bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 332: Establishment of animal model for metastasis of human lung cancers using NOJ/SCID mice.
Authors: Aragane Naoko , Sato Akemi , Kobayashi Naomi , Ide Masaru , Sueoka Eisaburo , Okada Seiji , Kimura Shinya 
Published: August 2014

Abstract
Lung cancer is the leading cause of cancer death among malignant tumors. Lymphogenic and distant metastases were found approximately 70% at the diagnosis, and most of recurrence after surgery occurs as distant metastasis. Therefore, it is important to establish the new strategy for control of metastatic lung cancers. Since metastasis consists of several steps such as loss of cell adhesion, intravasation, survival in circulation, exit into new tissues, and colonization in a distant site, a suitable animal model is indispensable for investigation of metastasis. In addition, recent investigations revealed that biological and molecular characteristics of cancer cells were different between primary and metastatic lesions, as well as among each metastatic site. To establish the animal model reflecting human metastatic lung cancers, we used NOD/Scid/Jak3null (NOJ) mice, which exhibit deficiencies in NK cell activity, macrophage and DC function, and complement activation as well as T and B cell deficiencies. This animal model enabled us to engraft various human cells more easily than previous immunodeficiency mouse models. At first, twenty kinds of human lung cancer cell lines were screened by expression pattern of E-cadherin and vimentin using western blot analysis, and two cell lines with high expression of vimentin and low expression of E-cadherin were selected. The cells (1×107/mouse) were injected into the dorsal flanks, and one cell line, H1975 showed lymphogenic metastasis to axillary and abdominal lymph nodes, resulting dissemination to pleura and diaphragm with ascites. Since H1975 cells carry epidermal growth factor receptor (EGFR) mutation, L858R and T790M, we can monitor the existence of cancer cells using a sensitive detection system, mutation-biased PCR and quenched probe system (MBP-QP), which were established in our laboratory. We have also tried to establish the animal model which lung cancer cells are inoculated into lung to mimic metastasis of human lung cancers. Using these model systems, we have been performing comprehensive analysis including mRNA expression, the mutation status of lung cancer related genes in each metastatic site, and compared the phosphorylation status of 38 kinds of molecules. Our goal of this study is to establish the animal model to reproduce phenotypes of metastasis of human lung cancers for screening of anti-metastatic agents.
Citation Format: Naoko Aragane, Akemi Sato, Naomi Kobayashi, Masaru Ide, Eisaburo Sueoka, Seiji Okada, Shinya Kimura. Establishment of animal model for metastasis of human lung cancers using NOJ/SCID mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 332. doi:10.1158/1538-7445.AM2013-332
Abstract 3324: The discovery and optimisation of small-molecule inhibitors of human 5’-tyrosyl DNA phosphodiesterase (Tdp2).
Authors: Jordan Allan M. , Depledge Paul , Hamilton Nicola , Hitchin James , Hopkins Gemma , Maguire Laura , McGonagle Alison , Mould Daniel , Raoof Ali , Rushbrooke Mathew , Smith James , Smilth Kate , Thomson Graeme , Turlais Fabrice , Waddell Ian , Watson Mandy , Ogilvie Donald 
Published: August 2014

Abstract
Topoisomerases (topo) regulate DNA topology by the transient cleavage and re-ligation of DNA during transcription and replication. Topo II poisons such as etoposide can induce abortive DNA strand breaks in which topo II remains covalently bound to a 5’ DNA strand terminus via a phosphotyrosyl linker. Tyrosyl DNA phosphodiesterase 2 (Tdp2, TTRAP, EAPII) is a recently discovered human 5’-tyrosyl DNA phosphodiesterase which repairs this topo-mediated DNA damage, therefore playing a central role in maintaining normal DNA topology in cells. Cellular depletion of Tdp2 has been shown to result in an increased susceptibility and sensitivity to topo II-induced DNA double strand breaks. It has therefore been proposed that selective pharmacological inhibition of Tdp2 may be a novel approach to overcome intrinsic or acquired resistance to topo II targeted drug therapy.
