bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1488: Association between microrna binding site polymorphism and susceptibility to colorectal cancer in Korean population
Authors: Kang Byung Woog , Jeon Hyosung , Chae Yee Soo , Lee Soo Jung , Kim Jong Gwang , Lee Hyunchul , Lee Eun-Jin , Lee Myung Hoon , Park Jae Yong , Choi Gyu Seog , Park Jun Seok
Published: May 2015

Background : MicroRNAs (miRNAs) play an important role in the colorectal cancer tumorigenesis by regulation of gene expression through binding to 3′ untranslational region (UTR) of target gene. Single nucleotide polymorphism (SNP) within 3′UTR of gene could be affected by miRNA binding. Docking protein 3 (DOK3) is adapter protein that acts as tumor suppressor through modulating tyrosine kinase signaling.

Methods : One hundred thirty five SNPs were selected in Silico analysis for the current study from 3′UTR SNPs of SNP database, which was based on several miRNA database, calculating miRNA bind efficiency on polymorphism, and HapMap database. Here independent two set of study was designed, 380 healthy controls and 371 colorectal adenocarcinoma patients were enrolled in discovery set, and 524 healthy controls and 521 colorectal adenocarcinoma patients were enrolled in replication set. The SNP genotyping was performed using the Sequenom MassARRAY

Result: In discovery set, 16 SNPs were significantly associated with colorectal cancer in any genetic model from 135 SNP genotyping. Among 16 SNPs, DOK3 rs2279398G>A was significantly associated in replication set recessive model (adjusted odds ratio (OR) = 0.65, 95% confidence interval (CI) = 0.44-0.97, P = 0.03). In combined analysis, DOK3 rs2279398G>A was associated with significantly decreased risk of colorectal cancer in codominant and recessive model (adjusted OR = 0.84, 95% CI = 0.73-0.96, P = 0.012, adjusted OR = 0.65, CI = 0.49-0.88, P = 0.004, respectively).

Conclusion : DOK3 rs2279398G>A may affect the expression of DOK3 by altering miRNA binding efficiency on miRNA-binding sites of the 3′-UTR of DOK3 and impact on colorectal cancer tumorigenesis. This finding suggests that DOK3 rs2279398G>A may be a useful biomarker for determining the susceptibility to colorectal cancer.

Citation Format: Byung Woog Kang, Hyosung Jeon, Yee Soo Chae, Soo Jung Lee, Jong Gwang Kim, Hyunchul Lee, Eun-Jin Lee, Myung Hoon Lee, Jae Yong Park, Gyu Seog Choi, Jun Seok Park. Association between microrna binding site polymorphism and susceptibility to colorectal cancer in Korean population. [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 1488. doi:10.1158/1538-7445.AM2014-1488
Abstract 1499: Expression and prognostic significance of microRNAs in Korean patients with myelodysplastic syndrome
Authors: Choi Yunsuk , Hur Eun-Hye , Moon Ju Hyun , Goo Bon-Kwan , Choi Dae R. , Lee Je-Hwan
Published: May 2015

Introduction: MicroRNAs (miRNAs) are short non-coding RNAs that are involved in post-transcriptional regulation of gene expression, which may result in translational inhibition or destabilization of the target mRNA. Various alterations of miRNA expression have been reported in myelodysplastic syndrome (MDS). We aimed to investigate the unique patterns and prognostic significance of miRNA expression in Korean patients with MDS.

Materials and Methods: Bone marrow mononuclear cells were collected from 10 healthy controls and 26 MDS patients, and miRNAs were isolated using a miRNeasy isolation kit (Qiagen) according to the manufacturer's instructions. Quantitative real-time PCR for selected miRNAs including miR-21, miR-124a, miR-126, miR-146b, miR-155, miR-200c, miR-342-5p, miR-708, and Let-7a was performed using a miScript SYBR green PCR kit (Qiagen). RNU6 was used to normalize the miRNA values of all RNA samples. Calculations were made using the comparative Ct (2-ΔΔCt ) method.

Results: Eleven patients with early stage MDS (2 RA, 1 RARS, and 8 RCMD), 10 patients with advanced MDS (3 RAEB-1 and 7 RAEB-2), 4 patients with unknown subtype of MDS, and 1 patient with chronic myelomonocytic leukemia were included in this study. Median age was 58.5 years (range, 13-76). MiR-124a, miR-155, miR-200c, miR-342-5p, and let-7a were significantly under-expressed in MDS patients compared to healthy controls. Levels of miR-124, miR-146b, and miR-155 transcripts were significantly lower in patients with early stage MDS than in those with advanced MDS. Secondary MDS patients (n=6) showed significantly under-expressed MiR-146b and miR-342-5p transcript levels than de novo MDS patients (n=19) (p=0.009 and p=0.043, respectively). We failed to find statistically significant survival differences according to the expression levels of miRNAs, although high expression of miR-124, miR-126, miR-155 and low expression of miR-342-5p tended to be associated with poor overall survival and leukemia free survival.

Conclusion: We first report under-expression of miR-200c in MDS patients. Expression of miRNAs appears to be different according to the stage of MDS. Further investigations are needed to evaluate genetic functions and prognostic significances of miRNAs in MDS.

