tbuckley/cabot-search huggingface dataset:
Nonsteroidal Anti-inflammatory Drugs and Endometrial Carcinoma Mortality and Recurrence
Authors: Theodore M Brasky, Ashley S Felix, David E Cohn, D Scott McMeekin, David G Mutch, William T Creasman, Premal H Thaker, Joan L Walker, Richard G Moore, Shashikant B Lele, Saketh R Guntupalli, Levi S Downs, Christa I Nagel, John F Boggess, Michael L Pearl, Olga B Ioffe, Kay J Park, Shamshad Ali, Louise A Brinton
Journal: JNCI Journal of the National Cancer Institute (2016)

Recent data suggest that the use of nonsteroidal anti-inflammatory drugs (NSAIDs) may be associated with reductions in endometrial cancer risk, yet very few have examined whether their use is related to prognosis among endometrial cancer patients.Study subjects comprised 4374 participants of the NRG Oncology/Gynecology Oncology Group 210 Study with endometrial carcinoma who completed a presurgical questionnaire that assessed history of regular prediagnostic NSAID use and endometrial cancer risk factors. Recurrences, vital status, and causes of death were obtained from medical records and cancer registries. Fine-Gray semiproportional hazards regression estimated adjusted subhazard ratios (HRs) and 95% confidence intervals (CIs) for associations of NSAID use with endometrial carcinoma-specific mortality and recurrence. Models were stratified by endometrial carcinoma type (ie, type I [endometrioid] vs type II [serous, clear cell, or carcinosarcoma]) and histology.Five hundred fifty endometrial carcinoma-specific deaths and 737 recurrences occurred during a median of five years of follow-up. NSAID use was associated with 66% (HR = 1.66, 95% CI = 1.21 to 2.30) increased endometrial carcinoma-specific mortality among women with type I cancers. Associations were statistically significant for former and current users, and strongest among former users who used NSAIDs for 10 years or longer (HR = 2.23, 95% CI = 1.19 to 4.18, two-sided P trend = .01). NSAID use was not associated with recurrence or endometrial carcinoma-specific mortality among women with type II tumors.In this study, use of NSAIDs was associated with increased endometrial carcinoma-specific mortality, especially in patients with type I tumors. Barring a clear biologic mechanism by which NSAIDs would increase the risk of cause-specific mortality, cautious interpretation is warranted.
139. Controllable Target-Specific Transgene Induction Using an Allosteric trans-Splicing Ribozyme
Authors: Ju-hyun Kim, Seonyeong Jeong, Alexis Kertsburg, Garrett A. Soukup, Seong-Wook Lee
Journal: Molecular Therapy (2015)

Gene therapeutic approaches are needed that can achieve inducible, adjustable, and well-controlled expression of therapeutic genes while simultaneously suppressing expression of disease-causing genes for maximization of their efficacy. In this study, we proposed a new approach to address one of the main challenges that current gene therapeutic techniques meet, which is based on group I intron ribozyme. To this end, we established an exogeneously and genetically controllable system through design of an allosteric ribozyme that comprises a Tetrahymena group I-based trans-splicing ribozyme as an active domain for RNA replacement, a small molecule-specific RNA aptamer as a sensor domain, and a communication module as an active transfer domain. This allosteric trans-splicing ribozyme can be used as a system that is able to specifically target, and thus reduce the level of, certain disease-specific RNAs, and simultaneously provides the ability to artificially control transgene expression through RNA replacement in therapeutic targets by using exogenous small molecules to activate the function of the ribozyme. The effectiveness of this approach was assessed by constructing various ribozymes in combination with a theophylline-binding aptamer to identify an allosteric ribozyme whose activity is controlled by theophylline both in vitro and in cell culture assays. Moreover, to confirm allosteric regulation of transgene expression, we constructed adenoviral vectors encoding the ribozymes and validated theophylline-dependent RNA replacement in target RNA-expressing cells. Results demonstrate that an allosteric trans-splicing ribozyme is an applicable RNA-based framework for engineering external ligand-controlled gene expression regulatory systems that exhibit adjustable regulation, design modularity (therefore, broad applicability), and target specificity. Such ribozyme could therefore be useful to monitor and/or modulate fate of any cells (such as cancer cells) using specific endogenous or exogenous cues.
391. Mutated Tumor Neoantigens Are Recognized by Tumor Infiltrating Lymphocytes from Metastatic Ovarian Cancer
Authors: Drew C. Deniger, Anna Pasetto, Todd D. Prickett, Jared J. Gartner, Mini Bharathan, Eric Tran, Paul F. Robbins, Steven A. Rosenberg
Journal: Molecular Therapy (2016)

