17 results on '"Jane Staunton"'
Search Results
2. Abstract CT142: TALAVE: Induction talazoparib (tala) followed by combined tala and avelumab in patients (pts) with advanced breast cancer (ABC)
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Filipa Lynce, Kenichi Shimada, Xue Geng, Edward T. Richardson, Candace Mainor, Mei Wei, Julie M. Collins, Paula P. Pohlmann, Arielle L. Heeke, Kelly F. Zheng, Madeline Townsend, Jane Staunton, Stuart J. Schnitt, Joan S. Brugge, Hongkun Wang, Claudine Isaacs, Geoffrey I. Shapiro, and Jennifer L. Guerriero
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Cancer Research ,Oncology - Abstract
Background: PARP inhibitors (PARPi) significantly extend progression-free survival (PFS) compared to chemotherapy in pts with BRCA1/2 mutated (BRCA1/2m) ABC, but responses are not durable. PARPi activate the cGAS-STING pathway leading to increased PD-L1 expression and cytotoxic T-cell recruitment, creating a tumor microenvironment (TME) that may be more vulnerable to immunotherapy. This study was conducted to evaluate the safety, efficacy and effects on the TME of the PARPi tala combined with the PD-L1 inhibitor avelumab in ABC, and to assess the impact of BRCA1/2 status on clinical outcomes. Methods: TALAVE was an open-label, multi-institutional trial (NCT03964532) for pts with HER2-negative ABC. Pts were enrolled in two cohorts: cohort 1 - BRCA1/2m and HER2-negative ABC; cohort 2 - BRCA1/2 wildtype TNBC. Pts received a 4-week induction of tala (1mg po daily D1-D28), followed by a combination of daily tala and avelumab (800mg IV D1, D15). The primary objective was the safety and tolerability of the combination. Secondary objectives included ORR, OS and PFS. Pts underwent serial biopsies to investigate molecular signatures associated with BRCA status or clinical benefit using multiple omics techniques: RNA profiling by NanoString PanCancer IO 360™ Panel, GeoMx® Digital Spatial Profiler (DSP) Whole Transcriptome Atlas (WTA) and protein spatial analysis by multiplex immunofluorescence (mIF) and Cyclic Immunofluorescence (CyCIF). Results: 12 pts were enrolled in each cohort. In cohort 1, 5 pts had gBRCA1, 6 had gBRCA2 and 1 had sBRCA2 mutation. The median age was 50 [IQR:43-59.5]; all pts were female, with median of 1 prior therapy for ABC [IQR: 0-2.5]. 42% pts had prior platinum. ORR was 42% (83% in cohort 1; 0% in cohort 2). There were 10 PRs, all in cohort 1. mPFS was 5.1 months (mo) (95% CI: 3.7-7.3 mo); 9.3mo in cohort 1 and 2.9mo in cohort 2. 5 out of 24 pts remain on treatment, all in cohort 1. Treatment related adverse events (TRAEs) included anemia 33%, neutropenia 25% (gr3+ 13%), thrombocytopenia 21% (gr3+ 13%), fatigue 33% and nausea 29%. Other TRAEs gr3+ included dyspnea (4%) and AST elevation (4%). There were no gr5 events. RNA analysis showed that in cohort 1, tala monotherapy disrupted MMEJ, induced antiproliferative effects and expression of genes in the cGAS-STING pathway, including TBK1-mediated IRF3 activation, with downstream induction of T-cell, dendritic cell and cytokine gene expression. These effects were not seen in biopsies post tala monotherapy in cohort 2. mIF analyses demonstrated T-cell and macrophage infiltration in BRCA1/2m tumors. Analyses of post-combination biopsies is ongoing. Conclusions: There were no new safety signals of PARPi combined with immunotherapy. Responses were limited to pts with BRCA1/2m. RNA and protein analyses indicate cGAS-STING activation and immune cell infiltration in BRCA1/2m tumors, validating murine preclinical findings. Citation Format: Filipa Lynce, Kenichi Shimada, Xue Geng, Edward T. Richardson, Candace Mainor, Mei Wei, Julie M. Collins, Paula P. Pohlmann, Arielle L. Heeke, Kelly F. Zheng, Madeline Townsend, Jane Staunton, Stuart J. Schnitt, Joan S. Brugge, Hongkun Wang, Claudine Isaacs, Geoffrey I. Shapiro, Jennifer L. Guerriero. TALAVE: Induction talazoparib (tala) followed by combined tala and avelumab in patients (pts) with advanced breast cancer (ABC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr CT142.
