bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1276: Pannexin 1 regulates rhabdomyosarcoma tumor growth: a potential novel therapeutic target
Authors: Xiang Xiao , Langlois Stephanie , St-Pierre Marie-Eve , Barre Jessica , Le Pham Tammy , Cowan Kyle N.
Published: July 2016

Rhabdomyosarcoma (RMS) is a skeletal muscle-derived soft tissue sarcoma for which a novel therapeutic strategy is greatly needed. RMS cells have lost the ability to terminally differentiate thus proliferating indefinitely. Promoting their redifferentiation to skeletal muscle tissue is thus a promising therapeutic approach. We have recently shown that levels of Pannexin 1 (Panx1) are below detectable limits in undifferentiated myoblasts, but become highly upregulated during their differentiation and indeed promote this process. We thus hypothesize that the levels of Panx1 are deregulated in RMS and that restoration of Panx1 levels may induce differentiation of RMS cells thereby alleviating their malignant properties. Briefly, our results demonstrate that both Panx1 transcript and protein are at very low levels in, both embryonal and alveolar RMS tumor specimens and patient-derived cell lines similar to that seen in fetal skeletal muscle tissue and undifferentiated myoblasts, respectively. In vitro functional analyses have revealed that while re-introduction of Panx1 in RMS cell lines led to their partial differentiation, it significantly inhibited their rate of proliferation and migration. Moreover, re-expression of Panx1 in RMS cell lines prevented both spheroid formation and growth, and induced RMS cell apoptosis. Based on this in vitro data, pre-clinical orthotopic xenograft studies are currently underway to assess the ability of Panx1 to inhibit tumor onset and growth, and induce tumor regression in vivo. Our preliminary results thus far have shown that Panx1 over-expression significantly suppresses RMS xenograft tumor growth. Taken together, these results indicate that restoration of Panx1 levels in RMS reduces its malignant properties. This, together with further investigation into the tumor-suppressive mechanism by which Panx1 mediates its effects, will establish Panx1 as a novel therapeutic target for RMS.

Citation Format: Xiao Xiang, Stephanie Langlois, Marie-Eve St-Pierre, Jessica Barre, Tammy Le Pham, Kyle N. Cowan. Pannexin 1 regulates rhabdomyosarcoma tumor growth: a potential novel therapeutic target. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1276.
Abstract 1278: Popdc proteins: novel targets in inhibiting breast cancer cell migration and proliferation
Authors: Amunjela Johanna N. , Tucker Steven J.
Published: July 2016

Tumour growth and metastasis are major challenges in cancer treatment and metastasis is the primary cause of death in more than 90% of cancer patients with solid tumours (Gupta & Massagué 2006). This necessitates identification and validation of clinically relevant anticancer therapeutic targets that result in maximum clinical efficacy with minimal side-effects when manipulated by drugs. Our research project has identified Popeye domain containing (Popdc) proteins as promising potential therapeutic targets in the treatment of breast cancer.

Popdc proteins are a class of membrane-tethered proteins encoded by the three members of the Popdc gene family Popdc1, Popdc2 and Popdc3. Their suppression has been correlated with disease progression and poor clinical outcomes in various cancers including breast cancer, gastric cancer, lung cancer, hepatocellular carcinoma and colorectal cancer (Kim et al., 2010, Luo et al., 2012, Williams et al., 2011, Wang et al., 2011). Loss of Popdc protein expression has been correlated with increased cancer cell migration, invasion, metastasis, drug-resistance and poor patient survival (Luo et al., 2012, Williams et al., 2011, Han et al., 2014). In breast cancer, Popdc1 suppression has been reported in breast carcinoma cells while upregulation of Popdc2 has been linked to the mechanism by which arsenic trioxide mediates breast cancer cell death (Williams et al., 2011, Wang et al., 2011).

Our use of various assays including single-cell migration assays, cell population migration assays, cell proliferation assays and fluorescent imaging strongly implicate Popdc proteins as drivers of cancer cell migration and proliferation. Expression patterns of Popdc proteins revealed high expression levels in non-malignant (HMepC) breast cells and low expression in breast cancer (MCF7) cells. This suggests that loss of Popdc protein expression correlates with an increasingly malignant phenotype. Furthermore, knocking down Popdc1, Popdc2 and Popdc3 in MCF7 breast cancer cells significantly promoted cell migration both at single cell and at cell population level while Popdc2 knockdown significantly promoted unstimulated MCF7 cell division. Taken together, this strong data set suggests that Popdc proteins play significant roles in the migration and proliferation of different cancer cell lines and that they indeed represent promising potential therapeutic targets in reducing breast cancer cell division and migration.