Citation Format: Yunsuk Choi, Eun-Hye Hur, Ju Hyun Moon, Bon-Kwan Goo, Dae Ro Choi, Je-Hwan Lee. Expression and prognostic significance of microRNAs in Korean patients with myelodysplastic syndrome. [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 1499. doi:10.1158/1538-7445.AM2014-1499
Abstract 1500: The difference of serum RNA profile: RNA detection method
Authors: Kondo Satoshi , Takizawa Satoko , Akiyama Hideo
Published: May 2015

Serum miRNA profiles are widely reported by many research groups, but they do not always coincide well. The reason of this difference is suggested to be, at least in some cases, due to a difference in the RNAdetection method. In this report, we compare the miRNA expression data obtained by different detection tools.. In these days, the whole miRNA expression profiling can be done by multiple qRT-PCR, microarrays, and Next Generation Sequencing. However, the feature of each method will reflect to the obtained data and it is no surprise that data obtained by different tools do not match perfectly. We will compare the serum miRNA expression profiles detected by (1) “TaqMan” Array MicroRNA Card (2) QIAGEN miScript miRNA PCR Array (3) “3D-Gene” Human miRNA oligo chip, (4)”HiSeq” Next Generation Sequencer (5) Nanostring “nCounter” Analysis system and discuss the base of method for serum miRNA profiling.

Citation Format: Satoshi Kondo, Satoko Takizawa, Hideo Akiyama. The difference of serum RNA profile: RNA detection method. [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 1500. doi:10.1158/1538-7445.AM2014-1500
Abstract 1519: Targeted enrichment and next generation sequencing of non-small cell lung carcinoma FFPE samples
Authors: Breuil Rusla du , Shvetsova Tatiana , Patel Kruti , Moore Troy , Fewell Gwen
Published: May 2015

Targeted enrichment is a powerful tool used for identifying genetic variations that underlie cancer and complex diseases. Current methodologies utilize hybridization probes or PCR to enrich for protein coding regions of a genome followed by deep sequencing. Most of these targeting methods fail to achieve the specificity, coverage and uniformity that are critical for accurate detection and analysis of genetic variability. Nested Patch PCR™ was developed to overcome these pitfalls with highly specific on-target sequence interrogation across hundreds of genomic regions in a single-tube format. This novel technology enables rapid detection of various genetic variations (i.e. SNPs, indels) across a large number of samples, including FFPE (formalin-fixed paraffin embedded) samples which are typically difficult to analyze due to low yield and quality of DNA. To show the utility of TargetRich technology, an enrichment panel was designed targeting NRAS, PTEN, KRAS, ALK, PIK3CA, KIT, BRAF and EGFR to interrogate 250 non-small cell lung carcinoma FFPE samples. The panel was designed against 19.8kb, 134 exons, with 99.9% coverage. A minimum of 50ng FFPE DNA were run for each sample on an Illumina Miseq and the resulting data was comparable to high molecular weight DNA in uniformity and performance; supporting TargetRich® as a sensitive and robust platform to analyze FFPE samples.

Citation Format: Rusla du Breuil, Tatiana Shvetsova, Kruti Patel, Troy Moore, Gwen Fewell. Targeted enrichment and next generation sequencing of non-small cell lung carcinoma 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 1519. doi:10.1158/1538-7445.AM2014-1519
Abstract 1528: Identification of FGFR3 as a potential therapeutic target gene for human clear cell ovarian cancer by global genomic analysis
Authors: Tsang Tsun Yee , Mohapatra Gayatry , Itamochi Hiroaki , Mok Samuel C. , Birrer Michael J.
Published: May 2015

Clear cell ovarian cancer (CCOC) is the second most common subtype of ovarian cancer after serous ovarian cancer. CCOC is more resistant to standard chemotherapy and has a poorer prognosis than serous and endometrioid histotypes. We attempted to identify genes that are responsible for the aggressive behavior of CCOC and could be potential therapeutic targets using genomic and expression profilings. To identify gene amplification, genome-wide DNA copy number alterations were measured in 13 CCOC cell lines using high-resolution oligonucleotide array CGH (Agilent 105k Human Genome CGH Microarray). GISTIC analysis identified 16 amplified regions containing 391 genes. After comparison to our database of differentially expressed genes between human CCOC specimens and normal ovarian surface epithelium specimen (Affymetrix U133 Plus 2.0 microarrays), 45 of the amplified genes showed mRNA overexpression in CCOC specimens. Among the 45 genes, fibroblast growth factor receptor 3 (FGFR3) was amplified in 9/13 CCOC cell lines and was overexpressed (12.9 folds) in CCOC specimens. Analysis using PathwayStudio software identified FGFR3 mediated pathways for cell migration and cell proliferation. Knockdown of FGFR3 by transfection with FGFR3 specific shRNAs suppressed cell migration in KOC-7c and HAC-2 cells. Moreover, knockdown of FGFR3 also suppressed anchorage-dependent cell proliferation of KOC-7c and HAC-2 cells. The results from the present study therefore suggested that FGFR3 may promote metastasis and growth of cancer cells and may serve as a potential target gene for treatment of human clear cell ovarian cancer.