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- 2023
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3. Identification of Synergistic Combinations of F508del Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Modulators
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Jane Staunton, Ping Zhu, Brenda Fung, Najib El Messadi, Jennifer Andersen, Stephen Lin, Jinliang Sui, Margaret S. Lee, Shakira Cotard, and Joseph Lehar
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Cystic Fibrosis ,Cell Culture Techniques ,Cystic Fibrosis Transmembrane Conductance Regulator ,Plasma protein binding ,medicine.disease_cause ,Cystic fibrosis ,Drug Discovery ,medicine ,Animals ,Humans ,Ion channel ,Sequence Deletion ,Mutation ,biology ,Cell Membrane ,Drug Synergism ,Epithelial Cells ,medicine.disease ,Rats, Inbred F344 ,Cystic fibrosis transmembrane conductance regulator ,High-Throughput Screening Assays ,Rats ,Cell biology ,Transport protein ,Luminescent Proteins ,Protein Transport ,Biochemistry ,Cell culture ,Luminescent Measurements ,biology.protein ,Molecular Medicine ,Function (biology) ,Protein Binding - Abstract
Cystic fibrosis (CF) is an inherited, life-threatening disease caused by mutations in the gene encoding cystic fibrosis transmembrane conductance regulator (CFTR), an ABC transporter-class protein and ion channel that transports ions across epithelial cell membranes. The most common mutation leads to the deletion of a single phenylalanine, and the resulting protein, F508del-CFTR, shows reduced trafficking to the membrane and defective channel gating. The ideal therapeutic approach would address both of these defects and restore channel function at the same time. We describe here the application of a combination high-throughput screening to search for synergistic modulators of F508del-CFTR. With the adapted Fischer rat thyroid-yellow fluorescent protein halide flux assay to the combination high-throughput screening platform, we identified many interesting single agents as CFTR modulators from a library of approved drugs and mechanistic probe compounds, and combinations that synergistically modulate F508del-CFTR channel function in Fischer rat thyroid cells, demonstrating the potential for combination therapeutics to address the defects that cause CF.
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- 2010
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- View/download PDF
4. Optimization of a Yellow Fluorescent Protein-Based Iodide Influx High-Throughput Screening Assay for Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Modulators
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Shakira Cotard, Jinliang Sui, Ping Zhu, Stephen Lin, Jane Staunton, Jennifer Andersen, and Margaret S. Lee
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Yellow fluorescent protein ,congenital, hereditary, and neonatal diseases and abnormalities ,Cystic Fibrosis ,High-throughput screening ,Cystic Fibrosis Transmembrane Conductance Regulator ,Phenylalanine ,Biology ,medicine.disease_cause ,Cystic fibrosis ,Cell Line ,Drug Discovery ,medicine ,Animals ,Humans ,Gene ,Cells, Cultured ,Mutation ,Iodides ,medicine.disease ,Molecular biology ,Cystic fibrosis transmembrane conductance regulator ,High-Throughput Screening Assays ,Rats ,Luminescent Proteins ,Protein Transport ,Luminescent Measurements ,biology.protein ,Molecular Medicine ,Plate reader - Abstract
Cystic fibrosis is an inherited, life-threatening disease associated with mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The most common mutation, F508del CFTR, is found in 90% of CF patients. The loss of a single amino acid (phenylalanine at position 508) results in malformed CFTR with defective trafficking to the plasma membrane and impaired channel function. A functional assay with cells expressing F508del CFTR has been previously described by others using genetically engineered halide-sensitive yellow fluorescent protein to screen for CFTR modulators. We adapted this yellow fluorescent protein assay to 384-well plate format with a high-throughput screening plate reader, and optimized the assay in terms of data quality, resolution, and throughput, with target-specific protocols. The optimized assay was validated with reference compounds from cystic fibrosis foundation therapeutics. On the basis of the Z-factor range (≥0.5) and the potential productivity, this assay is well suited for high-throughput screening. It was successfully used to screen for active single agent and synergistic combinations of single agent modulators of F508del CFTR from a library collection of current active pharmaceutical ingredients (supported by Cystic Fibrosis Foundation Therapeutics).