Citation Format: Johanna N. Amunjela, Steven J. Tucker. Popdc proteins: novel targets in inhibiting breast cancer cell migration and proliferation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1278.
Abstract 1280: The intersection of the PI3K/mTOR and HIPPO pathways: a potential therapeutic target for medulloblastoma
Authors: Ronecker Jennifer S. , Lee Paul , Sridhara Sudeepta , LaBagnara Michael , Murali Raj , Jhanwar-Uniyal Meena
Published: July 2016

Medulloblastoma (MB) is the most common malignant pediatric brain tumor; it is genetically defined into 4 subgroups which defines the clinical course of the disease. Despite current treatment modalities such as surgical resection, radiation, and chemotherapy, there are still significant morbidities and adverse side-effects. Recently defined genetic pathways could possibly offer a better management and treatment of patients with MB. The HIPPO signaling pathway has recently been shown to intersect with PI3K and Sonic Hedgehog (SHH) pathways to modulate mechanistic target of rapamycin (mTOR), an important regulator in cell growth and migration. The aim of this study is to elucidate the interaction of Hippo and Akt/mTOR signaling pathways with SHH pathways in MB, and define the potential downstream targets of therapy by using combined inhibitors. In order to achieve our goals, we evaluated the expression of activated AKT/mTOR in MB tumors using immunohistochemistry (IHC), along with the expression of transcriptional co-activators yes-associated protein (YAP), a down-stream effector of the HIPPO pathway. Scratch and chemotactic migration were used to assess the motility of MB cells following treatment with inhibitors of the aforementioned pathways. Cell cycle analysis and proliferation were measured using EdU and MTT techniques, respectively. YAP-SiRNA treatment was used to inhibit HIPPO pathway. MicroRNA-29, which mediates the mTOR pathway by modulating PTEN levels, was investigated. Results demonstrated: 1. A significant number of MBs expressed activated pAKTSer473 and pmTORSer2448, along with YAP, the downstream effector of the HIPPO pathway; 2. Treatment with inhibitors of PI3K/AKT (LY294002) and mTOR (rapamycin), given with EGF, significantly reduced cellular motility; 3. SHH inhibitor cyclopamine given with LY294002 or rapamycin significantly reduced cellular motility; 4. Cell proliferation was suppressed by inhibition of the HIPPO pathway using 3 unique 27mer duplexes of YAP1 siRNA treatments; 5. Combined treatment of cyclopamine with either LY294002 or rapamycin caused maximum suppression of cellular growth. In conclusion, these results demonstrate that the Hippo and PI3K/Akt/mTOR pathways are over-expressed in MB, and that the combined targeting of mTOR/HIPPO pathway with SHH inhibitor may provide alternative strategies for the treatment of MB, and offers a targeted therapy for improved prognosis.

Citation Format: Jennifer S. Ronecker, Paul Lee, Sudeepta Sridhara, Michael LaBagnara, Raj Murali, Meena Jhanwar-Uniyal. The intersection of the PI3K/mTOR and HIPPO pathways: a potential therapeutic target for medulloblastoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1280.
Abstract 1302: A comprehensive approach to delineate FGFR2 targeted therapy response in diffuse gastric cancer
Authors: Chen Jiamin , Lee Hojoon , Ji Hanlee
Published: July 2016

Dysregulation of fibroblast growth factor receptors (FGFRs) signaling has been associated with tumorigenesis and progression in various cancers. Preclinical models have shown that FGFR2 signaling activation due to FGFR2 amplification is essential driver for specific gastric cancers. Cancer cell lines or patient-derived gastric cancer xenograft models with FGFR2 amplification are highly sensitive to AZD4547, which is a small-molecule tyrosine kinase (TRK) inhibitor that selectively targets FGFR1-3. However, recent clinical trial studies failed to demonstrate the therapeutic potential of AZD4547 in patients with FGFR2 amplification. For this study, we identified critical genetic and molecular features that determine the response to FGFR2 inhibitors in gastric cancer.

To systematically delineate the pathways that modulate response to FGFR2 inhibitors, we identified CRISPR-library-based gene knockouts that result in resistance to AZD4547 in Kato III cells, a gastric cancer cell line with high sensitivity to this drug. Although majority of transduced Kato III cells undergo cell senescence upon AZD4547 treatment, a small population of resistant cells rendered by CRISPR knockout is enriched. Subsequent sequencing and pathway analysis reveal the alternative activated pathways that contribute to the cell growth in lieu of FGFR2 inhibition.

Furthermore, we applied the elastic-net regression analysis to identify genetic features associated with the drug sensitivity to FGFR2 inhibition in cancer cell lines. The cell line drug response data are derived from the Cancer Therapeutic Response Portal (CTRP v2) and the Cancer Cell Line Encyclopedia (CCLE). Notably, few cancer cell lines have high FGFR2 copy numbers (CN>4). The elastic-net algorithm simultaneously integrates mutation, copy number, gene expression data, and forms multiple linear regression to learn the coefficient weights associated with FGFR2 inhibitor sensitivity. Using the method, we identify the whole spectrum of genetic and molecular features that are potential drug response markers.

In summary, our study provides new insight into genetic and molecular features that implicate in the cellular response to FGFR2 inhibitors, which can facilitate the development of more effective strategies to treat patients with FGFR2 amplification.