Citation Format: Tsun Yee Tsang, Gayatry Mohapatra, Hiroaki Itamochi, Samuel C. Mok, Michael J. Birrer. Identification of FGFR3 as a potential therapeutic target gene for human clear cell ovarian cancer by global genomic analysis. [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 1528. doi:10.1158/1538-7445.AM2014-1528
Abstract 1534: Identification of LMX1B as a novel oncogene in human ovarian cancer
Authors: Mohapatra Gayatry , He Lei , Guo Lankai , Vathipadiekal Vinod , Sergent Petra , Growdon Whitfield , Rueda Bo , Engler David , Orsulic Sandra , Birrer Michael
Published: May 2015

Ovarian cancers are thought to result from the accumulation of multiple genetic aberrations that transform ovarian and/or fallopian tube surface epithelial cells, allowing for their abnormal growth, proliferation and metastasis. We carried out genome-wide copy-number analysis using array comparative genomic hybridization on a panel of mouse ovarian cancer (OVCA) cell lines previously established in our laboratory. We identified a recurrent focal amplification on mouse chromosomal region 2qB, which contains the LIM homeodomain containing transcription factor 1B (Lmx1b) gene. LMX1B is not expressed in normal human ovary, but is expressed in many human OVCA cell lines and primary tumors. High expression of LMX1B correlates with poor outcome. To clarify the role of LMX1B in ovarian carcinogenesis, we transduced LMX1B into a panel of mouse and human OVCA cell lines and demonstrated that LMX1B strongly promotes migration of cancer cells in culture and accelerates xenograft growth in nude mice. Conversely, knockdown of LMX1B in a human cell line with endogenous high expression of LMX1B inhibits cell migration in vitro and tumor growth in vivo. Microarray analysis of cells overexpressing LMX1B identified NF-κB pathway as a potential mediator of tumor progression and subsequent treatment of NF-κB inhibitor decreased the migratory capacity of these cells. Thus, our data demonstrate that LMX1B functions as an oncogene in OVCA pathogenesis.

Citation Format: Gayatry Mohapatra, Lei He, Lankai Guo, Vinod Vathipadiekal, Petra Sergent, Whitfield Growdon, Bo Rueda, David Engler, Sandra Orsulic, Michael Birrer. Identification of LMX1B as a novel oncogene in human 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 1534. doi:10.1158/1538-7445.AM2014-1534
Abstract 156: Autophagic degradation of granzyme B impairs NK-mediated killing of hypoxic tumor cells
Authors: Baginska Joanna , Viry Elodie , Berchem Guy , Poli Aurélie , Noman Muhammad Zaeem , Moer Kris van , Medves Sandrine , Mgrditchian Takouhie , Zimmer Jacques , Oudin Anais , Niclou Simone P. , Bleackley R. Chris , Chouaib Salem , Janji Bassam
Published: May 2015

Natural killer (NK) cells are effectors of the innate immune system, able to kill cancer cells through the release of the cytotoxic protease granzyme B. NK-based therapies have recently emerged as promising anticancer strategies. However, it is well established that hypoxic tumor microenvironment interferes with the antitumor function of immune cells and constitutes a major obstacle for defining cancer immunotherapies. Recent studies demonstrated that autophagy regulates the innate immune response by mechanisms which are not fully understood. In this study, we showed that hypoxia decreases breast cancer cell susceptibility to NK-mediated lysis by a mechanism involving the activation of autophagy in tumor cells. Targeting autophagy was sufficient to restore NK-mediated tumor cell killing. We showed that the resistance of hypoxic tumor cells was neither related to a defect in their recognition by NK cells, nor to a defect in the cytolytic function of NK cells toward hypoxic cells. We provided evidence that autophagy activation degrades NK-derived granzyme B in lysosomes of hypoxic cells making them less sensitive to NK-mediated killing. Genetic and pharmacological inhibition of autophagy restored granzyme B levels and reverted the resistance of hypoxic cells in vitro. Our results highlight autophagy as a critical factor in modulating NK-mediated anti-tumor immune response. We have validated this concept in vivo by showing that targeting autophagy significantly improved NK-mediated tumor shrinking in breast and melanoma models. This study provides a cutting-edge advance in our understanding of how hypoxia-induced autophagy impairs NK-mediated lysis and paves the way for formulating more effective NK-based antitumor therapy by combining autophagy inhibitors.

Citation Format: Joanna Baginska, Elodie Viry, Guy Berchem, Aurélie Poli, Muhammad Zaeem Noman, Kris van Moer, Sandrine Medves, Takouhie Mgrditchian, Jacques Zimmer, Anais Oudin, Simone P. Niclou, R. Chris Bleackley, Salem Chouaib, Bassam Janji. Autophagic degradation of granzyme B impairs NK-mediated killing of hypoxic 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 156. doi:10.1158/1538-7445.AM2014-156
Abstract 1575: ARF regulates the stability of p16 protein via REGγ-dependent proteasome degradation
Authors: Kobayashi Takashi , Ogawa Osamu , Abate-Shen Cory
Published: May 2015

The cell cycle regulatory gene INK4A-ARF (CDKN2A) has two alternative transcripts that produce entirely different proteins, namely p14ARF and p16, which have complementary functions as regulators of p53 and pRB tumor suppressor pathways, respectively. The unusual organization of INK4A-ARF has long led to speculation of a need for coordinated regulation of p14ARF and p16. We now show that p14ARF (ARF) regulates the stability of p16 protein in human cancer cell lines, as well as in mouse embryonic fibroblasts (MEFs). In particular, ARF promotes rapid degradation of p16 protein, which is mediated by the proteasome and, more specifically, by interaction of ARF with one of its subunits, REGγ. Furthermore, this ARF-dependent destabilization of p16 can be abrogated by knock-down of REGγ or by pharmacological blockade of its nuclear export. Thus our findings have uncovered a novel crosstalk of two key tumor suppressors mediated by a REGγ-dependent mechanism.