Somatic mutations expressed by the tumor can serve as neoantigens for autologous T cells. Tumor infiltrating lymphocytes (TIL) with varying degrees of neoantigen-reactivity infused for the treatment of melanoma resulted in 50% overall objective response and 20% complete response rates. A largely oligoclonal population of ERBB2IPE805G mutated neoantigen-specific T cells resulted in a long-term, ongoing partial regression of metastatic cholangiocarcinoma suggesting that infusion of selected mutation-reactive TIL could be efficacious in the treatment of common epithelial cancers. We now studied whether TIL obtained from metastatic ovarian cancer recognized tumor mutations. Exome and transcriptome sequencing was performed from resected metastatic ovarian cancer deposits in parallel with growth of TIL fragment cultures in interleukin-2. Long peptides and tandem minigenes encompassing all mutations were synthesized, introduced into autologous antigen presenting cells, co-cultured with individual TIL fragments and T-cell reactivity was determined by interferon-y ELISPOT and surface expression of 41BB. In the 8 ovarian tumors evaluated, there was a median of 227 mutations (range: 63-332) and an average of 94% of 24 fragments initiated (range: 58% - 100%) available for testing. Six of eight (75%) patients had T cell responses to mutated neoantigens at >5% of the fragment culture. Antigens identified to date were unique to each patient, i.e. no overlapping mutations between patients, and both CD4 and CD8 responses have been detected. One patient had a CD4 T cell response to p53G245S hotspot mutation, which opens opportunities for treatment of other cancer patients with TCR-transduced T cells because this mutation is present in 2.8% of all cancers. The average time from resection to identification of mutation-reactive T cell fragment culture was 8 weeks indicating that this strategy could be used for prospective therapy. In summary, mutation-specific T-cell responses were found in 6 of 8 patients with metastatic ovarian cancer, which opens the opportunity to use these cells for adoptive T cell treatment of advanced ovarian cancer.
Combination of ofatumumab and reduced-dose CHOP for diffuse large B-cell lymphomas in patients aged 80 years or older: an open-label, multicentre, single-arm, phase 2 trial from the LYSA group
Authors: Frédéric Peyrade, Serge Bologna, Vincent Delwail, Jean François Emile, Laurent Pascal, Christophe Fermé, Jean-Marc Schiano, Bertrand Coiffier, Bernadette Corront, Hassan Farhat, Christophe Fruchart, Herve Ghesquieres, Margaret Macro, Hervé Tilly, Bachra Choufi, Richard Delarue, Olivier Fitoussi, Jean Gabarre, Corinne Haioun, Fabrice Jardin
Journal: The Lancet Haematology (2016)
Astrocytic calcium release mediates peri-infarct depolarizations in a rodent stroke model
Authors: Cordula Rakers, Gabor C. Petzold
Journal: Journal of Clinical Investigation (2016)

Stroke is one of the most common diseases and a leading cause of death and disability. Cessation of cerebral blood flow (CBF) leads to cell death in the infarct core, but tissue surrounding the core has the potential to recover if local reductions in CBF are restored. In these areas, detrimental peri-infarct depolarizations (PIDs) contribute to secondary infarct growth and negatively affect stroke outcome. However, the cellular pathways underlying PIDs have remained unclear. Here, we have used in vivo multiphoton microscopy, laser speckle imaging of CBF, and electrophysiological recordings in a mouse model of focal ischemia to demonstrate that PIDs are associated with a strong increase of intracellular calcium in astrocytes and neurons. We found that astroglial calcium elevations during PIDs are mediated by inositol triphosphate receptor type 2-dependent (IP3R2-dependent) release from internal stores. Importantly, Ip3r2-deficient mice displayed a reduction of PID frequency and overall PID burden and showed increased neuronal survival after stroke. These effects were not related to local CBF changes in response to PIDs. However, we showed that the release and extracellular accumulation of glutamate during PIDs is strongly curtailed in Ip3r2-deficient mice, resulting in ameliorated calcium overload in neurons and astrocytes. Together, these data implicate astroglial calcium pathways as potential targets for stroke therapy.
667. Natural Selection of a Neurovirulent Herpes Simplex Virus Type1 Strain via Passage in Human Glioma Xenografts
Authors: Kentaro Fujii
Journal: Molecular Therapy (2016)

