identified as a defining marker of TNBC by molecular profiling using deep sequencing. Previous reports indicate that miR-145 may function as a growth suppressor in some breast cancers. However, we find that in TNBC miR-145 does not regulate proliferation or apoptosis but instead, miR-145 regulates tumor cell invasion. Since metastatic breast cancer remains an incurable disease we investigated miR-145 regulated pathways to identify new potential biomarkers or therapeutic targets. Here we identify a novel target of miR-145, the small GTPase ADP-ribosylation factor 6 (Arf6). We find that ARF6, a known regulator of breast tumor cell invasion, is dramatically upregulated in triple-negative and metastatic breast tumors. We find that ARF6 can regulate E-cadherin localization and impact cell-cell adhesion. Finally, we examined the molecular mechanism behind miR-145 down-regulation in TNBC. We find that when compared to ER+ disease, TNBC shows dramatic downregulation of mature miR-145, but not pri-miR-145 or pre-miR-145, indicating that loss of miR-145 is occurring post-transcriptionally and following Drosha and Dicer processing. We find that overexpression of the long non-coding RNA, lincRNA-ROR, functions as a competitive endogenous RNA sponge in TNBC. We find that lincRNA-ROR is dramatically upregulated in TNBC and in metastatic disease. Finally, we find that knockdown of lincRNA-ROR restores miR-145 expression and reduces breast tumor cell invasion. These results reveal a lincRNA-ROR / miR-145 / ARF6 pathway that is a critical regulator of invasion in TNBCs.

Citation Format: Gabriel L. Eades, Qun Zhou. Long non-coding RNA RoR and microRNA-145 regulate tumor cell invasion in triple-negative breast cancer via targeting of ADP-ribosylation factor 6. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1463. doi:10.1158/1538-7445.AM2014-1463
Abstract 1466: Regulation of colon cancer metastasis by sprouty-2
Authors: Zhang Qiong , Vaish Vivek , Khare Sharad
Published: May 2015

Introduction: We earlier established that sprouty-2 is overexpressed in human colon cancer and up-regulates c-met and thereby increases the growth phenotype and metastatic potential of colon cancer cells (Oncogene, 29: 5241-53, 2010). To our knowledge, this was the first study to demonstrate the tumor oncogene phenotype of sprouty-2 in colon cancer metastasis. On the contrary, cancer protective role of sprouty-2 was noted in breast, liver and prostate cancers. However, a recent report on non-small cell lung cancer (NSCLC) discovered that silencing of sprouty-2 reduced c-met expression and thus confirmed our findings in colorectal cancer (PNAS, 110: 8573-78, 2013). We further extended our studies to demonstrate that this pathway modulates gastrointestinal specific microRNAs (miRNAs) and regulates various oncogenes and tumor suppressor genes in colon cancer cells. Methods: RNA from control and shRNA sprouty-2 transfected cells were labeled with Hy5 and Hy3 respectively and hybridized to miRCURY LNA arrays. Unsupervised miRNA clustering was carried out and a heat map prepared with Java TreeView. MicroRNA targets were identified by Real-time PCR, western blotting and 3′UTR luciferase assays. Migratory phenotype of colon cancer cells was confirmed by cell migration and invasion assays in trans-well chambers. Cancer cells were transfected by miRNA mimics to assess the expression levels of target genes. Human colon cancer tissue microarrays were utilized to assess the expression of miRNA target genes. Human tumor sections were evaluated by immunofluorescence and immunohistochemical staining. Results: Sprouty-2 shRNA transfected stable cell lines demonstrated upregulation of tumor suppressive miRNAs and down-regulation of oncogenic miRNAs. Overexpression of gastrointestinal specific miRNA mimics inhibits colon cancer cells proliferation and metastasis. On the contrary, overexpression of miRNA inhibitor in colon cancer cells promotes proliferation and colonization. Studies further indicated that miRNA down-regulates expression of CDK7, RB1, AKT, FASL, IGF1R, EGFR and N-cadherin and up-regulates expression of P21 and E-cadherin. MiRNA regulated genes were responsible for cell cycle arrest, enhanced apoptosis and decreased metastasis. AKT was upregulated whereas E-cadherin was downregulated at the invasive fronts in human colon tumors. Conclusion: Sprouty-2/miRNA expression levels could be used to risk-stratify patients for CRC recurrence and sprouty-2 axis regulation may lead to the development of therapeutics in colon and lung cancer.