Citation Format: Takashi Kobayashi, Osamu Ogawa, Cory Abate-Shen. ARF regulates the stability of p16 protein via REGγ-dependent proteasome degradation. [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 1575. doi:10.1158/1538-7445.AM2014-1575
Abstract 1594: Transgenic overexpression of NanogP8 in the mouse prostate does not initiate tumorigenesis nor promotes tumor development in the Hi-Myc mouse model
Authors: Gong Shuai , Liu Bigang
Published: May 2015

Nanog1 is a homeobox transcription factor critical for the self-renewal and pluripotency of ES cells. NanogP8 is a retrotransposed homolog of Nanog1 preferentially expressed in somatic cancer cells. Work from our lab has shown that shRNA-mediated knockdown of NanogP8 in prostate, breast, and colon cancer cells inhibits tumor regeneration whereas inducible overexpression of NanogP8 in somatic cancer cells promotes cancer stem cell phenotypes and properties. A key unanswered question is whether tissue-specific overexpression of NanogP8 is sufficient to promote tumor development in vivo. To address this question, we generated a NanogP8 transgenic mouse model in which the NanogP8 cDNA derived from a primary prostate cancer was driven by the ARR2Pb promoter to direct prostate-specific expression. IHC analysis revealed that human NanogP8 was expressed mainly in the mouse prostate and the expression levels varied from lobe to lobe, with the highest expression in the lateral and ventral lobes. The ARR2Pb-NanogP8 transgenic mice were viable, developed normally, and did not form spontaneous tumors during the observation period of >2 years. In addition, both wild-type and ARR2Pb-NanogP8 transgenic prostates responded similarly to castration and regeneration and did not develop any tumors. These results indicate that overexpression of human NanogP8 in the mouse prostate is insufficient to initiate tumorigenesis in both androgen intact and deprived conditions. We then crossed the ARR2Pb-NanogP8 transgenic mice with ARR2Pb-Myc (i.e., Hi-Myc) mice and found that the double transgenic (i.e., ARR2Pb-NanogP8; Hi-Myc) mice showed similar tumor incidence and histology to the Hi-Myc mice. Unexpectedly, we found white dots in the ventral lobes of the double transgenic mice, which are currently being characterized. Taken together, our present work demonstrates that transgenic overexpression of NanogP8 in the mouse prostate does not initiate tumorigenesis nor promotes tumor development in the Hi-Myc mouse model. These results are consistent with our recent findings that NanogP8 overexpression fails to initiate tumorigenesis in a K14-NanogP8 transgenic model. Our results are also consistent with others' recent work showing that NanogP8 overexpression in transgenic animals is insufficient to initiate mammary tumor development.

Citation Format: Shuai Gong, Bigang Liu. Transgenic overexpression of NanogP8 in the mouse prostate does not initiate tumorigenesis nor promotes tumor development in the Hi-Myc mouse model. [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 1594. doi:10.1158/1538-7445.AM2014-1594
Abstract 1635: Discovery of novel SMAC mimetics as selective IAP inhibitors
Authors: Wang Jin , Li Wei
Published: May 2015

Inhibitor of apoptosis (IAP) proteins are widely considered as promising cancer drug targets, especially for drug-resistant tumors. Mimicking the IAP-binding motif of second mitochondria-derived activator of caspases (SMAC) is a rational strategy to design potential IAP inhibitors. In this report, we used the bioactive conformation of AVPI tetrapeptide in the N-terminus of SMAC as a template and performed a shape-based virtual screening against a drug-like compound library to identify novel IAP inhibitors. Top hits were subsequently docked to available IAP crystal structures as a secondary screening followed by validation using in vitro biological assays. Four novel hit compounds were identified to potently inhibit cell growth in two human melanoma (A375 and M14) and two human prostate (PC-3 and DU145) cancer cell lines. The best compound, UC-112, has IC50 values ranging from 0.7 to 3.4 µM. UC-112 also potently inhibits the growth of P-glycoprotein (P-gp) overexpressed multidrug-resistant cancer cells, strongly activates capase-3/7 and caspase-9 activities, and selectively down-regulates survivin level at a concentration as low as 1 µM. Co-incubation of UC-112 with a known proteasome inhibitor (MG132) rescued survivin inhibition, consistent with the anticipated mechanism of action for UC-112. As a single agent, UC-112 strongly inhibits tumor growth and reduces both XIAP and survivin levels in an A375 human melanoma xenograft model. Overall, our study identified novel scaffolds, especially UC-112, as new platforms on which potent and selective IAP antagonists can be developed.