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- 2010
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- View/download PDF
5. V(D)J Recombination: Double-Strand Break Repair Gene Products Used in the Joining Mechanism
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Jane Staunton, Kathryn T. Hall, Nikolai V. Boubnov, David R. Weaver, and Zachary P. Wills
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DNA Repair ,Molecular Sequence Data ,CHO Cells ,Mice, SCID ,General Biochemistry, Genetics and Molecular Biology ,Mice ,History and Philosophy of Science ,Cricetinae ,Consensus Sequence ,Animals ,Humans ,Ku Autoantigen ,VDJ Recombinases ,Gene ,Base Sequence ,Chemistry ,General Neuroscience ,V(D)J recombination ,DNA Helicases ,Nuclear Proteins ,Antigens, Nuclear ,DNA ,Double Strand Break Repair ,Cell biology ,DNA-Binding Proteins ,DNA Nucleotidyltransferases ,Mechanism (sociology) ,Chromosomes, Human, Pair 8 ,HeLa Cells - Published
- 2008
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6. Gene expression signatures define novel oncogenic pathways in T cell acute lymphoblastic leukemia
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Christine Huard, Fred G. Behm, Jane Staunton, Todd R. Golub, Adolfo A. Ferrando, James R. Downing, Eric S. Lander, A. Thomas Look, D. Gary Gilliland, Mignon L. Loh, Susana C. Raimondi, Ching-Hon Pui, and Donna Neuberg
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LMO2 ,0303 health sciences ,Cancer Research ,T cell ,Cellular differentiation ,Cell Biology ,Biology ,Gene expression profiling ,03 medical and health sciences ,Thymocyte ,0302 clinical medicine ,medicine.anatomical_structure ,LYL1 ,Oncology ,030220 oncology & carcinogenesis ,T-Cell Receptor Gene ,Cancer research ,medicine ,030304 developmental biology ,TAL1 - Abstract
Human T cell leukemias can arise from oncogenes activated by specific chromosomal translocations involving the T cell receptor genes. Here we show that five different T cell oncogenes (HOX11, TAL1, LYL1, LMO1, and LMO2) are often aberrantly expressed in the absence of chromosomal abnormalities. Using oligonucleotide microarrays, we identified several gene expression signatures that were indicative of leukemic arrest at specific stages of normal thymocyte development: LYL1+ signature (pro-T), HOX11+ (early cortical thymocyte), and TAL1+ (late cortical thymocyte). Hierarchical clustering analysis of gene expression signatures grouped samples according to their shared oncogenic pathways and identified HOX11L2 activation as a novel event in T cell leukemogenesis. These findings have clinical importance, since HOX11 activation is significantly associated with a favorable prognosis, while expression of TAL1, LYL1, or, surprisingly, HOX11L2 confers a much worse response to treatment. Our results illustrate the power of gene expression profiles to elucidate transformation pathways relevant to human leukemia.