Citation Format: Jiamin Chen, Hojoon Lee, Hanlee Ji. A comprehensive approach to delineate FGFR2 targeted therapy response in diffuse gastric cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1302.
Abstract 1311: High order drug combinations are required to effectively kill colorectal cancer cells
Authors: Horn Thomas , Ferretti Stephane , Ebel Nicolas , Tam Angela , Ho Samuel , Harbinski Fred , Farsidjani Ali , Zubrowski Matt , Sellers William R. , Schlegel Robert , Porter Dale , Morris Erick , Wuerthner Jens , Jeay Sebastien , Greshock Joel , Halilovic Ensar , Garraway Levi A. , Caponigro Giordano , Lehar Joseph
Published: July 2016

Tumors are complex biological systems that often retain proliferative capacity even when challenged with drug treatment. Given this resiliency, drug combinations may provide greater therapeutic benefit, however, which molecules to combine and how many to include in combinations for effective responses is not clear yet. Using image-based proliferation and apoptosis assays in colorectal cancer cell lines we systematically investigated combinations that ranged in number from two to six drugs and targeted critical oncogenic pathways. Drug pairs targeting key signaling pathways resulted in synergies across a broad spectrum of genetic backgrounds, but often yielded only cytostatic responses. Enhanced cytotoxicity was observed when additional processes including apoptosis and cell cycle were targeted as part of the combination. In many cases, where cell lines were resistant to two- and three-way drug combinations, increased expression of anti-apoptotic proteins was observed and induction of cytotoxic responses required up to fourth-order combinations. Our results demonstrate that high-order drug combinations might be needed to kill cancers and show how systematic drug combination screening together with a molecular understanding of drug responses can guide their identification.

Citation Format: Thomas Horn, Stephane Ferretti, Nicolas Ebel, Angela Tam, Samuel Ho, Fred Harbinski, Ali Farsidjani, Matt Zubrowski, William R. Sellers, Robert Schlegel, Dale Porter, Erick Morris, Jens Wuerthner, Sebastien Jeay, Joel Greshock, Ensar Halilovic, Levi A. Garraway, Giordano Caponigro, Joseph Lehar. High order drug combinations are required to effectively kill colorectal cancer cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1311.
Abstract 1314: Evaluating payload delivery to the CNS from surface-modified nanoparticles
Authors: Householder Kyle T. , DiPerna Danielle , Ghaffari Layla , Sirianni Rachael
Published: July 2016

The blood-brain barrier (BBB) remains a major obstacle to treatment of intracranial tumors. Most drugs are poorly bioavailable in the brain, and peripheral toxicity can severely limit administerable dose. Drug loaded nanoparticles are capable of accumulating preferentially in the core of angiogenic tumors by a mechanisms known as the enhanced permeation and retention (EPR) effect, which can improve therapeutic efficacy of encapsulated molecules compared to free form. However, to achieve complete tumor kill, chemotherapy must target both the leaky tumor core and cells that reside behind an intact blood-brain barrier.

The goal of this work was to evaluate payload delivery to the CNS from nanoparticles modified with one of four targeting ligands: TAT, antennapedia (AP), angiopep2 (Ang2) or Tet-1. Poly(lactic acid)-co- poly(ethylene glycol) nanoparticles (NPs) encapsulating Nile Red, a small fluorescent dye, were produced by solvent-evaporation technique. Peptides were covalently attached to the nanoparticle surface by a maliemide-thiol reaction. The effect of targeting ligands on NP payload delivery to healthy brain and spinal cord compared to unmodified NPs was evaluated in mice at 2 and 6 hours post injection. Mice were perfused with saline, and the brains and spinal cord were removed, homogenized and payload delivery quantified by fluorescence.

All targeting ligands evaluated significantly increased payload delivery to the brain at both 2 and 6 hours, with the cell-penetrating peptides, TAT and AP, producing the greatest effect of a 4-fold increase over control. Additionally, the cell-penetrating peptides significantly increased payload delivery to the spinal cord at 2 hours but had no significant effect on delivery at 6 hours. Interestingly, the receptor-mediated peptides, Ang2 and Tet-1, did not significantly change spinal cord delivery at 2 hours but significantly increased payload concentration at 6 hours. These data suggest both receptor and nonspecific targeting ligands affect payload delivery and kinetics in a regionally specific manner. Future work is focused on evaluating additional ligands to better understand how the type of cell-nanoparticle interactions affects payload distribution.

Citation Format: Kyle T. Householder, Danielle DiPerna, Layla Ghaffari, Rachael Sirianni. Evaluating payload delivery to the CNS from surface-modified nanoparticles. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1314.
Abstract 1353: Validation and implementation of screens for partial agonists and antagonists of the ErbB4/HER4 receptor tyrosine kinase: Targeted melanona drug discovery
Authors: Cullum Richard , David Allan , Riese David J.
Published: July 2016

Introduction: Gain-of-function mutations in the gene encoding the ErbB4 (HER4) receptor tyrosine kinase have been detected in melanoma cell lines and these cell lines are dependent on ErbB4 expression for proliferation. Consequently, ErbB4 antagonists hold promise as targeted melanoma therapeutics and we have sought to identify small molecule ErbB4 antagonists. Our approach is based on the observation that the Q43L mutant of the ErbB4 peptide growth factor agonist Neuregulin 2beta (NRG2b) functions as a partial agonist at ErbB4; NRG2b/Q43L stimulates ErbB4 tyrosine phosphorylation, fails to stimulate ErbB4 coupling to cell proliferation, antagonizes stimulation of cell proliferation by ErbB4 full agonists, and causes ErbB4 down-regulation.