Citation Format: Jin Wang, Wei Li. Discovery of novel SMAC mimetics as selective IAP inhibitors. [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 1635. doi:10.1158/1538-7445.AM2014-1635
Abstract 1637: Discovery of ATR kinase inhibitors from natural products
Authors: Wang Hui-Chun , Lee Alan Yueh-Luen , Wu Chin-Chung , Wu Yang-Chang
Published: May 2015

Inhibition of the DNA damage checkpoint and repair functions is a promising approach to improve chemosensitivity. Ataxia telangiectasia and Rad3-related protein (ATR) is an important checkpoint kianse responsible for organizing both normal and stressful DNA replication. In previous, we discovered a natural compound protoapigenone (WYC02) and its synthetic derivate WYC0209 are able to inhibit ATR-mediated Chk1 phosphorylation, impair the G2/M checkpoint, and improve cancer sensitivity to cisplatin both in vitro and in vivo. Although all evidences convince ATR signaling is disrupted by WYC compounds, the effect of compounds on ATR kinase activity is still unknown. In this study, we further provide evidences to prove that WYC02 and WYC0209 are undoubted ATR inhibitors by using of in vitro kinase assay. The data showed WYC0209 inhibits ATR kinase activity at least 4 times greater than WYC02, and enhances MDA-MB-231cells sensitivity to DNA replication-affecting agents such as doxorubicin, mitomycin C, and etoposide, but has little or no effect to mitosis-affecting microtubule inhibitor palcitaxol. The result of this study proposes a strategy to improve most of current chemotherapeutics in which aim to selective disrupting DNA replication in fast growing cancers.

Citation Format: Hui-Chun Wang, Alan Yueh-Luen Lee, Chin-Chung Wu, Yang-Chang Wu. Discovery of ATR kinase inhibitors from natural products. [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 1637. doi:10.1158/1538-7445.AM2014-1637
Abstract 1641: Structure-based design and development of pyrazolopyridine-based inhibitors of Mcl-1
Authors: Abulwerdi Fardokht A. , Mady Ahmed S.A. , Perdih Andrej , Stuckey Jeanne A. , Showalter Hollis D. , Nikolovska-Coleska Zaneta
Published: May 2015

The anti-apoptotic myeloid cell leukemia protein Mcl-1, a member of the Bcl-2 family proteins, has emerged as a promising therapeutic target. Mcl-1 is overexpressed in many human cancers which has been associated with inferior survival, poor prognosis and resistance to chemotherapies. Disruption of Mcl-1 interaction with its pro-apoptotic partners through small molecules is a viable strategy to overcome Mcl-1 mediated resistance to apoptosis. Targeting Mcl-1 protein represents a promising strategy either alone or in combination with other therapies.

Applying an integrated screening approach through combining high throughput and virtual screenings, several novel chemical classes were identified as Mcl-1 inhibitors. Compound 38 with a pyrazolopyridine scaffold was selected as a promising high throughput lead for medicinal chemistry efforts. Using reported chemistry a focused library of analogs of 38 was generated. Through HSQC protein-observed NMR studies and chemical shift mapping, the binding of the lead compound 38 was characterized and confirmed to be the BH3 binding pocket of Mcl-1. Computational modeling guided by NMR studies was applied in lead optimization and rational design of more potent analogs. Structure-activity relationship was established utilizing two different competitive platforms of fluorescent polarization and surface plasmon resonance, and confirmed by HSQC NMR spectroscopy. The binding affinity of this class of compounds was improved more than twenty fold in comparison with the lead compound 38. In vitro binding, functional and cell-based assays were performed in order to determine selectivity profile against five members of Bcl-2 family, mechanism of action and cellular activity of analogs with improved potency. The obtained results show promise for further chemical modification and development of pyrazolopyridine-based inhibitors of Mcl-1.

Citation Format: Fardokht A. Abulwerdi, Ahmed S.A. Mady, Andrej Perdih, Jeanne A. Stuckey, Hollis D. Showalter, Zaneta Nikolovska-Coleska. Structure-based design and development of pyrazolopyridine-based inhibitors of Mcl-1. [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 1641. doi:10.1158/1538-7445.AM2014-1641
Abstract 1643: BMS-983970, an oral pan-Notch inhibitor for the treatment of cancer
Authors: Gavai Ashvinikumar V. , Zhao Yufen , O'Malley Daniel , Fink Brian , Quesnelle Claude , Norris Derek , Chen Libing , Kim Soong-Hoon , Han Wen-Ching , Gill Patrice , Shan Weifang , Balog Aaron , Tebben Andrew , Rampulla Richard , Wu Dauh-Rurng , Zhang Yingru , Mathur Arvind , Wang Haiqing , Yang Zheng , Ruan Qian , Moore Robin , Rodrigues David , Ranasinghe Asoka , D'Arienzo Celia , Tye Ching Kim , Su Ching , Everlof Gerry , Yarde Melissa , Cvijic Mary Ellen , Menard Krista , Wen Mei-Li , Trainor George , Fischer Bruce , Hunt John , Vite Gregory , Westhouse Richard , Lee Francis
Published: May 2015

Deregulation of the Notch pathway has been shown to be oncogenic in numerous tissue types including T-cell acute lymphoblastic leukemia (T-ALL), breast cancer, non-small cell lung cancer, and colorectal carcinoma. Notch signal activation can cause uncontrolled proliferation, restrict differentiation leading to increased self-renewal capacity, evasion of apoptosis, and enhancement of angiogenesis and metastasis. There is increasing evidence that Notch plays a role in the maintenance and survival of cancer stem cells. γ-Secretase mediates the Notch signaling pathway by releasing the Notch intracellular domain (NICD) which translocates to the nucleus and binds to the transcription factor CSL to activate transcription of various target genes. BMS-906024 is a potent pan-Notch inhibitor that demonstrated robust anti-tumor activity at tolerated doses in multiple tumor xenograft models. It is being evaluated in Phase 1 clinical studies. BMS-906024 is being administered IV (once weekly) in the clinic and the projected human efficacious dose is 4 - 6 mg. Based on the preclinical data, the projected human half-life of BMS-906024 is in the 37 h - 124 h range.

