HTS for FADD kinase inhibitors using molecular imaging

使用分子成像对 FADD 激酶抑制剂进行 HTS

基本信息

批准号：
7682117
负责人：
Alnawaz Rehemtulla
金额：
$ 20.86万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-03 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7682117
关键词：
Adaptor Signaling Protein Adult Amino Acids Antibodies Apoptotic Biochemical Biological Assay Biological Factors Biological Markers Bioluminescence Brain C-terminal Cancer Patient Cancer cell line Cell Cycle Progression Cells Cessation of life Chemicals Clinical Collection Complex Cyclin D1 Data Death Domain Development Diagnosis Disease Distress Dose Embryonic Development Ensure Environment Enzymes Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Erlotinib Event FDA approved Family Fluorescent Dyes G2/M Transition Gene Mutation Genetically Engineered Mouse Genomics Germ cell tumor Goals Head and Neck Cancer Head and neck structure Human Image Imaging Device Immunohistochemistry In Vitro Inhibitory Concentration 50 Knockout Mice Lead Letters Libraries Luciferases Lung Malignant Neoplasms Malignant neoplasm of lung Measures Mediator of activation protein Methodology Methods Michigan Microarray Analysis Molecular Profiling Molecular and Cellular Biology Monitor Noise Nuclear Operative Surgical Procedures Outcome Pan Genus Peptides Permeability Persons Phenotype Phosphorylation Phosphotransferases Play Prevalence Probability Protein Kinase Proteins Publishing Quantitative Evaluations Radiation therapy Regulation Relative (related person)Reporter Reporting Resort Role Science Screening procedure Sensitivity and Specificity Series Serine Signal Transduction Signal Transduction Pathway Solubility Specificity Stream Survival Rate T-Cell Proliferation Technology Time Tissue Microarray Toxic effect Two-Dimensional Gel Electrophoresis Universities Western Blotting aggressive therapy base cancer cell cancer initiation cancer therapy chemotherapeutic agent clinically significant cyclin B1 cytotoxic data mining enzyme activity high throughput screening inhibitor/antagonist inorganic phosphate kinase inhibitor male molecular imaging mortality mutant neoplastic cell novel novel therapeutics overexpression prognostic research study small molecule libraries success therapy resistant tumor two-dimensional

项目摘要

DESCRIPTION (provided by applicant): Using differential expression profiling, quantitative two-dimensional (2-D) gel electrophoresis and data mining we recently identified a new prognostic biomarker, Fas-associated death domain (FADD), which is overexpressed in a number of human malignancies such as lung, head and neck, brain and adult male germ cell tumors. Studies in lung cancer revealed that overexpression of FADD significantly associated with poor clinical outcome. Immunohistochemistry-based tissue microarray analysis confirmed the association between FADD over-expression and the poor outcome, and also revealed the presence of nuclear localized phosphorylated FADD (p-FADD). Tumors with increased p-FADD expression also showed elevated NF-?B activation. Taken together, published results from our lab and others suggest a causal relationship between the phosphorylation of FADD and NF-?B activation, a hallmark of an aggressive therapy resistant cancer phenotype. Thereby, we hypothesize that inhibiting FADD phosphorylation in tumor cells may sensitize cancer cells to chemotherapeutic agents. To aid in experimentation of this hypothesis we have resorted to molecular imaging tools and developed a pan FADD kinase reporter (FKR) which non-invasively senses FADD-kinase activity in real time. In Specific Aim 1, we will characterize the sensitivity and specificity of FKR. In Specific Aim 2A we will perform a high throughput screen to identify molecules from a diverse set of compound libraries that target FADD phosphorylation. Utilizing secondary screens with cells expressing either mutant FKR or luciferase, the toxic and less sensitive lead molecules will be eliminated. In Specific Aim 2B we will evaluate the relative efficacy of the candidate molecules by quantifying IC50 of the top leads. In Specific Aim 2C the specificity of candidate molecules in inhibiting FADD kinases will be investigated using western blotting and protein kinase arrays. The utility of these compounds and their derivatives in the treatment of cancers will be investigated in subsequent years.

描述（由申请人提供）：使用差分表达谱，定量二维（2-D）凝胶电泳和数据挖掘，我们最近确定了一种新的预后生物标志物，FAS相关的死亡结构域（FADD），这些死亡结构域（FADD）在许多人类恶性肿瘤中过表达的人类恶性肿瘤，例如肺，脑和脑，大脑和成人男性细胞和成人男性细胞和成人男性细胞。对肺癌的研究表明，FADD的过表达与临床结果不良显着相关。基于免疫组织化学的组织微阵列分析证实了FADD过表达与不良预后之间的关联，并且还揭示了存在核局部磷酸化的FADD（P-FADD）。 P-FADD表达增加的肿瘤还显示NF-？B激活升高。综上所述，我们实验室和其他实验室的结果表明，FADD的磷酸化与NF-？B激活之间存在因果关系，这是抗侵略性治疗癌症表型的标志。因此，我们假设抑制肿瘤细胞中的FADD磷酸化可能会使癌细胞对化学治疗剂敏感。为了帮助实验这一假设，我们诉诸于分子成像工具，并开发了PAN FADD激酶报告基因（FKR），该基因酶报道器（FKR）无侵入性地实时感知FADD-激酶活性。在特定目标1中，我们将表征FKR的灵敏度和特异性。在特定的目标2a中，我们将执行高吞吐量屏幕，以鉴定针对针对FADD磷酸化的各种化合物库中的分子。使用表达突变体FKR或荧光素酶的细胞利用次级筛选，将消除有毒和敏感的铅分子。在特定的目标2B中，我们将通过量化顶级铅的IC50来评估候选分子的相对功效。在特定的目标2C中，将使用蛋白质印迹和蛋白激酶阵列研究候选分子在抑制FADD激酶中的特异性。这些化合物及其在癌症治疗中的效用将在随后的几年中进行研究。