Experimental procedures: We have developed a high-throughput screening (HTS) strategy that can be used to identify ErbB4 antagonists. The primary screen identifies molecules that stimulate ErbB4 tyrosine phosphorylation. The secondary screen distinguishes between molecules that stimulate and fail to stimulate ErbB4-dependent proliferation. The tertiary screen identifies molecules that antagonize agonist stimulation of ErbB4-dependent proliferation.

Results: An phospho-ErbB4 sandwich ELISA assay identifies molecules that stimulate ErbB4 tyrosine phosphorylation with high sensitivity and fidelity (Z’ >0.5). MTT assays using a cell line that displays ErbB4-dependent proliferation distinguish between molecules that stimulate and fail to stimulate ErbB4-dependent proliferation (Z’>0.5) and identify molecules that antagonize agonist stimulation of ErbB4 dependent proliferation (Z’>0.5). These assays have been used to identify small molecules that stimulate ErbB4 tyrosine phosphorylation. Efforts to determine whether these hits function as ErbB4 full agonists or partial agonists (antagonists) are underway and will be reported. Structures of these small molecule ErbB4 full and partial agonists may be reported, pending submission of a provisional patent application.

Conclusions: We have validated an HTS strategy for identifying ErbB4 partial agonists that function as ErbB4 antagonists and deployment of that strategy has led to the identification of several hits. Such molecules may hold promise as targeted therapeutics for melanoma and other ErbB4-dependent tumors.

Citation Format: Richard Cullum, Allan David, David J. Riese. Validation and implementation of screens for partial agonists and antagonists of the ErbB4/HER4 receptor tyrosine kinase: Targeted melanona drug discovery. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1353.
Abstract 1355: A novel derivative of benzoxazole enhances radiosensitivity of human colon cancer cells through p53-mediated G2/M arrest and mitochondrial apoptotic pathway
Authors: Ryu Hwani , Seo Bitna , Kim Jooyoung , Jung In-Sung , Song Jie-Young , Ahn Jiyeon
Published: July 2016

The tumor suppressor p53 plays a critical role in cell cycle regulation and apoptotic cell death, and its activation can sensitize cancer cells to radio- or chemotherapy. To identify small molecules that induce apoptotic cell death via increasing p53 transcriptional activity, we designed and synthesized 44 benzoxazole derivative small molecule compounds. Using a cell-based screening with p53-responsive luciferase reporter assay system of benzoxazole derivatives, we found AU14022, which significantly increased p53 transcriptional activity in a concentration dependent manner. Treatment of AU14022 increased p53 protein expression, its phosphorylation on serine 15 and its target p21 gene/protein expression and induced apoptotic cell death measured by FACS analysis using Annexin V/PI double staining in wild-type p53 HCT116 human colon cancer cells but not in p53 knockout HCT116 cells. Furthermore, treatment of HCT116 cells with AU14022 exhibited mitochondrial dysfunction including bcl-2 family modulation and cytochrome c release. Combination treatment of AU14022 and ionizing radiation (IR) synergistically induced apoptotic cell death, compared to IR or AU14022 alone. Further investigation demonstrated that cell cycles are significantly arrested at G2/M phase, and marked inductions of p21 protein expression and phosphorylation of Chk1 protein and a decrease of cyclin B1 protein expression were observed by treatment of AU14022 alone and combination treatment with AU14022 and IR. Our findings indicate that AU14022 induced apoptosis through p53-mediated cell cycle arrest with mitochondrial dysfunction, leading to enhanced radiosensitivity of colon cancer cells. These results provide a basis for the further investigations required for its assessment as a promising anticancer agent.

Citation Format: Hwani Ryu, Bitna Seo, Jooyoung Kim, In-Sung Jung, Jie-Young Song, Jiyeon Ahn. A novel derivative of benzoxazole enhances radiosensitivity of human colon cancer cells through p53-mediated G2/M arrest and mitochondrial apoptotic pathway. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1355.
Abstract 1360: Genistein-AR antagonist conjugate potently induce growth arrest in both androgen dependent and independent prostate cancer cell lines
Authors: Raji Idris O. , George Alex , Kucuk Omer , Oyelere Adegboyega K.
Published: July 2016

The ability of prostate cancer to transition from androgen dependent- to androgen independent state (castration resistant), during androgen deprivation therapy, requires that new chemotherapeutic agents be introduced that will retain activity in the two prostate cancer types. Though described as androgen independent, castration resistant prostate cancer (CRPC) still relies heavily on androgen receptor (AR) signaling for survival. Enzalutamide, an AR antagonist, is probably that most effective drug used in androgen deprivation therapy for CRPC and AR dependent prostate cancer. Despite its clinical success, there is the fear of resistant to enzalutamide due to mutation in the ligand binding domain of AR. This poses a challenge that requires developing new agents devoid of resistance, in the event of mutation. One of the approaches being proposed is to develop new agents that can selectively induce AR degradation, an approach that was successfully used to develop the selective estrogen receptor degrader, fulvestrant.