This presentation will describe further structure-activity relationships in the 1,4-benzodiazepinone series that culminated in the identification of BMS-983970 as an oral-pan-Notch inhibitor. Pharmacokinetic properties and in vivo evaluation of BMS-983970 in T-ALL and solid tumor xenograft models will be presented.

Citation Format: Ashvinikumar V. Gavai, Yufen Zhao, Daniel O'Malley, Brian Fink, Claude Quesnelle, Derek Norris, Libing Chen, Soong-Hoon Kim, Wen-Ching Han, Patrice Gill, Weifang Shan, Aaron Balog, Andrew Tebben, Richard Rampulla, Dauh-Rurng Wu, Yingru Zhang, Arvind Mathur, Haiqing Wang, Zheng Yang, Qian Ruan, Robin Moore, David Rodrigues, Asoka Ranasinghe, Celia D'Arienzo, Ching Kim Tye, Ching Su, Gerry Everlof, Melissa Yarde, Mary Ellen Cvijic, Krista Menard, Mei-Li Wen, George Trainor, Bruce Fischer, John Hunt, Gregory Vite, Richard Westhouse, Francis Lee. BMS-983970, an oral pan-Notch inhibitor for the treatment of 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 1643. doi:10.1158/1538-7445.AM2014-1643
Abstract 1645: Discovery of AMG 232, an inhibitor of the MDM2-p53 interaction: From lead to a clinical candidate
Authors: Li Zhihong
Published: May 2015

The p53 tumor suppressor is controlled by MDM2, which binds p53 and negatively regulates its transcriptional activity and stability. Many tumors overproduce MDM2 to impair p53 function. Therefore, restoration of p53 activity by inhibiting the p53-MDM2 binding represents an attractive novel approach to cancer therapy. We previously reported the discovery AM-8553, a potent and selective piperidinone inhibitor of the MDM2-p53 interaction (Rew et al. J. Med. Chem. 2012, 55, 4936). We report here continued research on the N-alkyl substituent of this series, focused in particular on a previously underutilized interaction in a shallow cleft on the MDM2 surface that led to the discovery of variety of sulfonamides and sulfones which gave rise to substantial improvements in biochemical and cellular potency. Further work led to discovery of AMG 232, which is currently being evaluated in human clinical trials for the treatment of cancer. AMG 232 is an extremely potent MDM2 inhibitor (SPR KD = 0.045 nM, SJSA-1 EdU IC50 = 9.1 nM), with remarkable pharmacokinetic properties. The compound interacts specifically with the p53-binding pocket of MDM2 and releases the p53 protein from negative control. Treatment of cancer cells expressing wild-type p53 with AMG 232, stabilizes p53 and activates the p53 pathway, leading to cell cycle arrest and apoptosis. The compound showed excellent efficacy in a number of mouse tumor xenograft models and was well tolerated in preclinical toxicology studies at therapeutically relevant dosages.

Citation Format: Zhihong Li. Discovery of AMG 232, an inhibitor of the MDM2-p53 interaction: From lead to a clinical candidate. [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 1645. doi:10.1158/1538-7445.AM2014-1645
Abstract 1649: Extent and location of tumor infiltrating lymphocytes in colon cancer predicts outcome to adjuvant active specific immunotherapy
Authors: Hooijberg Erik , Turksma Annelies W. , Shamier Marc , Lam Kevin , Coupe Veerle MH , Weger Vincent A. de , Belien Jeroen AM , Eertwegh Alfons J. van den , Meijer Gerrit A. , Meijer Chris JLM
Published: May 2015

Purpose: We investigated the prognostic and predictive value of tumor infiltrating lymphocytes (TIL) in colorectal cancer in a cohort of patients who previously took part in a trial on adjuvant Active Specific Immunotherapy (ASI).

Experimental Design: We determined the numbers of CD3+ and CD8+ cells in archival tumor samples of 110 colorectal cancer patients as total infiltrating TIL, intraepithelial TIL and stromal TIL. Survival curves were used to determine the prognostic and predictive value of TIL.

Results: Patients with microsatellite instable (MSI) tumors had higher numbers of intraepithelial CD3+ and CD8+ TIL compared to patients with microsatellite stable (MSS) tumors. Patients with MSI tumors had a good prognosis. Patients with MSS cancers with high intraepithelial or stromal TIL numbers had a better prognosis compared to those with low TIL numbers. Significant survival benefits were found for the disease specific survival (DSS) and recurrence free interval (RFI) for patients with high infiltrates of stromal CD3+ or intraepithelial CD8+ cells. Additionally high numbers of total CD8+ and high stromal CD8+ cells gave a significant difference in RFI compared to low TIL numbers. The numbers of TIL found in MSS tumors also have predictive value with respect to response to adjuvant ASI treatment. Compared to controls significance was reached in ASI treated patients with high total CD3, high stromal CD3 or high stromal CD8 numbers in DSS as well as RFI.

Conclusion: ASI therapy is effective in patients with MSS colorectal tumors harboring high numbers of preexisting stromal CD3 or CD8 positive TIL.

