Core:1 Biological Testing

核心：1 生物检测

基本信息

批准号：
10410430
负责人：
Joanna E Burdette
金额：
$ 16.45万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10410430
关键词：
3-Dimensional Animal Experiments Animal Model Animals Antineoplastic Agents Autophagocytosis Biological Biological Assay Biological Testing Biometry Cancer Model Cancer cell line Cell Line Cells Cellular Assay Chemicals Collaborations Complex Mixtures Crude Extracts Cyanobacterium Data Development Dose Drug Kinetics Evaluation Fingerprint Formulation Foundations Genetic Transcription Goals Gold Growth Human Immune system In Vitro Individual Industrialization Lead Lichen - organism Malignant neoplasm of ovary Mass Spectrum Analysis Materials Testing Modeling Molecular Molecular Profiling Mus National Center for Advancing Translational Sciences Natural Products Nude Mice Nutrient Pathway interactions Penetration Pharmaceutical Preparations Phenotype Plants Proteins Proteomics Reporting Research Personnel Role Route Solid Solvents Source Structure Structure-Activity Relationship System Testing Toxic effect United States National Institutes of Health Work Xenograft procedure analog anti-cancer anticancer research base cancer therapy chemotherapy cytotoxic cytotoxicity cytotoxicity test drug candidate efficacy study experimental study fungus high throughput screening improved in vivo in vivo evaluation novel plant fungi programs response screening transcriptome sequencing transcriptomics tumor

项目摘要

Core 1 will focus on the biological evaluation of anticancer natural products that are identified from complex extracts, and after subsequent isolation, confirmed to be the active constituent. This strategy, called bioassay-guided isolation, facilitates the identification of biological activity within complex mixtures from plants, fungi, and cyanobacteria. Extracts and compounds that display cytotoxicity and modulation of autophagy will be prioritized based on high-content phenotypic screening and the ability to inhibit 3D spheroid growth by Core 1. Hits will be further purified by Projects 1-3 (OSU, UIC, and UNCG, respectively) and the purified fractions interrogated until pure compounds emerge. Promising pure compounds will ultimately be evaluated in animal models to assess efficacy and toxicity. Core 1 will work closely with Core 2, led by Dr. Fuchs, to evaluate structure-activity relationships and to establish optimal formulation strategies and determine pharmacokinetics profiles prior to in vivo testing in Core 1. This approach will help maximize the possibility of successful animal experiments based on dose and route of administration. Mechanism of action studies will be conducted to find protein interacting partners and pathways based on transcriptomics. Dr. Burdette of Core 1 and her Co-Investigator, Dr. Aldrich, have well-established collaborations with all of the Project Leaders and this application includes preliminary data indicating our ability to successfully integrate compounds from all of the projects and cores. In Aim 1, we will evaluate all extracts and compounds using a cytotoxicity and autophagy high throughput screen. In Aim 2, we will prioritize compounds using a high content molecular fingerprinting assay to evaluate whether compounds likely have unique mechanisms of action as compared to standard chemotherapies used as controls. We will also determine the potency of compounds against 3D spheroids, which typically helps to identify lead compounds that have potency in vivo. The prioritized compounds will be evaluated in vivo in both a human OVCAR8-RFP xenograft in athymic nude mice and using a newly developed syngeneic model with an intact immune system. Finally, in vivo active compounds will be optimized in collaboration with Core 2 to develop more active synthetic analogues and to develop photo-affinity probes and RNA sequencing for target and pathway identification. Overall, Core 1 tests all compounds from the Projects as well as those made in Core 2 for in vitro and in vivo activity. Data are then integrated fully with Core A for biostatistical analysis.

核心1将重点介绍从复杂的抗癌天然产品的生物学评估提取物以及随后的分离后，证实为活性成分。这种策略，称为生物测定指导隔离，促进植物，真菌和真菌复杂混合物中生物活性的鉴定蓝细菌。将优先考虑显示细胞毒性和调节的提取物和化合物基于高含量的表型筛选以及抑制3D球体生长的能力。由项目1-3（分别为OSU，UIC和UNCG）进一步纯化，并质疑纯化的部分纯化合物出现。有希望的纯化合物最终将在动物模型中评估以评估功效和毒性。 Core 1将与Fuchs博士领导的Core 2紧密合作，以评估结构活动关系并建立最佳配方策略，并在此之前确定药代动力学概况核心1中的体内测试。这种方法将有助于最大化基于动物实验的成功可能性关于剂量和行政途径。将进行作用研究机理以发现蛋白质相互作用基于转录组学的合作伙伴和途径。 Core 1的Burdette博士和她的共同研究员Aldrich博士，与所有项目负责人进行了完善的合作，该应用程序包括初步数据表明我们能够成功整合所有项目和核心的化合物的能力。在AIM 1中，我们将使用细胞毒性和自噬高吞吐量屏幕评估所有提取物和化合物。在AIM 2中，我们将使用高含量的分子指纹测定法来评估化合物是否优先考虑化合物与用作对照的标准化学疗法相比，可能具有独特的作用机理。我们将还可以确定化合物对3D球体的效力，这通常有助于识别铅化合物在体内具有效力。优先化的化合物将在人体OVCAR8-RFP中在体内进行评估无胸腺裸鼠中的异种移植物，并使用具有完整免疫系统的新开发的同步模型。最后，体内活性化合物将与Core 2合作进行优化，以开发更多的活性合成类似物并开发用于目标和途径识别的光亲和力探针和RNA测序。总体而言，核心1测试了项目的所有化合物，以及在体外和体内核心2中制造的化合物活动。然后将数据与核心A完全集成以进行生物统计分析。