The strategy of using oncolytic viruses (OVs) in cancer therapy is based on their replicative selectivity in tumor compared to normal tissue. In general, these tumor-tropic OVs are genetically engineered to be attenuated to minimize toxicity and for retargeting towards tumors. Directed genetic manipulation of viruses depends on knowledge of the host-virus interactions at the molecular level, which can be highly complex and context-dependent. Another approach is to utilize natural selection and evolution to adapt a virus to be more selective for tumor cells. We thus obtained a wild-type HSV1 strain harvested from a human patient with HSV1 encephalitis patient. This HSV1 was repeatedly passaged in nude mice with human glioblastoma xenografts. After 10 rounds of serial passage, we propagated and purified the passaged virus for subsequent experiments. To assay for phenotypic differences, the parent virus and passaged virus were compared in their replicative efficiency in glioma cells in vitro and in vivo, as well as in toxicity by injection in nude mouse brains. There was a statistical significant increase in the replication yield of the passaged virus compared to the parental virus. Although there was also a trend for enhanced anticancer efficacy of the paasaged virus compared to the parental virus, it did not reach a p value less than 0.05. There was no difference in the toxicity of either virus in nude mouse brains. These results suggest that natural evolution by serial passage may lead to a more efficacious oncolytic HSV1, but that more passages may be required to further evolve this strain.
301. Efficient Delivery of Nuclear and Cytoplasmic Proteins Fused to HIV-1 Gag Polypeptide By Means of Virus-Like Particles
Authors: Marc-André Robert, Rénald Gilbert, Bruno Gaillet
Journal: Molecular Therapy (2015)

The expression of the retroviral polypeptide Gag can induce the formation of virus-like particles (VLP). Upon expression, the polypeptide is targeted to the cell membrane and incorporated in the VLP during membrane budding. Chimeric proteins consisting of Gag polypeptide fused to different proteins were engineered and the VLP produced were used as vehicles for protein delivery. One major advantage of this approach is the low mutation risk due to the absence of DNA transfer and genomic integration. In this study, the C-Terminus of Gag from HIV-1 was fused to the green fluorescent protein (GFP), a chimeric transactivator (cTA) and a reprogamming factor (KLF4). VLP were produced by transfection using a stable cell line (293SF-pacLV) that expresses VSVg and Gag-pol. Analysis of the supernatants from producing cells by western blot confirmed the presence of Gag-GFP, -cTA and -KLF4. Confocal microscopy showed that the vast majority of the cells (> 90%) treated with VLP-GFP/polybrene complexes was successfully transduced. The cells also displayed a GFP signal almost exclusively localized inside the cytoplasm. Additional VSVg expression during the production facilitated the endosomal escape of VLP-GFP in transduced cells. The insertion of a nuclear localisation signal (NLS) shifted the localization of the GFP to the cell nucleus demonstrating that a nuclear protein could be successfully delivered by VLP. The experiment was thus repeated using two transcription factors. Lentiviral vectors were used to make two stable pools of HEK293 cells each containing a specific GFP reporter cassette. The GFP gene was regulated either by the CR5 promoter (specifically activated by the cTA) or by a minimal promoter fused to KLF4 transcription response elements (TRE). Transduction of the CR5-GFP pool with VLP-cTA/polybrene complexes showed a powerful activation of the reporter (365-fold compared to the negative control) as measured by flow cell cytometry two days post-transduction. Surprisingly, no activation was observed in the TRE-GFP pool three days after transduction by VLP-KLF4/polybrene complexes. Evidence obtained by transfection suggested that the Gag fusion inhibits KLF4 activity. To augment activity, the activation domain of VP16 was fused to KLF4. Transfection of a plasmid encoding Gag-VP16KLF4 strongly activated the reporter by a factor of 126-fold (6-fold higher than wild type KLF4). The ability of VLP produced with Gag-VP16KLF4 to activate transcription is currently under investigation. In summary, VLP based on HIV-1 Gag can deliver nuclear and cytoplasmic proteins directly into cells with a low mutation risk. Therefore, our platform for VLP production could be useful for several applications including cell reprogramming and genome editing oriented toward cell therapies of diseases like Duchenne muscular dystrophy.
Abstract 4905: Comprehensive genomic analyses of oral squamous cell carcinoma tissues by semiconductor-based next-generation sequencing
Authors: Takafumi Nakagaki, Yasushi Sasaki, Kenta Kobashi, Kousuke Takeda, Miyuki Tamura, Tomoko Ohashi, Kazuhiro Ogi, Masashi Idogawa, Hiroyoshi Hiratsuka, Takashi Tokino
Journal: Cancer Research (2015)