Citation Format: Erik Hooijberg, Annelies W. Turksma, Marc Shamier, Kevin Lam, Veerle MH Coupe, Vincent A. de Weger, Jeroen AM Belien, Alfons J. van den Eertwegh, Gerrit A. Meijer, Chris JLM Meijer. Extent and location of tumor infiltrating lymphocytes in colon cancer predicts outcome to adjuvant active specific immunotherapy. [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 1649. doi:10.1158/1538-7445.AM2014-1649
Abstract 1659: Characterizing cytokine secretion in response to dsRNA treatment in ovarian cancer cells
Authors: Muccioli Maria , Pate Michelle , Benencia Fabian
Published: May 2015

Ovarian cancer exhibits the lowest long-term survival rates amongst all gynecological cancers, warranting more research directed at finding novel therapies, especially for late-stage disease. Inflammation is known to affect cancer progression, with tumor-promoting or repressive effects possible in different contexts, where the types of leukocytes at the tumor site are heavily implicated in disease prognosis. Tumor-associated macrophages and T-regulatory cells, for example, are known to promote tumor growth by inhibiting the anti-cancer response and by secreting pro-angiogenic factors. Importantly, the specific processes and cellular signaling pathways leading up to leukocyte infiltration in different cancer types remain to be fully elucidated. We hypothesize that dsRNA in the vicinity of ovarian cancer cells (such as from a viral infection or cellular debris) can activate pro-inflammatory regulators in tumor cells, in particular the NF-KB transcription factor, the aberrant over-activation of which is implicated in many cancers. NF-KB activation results in increased production and secretion of numerous cytokines that can in turn recruit pro-angiogenic leukocytes to the tumor microenvironment, thus favoring cancer progression. Herewith, we evaluated the production of numerous cytokines after dsRNA treatment in mouse and human ovarian cancer cells. Additionally, we determined which of the four known dsRNA receptors contributes most to this process. Consistently with our hypothesis, we have shown an increase in several pro-inflammatory cytokines (eg. CCL5 and IL6) in response to dsRNA treatment in both human and mouse ovarian cancer cell lines. We have also determined that the dsRNA receptor MDA5 contributes the most to dsRNA-induced cytokine secretion in both human and mouse ovarian cancer cell lines employing siRNA-directed knock-down of each receptor. Altogether, these data suggest that dsRNA-induced signaling in ovarian cancer cells may facilitate a tumor-favoring environment; thus dsRNA-sensing receptors may constitute attractive therapeutic targets for cancer immunotherapy. Further studies in our lab will determine the specific signaling pathways responsible for the upregulated cytokines as well as use a mouse model to examine the effect of dsRNA receptor knock-down in the cancer cells on tumor growth, angiogenesis, and the leukocyte infiltration profile.

Citation Format: Maria Muccioli, Michelle Pate, Fabian Benencia. Characterizing cytokine secretion in response to dsRNA treatment in ovarian cancer 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 1659. doi:10.1158/1538-7445.AM2014-1659
Abstract 1673: Alternative NF-κB signaling promotes inflammatory cell recruitment and lung tumor formation
Authors: Ausborn Jamie Alicyn , Cheng Dong-Sheng , Polosukhin Vasiliy , Han Wei , Yull Fiona , Blackwell Timothy
Published: May 2015

Nuclear factor-κB (NF-κB) is a transcription factor that can be activated through either the classical or alternative signaling pathways. In lung tumors, high levels of classical and alternative pathway components have been observed. While many studies have demonstrated that classical NF-κB signaling is a central pathway regulating inflammation and tumorigenesis, the role of alternative NF-κB signaling remains undefined. We hypothesize that alternative NF-κB signaling in the lung epithelium plays a key role in inflammation and lung tumor formation through paracrine signaling to the inflammatory microenvironment. To investigate alternative NF-κB signaling in the lung epithelium, we generated a transgenic mouse model with doxycycline (dox)-inducible expression of the alternative pathway component p52 in airway epithelial cells (designated ALTA for alternative pathway trans-activated). When placed on dox, nuclear p52 is increased in the lungs of ALTA mice with no effect on activation of other NF-κB components. Surprisingly, alternative pathway activation alone did not cause a significant increase in inflammatory cell recruitment. However, in conjunction with the inflammatory stimulus lipopolysaccharide (LPS), alternative pathway activation caused increased mortality, which was associated with a significant increase in inflammation and increased lung permeability. To determine the effect of alternative pathway activation on lung tumor formation, mice were injected with the lung carcinogen urethane. Six weeks after urethane administration, ALTA mice demonstrated an increased number of bronchoalveolar lavage neutrophils, and by 6 months post-urethane, ALTA mice had an increased number of tumors, increased tumor size, and more advanced lesions. Together, these data suggest that alternative NF-κB signaling plays an important role in stimulus-induced inflammatory cell recruitment and that alternative pathway activation can promote lung tumor formation and progression. By defining the role of alternative NF-κB signaling in tumor formation, we hope to gain insight into the role of inflammation in lung carcinogenesis and to identify novel pathways and signaling targets that could potentially serve as therapeutic targets.