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- 2002
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7. Classification of human lung carcinomas by mRNA expression profiling reveals distinct adenocarcinoma subclasses
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Griffin M. Weber, Javad Beheshti, Raphael Bueno, William G. Richards, Wing Hung Wong, Priya Vasa, David J. Sugarbaker, Massimo Loda, Arindam Bhattacharjee, Christine Ladd, Todd R. Golub, Eugene J. Mark, Eric S. Lander, Cheng Li, Bruce E. Johnson, Matthew Meyerson, Jane Staunton, Michael A. Gillette, and Stefano Monti
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Lung Neoplasms ,Time Factors ,Gene Expression ,Adenocarcinoma ,Biology ,Bioinformatics ,Gene expression ,medicine ,Carcinoma ,Humans ,RNA, Messenger ,RNA, Neoplasm ,Carcinoma, Small Cell ,Neoplasm Metastasis ,Lung cancer ,Gene ,Survival rate ,Retrospective Studies ,Multidisciplinary ,Lung ,Gene Expression Profiling ,Smoking ,Biological Sciences ,medicine.disease ,Survival Rate ,Gene expression profiling ,medicine.anatomical_structure ,Carcinoma, Squamous Cell ,Disease Progression ,Cancer research - Abstract
We have generated a molecular taxonomy of lung carcinoma, the leading cause of cancer death in the United States and worldwide. Using oligonucleotide microarrays, we analyzed mRNA expression levels corresponding to 12,600 transcript sequences in 186 lung tumor samples, including 139 adenocarcinomas resected from the lung. Hierarchical and probabilistic clustering of expression data defined distinct subclasses of lung adenocarcinoma. Among these were tumors with high relative expression of neuroendocrine genes and of type II pneumocyte genes, respectively. Retrospective analysis revealed a less favorable outcome for the adenocarcinomas with neuroendocrine gene expression. The diagnostic potential of expression profiling is emphasized by its ability to discriminate primary lung adenocarcinomas from metastases of extra-pulmonary origin. These results suggest that integration of expression profile data with clinical parameters could aid in diagnosis of lung cancer patients.
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- 2001
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8. Chemosensitivity prediction by transcriptional profiling
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Jill P. Mesirov, John N. Weinstein, William O. Reinhold, Pablo Tamayo, Todd R. Golub, Donna K. Slonim, Jane Staunton, Eric S. Lander, Johnny Park, Uwe Scherf, Michael Angelo, Hilary A. Coller, and Jae K. Lee
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Genetics ,Multidisciplinary ,Transcription, Genetic ,Gene Expression Profiling ,Genomics ,Computational biology ,Biological Sciences ,Biology ,Gene expression profiling ,Oligonucleotide Microarrays ,Drug Resistance, Neoplasm ,Predictive Value of Tests ,Cell culture ,Transcription (biology) ,Neoplasms ,Gene expression ,Tumor Cells, Cultured ,Humans ,Gene ,Human cancer ,Oligonucleotide Array Sequence Analysis - Abstract
In an effort to develop a genomics-based approach to the prediction of drug response, we have developed an algorithm for classification of cell line chemosensitivity based on gene expression profiles alone. Using oligonucleotide microarrays, the expression levels of 6,817 genes were measured in a panel of 60 human cancer cell lines (the NCI-60) for which the chemosensitivity profiles of thousands of chemical compounds have been determined. We sought to determine whether the gene expression signatures of untreated cells were sufficient for the prediction of chemosensitivity. Gene expression-based classifiers of sensitivity or resistance for 232 compounds were generated and then evaluated on independent sets of data. The classifiers were designed to be independent of the cells' tissue of origin. The accuracy of chemosensitivity prediction was considerably better than would be expected by chance. Eighty-eight of 232 expression-based classifiers performed accurately (with P < 0.05) on an independent test set, whereas only 12 of the 232 would be expected to do so by chance. These results suggest that at least for a subset of compounds genomic approaches to chemosensitivity prediction are feasible.
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- 2001
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9. aex-3 Encodes a Novel Regulator of Presynaptic Activity in C. elegans
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Jane Staunton, Michael L. Nonet, James H. Thomas, Kouichi Iwasaki, and Owais Saifee
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rab3 GTP-Binding Proteins ,Neuroscience(all) ,Molecular Sequence Data ,Mutant ,Presynaptic Terminals ,Regulator ,Neurotransmission ,Biology ,Synaptic Transmission ,Synaptic vesicle ,03 medical and health sciences ,0302 clinical medicine ,GTP-Binding Proteins ,Animals ,Nervous System Physiological Phenomena ,Tissue Distribution ,Amino Acid Sequence ,Caenorhabditis elegans ,Peptide sequence ,Gene ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,Behavior, Animal ,General Neuroscience ,Chromosome Mapping ,Cell biology ,Amino acid ,Electrophysiology ,Genes ,chemistry ,Mutation ,Synapses ,Synaptic plasticity ,Pharynx ,Cholinesterase Inhibitors ,Aldicarb ,030217 neurology & neurosurgery - Abstract
C. elegans aex-3mutations cause pleiotropic behavioral defects that are suggestive of reduced synaptic transmission. aex-3mutations also show strong genetic interactions with mutations in unc-31and unc-64, two other genes implicated in synaptic transmission. Physiological and pharmacological studies indicate that aex-3defects are presynaptic. In aex-3mutants, the synaptic vesicle–associated RAB-3 protein aberrantly accumulates in neuronal cell bodies and is reduced in synapse-rich axons. This localization defect is specific to RAB-3, since other synaptic proteins are localized normally in aex-3mutants. aex-3encodes a 1409 amino acid protein with strong homology to DENN, a human protein of unknown function. In C. elegans, aex-3is expressed in all or nearly all neurons. These results suggest that AEX-3 is a novel regulator of presynaptic activity that interacts with RAB-3 to regulate synaptic vesicle release.