To date, no known drug-like inhibitors of Tdp2 have been identified. We have recently reported a robust ‘mix and read’ HTS compatible assay and this was used to screen a diverse chemical library of approximately 92,000 compounds. From this endeavour, 2 distinct hit series have been identified. Following further chemical exploration of the original hit compounds, small molecule inhibitors of Tdp2 with sub-100nM potencies have been identified. This poster will describe our biological and chemical progress in this area, detailing SAR and some lessons learnt during investigation of this target.
Citation Format: Allan M. Jordan, Paul Depledge, Nicola Hamilton, James Hitchin, Gemma Hopkins, Laura Maguire, Alison McGonagle, Daniel Mould, Ali Raoof, Mathew Rushbrooke, James Smith, Kate Smilth, Graeme Thomson, Fabrice Turlais, Ian Waddell, Mandy Watson, Donald Ogilvie. The discovery and optimisation of small-molecule inhibitors of human 5’-tyrosyl DNA phosphodiesterase (Tdp2). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3324. doi:10.1158/1538-7445.AM2013-3324
Abstract 3326: TRAIL induces apoptosis preferentially in cell cycle arrested tumor cells, e.g., in tumor cells from children with acute lymphoblastic leukemia upon knockdown of cyclinE.
Authors: Ehrhardt Harald , Wachter Franziska , Grunert Michaela , Jeremias Irmela 
Published: August 2014

Abstract
Resting tumor cells represent a huge challenge during anti-cancer therapy due to their increased treatment resistance. Example given, during minimal residual disease remaining tumor cells might be non-cycling. TNF-related apoptosis-inducing ligand (TRAIL) is a putative future anti-cancer drug currently in phase I and II clinical studies. We recently showed that TRAIL is able to target leukemia stem cell surrogates (Blood 2012,119,4224). Here we tested the ability of TRAIL to target cell cycle arrested tumor cells.
Primary tumor cells from children with acute lymphoblastic leukemia were amplified in immuno-compromised mice and used for in vitro knockdown studies, besides tumor cell lines. Cell cycle arrest was induced in G0, G1 or G2 using cytotoxic drugs, phase-specific inhibitors or RNA interference against cyclinB and E. Cell cycle was visualized by flowcytometry analysis including phosphorylation of Serine 10 in Histone H3 to discriminate G2 from M phase.
Biochemical or molecular arrest at any point of the cell cycle increased TRAIL-induced apoptosis, independently from the tumor cell type, the cell cycle inhibitor used or the check point addressed. In contrast, when cell cycle arrest was disabled by addition of caffeine, the anti-tumor activity of TRAIL was reduced. Most importantly for clinical translation, tumor cells from three children with B-precursor or T-cell acute lymphoblastic leukemia showed increased TRAIL-induced apoptosis upon knockdown of either cyclinB or cyclinE arresting the cell cycle in G2 or G1, respectively.
Taken together and in contrast to most conventional cytotoxic drugs, TRAIL exerts enhanced anti-tumor activity against cell cycle arrested tumor cells. Therefore, TRAIL might represent an interesting drug to treat static tumor disease, e.g., during minimal residual disease.
Citation Format: Harald Ehrhardt, Franziska Wachter, Michaela Grunert, Irmela Jeremias. TRAIL induces apoptosis preferentially in cell cycle arrested tumor cells, e.g., in tumor cells from children with acute lymphoblastic leukemia upon knockdown of cyclinE. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3326. doi:10.1158/1538-7445.AM2013-3326
Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 3327: N-terminal domain of Tyrosyl-DNA phosphodiesterase I (Tdp1) is critical for its cellular function.
Authors: Cuya Selma M. , Wanzeck Keith C. , Comeaux Evan Q. , van Waardenburg Robert C. 