Citation Format: Jamie Alicyn Ausborn, Dong-Sheng Cheng, Vasiliy Polosukhin, Wei Han, Fiona Yull, Timothy Blackwell. Alternative NF-κB signaling promotes inflammatory cell recruitment and lung tumor formation. [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 1673. doi:10.1158/1538-7445.AM2014-1673
Abstract 1688: Overcoming drug resistance to gemcitabine in pancreas cancer cells by targeting activated PI3K/mTOR pathway
Authors: Murakami Yuichi , Shinoda Ai , Watari Kosuke , Nabeshima Hiroshi , Kawahara Akihiko , Nagayama Daisuke , Naito Yoshiki , Higaki Koichi , Inoue Masako , Kuwano Michihiko , Ono Mayumi
Published: May 2015

Pancreas cancer is one of the most difficult neoplasms to treat curatively and 5-year survival rate is only 5%. Gemcitabine, a difluorinated deoxycitidine, has been first reported to improve pancreas cancer both response rate and median overall survival of patients with pancreas cancer. Treatment with combination of gemcitabine and erlotinib was reported to prolong survival compared with gemcitabine alone against pancreas ductal adenocarcinoma, but its benefit is modest. Further development of potent combination therapeutics with gemcitabine should be very useful for treatment of pancreas cancer.

In our present study, we examined whether the cytotoxic effect of gemcitabine could be potentiated when combined with other anticancer drugs including molecular targeted drugs and cytotoxic anticancer agents, and also whether such combination could overcome drug resistance to gemcitabine. We observed following findings [1]We first screened various anticancer agents that could synergistically potentiate cytotoxic effect of gemcitabine against pancreas cancer cell lines in culture, and observed that mTOR inhibitors, such as rapamycin, everolimus and AZD8055, synergistically enhanced the anti-proliferative effect of gemcitabine. [2] PI3K/mTOR/S6K-driven signaling pathway for de novo pyrimidine biosynthesis was markedly blocked when combined with gemcitabine and mTOR inhibitors. [3] We also observed that mTOR/S6K-driven de novo pyrimidine synthetic pathway was markedly activated in drug-resistant cancer cells as compared to their parental cell line. [4] Immunohistochemical analysis of clinical samples derived from resected pancreas tumors showed that mTOR was highly expressed in progressive pancreas cancer. Combination of gemcitabine and drugs targeting mTOR-driven signaling pathway will further provide potent therapeutics against both gemcitabine-sensitive and -resistant pancreas cancer. Furthermore, personalized therapeutics against pancreas cancer patients by this combination will be discussed based on expression levels of activated PI3K/mTOR/S6K in tumors.

Citation Format: Yuichi Murakami, Ai Shinoda, Kosuke Watari, Hiroshi Nabeshima, Akihiko Kawahara, Daisuke Nagayama, Yoshiki Naito, Koichi Higaki, Masako Inoue, Michihiko Kuwano, Mayumi Ono. Overcoming drug resistance to gemcitabine in pancreas cancer cells by targeting activated PI3K/mTOR pathway. [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 1688. doi:10.1158/1538-7445.AM2014-1688
Abstract 1689: Synergistic effect of JQ1 and rapamycin for treatment of human osteosarcoma
Authors: Lee Dhong Hyun Tony , Qi Jun , Bradner James , Said Jonathan , Doan Ngan , Forscher Charles , Koeffler H Phillip
Published: May 2015

Bromodomain and extra terminal domain (BET) proteins are important epigenetic regulators facillitating the transcription of genes in chromatin areas linked to acetylated histones. JQ1, a BET protein inhibitor, has antiproliferative activity againts many hematological and solid cancers, mainly through inhibition of c-myc and upregulation of p21. In this research, we investigated the use of JQ1 for human osteosarcoma (OS) treatment, due to the frequent copy number gain and overexpression of c-myc in OS cells. JQ1 significantly inhibited the proliferation and survival of seven OS cells (U2OS, G292, MG-63, HT-161, MNNG/HOS, SAOS-2, SJSA), at a relatively higher dose than other cancers (median IC50 = 7.35 μM). JQ1 induced G1 cell cycle arrest and premature senescence of OS cells, but produced little apoptosis. Interestingly, c-myc protein levels in JQ1-treated cells remained unchanged, whereas the upregulation of p21 protein was still observable, as noted by Western blots. Although effective in vitro, JQ1 failed to reduce the size of the MNNG/HOS xenografts in immunocompromised mice in vivo.

To overcome the resistance of OS cells to JQ1 treatment, we tested JQ1 in combination with rapamycin. Rapamycin inhibits the assembly of mTOR complexes which is one of the important growth signaling pathways in many cancers including OS. JQ1 and rapamycin synergistically inhibited the growth and survival of OS cells in vitro. The drug combination nearly abrogated expression of c-myc protein, enhanced levels of p21, and induced apoptosis in OS cells in vitro. After 24 days of treatment in vivo, JQ1 (50mg/kg body weight) or rapamycin (1.5 mg/kg body weight) alone showed little effect on the size of human OS xenografts in nude mice. However, combination of both drugs significantly inhibited tumor growth by 85% and 73% (p<0.05 for both) compared to either JQ1 or rapamycin alone, respectively.

In conclusion, JQ1 inhibited proliferation and survival of OS cells in vitro but failed to reduce growth of OS xenografts in immunodeficient mice. When combined with rapamycin, JQ1 synergistically inhibited proliferation of OS cells both in vitro and in vivo.

Citation Format: Dhong Hyun Tony Lee, Jun Qi, James Bradner, Jonathan Said, Ngan Doan, Charles Forscher, H Phillip Koeffler. Synergistic effect of JQ1 and rapamycin for treatment of human osteosarcoma. [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 1689. doi:10.1158/1538-7445.AM2014-1689