Abstract Somatic mutation analysis is standard of practice for human cancers in order to identify therapeutic sensitizing and resistance mutations. We performed comprehensive genomic analyses that use PCR target enrichment and next-generation sequencing on the Ion Torrent Personal Genome Machine (PGM). We first validated a multigene sequencing screen interrogating 2855 mutational hotspots in 50 cancer-related genes using the Ion Torrent AmpliSeq cancer panel v2. Ten nanogram of DNA was used as template to amplify 207 regions in oral squamous cell carcinoma (OSCC). Approximately 1000 average coverage was obtained with more than 90% of target bases having at least 100 sequence reads. We detected mutations of the TP53 gene in 96% (22 of 23) of OSCC cell lines. Of the 62 OSCC specimens, 42 presented at least one somatic mutation among the 50 investigated genes, and 21 of these showed multiple gene somatic mutations. The most frequent mutations were detected on TP53 (26 of 62; 41.9%). CDKN2A mutations were identified in 9 cases; STK11 mutations were observed in 8 cases; and PIK3CA were mutated 5 cases. In addition, we sequenced 409 cancer-related genes in matched tumor and normal DNA from OSCC patients using the Ion Ampliseq Comprehensive Cancer Panel. This panel targets all exons of 409 tumor suppressor genes and oncogenes frequently cited and frequently mutated (covered regions: 95.4% of total), and 15,992 amplicons amplify more than 1.2 megabases of target sequence. We piloted the use of this platform to identify somatic mutations in 37 OSCC specimens including formalin-fixed paraffin-embedded (FFPE) samples. We also detected copy number variations (CNVs) in which segments of the genome can be duplicated or deleted from sequencing data. This targeted next generation sequencing using low amounts of FFPE DNA is a valuable tool for high-throughput genetic testing in research and clinical settings. Citation Format: Takafumi Nakagaki, Yasushi Sasaki, Kenta Kobashi, Kousuke Takeda, Miyuki Tamura, Tomoko Ohashi, Kazuhiro Ogi, Masashi Idogawa, Hiroyoshi Hiratsuka, Takashi Tokino. Comprehensive genomic analyses of oral squamous cell carcinoma tissues by semiconductor-based next-generation sequencing. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4905. doi:10.1158/1538-7445.AM2015-4905
An Observational Multicenter Study to Assess the Cost of Illness and Quality of Life in Patients with Myelodysplastic Syndromes in Italy
Authors: Valeria Santini, Alessandro Sanna, Alberto Bosi, Giuliana Alimena, Giuseppina Loglisci, Alessandro Levis, Flavia Salvi, Carlo Finelli, Cristina Clissa, Giorgina Specchia, Alessandra Ricco, Agostino Cortelezzi, Valeria Ferla, Mariarita Sciumé, Francesco Nobile, Esther Oliva, Carlo Lazzaro, Elisa Martelli
Journal: Blood (2011)
Abstract 5457: MAD2γ, a new MAD2 isoform, is ubiquitously expressed in distinct cell lines, reduces mitotic arrest, and associates with resistance to cisplatin-based chemotherapy in testicular germ cell tumors
Authors: Alejandro Lopez-Saavedra, Rodrigo Caceres, Fernando Luna, Irwin Hernandez, Luis Alonso Herrera
Journal: Cancer Research (2015)