Published: August 2014

Abstract
Tyrosyl-DNA phosphodiesterase I (Tdp1) is a highly conserved eukaryotic DNA repair enzyme that catalyzes the resolution of 3’ and 5’ phospho-DNA adducts. Tdp1 has been implicated in the repair of DNA topoisomerase I (Top1)-DNA covalent complexes reversibly stabilized by camptothecins (CPTs) such as the FDA approved CPT derivatives topotecan and irinotecan. Tdp1 contains two HKD-motifs that provide two catalytic histidines that function as a nucleophile and an acid-base residue. A mutation of the acid-base His to Arg (H493R) in human Tdp1 is associated with the rare recessive ataxia SCAN1. hTdp1H493R and the analogous yeast mutant (Tdp1H432R) enhances cell sensitivity to CPT. In addition, the toxicity induced by this mutant is caused by the formation of a more stable Tdp1-DNA covalent intermediate, a rare characteristic for a DNA repair enzyme. However, this His to Arg substitution induces a minor toxic phenotype compared to other substitutions, such as the His432 to Asn substitution, which induces a Top1 dependent cellular lethality. A band depletion assay suggests that in vivo/cell Tdp1His432Asn remains in complex with Top1 on the DNA, which was not observed in a biochemical in vitro assay.
Biochemical studies revealed that Tdp1 catalysis is independent of the N-terminal domain. Among Tdp1 proteins, the N-terminal domain is poorly conserved in sequence and size (∼80aa for yeast and 140aa for human Tdp1).
We investigated the role of the N-terminal domain for Tdp1 activity in the cell. The N-terminal truncated proteins showed similar cellular distribution as the full-length proteins. Interestingly, the N-terminal truncated proteins did not display the toxicity that was observed with the full-length Tdp1 mutant proteins. This suggests that the N-terminal domain is a critical determinate of Tdp1 cellular function.
Preliminary results from our human cell line model shows similar results implying that the function of the N-terminal domain is conserved among Tdp1 proteins although it is poorly conserved. Further studies are necessary to ensure that these constructs are properly distributed. Moreover, the N-terminal domain of hTdp1 is post-translational modified, while our preliminary results suggest that this domain is important for protein-protein interaction and Tdp1 recruitment to its substrates. Understanding Tdp1 substrate and protein-interactions are important in the development of Tdp1 as therapeutic target.
This work is in part supported by the ADDA.
Citation Format: Selma M. Cuya, Keith C. Wanzeck, Evan Q. Comeaux, Robert C. van Waardenburg. N-terminal domain of Tyrosyl-DNA phosphodiesterase I (Tdp1) is critical for its cellular function. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3327. doi:10.1158/1538-7445.AM2013-3327
Abstract 3332: An artificial zinc finger glycosylase causes site-directed DNA damages and specifically kills the targeted cancer cells via synthetic lethality.
Authors: Fung Hua , Gerson Stanton L. , Barbas Carlos F. 
Published: August 2014

Abstract
Synthetic lethality is a new developed therapy, in which PARP inhibitor selectively kills BRCA-deficient cancer cells. However, BRCA-deficiency only accounts for a small fraction of cancers. We are interesting in applying synthetic lethal approach to target all cancers regardless BRCA statute. As single strand break (SSB) is the start point of the synthetic lethal, we hypothesized the targeted induction of persistent SSB at tumor unique sequences (cancer somatic mutation sequences) with assistance of synthetic lethality would specifically kill cancer cells. To test this principle we have constructed a six-zinc finger/mutant uracil glycosylase fusion protein (zinc finger glycosylase, ZFG). Upon binding to a specific 18-bp sequence by ZFG six-zinc binding domain, the mutated uracil glycosylase will remove a cytosine residue leaving an apyrimidinic site, which is subsequently convert to SSB by AP-endonuclease cleavage. The SSB lesion is capable of being repaired, however, upon repair, the zinc finger binding site is restored and the site-specific ZFG will again initiate a lesion leading to a persistent SSB. We have transfected ZFGs into BRCA2-deficient human breast cancer MCF7 and lung cancer A549 cells, the cells were then treated with PARP inhibitors. We observed increased killing of the ZFG-transfected cells compare to mock-transfected controls. Since BRCA2 deficiency causes homologous recombination (HR) repair defect, we next sought to determine if inhibitors of HR is a suitable substitute for loss of BRCA2. We are testing this concept by transfection of ZFG to BRCA2-proficient MCF7 cells with the treatment of combination of PARP and HR inhibitors, to determine whether 1. Combination of PARP and HR inhibitors treatments can achieve synthetic lethality; 2. ZFG constructs can enhance synthetic lethality. We conclude ZFG coupled with synthetic lethality is a promising direction towards the development of cancer specifically treatment.