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- 1997
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10. seid Cells Efficiently Integrate Hairpin and Linear DNA Substrates
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David T. Weaver and Jane Staunton
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Exodeoxyribonuclease V ,T-Lymphocytes ,Restriction Mapping ,Context (language use) ,Mice, SCID ,Biology ,Transfection ,medicine.disease_cause ,Cell Line ,Mice ,chemistry.chemical_compound ,Restriction map ,medicine ,Animals ,DNA Integration ,Molecular Biology ,Southern blot ,Gene Rearrangement ,Recombination, Genetic ,RecBCD ,B-Lymphocytes ,Mutation ,3T3 Cells ,DNA ,Cell Biology ,Gene rearrangement ,Molecular biology ,Clone Cells ,Blotting, Southern ,Exodeoxyribonucleases ,chemistry ,Nucleic Acid Conformation ,Research Article - Abstract
The scid mouse mutation affects V(D)J rearrangement and double-strand break repair. scid V(D)J rearrangement is characterized by defective coding joint formation which prevents the development of mature B and T cells. Hairpin DNA has been implicated in the formation of V(D)J coding joints. We found scid cells to be proficient in hairpin processing in the context of DNA integration. In addition, we found that the scid defect did not impair integration of linear DNA via nonhomologous recombination. Therefore, hairpin processing and integration of DNA into the genome are distinct from hypersensitivity to ionizing radiation and the defect in V(D)J recombination.
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- 1994
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11. Synergistic drug combinations tend to improve therapeutically relevant selectivity
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Joseph Lehar, Richard Rickles, Andrew S Krueger, Grant R. Zimmermann, Margaret S. Lee, Alexis Borisy, E. Roydon Price, Xiaowei Jin, Jane Staunton, Lisa M. Johansen, William Avery, Adrian Heilbut, and Glenn F. Short
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Drug ,Male ,Drug-Related Side Effects and Adverse Reactions ,media_common.quotation_subject ,Rat model ,Biomedical Engineering ,Bioengineering ,Computational biology ,Pharmacology ,Biology ,Applied Microbiology and Biotechnology ,Models, Biological ,Article ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Drug Discovery ,Escherichia coli ,Animals ,Humans ,Single agent ,Differential expression ,030304 developmental biology ,media_common ,0303 health sciences ,Drug discovery ,Reproducibility of Results ,Drug Synergism ,3. Good health ,Rats ,Sprague dawley ,Disease Models, Animal ,Drug activity ,Pharmaceutical Preparations ,030220 oncology & carcinogenesis ,Molecular Medicine ,Context specificity ,Drug Therapy, Combination ,Biotechnology - Abstract
Prevailing drug discovery approaches focus on compounds with molecular selectivity, inhibiting disease-relevant targets over others in vitro. However in vivo, many such agents are not therapeutically selective, either because of undesirable activity at effective doses or because the biological system responds to compensate. In theory, drug combinations should permit increased control of such complex biology, but there is a common concern that therapeutic synergy will generally be mirrored by synergistic side-effects. Here we provide evidence, from 94,110 multi-dose combination experiments representing diverse disease areas and large scale flux balance simulations of inhibited bacterial metabolism, that multi-target synergies are more specific than single agent activities to particular cellular contexts. Using an anti-inflammatory combination, we show how multi-target synergy can achieve therapeutic selectivity in animals through differential target expression. Synergistic combinations can increase the number of selective therapies using the current pharmacopeia, and offer opportunities for more precise control of biological systems.