Citation Format: Qiong Zhang, Vivek Vaish, Sharad Khare. Regulation of colon cancer metastasis by sprouty-2. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1466. doi:10.1158/1538-7445.AM2014-1466
Abstract 1477: Evaluation of microRNA-10b prognostic significance in a prospective cohort of breast cancer patients
Authors: Parrella Paola , Barbano Raffaela , Pasculli Barbara , Fontana Andrea , Copetti Massimiliano , Valori Vanna Maria , Poeta Maria Luana , Perrone Giuseppe , Coco Michelina , Balsamo Teresa , Pellegrini Fabio , Muda Andrea Onetti , Maiello Evaristo , Murgo Roberto , Fazio Vito Michele
Published: May 2015

Several prognostic factors are currently used to stratify breast cancer patients based on the risk of disease progression. However, even patients classified as at low risk can progress and die for the disease. MicroRNA-10b (miR-10b) has a prominent role in regulating tumor invasion and metastasis by targeting the HOXD10 transcriptional repressor and has been found up-regulated in several tumor types including breast cancer.

We evaluated the expression of miR-10b in paired tumor and normal specimens obtained from a prospective cohort of breast cancer patients with at least 36 months follow-up enrolled according to the REMARK guidelines (n=150). RNA quality was measured and only samples with RNA Integrity Number (RIN) ≥7.0 were analyzed. The relative expression of miR-10b in tumor as compared to its normal counterpart (RER) was determined by RT-qPCR. miR-10b RERs were higher in the subgroup of patients with synchronous metastases (n=11, Median 0.25; IQR 0.11-1.02) as compared with patients without metastases (n=90, Median 0.09; IQR 0.04-0.29) (p=0.028). In the subgroup of patients without synchronous metastases (n=90), higher miR-10b RERs were associated with increased risk of disease progression and death in both univariable (HR 4.35, p=0.021 and HR 6.02, p=0.015 respectively) and multivariable (HR 13.37, p<0.001, and HR 15.39, p=0.003 respectively) Cox regression models. The addition of miR-10b RERs to the Nottingham Prognostic Index (NPI) provided an improvement in discrimination power and risk reclassification abilities for the clinical outcomes at 36 months. Survival C-indices significantly increased from 0.849 to 0.889 (p=0.009) for OS and from 0.735 to 0.767 (p=0.050) for DFS.

Our results provide evidences that the addition of miR-10b RERs to the prognostic factors used in clinical routine could improve the prediction abilities for both overall mortality and disease progression in breast cancer patients.

Citation Format: Paola Parrella, Raffaela Barbano, Barbara Pasculli, Andrea Fontana, Massimiliano Copetti, Vanna Maria Valori, Maria Luana Poeta, Giuseppe Perrone, Michelina Coco, Teresa Balsamo, Fabio Pellegrini, Andrea Onetti Muda, Evaristo Maiello, Roberto Murgo, Vito Michele Fazio. Evaluation of microRNA-10b prognostic significance in a prospective cohort of breast cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1477. doi:10.1158/1538-7445.AM2014-1477
Abstract 1481: miR-296 as prognostic and predictive molecular marker for recurrence in early-stage laryngeal carcinoma treated with definitive radiotherapy
Authors: Maia Danielle C C , Carvalho Ana Carolina de , Horst Maria Aderuza , Neto Cristovam Scapulatempo , Carvalho Andre Lopes , Vettore Andre L.
Published: May 2015

Early-stage laryngeal cancer has cure rates above 80% with exclusive radiotherapy. However, when salvage treatment is needed, surgery is the main option and despite efforts to conservative procedures to preserve laryngeal function, up to 50% will require total laryngectomy at diagnosis of recurrence or future. So, the use of microRNAs as molecular markers is a method to be tested with the aim of predicting the chances of success with radiotherapy, lowering tumor relapses and turning possible to reduce the need for salvage procedures that would affect speech and impact in quality of life. Objectives: To identify microRNAs capable of segregating radioresistant from radiosensitive tumors, to determine prognosis in pre-treatment tumors and to predict failure with radiotherapy. Methods: Molecular Screening of 667 microRNAs using taqman low density array (TLDA) for target selection and then validation in 34 patients, being 20 radioresistant and 14 radiosensitive by RT-PCR with individual assays. Statistical analysis was performed by Chi-square and Fisher exact test for associations between variables, and log-rank method of Kaplan-Meier for recurrence free survival between molecular, clinical, demographic and treatment variables. Also to verify differences in distribution between the expression levels in radioresistant and radiosensitive the Mann-Whitney test was performed. Results: Four microRNAs were selected for validation, miR-296, miR-452, miR-183*, miR-200c based on fold-change and literature review and those were analyzed with respect to age, gender, TNM, laryngeal subsite, radiation dose and treatment interval. Among clinical, demographic, treatment and molecular variables, only miR-296 was shown to present different distribution with statistical significance between radioresistant and radiosensitive (p = 0.002), also demonstrated association with recurrence (p = 0.01) with an odds ratio (OR) of 8.6, confidence interval (CI) between 1.7 and 42.2. Conclusions: In our study, only miR-296 demonstrated the ability to predict patients most likely to fail radiotherapy, also showing a worse prognosis for tumors with high expression level. miR-296 is a molecular predictive and prognostic marker to be considered prior to treatment selection for early-stage laryngeal tumors.