Citation Format: Hua Fung, Stanton L. Gerson, Carlos F. Barbas. An artificial zinc finger glycosylase causes site-directed DNA damages and specifically kills the targeted cancer cells via synthetic lethality. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3332. doi:10.1158/1538-7445.AM2013-3332
Abstract 334: PIK3CA overexpression induces epithelial-mesenchymal transition and progression of head and neck squamous cell carcinonoma.
Authors: Du Li , Chen Xi , Shen Jingping , Zhang Fang , Lu Shi-Long 
Published: August 2014

Abstract
Head and neck squamous cell carcinoma (HNSCC) patients have a poor prognosis and invasion and metastasis are the major causes of mortality. However, the molecular mechanisms of HNSCC invasion and metastasis remain obscure. The phosphatidylinositol 3-kinase (PI3K) pathway regulates a wide range of cellular processes crucial for tumorigenesis. Amplification and mutation of PIK3CA, the gene coding for the catalytic subunit of PI3K, are among the most common genetic alterations in human HNSCC. To delineate the in vivo roles of PIK3CA during head and neck tumorigenesis, we developed a PIK3CA genetically engineered mouse model (GEMM), in which PIK3CA overexpression is specifically induced in the head and neck epithelia.
While overexpression of PIK3CA alone resulted in epithelial hyperplasia, and carcinoma in situ, it was not sufficient to induce visible tumor formation. In contrast, when we applied 4NQO, a DNA adduct-forming agent widely used as a tobacco surrogate together with PIK3CA overexpression, the PIK3CA-GEMM developed ∼50% poorly differentiated HNSCC compared to ∼10% of those in the control mice. PIK3CA tumors also had increased numbers of infiltrated leukocytes and angiogenesis. In addition, either regional lymph node metastases or lung metastases was observed in ∼40% of the PIK3CA-GEMM compared to no metastases in control mice. Characterizations of PIK3CA tumors revealed that PIK3CA tumors underwent epithelial and mesenchymal transition, and formed poorly differentiated metastatic tongue SCC in an orthotopical mouse model. Molecular analysis of the PIK3CA tumors suggests that rather than AKT, PDK1 facilitates progression of PI3K-driven HNSCC, and enhanced TGFbeta signaling with increased TGFbeta1 ligand and Smad3 may further contribute to this process. Lastly, overexpression of PIK3CA is associated with HNSCC progression and metastasis, and positively correlated with PDK1 expression but negatively with AKT activation, further verified the clinical relevance of our GEMM study. In summary, our results suggest that PIK3CA oncogene drives invasion and metastasis of HNSCC through PDK1 and TGFbeta signaling, and combined targeting of these pathways will reduce tumor progression in advanced HNSCC patients with PIK3CA alterations, which comprise about ∼40% of all HNSCC patients.
Citation Format: Li Du, Xi Chen, Jingping Shen, Fang Zhang, Shi-Long Lu. PIK3CA overexpression induces epithelial-mesenchymal transition and progression of head and neck squamous cell carcinonoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 334. doi:10.1158/1538-7445.AM2013-334
Abstract 3348: Inhibition of MDMX by XI-011 induces p53-mediated apoptosis in head and neck cancer.
Authors: Ryu In Sun , Ryu Chang Hwan , Park Hyun Bae , Kim Eunhye , Roh Jong-Lyel 
Published: August 2014

Abstract
Overexpression of MDM2 or MDMX inactivates the tumor-suppressive function of p53. Restoration of p53 function counteracting these p53 repressors can lead to in vivo tumor regression. Therefore, we evaluated whether the restoration of p53 function by a small-molecule p53 activator XI-011, a pseudouria derivative NSC146109, induce apoptosis in head-and-neck cancer (HNC). The effects of XI-011 treatment in four HNC cell lines were tested, individually or in combination with nutlin-3a, an inhibitor of MDM2-p53 interactions, and its growth suppression, cell-cycle arrest, and apoptosis were assessed. XI-011 induced accumulation and reactivation of p53 in a wild-type TP53-bearing HNC cell line. XI-011 showed maximal growth suppression in tumor cells with overexpression of MDMX and MDMX-dependent p53 degradation. With down-regulation of MDMX mRNA and protein levels, XI-011 upregulated expression of p21 and proapoptotic genes and promoted apoptosis in a dose-dependent manner. Notably, the apoptotic response was blocked by inhibition of p53 or MDMX gene expression, demonstrating the p53 and MDMX dependence of XI-011 effects. XI-011 decreased the viability of HNC cells expression low levels of MDMX in a less-efficient manner. Interestingly, in combination therapy, XI-011 synergistically acted with nutlin-3a to inhibit growth of HNC cells. Our data suggest that the MDMX inhibition and p53 functional restoration by XI-011 effectively induced cytotoxicity and apoptosis in cancer cells, an action that may offer a promising strategy for treating HNC.