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- 2009
12. Rabphilin potentiates soluble N-ethylmaleimide sensitive factor attachment protein receptor function independently of rab3
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Michael L. Nonet, Jane Staunton, and Barry Ganetzky
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Synaptosomal-Associated Protein 25 ,Synaptobrevin ,rab3 GTP-Binding Proteins ,Mutant ,Molecular Sequence Data ,Vesicular Transport Proteins ,Nerve Tissue Proteins ,Biology ,Protein structure ,Physical Stimulation ,Syntaxin ,Animals ,ARTICLE ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Adaptor Proteins, Signal Transducing ,Sequence Deletion ,Behavior, Animal ,Sequence Homology, Amino Acid ,General Neuroscience ,Signal transducing adaptor protein ,Membrane Proteins ,Long-term potentiation ,Helminth Proteins ,Cell biology ,Protein Structure, Tertiary ,Electrophysiology ,Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins ,Phenotype ,rab GTP-Binding Proteins ,Synapses ,Mutagenesis, Site-Directed ,Rab ,Guanosine Triphosphate ,Sleep Stages ,Synaptic Vesicles ,Carrier Proteins ,SNARE Proteins ,Biomarkers ,Locomotion - Abstract
Rabphilin, a putative rab effector, interacts specifically with the GTP-bound form of the synaptic vesicle-associated protein rab3a. In this study, we define in vivo functions for rabphilin through the characterization of mutants that disrupt the Caenorhabditis elegans rabphilin homolog. The mutants do not display the general synaptic defects associated with rab3 lesions, as assayed at the pharmacological, physiological, and ultrastructural level. However, rabphilin mutants exhibit severe lethargy in the absence of mechanical stimulation. Furthermore, rabphilin mutations display strong synergistic interactions with hypomorphic lesions in the syntaxin, synaptosomal-associated protein of 25 kDa, and synaptobrevin soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) genes; double mutants were nonresponsive to mechanical stimulation. These synergistic interactions were independent of rab3 function and were not observed in rab3-SNARE double mutants. Our data reveal rab3-independent functions for rabphilin in the potentiation of SNARE function.
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- 2001
13. Caenorhabditis elegans rab-3 mutant synapses exhibit impaired function and are partially depleted of vesicles
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Jane Staunton, Michael P. Kilgard, Erik M. Jorgensen, Tim Fergestad, Erika Hartwieg, Michael L. Nonet, Barbara J Meyer, and H. Robert Horvitz
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rab3 GTP-Binding Proteins ,Endocytic cycle ,Molecular Sequence Data ,Neuromuscular Junction ,Nerve Tissue Proteins ,Biology ,Neurotransmission ,Synaptic vesicle ,Polymerase Chain Reaction ,Synaptic Transmission ,Exocytosis ,Mice ,Species Specificity ,GTP-Binding Proteins ,medicine ,Animals ,Paralysis ,Active zone ,Amino Acid Sequence ,Axon ,Cloning, Molecular ,Caenorhabditis elegans ,Genes, Helminth ,Motor Neurons ,Sequence Homology, Amino Acid ,General Neuroscience ,Vesicle ,Chemotaxis ,Helminth Proteins ,Articles ,Cell biology ,medicine.anatomical_structure ,Drosophila melanogaster ,Synapses ,Cattle ,Rab ,Cholinesterase Inhibitors ,Synaptic Vesicles ,Sequence Alignment ,Aldicarb - Abstract
Rab molecules regulate vesicular trafficking in many different exocytic and endocytic transport pathways in eukaryotic cells. In neurons, rab3 has been proposed to play a crucial role in regulating synaptic vesicle release. To elucidate the role of rab3 in synaptic transmission, we isolated and characterizedCaenorhabditis elegans rab-3mutants. Similar to the mouse rab3A mutants, these mutants survived and exhibited only mild behavioral abnormalities. In contrast to the mouse mutants, synaptic transmission was perturbed in these animals. Extracellular electrophysiological recordings revealed that synaptic transmission in the pharyngeal nervous system was impaired. Furthermore,rab-3animals were resistant to the acetylcholinesterase inhibitor aldicarb, suggesting that cholinergic transmission was generally depressed. Last, synaptic vesicle populations were redistributed inrab-3mutants. In motor neurons, vesicle populations at synapses were depleted to 40% of normal levels, whereas in intersynaptic regions of the axon, vesicle populations were elevated. On the basis of the morphological defects at neuromuscular junctions, we postulate that RAB-3 may regulate recruitment of vesicles to the active zone or sequestration of vesicles near release sites.