Abstract The mitotic spindle assembly checkpoint (SAC) signaling has been emerging as an important promising mechanism in cancer therapy. A prolonged mitotic arrest in response to cancer chemotherapy induces apoptosis, mitotic catastrophe, mitotic slippage and senescence, depending on what cell line and/or inhibitor is studied. In consequence, a weakened or disturbed SAC signaling in human cancers might contribute to drug resistance both to spindle inhibitors and to DNA damaging agents. In several studies the mitotic arrest deficient 2 (MAD2α) has been implicated in the association between failure in the SAC signaling and chemotherapy response. In this study, we identified a novel splicing variant of MAD2, designated as MAD2γ. This isoform was ubiquitously expressed in several cancer cell lines and non-cancerous primary foreskin fibroblasts. When it was ectopically expressed, MAD2γ localized in the nucleus. Its overexpression in a fully functional SAC-competent cancer cell line, HCT116, reduced the mitotic index, suggesting SAC impairment. Furthermore, the endogenous overexpression of MAD2γ in testicular germ cell tumor patients was associated with a resistant response to cisplatin-based chemotherapy, whereas expression of MAD2α was not significantly different between resistant and sensitive patients. In addition, HCT116 raised significantly its endogenous expression of MAD2γ only in response to cisplatin, but not that of MAD2α. Our data suggest that MAD2γ may have an opposing role to MAD2α in the activation of the SAC signaling, and that its expression is associated to a resistant response to DNA-damaging agents, such as cisplatin-based chemotherapy. This highlight the importance to study alternative splicing that may compromise the SAC function, which has been regarded as a promise for cancer therapy. Citation Format: Alejandro Lopez-Saavedra, Rodrigo Caceres, Fernando Luna, Irwin Hernandez, Luis Alonso Herrera. MAD2γ, a new MAD2 isoform, is ubiquitously expressed in distinct cell lines, reduces mitotic arrest, and associates with resistance to cisplatin-based chemotherapy in testicular germ cell tumors. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5457. doi:10.1158/1538-7445.AM2015-5457
Tweak up-regulates endothelin-1 system in mouse and human endothelial cells
Authors: Patricia Martínez-Miguel, Diana Medrano-Andrés, Mercedes Griera-Merino, Alberto Ortiz, Manuel Rodríguez-Puyol, Diego Rodríguez-Puyol, Susana López-Ongil
Journal: Cardiovascular Research (2016)

To analyse the ability of TWEAK to modify the endothelin system, particularly endothelin-1 (ET-1) and endothelin-converting enzyme-1 (ECE-1), studying the intracellular mechanisms implied. TNF-like weak inducer of apoptosis (TWEAK) is a member of TNF superfamily; it has different biological functions such as inflammation, angiogenesis, proliferation, and apoptosis. TWEAK and fibroblast growth-factor-inducible 14 are expressed in different cell types, including endothelial and smooth muscle cells. Despite their presence in endothelial cells, the effect of TWEAK on endothelial function is incompletely defined.In cells, TWEAK induced protein (Western blot) and mRNA (quantitative polymerase chain reaction) expression of ECE-1. Results were related to transcriptional changes, as ECE-1 promoter activity (transfection assays) was also increased. Transfections with serial deletions of ECE-1 promoter suggest a potential role for AP-1 and NFkB, which were confirmed by electrophoretic mobility shift assays. When AP-1 or NFkB activations were inhibited by specific inhibitors of AP-1, PD-98059 (Erk1/2 inhibitor), or SP-600125 (JNK inhibitor), and also with an inhibitor of NFKB and PDTC, TWEAK effect was partially blocked in both cases, suggesting that both transcription factors are implied in ECE-1 regulation. Moreover, the endothelial changes induced by TWEAK were also tested in vivo, using 3-month-old male CD-1 mice treated with TWEAK 10 µg/kg body weight for 24 h, finding similar effects, a rise in ET-1 production (enzyme-linked immunosorbent assay), and ECE-1 expression in aorta and lung tissues. Mice showed slight hypertension after 4 h of treatment, which disappeared at 24 h.In pathological situations such as chronic inflammation, TWEAK could be more harmful through this effect at endothelial level. Pharmacological blockade of this cytokine could prevent the haemodynamic and structural changes related to an increased ET-1 synthesis.
Neurological Toxicity Profile of Weekly Vincristine Sulfate Liposomes Injection (Marqibo®) 2.25 mg/m2 Compared to Historical Standard Vincristine Sulfate 1.4 mg/m2, 2 mg, and 4 mg Administered Less Frequently
Authors: Lloyd E. Damon, Stuart L. Goldberg, Leonard T. Heffner, John Lister, Gary J. Schiller, Steven R. Deitcher
Journal: Blood (2011)