Citation Format: In Sun Ryu, Chang Hwan Ryu, Hyun Bae Park, Eunhye Kim, Jong-Lyel Roh. Inhibition of MDMX by XI-011 induces p53-mediated apoptosis in head and neck cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3348. doi:10.1158/1538-7445.AM2013-3348
Abstract 3356: Comparative plasma and cerebrospinal (CSF) pharmacokinetics (PK) of vincristine sulfate standard preparation (VCR) and vincristine sulfate liposomes injection (VSLI) in a non-human primate (NHP) model.
Authors: Cole Diane E. , Lester-McCully Cynthia M. , Shah Nirali N. , Warren Katherine E. , Wayne Alan S. , Widemann Brigitte C. 
Published: August 2014

Abstract
Background: Vincristine has been used as standard therapy for adult and pediatric malignancies for 50 years. Cumulative neurotoxicity often requires capping the maximum single and cumulative VCR dose. VSLI is a novel preparation of VCR encapsulated in sphingomyelin/cholesterol liposomes. Following clinical trials that demonstrated safety, tolerability and efficacy in adults with relapsed/refractory acute lymphoblastic leukemia, VSLI has been granted FDA accelerated approval at a dose of 2.25 mg/m2given once weekly for that population. A pediatric Phase I trial is ongoing. We evaluated plasma and CSF PK of intravenously (IV) administered VCR and total VSLI (liposomal encapsulated and non-encapsulated) in our NHP model.
Methods: 0.1 mg/kg (1.2 mg/m2 human-equivalent dose) VCR over 1 minute or VSLI over 15 minutes was administered on separate occasions to each of 3 adult rhesus monkeys (Macaca mulatta). Serial paired plasma and CSF samples were collected for up to 48 hrs. Drug concentrations were quantified using HPLC/tandem mass spectrometry assays. For plasma samples, VCR standard curves were linear over a range of .0025-10.0 μg/ml (lower limit of quantification, .0025 μg/ml). Lower limit of detection for VCR in NHP CSF was .001 μg/ml. PK parameters were estimated using non-compartmental methods.
Results:
Plasma PK parameters (median and range) for VCR and VSLI Cmax (μg/ml) AUC0-infinity (μg*hr/ml) Cl (ml/min) Terminal T1/2 (hours) VCR 1.07 (0.99–2.1) 0.614 (0.336–0.676) 33.8 (29.6–54) 24.3 (14.2–54.4) VSLI 4.8 (3.7–7.3) 195 (163–312) 0.089 (0.073–0.113) 17.9 (13.9–21.5) Cmax = maximal concentration, AUC = area under the concentration x time curve, Cl = clearance, T1/2 = half life
VCR demonstrated a multi-exponential and VSLI a near-monoexponential plasma concentration versus time curve. The ratios ClVCR: ClVSLI for the individual NHP were 300, 463 and 477. Vincristine was not detected in any CSF sample.
Conclusions: The NHP model allowed for a direct comparison of VCR and VSLI PK. VCR Cl exceeds that of VSLI by approximately 400-fold. No penetration into the CSF was observed for either preparation.
Citation Format: Diane E. Cole, Cynthia M. Lester-McCully, Nirali N. Shah, Katherine E. Warren, Alan S. Wayne, Brigitte C. Widemann. Comparative plasma and cerebrospinal (CSF) pharmacokinetics (PK) of vincristine sulfate standard preparation (VCR) and vincristine sulfate liposomes injection (VSLI) in a non-human primate (NHP) model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3356. doi:10.1158/1538-7445.AM2013-3356