- Published
- 1997
14. Optimization of a Yellow Fluorescent Protein-Based Iodide Influx High-Throughput Screening Assay for Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Modulators.
- Author
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Jinliang Sui, Shakira Cotard, Jennifer Andersen, Ping Zhu, Jane Staunton, Margaret Lee, and Stephen Lin
- Subjects
BIOLOGICAL assay ,PROCESS optimization ,IODIDES ,CYSTIC fibrosis ,GENETIC mutation ,AMINO acids ,CELL membranes ,PHENYLALANINE - Abstract
AbstractCystic fibrosis is an inherited, life-threatening disease associated with mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The most common mutation, F508del CFTR, is found in 90% of CF patients. The loss of a single amino acid (phenylalanine at position 508) results in malformed CFTRwith defective trafficking to the plasma membrane and impaired channel function. A functional assay with cells expressing F508del CFTRhas been previously described by others using genetically engineered halide-sensitive yellow fluorescent protein to screen for CFTRmodulators. We adapted this yellow fluorescent protein assay to 384-well plate format with a high-throughput screening plate reader, and optimized the assay in terms of data quality, resolution, and throughput, with target-specific protocols. The optimized assay was validated with reference compounds from cystic fibrosis foundation therapeutics. On the basis of the Z-factor range (≥0.5) and the potential productivity, this assay is well suited for high-throughput screening. It was successfully used to screen for active single agent and synergistic combinations of single agent modulators of F508del CFTRfrom a library collection of current active pharmaceutical ingredients (supported by Cystic Fibrosis Foundation Therapeutics). [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
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15. Identification of Synergistic Combinations of F508del Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Modulators.
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Stephen Lin, Jinliang Sui, Shakira Cotard, Brenda Fung, Jennifer Andersen, Ping Zhu, Najib El Messadi, Joseph Lehar, Margaret Lee, and Jane Staunton
- Subjects
CYSTIC fibrosis ,CHEMICAL inhibitors ,GENETIC mutation ,EPITHELIAL cells ,ATP-binding cassette transporters ,HALIDES ,BIOLOGICAL assay - Abstract
AbstractCystic fibrosis (CF) is an inherited, life-threatening disease caused by mutations in the gene encoding cystic fibrosis transmembrane conductance regulator (CFTR), an ABC transporter-class protein and ion channel that transports ions across epithelial cell membranes. The most common mutation leads to the deletion of a single phenylalanine, and the resulting protein, F508del-CFTR, shows reduced trafficking to the membrane and defective channel gating. The ideal therapeutic approach would address both of these defects and restore channel function at the same time. We describe here the application of a combination high-throughput screening to search for synergistic modulators of F508del-CFTR. With the adapted Fischer rat thyroid-yellow fluorescent protein halide flux assay to the combination high-throughput screening platform, we identified many interesting single agents as CFTR modulators from a library of approved drugs and mechanistic probe compounds, and combinations that synergistically modulate F508del-CFTR channel function in Fischer rat thyroid cells, demonstrating the potential for combination therapeutics to address the defects that cause CF. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
16. Adult male-specific and neonatally programmed rat hepatic P-450 forms RLM2 and 2a are not dependent on pulsatile plasma growth hormone for expression
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Jane Staunton, Joseph J. Morrissey, Gerald A. LeBlanc, David J. Waxman, and David P. Lapenson
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medicine.medical_specialty ,Hypophysectomy ,medicine.drug_class ,medicine.medical_treatment ,Developmental induction ,Alpha (ethology) ,Cell Biology ,Biology ,Biochemistry ,Steroid ,Endocrinology ,Estrogen ,Internal medicine ,medicine ,Phenobarbital ,Molecular Biology ,Testosterone ,Hormone ,medicine.drug - Abstract