Citation Format: Danielle C C Maia, Ana Carolina de Carvalho, Maria Aderuza Horst, Cristovam Scapulatempo Neto, Andre Lopes Carvalho, Andre L. Vettore. miR-296 as prognostic and predictive molecular marker for recurrence in early-stage laryngeal carcinoma treated with definitive radiotherapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1481. doi:10.1158/1538-7445.AM2014-1481
Abstract 1484: CD57 defines a novel maker of glioblastoma stem cells that have greater invasive potential than CD133+ tumor cells
Authors: Qi Lin , Huang Yu-lun , Kogiso Mari , Mao Hua , Baxter Patricia , Su Jack MF , Perlaky Laszlo , Lau Ching C. , Chintagumpala Murali , Li Xiao-Nan
Published: May 2015

Glioblastoma multiforme (GBM) is the most aggressive and lethal brain tumor that occurs both in children and adults. Diffuse invasion into normal brain tissue is one of the important biologic features that make GBM refractory to conventional therapies. While existing studies on GBM invasion are primarily conducted using tumor core tissues from surgical resections, it is unclear whether unresectable, infiltrative GBM cells would be more informative for studying their invasive nature compared to those in the resected tumor cores. More importantly, little is known if and which cancer stem cell populations are driving glioma invasion. To address these fundamental issues, we utilized our panel of 7 (6 pediatric and 1 adult) patient tumor-derived orthotopic xenograft mouse models of GBM to isolate invasive GBM cells (infiltrating normal mouse brain parenchyma) and tumor core GBM cells and directly compared their biological differences. Our result showed that the invasive cells have stronger neurosphere forming efficiency in vitro in a serial dilution assay and increased tumorigenic capacity after in vivo transplantation (particularly at 100 cells/mouse) compared to the tumor core cells. A screening of putative cancer stem cell markers (CD133, CD15, CD24/CD44, CD57 and CD117) showed that invasive GBM cells are enriched (>2 folds) with CD57+ cells compared to the tumor core cells, and these infiltrating cells were predominantly CD57+/CD133-. Even the CD133+ cells were frequently dual-positive with CD57 (CD33+CD57+), not only in the xenograft tumors but also in a separate set of patient GBM samples. Mechanistically, we found that CD57+ cells expressed high levels of self-renewal genes and tend to stay in G0/G1 phases. In conclusion, we showed that invasive GBM cells are biologically deferent from the matched tumor core cells and identified CD57 as a novel stem cell marker that is associated with GBM infiltration. Our findings suggest that new anti-invasion therapies should target CD57+ cells in addition to CD133+ cells in GBM.

Citation Format: Lin Qi, Yu-lun Huang, Mari Kogiso, Hua Mao, Patricia Baxter, Jack MF Su, Laszlo Perlaky, Ching C. Lau, Murali Chintagumpala, Xiao-Nan Li. CD57 defines a novel maker of glioblastoma stem cells that have greater invasive potential than CD133+ tumor cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1484. doi:10.1158/1538-7445.AM2014-1484
Abstract 1492: Prediction of cancer progression in Barrett's esophagus patients using miRNA profiling
Authors: Srovnal Josef , Slaby Ondrej , Ehrmann Jiri , Gregar Jan , Radova Lenka , Sedlackova Michaela , Hajduch Marian
Published: May 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 histopathological examination. Approximately 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 68 patients (10 EAC, 4 high-grade and 21 low-grade dysplasia of esophagus, and 33 BE). There were 49 males and 19 females; median age was 58 years, range 27 - 97 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, 68 GeneChip miRNA 3.0 Arrays were performed in 68 patients (8 poor quality arrays were excluded). Expression profiles of 1733 human miRNAs, 1658 human pre-miRNAs and 2216 human sno-RNAs and sca-RNAs were analyzed. In supervised clustering analysis we found 29 differentially expressed miRNAs, 7 pre-miRNAs and 9 sno-RNAs (p<0.001 or logFC <-2 or logFC >2) in EAC patients compared to BE patients. One of the dominant findings was decreased expression of miR-22 in EAC samples, which is known to function as a tumor suppressor and histone deacetylase 4 regulator.