In our study, we incorporate genistein, a soy isoflavone known to perturb multiple biochemical pathways relevant for cancer survival, into an AR binding template to give AG-1-33. AG-1-33 was evaluated for growth inhibitory activity in LNCaP (AR dependent) and DU-145 (AR independent) prostate cancer cell lines. This compound retained its AR antagonist activity and also showed low micromolar anti-proliferation activity in the two prostate cancer cell lines. We also evaluated the effect of AG-1-33 on AR expression, as well as its ability to act as a selective AR degrader in LNCaP.

Citation Format: Idris O. Raji, Alex George, Omer Kucuk, Adegboyega K. Oyelere. Genistein-AR antagonist conjugate potently induce growth arrest in both androgen dependent and independent prostate cancer cell lines. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1360.
Abstract 1370: A new RAS G12D specific rabbit polyclonal antibody for immunohistochemical FFPE application
Authors: Chen. Chi-Kuan
Published: July 2016

Background: Determining RAS mutation status is considered as prognostic factor for colorectal cancer patients in anti-epidermal growth factor receptor (EGFR) antibody therapy. Because genetic testing remains costly and time-consuming, it is necessary to develop mutation specific antibody for immunological analysis. However RAS G12D mutation antibody for immunohistochemical (IHC) staining is not available.

Results: Here we report a novel RAS G12D rabbit polyclonal antibody (GTX132407) for immunochemistry (IHC) testing, which was developed by cooperating with GeneTex In. Based on analyzing 15 clinical colorectal cancer specimens, the result of IHC staining using RAS G12D mutation specific antibody (GTX132407) was correlated with genetic testing of RAS G12D mutation using ultra-sensitive total RAS mutation screen kit [Hong Jing Biotechnology Inc. (FemtoPath)].

Conclusion: In summary, the result showed that IHC testing with RAS G12D specific mutation antibody could be a reliable tool for predicting RAS genetic mutation status.

Citation Format: Chi-Kuan Chen. A new RAS G12D specific rabbit polyclonal antibody for immunohistochemical FFPE application. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1370.
Abstract 1395: Development and validation of NGS-based clinical cancer panels for precision medicine in Asian and Caucasian adult cancers
Authors: Kuo Winston Patrick , Wang Amy , Cheng Tak , Du Pan , Jia Shidong
Published: July 2016

In the new era of precision medicine, comprehensive cancer panels are needed to identify actionable cancer genes and match patients to personalized therapies. Here, we report a Pan-Cancer NGS Panel designed by leaders in the field that comprises of all exon coding and selected hotspots of critical cancer genes in the most common cancer types for both Asian and Caucasian cohorts, including but not limited to lung, colon, breast and prostate cancer. The panel allows detection of known and novel variants in indication-specific signaling pathways, disease biology, DNA repair, and in particular drug resistance, to advance research into personalized cancer treatment and for use in clinical trials to help the development of new targeted therapies. Technical validation was performed to characterize point mutations, indels, copy number variations and fusions across clinically relevant, actionable cancer genes in both liquid biopsies and formalin-fixed paraffin-embedded (FFPE) clinical specimens. Assay sensitivity, specificity and accuracy were tested using reference samples with known genetic profiling and further validated on a variety of orthogonal platforms such as digital PCR, allele-specific PCR and Sanger sequencing. To the best of our knowledge, this is the first ethnic-specific NGS diagnostic test designed specifically for Asian and Caucasian adult cancer patients.

Citation Format: Winston Patrick Kuo, Amy Wang, Tak Cheng, Pan Du, Shidong Jia. Development and validation of NGS-based clinical cancer panels for precision medicine in Asian and Caucasian adult cancers. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1395.
Abstract 1426: Racial variation in annexin A2(AnxA2) gene expression and poor outcome in triple-negative breast cancer
Authors: Gibbs Lee D. , Vishwanatha Jamboor K.
Published: July 2016

Background: Triple-negative breast cancer (TNBC) is identified by the absence of three major receptors that drive most breast cancers. The incidence of TNBC is also associated with health disparity due to its disproportionate occurrence in African American (AA) women. Our previous studies and others have shown that AnxA2 is overexpressed in TNBC, but its association with racial variation and outcomes is unknown.

Methods: AnxA2 gene expression was evaluated in breast cancer subtypes from The Cancer Genome Atlas (TCGA) Breast Invasive Carcinoma (BRCA) mRNA database that include 1,094 patients. Associations between clinical outcomes and AnxA2 gene expression were tested in a genome wide association study of combined publicly available datasets that include 4,147 patients.