Rat hepatic cytochrome P-450 form RLM2 is a testosterone 15 alpha-hydroxylase reported to be male-specific on the basis of purification studies (Jansson, I., Mole, J., and Schenkman, J. B. (1985) J. Biol. Chem. 260, 7084-7093). The sex dependence, developmental regulation, xenobiotic induction, and hormonal control of P-450 RLM2 expression were studied using P-450 form-specific immunochemical and catalytic assays. Polyclonal antibodies raised to rat hepatic P-450 3 (P-450 gene IIA1) were found to cross-react strongly with P-450 RLM2, but not with 10 other rat P-450 forms, suggesting that P-450 3 and P-450 RLM2 are highly conserved in primary structure. Western blotting of liver microsomes under conditions where P-450s 3 and RLM2 are resolved electrophoretically revealed that P-450 RLM2 is markedly induced at puberty in male rats, with no protein detected (less than or equal to 5% of adult male levels) in adult females or immature animals of either sex. A similar developmental dependence was observed for hepatic microsomal testosterone 15 alpha-hydroxylase activity, which was found to be catalyzed primarily by P-450 RLM2. P-450 RLM2 was resistant to induction by several xenobiotics and in the case of phenobarbital and beta-naphthoflavone, was suppressed by 50-60%. Studies on the steroid hormonal regulation of P-450 RLM2 revealed that its adult male-specific expression is imprinted (programmed) in response to neonatal testosterone exposure. Ovariectomy studies demonstrated that suppression by estrogen does not contribute significantly to the absence of P-450 RLM2 in adult female rats. Although the male-specific developmental induction of P-450 RLM2 in response to neonatal testosterone is strikingly similar to that of P-450 2c (testosterone 2 alpha/16 alpha-hydroxylase; gene IIC11), P-450 RLM2 expression is not dependent on the pulsatile pituitary growth hormone secretion required for P-450 2c synthesis. Rather, hypophysectomy of adult male rats increased P-450 RLM2 and its associated testosterone 15 alpha-hydroxylase activity by 50-100%.(ABSTRACT TRUNCATED AT 400 WORDS)
- Published
- 1988
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17. Improving Autobiographical Memory and Social Interaction in Children with Autism Spectrum Disorder Through Reminiscence
- Author
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Gaelen, Jane Staunton
- Subjects
- Psychology, Educational psychology
- Abstract
Background: It is hypothesized that with improved autobiographical memory access, children with Autism Spectrum Disorder may be able to implement better Theory of Mind skills through shared conversations about personal past events. Despite increasing research into the correlations between autobiographical memory, Theory of Mind, and autism spectrum symptoms, no studies have examined the influence of reminiscing practices that might enhance or diminish individual’s participation in conversations about personal past events. It was hypothesized that the quality and quantity of a conversational partner’s elaborations would relate positively to children’s autobiographical memory recall and Theory of Mind abilities. It was expected that better episodic memory performance would be associated with fewer symptoms. This study examined correlations between the researcher’s reminiscing style, children’s recall of personal past experiences, and autism specific behaviors. Twelve children diagnosed with Autism Spectrum Disorder completed two semi-structured reminiscing interview tasks. The dependent measure was the number of memory responses. Children completed the Origins of Knowledge Task and the False Belief Task as measures of social cognition. Parents completed the CARS2QPC and the investigator completed the CARS2-HF as measures of symptom severity. The study used the Indirect-Direct Experiences Tasks and the narrative measure to identify associations with autobiographical memory, Theory of Mind and, autism behaviors. Data showed significant positive correlations between the frequencies of the investigator’s comments and questions and free and cued memory responses. For all children, enhanced free recall performance was related to lower levels of social problems. These associations were not accounted for by performance on the Theory of Mind tasks. Overall, children showed increased processing of autobiographical memory during reminiscing associated with both comments and questions. Higher measures of indirect and direct experience memory task accuracy revealed higher levels of performance on tasks measuring social cognition. Individual differences in autobiographical memory abilities may be associated with Theory of Mind and the expression of social symptoms in children with ASD.
- Published
- 2021
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