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

Acknowledgment: This work was financially supported by IGA MZ CR NT13585-3/202, IGA UP LF 2013_015, 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, Michaela Sedlackova, Marian Hajduch. Prediction of cancer progression in Barrett's esophagus patients using miRNA profiling. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1492. doi:10.1158/1538-7445.AM2014-1492
Abstract 1508: A highly sensitive and reliable methodology (ARMS-PM system) for detection of K-ras mutations in FFPE samples
Authors: Duan Wei-Tao , Wang Xiujuan , Hu Lihong , Zhu Feng , Yu Dehua
Published: May 2015

The association between the activating mutations of K-ras gene and poor clinical response to current targeted therapies against epidermal growth factor receptor (EGFR ) such as cetuximab (Erbitux®) and panitumumab (Vectibix®) in colorectal cancer (CRC) has been well documented. Therefore detection of K-ras mutation status will guide effective therapeutical options for patients with CRC. Here we developed a new method for the rapid and reliable detection of K-ras mutations by using a combination of the modified mutation-specific ARMS primers, peptide nucleic acid (PNA) clamping and taqman-MGB probes, named ARMS-PM system. In the ARMS-PM system, the wild-type K-ras DNA is completely blocked by PNA at a higher annealing temperature, and the mutant gene is selectively amplified by the modified ARMS primers and the amplicon extending signals are collected by the taqman-MGB probe at the same time. The ARMS-PM system can detect approximately 10 copies of the mutant gene stably in somatic cells at the background of 10ng genomic DNA. In addition, the premixed reaction was introduced into this system to allow the assay to be completed in only one step of sample loading, which minimizes the potential contamination significantly.

By using this assay, we analyzed the K-ras mutations in 1014 Chinese CRC and 205 Chinese lung cancer FFPE samples in comparison with Sanger sequencing. Our results showed that the positive rates were 38.6% and 10.2% 38.0% in CRC and lung cancer samples respectively, which were higher than that measured by Sanger sequencing (38.0% and 7.1%). The coincidence rates of the two methods were 97.3% for the positive samples and 97.7 for the negative samples. The total coincidence rate was 97.8%. In conclusion, our study provides a basis for the detection of somatic mutations in K-ras in a routine clinical setting. The ARMS-PM assay allows the detection of the activating K-ras mutations in FFPE samples with better sensitivity and feasibility than the current assays used in clinic.

Citation Format: Wei-Tao Duan, Xiujuan Wang, Lihong Hu, Feng Zhu, Dehua Yu. A highly sensitive and reliable methodology (ARMS-PM system) for detection of K-ras mutations in FFPE samples. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1508. doi:10.1158/1538-7445.AM2014-1508
Abstract 1509: A ARMS-PNA PCR assay for detection of EGFR mutations in FFPE tumor samples
Authors: Zhao Shanshan , Guo Xiao , Long Ziwen , Yu Dehua
Published: May 2015

The activating mutations of Epidermal growth factor receptor (EGFR) are the predictive markers for response to tyrosine kinase inhibitors gefitinib and erlotinib in non-small cell lung cancer(NSCLC). Therefore detection of EGFR mutations will guide effective therapeutical options for patients with NSCLC. Here we developed a new method for the rapid and reliable detection of EGFR mutations by using a combination of modified mutation-specific ARMS primers and peptide nucleic acid (PNA) clamping system, named ARMS-PNA assay. In this real-time PCR system, the wild-type EGFR DNA is completely blocked by PNA with a higher annealing temperature, and the mutant gene is selectively amplified by the modified ARMS primers and the amplicon extending signals are collected by the taqman-probe at the same time. The ARMS-PNA assay could detect 29 mutations on EGFR gene with a sensitivity of 0.5% under the background of a total of 10ng wild-type genomic DNA. By using this assay, we analyzed the EGFR mutations in 1033 Chinese NSCLC FFPE samples in comparison with Sanger sequencing. Our results showed that the positive rates were 50.1%, which were higher than that measured by Sanger sequencing (48.3%). The coincidence rates of the two methods were 99.0% for the positive samples and 95.5% for the negative samples. The total coincidence rate was 97.2%. In conclusion, our study provides a basis for the detection of somatic mutations in EGFR in a routine clinical setting. The ARMS-PNA PCR assay allows the detection of the activating EGFR mutations in FFPE samples with better sensitivity and feasibility than the current assays used in clinic.