Results: AnxA2 gene expression was significantly increased in TNBC in comparison to other breast cancer subtypes. Furthermore, AnxA2 gene expression was significantly elevated in AA women in comparison with Caucasian (CA) women and was associated with the disproportionate occurrence of TNBC in AA women. High expression levels of AnxA2 was associated with reduced overall survival (hazard of death = 2.66; 95% confidence interval {CI] = 1.14 - 6.25, P = 0.0192), reduced relapse free survival (hazard of relapse = 1.45; 95% confidence interval {CI] = 1.12 – 1.89, P = 0.0051), and reduced distant metastasis free survival (hazard of metastasis = 1.7; 95% confidence interval {CI] = 1.00 – 2.91, P = 0.048). AnxA2 gene expression was not associated with poor outcome in other intrinsic breast cancer subtypes, such as Luminal A, Luminal B, and Her-2, indicating the specific association of AnxA2 with the aggressive behavior of TNBC.

Conclusion: AnxA2 overexpression is associated with racial variation and is a potential prognostic candidate for TNBC.

Impact: AnxA2 has potential prognostic value, implicating a role for AnxA2 in the

aggressive biology of TNBC in AA women.

Citation Format: Lee D. Gibbs, Jamboor K. Vishwanatha. Racial variation in annexin A2(AnxA2) gene expression and poor outcome in triple-negative breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1426.
Abstract 1435: Psychoimmunological effect of depression on cervical carcinoma: Genome-wide long noncoding RNA implications
Authors: Dai Wanjun , Yu Yinhua , Kang Yu
Published: July 2016

Objective: Cervical cancer is the second most commonly diagnosed cancer and the third leading cause of cancer death among females in developing countries. Depression is a vital psychological factor for tumor progression. Long noncoding RNAs(LncRNAs) are a novel group of gene regulators and may contribute to the disease development. Herein, the study aims to discover the mechanisms of psychoimmunological effect of depression on cervical carcinoma from the perspective of long noncoding ribonucleic acid.

Methods: Six tumor tissues were obtained from Ib-IIa invasive squamous cell cervical carcinoma patients during laproscopic surgeries. They were divided into high-depressed group and low-depressed group according to the scoring system. LncRNA microarray experiments were performed. Expression levels of six lncRNA-mRNA pairs were confirmed by quantitative real-time PCR in other 20 patients’ materials.

Results: We found that 1621 lncRNA transcripts are differently expressed between high-depressed and paired low-depressed samples, with 510 up-regulated and 1111 down-regulated (fold-change>2.0). Most of lncRNAs were found trans-regulated by three TFs with most credentiality: BCL11A,MEF2A and TBP. In the KEGG analysis, the most down-regulated mRNA involved was systemic lupus erythematosus (Maximun transcriptional domain coverage: 25.5%). In the LncRNA and mRNA Coexpression, the function of lncRNAs with most differently expressions was predicted in relationship with interleukin-1 receptor activity (Number of lncRNAs annotated: 15).

Conclusions: Our study identified that genome-wide LncRNA profiles are significantly altered in cervical carcinoma patients with worse depressed profiles, and the psychoimmunological effect of tumor progression needs further investigation.

Keywords: Depression, Cervical Cancer, Immunology, Long-noncoding RNA

Citation Format: Wanjun Dai, Yinhua Yu, Yu Kang. Psychoimmunological effect of depression on cervical carcinoma: Genome-wide long noncoding RNA implications. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1435.
Abstract 1456: Dexamethasone suppresses cytokine-induced dual oxidase 2 (Duox2) and VEGF-A expression in human pancreatic cancer cells in vitro and pancreatic cancer growth in xenografts
Authors: Wu Yongzhong , Antony Smitha , Meitzler Jennifer L. , Lu Jiamo , Juhasz Agnes , Jiang Guojian , Roy Krishnendu , Doroshow James H.
Published: July 2016

Chronic pancreatic inflammation is strongly associated with pancreatic cancer. We previously demonstrated that inflammatory cytokines interact to produce a Duox2 (one of 7 members of the NADPH Oxidase family)-dependent, reactive oxygen species (ROS)-related, pro-oxidant milieu that could increase the pathologic potential of pancreatic inflammation, enhancing malignant transformation. Furthermore, we have shown that Duox2 expression is upregulated in patients with chronic pancreatitis as well as pancreatic ductal adenocarcinoma (PDAC). Dexamethasone (Dex) has been reported to inhibit PDAC invasiveness, the formation of pancreatic intraepithelial neoplasia in genetically-engineered mouse models of pancreatic cancer, as well as epithelial to mesenchymal transition, and local tumor recurrence and metastasis in vivo. Using cultured human pancreatic cancer cell lines, we found that in a dose- and time-dependent fashion Dex inhibited cytokine (IFN-γ/LPS, IL-4, and IL-17)-mediated up-regulation of Duox2 and VEGF-A expression in several human pancreatic cancer cell lines, including BxPC-3, ASPC-1, and CFPAC-1. The effects of Dex were abolished by pre-treatment with the Dex antagonist RU-486. As expected, we did not observe anti-proliferative effects of Dex on PDAC cells in vitro. However, Dex strongly repressed Duox2 mRNA and protein expression as well as the growth of xenografts initiated from BxPC-3 cells; in contrast, for the MIA-Paca line that is unresponsive to cytokines in culture, Dex produced no effect on Duox2 expression and tumor growth when these cells were grown as xenografts. Examination of the human Duox2 promotor in silico revealed a putative negative glucocorticoid receptor (GR) binding element. Western analysis, using nuclear extracts from pancreatic cancer cells treated with Dex, revealed that both activated glucocorticoid receptor and certain co-repressors, such as NCOR-1/2 and histone deacetylases (HDAC1, 2, and 3) exist in human pancreatic cancer cell nuclei. Our ongoing experiments are focused on understanding the molecular mechanism of Dex-mediated Duox2 repression both in vitro and in vivo. In summary, these studies suggest that cytokine-related oxidant stress, generated by Duox2, could play a role in the progression of pancreatic cancer.