Citation Format: Shanshan Zhao, Xiao Guo, Ziwen Long, Dehua Yu. A ARMS-PNA PCR assay for detection of EGFR mutations in FFPE tumor samples. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1509. doi:10.1158/1538-7445.AM2014-1509
Abstract 1512: Detection of somatic alterations in plasma from lung cancer patients
Authors: Costa Jose L. , Justino Ana , Barroso Ana , Parente Barbara , Machado Jose C.
Published: May 2015

Tumor-specific (somatic) mutations in plasma can serve as biomarkers for tumor detection, monitor tumor response to specific therapies, detect residual disease after surgery, and long-term follow-up. The intrinsic low abundance of circulating cell-free tumor DNA (cfDNA) makes the detection and quantification of such mutations in plasma a challenging task. This small scale study aims to establish a comprehensive strategy to be used in the detection of clinical relevant somatic alterations in plasma of lung cancer patients.

Plasma samples obtained at different stages of disease progression and/or treatment were collected from a group of 11 lung cancer patients and used to isolate cfDNA. Genetic alterations in the EGFR gene (p.E746_A750del and p.L858R) identified at diagnosis in tumor biopsies were used as surrogate markers to optimize and validate the next generation sequencing strategy and data analysis workflow. The Ion AmpliSeq Colon and Lung cancer panel was used to analyze hotspot and targeted regions of 22 genes implicated in colon and lung cancers, including the EGFR gene mutations mentioned. The amplified products were used to prepare libraries and were sequenced using the Ion PGM system. Quantitative real time PCR and digital PCR assays were used to confirm selected results.

Tumor derived genetic alterations could be identified in as little as 10ng of cfDNA. EGFR alterations identified in cfDNA mirrored the alterations identified in all tumor biopsies. EGFR alterations with allelic frequencies as low as 3% could be detected in cfDNA. Additionally, in case of samples collected after therapy, a clear decrease in the cfDNA allelic frequency of the EGFR mutation, that made the patient eligible for therapy, was observed. Furthermore, the screening of a larger panel of genes allowed the identification in two cases of additional gene mutations (e.g. MET and KRAS) that may have impact in the clinical management of patients.

In this study, we demonstrate the capacity to identify clinically relevant somatic EGFR mutations in plasma. The possibility to assess information from a larger panel of genes makes this strategy attractive for further optimization of treatment options. The strategy is now being extended to a larger cohort of patients to push forward the concept of liquid biopsy in the clinical management of cancer patients.

Citation Format: Jose L. Costa, Ana Justino, Ana Barroso, Barbara Parente, Jose C. Machado. Detection of somatic alterations in plasma from lung cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1512. doi:10.1158/1538-7445.AM2014-1512
Abstract 1515: Simultaneous detection of KRAS and TP53 mutations in human cancer cell lines using multiplex qPCR
Authors: Patel Sunali , Woo Cora , Wilde Joyce , Phanse Sundiep
Published: May 2015

Genetic alterations in KRAS and TP53 genes are common in many cancers and the understanding of their role in progression of cancer is vital for the design of successful treatment approaches. Studies suggest that mutations in KRAS and TP53 genes independently hindered the use of EGFR inhibiting drugs like Cetuximab (Erbitux) for the treatment of colorectal cancer (1). The genetic alterations in these genes are now routinely checked before starting any expensive treatments. Here, we successfully demonstrated use of duplex mutli-color SNP genotyping assays to detect KRAS and TP53 mutation in human colon cancer cell lines. This method can be potentially used to analyze samples with limited quantity and get as much information as possible from a single sample. To study the impact of these mutations, we generated gene expression profiles of 672 genes related to signaling pathways by using the Human signal transduction medium-throughput OpenArray® Panel. Selected cancer genes like BRAC1, BAX, CFLAR, MYC and others were evaluated using multiplexed single-tube PCR simultaneously detecting four different targets in the same sample. These results demonstrate that duplex SNP genotyping and multiplexed gene expression profiling from a single sample increases research capacity in situations when sample is limited.

Reference:

1. A Oden-Gangloff, F Di Fiore, F Bibeau, A Lamy, G Bougeard, F Charbonnier, F Blanchard, D Tougeron, M Ychou, F Boissière, F Le Pessot, J-C Sabourin, J-J Tuech, P Michel and T Frebourg.TP53 mutations predict disease control in metastatic colorectal cancer treated with cetuximab-based chemotherapy. British Journal of Cancer (2009) 100, 1330-1335

Citation Format: Sunali Patel, Cora Woo, Joyce Wilde, Sundiep Phanse. Simultaneous detection of KRAS and TP53 mutations in human cancer cell lines using multiplex qPCR. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1515. doi:10.1158/1538-7445.AM2014-1515