Citation Format: Yongzhong Wu, Smitha Antony, Jennifer L. Meitzler, Jiamo Lu, Agnes Juhasz, Guojian Jiang, Krishnendu Roy, James H. Doroshow. Dexamethasone suppresses cytokine-induced dual oxidase 2 (Duox2) and VEGF-A expression in human pancreatic cancer cells in vitro and pancreatic cancer growth in xenografts. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1456.
Abstract 1483: MT-3724, an engineered toxin body targeting CD20 for non-Hodgkin's lymphoma
Authors: Robinson Garrett L. , Rajagopalan Sangeetha , Brieschke Brigitte , Erdman Jennifer , Neill Jane , Flores-Lefranc Rodney E. , Foree Julia , Null William , Liu Jensing , Higgins Jack P. , Willert Erin K.
Published: July 2016

Molecular Templates has developed engineered toxin bodies (ETBs), potent recombinant immunotoxins that combine the specificity of an antibody fragment with the powerful direct cytotoxicity of the Shiga-like toxin A subunit to specifically destroy target expressing cells. The use of other immunotoxins and antibody-drug conjugates is limited to targets with tumor-specific cell surface expression that efficiently internalize. The ETB scaffold has been designed to overcome this limitation and promote forced internalization of ETBs, a property that expands the potential targets to receptors with poor internalization kinetics. An additional benefit to ETBs is the mechanism of action (MOA) which is unique to that of other oncology treatments.

CD20, a receptor that does not undergo rapid internalization, is a well characterized target for agents used to treat non-Hodgkin's lymphoma (NHL). Thus, successful CD20-targeted clinical strategies include monoclonal antibodies (mAbs) and radiolabeled mAbs that act on the cell surface. Molecular Templates’ lead compound, MT-3724, is a CD20-targeted ETB that has been engineered to force the rapid internalization of the immunotoxin after binding to CD20. MT-3724 has potent direct cell kill activity on CD20 positive lymphoma cells and is the first immunotoxin to CD20 to enter the clinic. The on-going phase I study being conducted at Memorial Sloan-Kettering Cancer Center, MD Anderson Cancer Center, and New York University Langone Medical Center has shown promising safety and efficacy in highly refractory NHL patients.

Once MT-3724 is delivered to appropriate cells, the Shiga toxin A subunit inhibits protein synthesis and promotes apoptosis of tumor cells. The difference in MOA allows for activity in the refractory setting, where resistance to other treatments has emerged, and may also allow for combination therapy. Pre-clinical studies with immunomodulatory drugs (IMiDs), PI3K and Bcl-2 inhibitors that are used in treatment of NHL have shown promising results when combined with MT-3724. MT-3724 is a promising novel agent in the treatment of NHL and other CD20 positive hematological malignancies. Data around the internalization kinetics, enzymatic activity, and combination with other agents will be presented.

Citation Format: Garrett L. Robinson, Sangeetha Rajagopalan, Brigitte Brieschke, Jennifer Erdman, Jane Neill, Rodney E. Flores-Lefranc, Julia Foree, William Null, Jensing Liu, Jack P. Higgins, Erin K. Willert. MT-3724, an engineered toxin body targeting CD20 for non-Hodgkin's lymphoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1483.
Abstract 149: Integrated analysis of intratumor heterogeneity with genetic and epigenetic aberrations during colorectal cancer evolution
Authors: Kidogami Shinya , Niida Atsushi , Uchi Ryutaro , Takahashi Yusuke , Mori Masaki , Miyano Satoru , Mimori Koshi
Published: July 2016

[background] Intratumor heterogeneity (ITH) is presumably generated by branching clonal evolution of cancer cells. Recently, a multiregional sequencing approach, which sequences DNA sampled from geographically separated regions of a single tumor, has revealed branched evolution and ITH. In this study, we present genetic and epigenetic analysis of ITH in a series of nine colorectal cancers

[material and method] We conducted analysis of samples from geographically separated regions from nine colorectal tumors. From each of the nine tumor, we obtained 5-21 multiregional samples, which were 75 samples in total, together with 9 paired normal mucosa samples. For two cases, samples from liver metastases were obtained. We performed exome sequencing and copy number (CN), methylation, and mRNA expression array profiling.

[Result] Each of the multiregional mutation profiles harbored founder and progressor mutations; founder mutations are shared by all regions while progressor mutations are not. We found that mutations in well-known driver genes such as APC, KRAS, and FBWX7 were acquired as founder mutations, while mutations in PIK3CA recurrently occurred as progressor mutations, suggesting that PIK3CA mutations are a late event in the evolution of colorectal cancer. We counted each category of mutation and then identified a correlation between the number of founder mutations and the age of the patients. Our analysis showed that C > T transitions at CpG sites are more prominent in founder mutations than in progressor mutations. Then, the multiregional CN profiles showed that amplifications of 7p, 13q, 10q, 20p, and 20q frequently occurred across all samples in multiple tumors, namely as founder CN alterations. Finally, our data showed that hypermethylations in CpG islands were more prominent in founder methylations than in progressor methylations, suggesting that CpG island hypermethylations mainly occur in the early phase of colorectal cancer evolution.

[Conclusion] In this study, our integrated analysis revealed not only extensive ITH, but also the evolutionary histories of the nine tumors. In particular, by focusing on founder and progressor mutations, we identified clues for decoding the life history of the tumors.

Citation Format: Shinya Kidogami, Atsushi Niida, Ryutaro Uchi, Yusuke Takahashi, Masaki Mori, Satoru Miyano, Koshi Mimori. Integrated analysis of intratumor heterogeneity with genetic and epigenetic aberrations during colorectal cancer evolution. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 149.
Abstract 1502: Three intrinsic subtypes of prostate cancer with distinct pathway activation profiles differ in prognosis and treatment response
Authors: You Sungyong , Knudsen Beatrice , Erho Nicholas , Alshalalfa Mohammed , Takhar Mandeep , Ashab Hussam Al-deen , Davicioni Elai , Karnes Robert Jeffrey , Klein Eric A. , Den Robert B. , Ross Ashley E. , Schaeffer Edward M. , Garraway Isla P. , Kim Jayoung , Freeman Michael R.
Published: July 2016

In the era of precision medicine, genomic classifications of prostate cancer (PC) have not broadly impacted patient care. Here we present an integrative transcriptome analysis of 14 disease-related pathways in over 4,600 clinical specimens and 25 PC preclinical models. This approach provides a novel classification scheme, inclusive of 3 PC subtypes (PCS1-3), which predict disease progression and drug resistance. The PCS1 and PCS2 categories possess characteristics of luminal cells, while PCS3 exhibits basal features. Castration-resistant and metastatic cancers are over-represented in PCS1 and PCS3 tumors. All 3 subtypes are represented in commonly used PC cell lines; however, none of the mouse models we analyzed resembles PCS3. PCS classification using RNA expression data appears to be stable across primary and metastatic human tumors, cell lines, xenografts and mouse models. This new subtyping method provides novel opportunities for patient stratification that reflect specific features of tumor biology and may influence therapeutic decisions.

Citation Format: Sungyong You, Beatrice Knudsen, Nicholas Erho, Mohammed Alshalalfa, Mandeep Takhar, Hussam Al-deen Ashab, Elai Davicioni, Robert Jeffrey Karnes, Eric A. Klein, Robert B. Den, Ashley E. Ross, Edward M. Schaeffer, Isla P. Garraway, Jayoung Kim, Michael R. Freeman. Three intrinsic subtypes of prostate cancer with distinct pathway activation profiles differ in prognosis and treatment response. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1502.
Abstract 1503: Discovery of new breast cancer relevant genes using the cancer genomics public database
Authors: Samant Ritika R. , Samant Rajeev S.
Published: July 2016

We used the cBio portal to query the importance of key players in breast cancer. Telomerase (TERT) is an important protein that ensures successful immortalization of cancer cells. Immortalization is a key step to establish uncontrolled cell division. MYC is an oncogene that is very relevant in breast cancer. Our searches showed that both genes (TERT and MYC) are amplified in a high percent of patients. We wanted to explore if there are genetic events that co-occur with telomerase amplification. We found that NONO gene mutation had a high tendency of co-occurrence with telomerase amplification. We wanted to understand if this subset of tumors with mutation of NONO reveals any additional correlations. Our query revealed that PDE4DIP (Phosphodiesterase 4D interacting protein) is amplified concurrent with the NONO mutation. There is no notable cancer related data on PDE4DIP in publicly available peer-reviewed publication records (PubMed). However, when we queried PDE4DIP, we discovered that it is amplified in a large majority of the breast cancer cases. Thus, it is surprisingly more common than TERT or MYC amplification. We propose that it is very important to understand the relevance of PDE4DIP and NONO to breast cancer.

Citation Format: Ritika R. Samant, Rajeev S. Samant. Discovery of new breast cancer relevant genes using the cancer genomics public database. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1503.