BIOMOLECULAR ANALYSIS USING LIQUID CRYSTALS

使用液晶进行生物分子分析

基本信息

批准号：
8107873
负责人：
NICHOLAS L ABBOTT
金额：
$ 39.9万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-03 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8107873
关键词：
Address Agriculture Amplifiers Animal Cancer Model Animal Model Animals Antibodies Antineoplastic Agents Area Basic Cancer Research Basic Science Binding Biochemistry Biological Biological Assay Biological Sciences Biology Biomedical Engineering Biopsy Buffers Cancerous Cell Count Cell Culture Techniques Cell Extracts Cells Cellular biology Characteristics Chemicals Chemistry Clinic Clinical Clinical Research Clinical Trials Clinical assessments Complex Cultured Cells Data Deletion Mutation Development Doctor of Medicine EGF gene EGFR Protein Overexpression Engineering Ensure Epidermal Growth Factor Receptor Epitopes Evaluation Event Exhibits Faculty Family Fluorescence Foundations Future Glioblastoma Glioma Glycoproteins Goals Grant Human Image Immobilization Immunoblotting Impact evaluation In Vitro Individual Interfacial Phenomena Investigation Knowledge Laboratory Research Letters Ligand Binding Domain Ligation Link Malignant Neoplasms Malignant neoplasm of lung Mammary gland Measures Mediating Membrane Methodology Methods Microfabrication Microfluidic Microchips Microfluidics Molecular Molecular Analysis Molecular Biology Mutation Non-Small-Cell Lung Carcinoma Oncogenic Outcomes Research Pathway interactions Peptides Pharmaceutical Preparations Phase I Clinical Trials Philosophy Phosphorylation Phosphotransferases Preparation Process Progress Reports Prostate Protein Chemistry Protein Engineering Protein Tyrosine Kinase Proteins Reagent Receptor Signaling Regulatory Pathway Reporter Reporting Research Research Personnel Role Sampling Schools Science Screening procedure Sensitivity and Specificity Signal Transduction Signaling Molecule Signaling Protein Site Specificity Squamous cell carcinoma Staging Surface System Testing Therapeutic Agents Tissues Tumor Tissue Universities Validation Wisconsin Work analytical method analytical tool anticancer activity anticancer research base carcinogenesis cell preparation clinically relevant college drug sensitivity extracellular genetic regulatory protein immunocytochemistry in vitro activity in vivo inhibitor/antagonist kinase inhibitor liquid crystal medical schools meetings member metaplastic cell transformation method development mutant nano nanoscale nanostructured novel overexpression pancreatic neoplasm professor public health relevance receptor receptor expression small molecule success tool tool development

项目摘要

DESCRIPTION (provided by applicant): The University of Wisconsin Bioengineering Research Partnership (UW-BRP) is developing new molecular analysis tools to identify and validate biological endpoints whereby the activity of novel anti-cancer agents can be more accurately and rapidly evaluated as to their molecular mechanism(s) and clinical relevance. The work is focused on epidermal growth factor receptor (EGFR), given that its overexpression and mutation has been closely-associated with some of the most incurable cancers. However, the tools are sufficiently versatile to be applicable to other key signaling molecules. Whereas the basis of many existing screens restricts their application to in vitro molecular analyses of enriched preparations of receptors or other signaling molecules, the UW-BRP seeks to establish principles for tools that can also be applied to the analysis of samples from cultured cells, and from biopsies of xenographs and spontaneous tumor tissues. This capability will ultimately enable a fundamental approach that will span the molecular, cellular and tissue levels and will be used in both basic research and in animal and human clinical trials. Over the past 4 years, the UW-BRP has defined key analytic characteristics of liquid crystal-based tools for molecular analysis, and demonstrated reporting of EGFR expression and activation (phosphorylation) as well as inhibition of EGFR tyrosine kinase activity by small molecules (in complex samples such as membrane extracts and cell lysates). The present proposal seeks continued support for the multi-disciplinary team of researchers with diverse expertise in chemical and biological engineering, chemistry and biochemistry, and the biomolecular and biomedical sciences to develop further this broadly-applicable bioanalytical approach via integration of advances in the following areas: a) identification and optimization of uniformly immobilized, single chain antibodies that recognize epitopes of key EGFR mutants in cell lysates, b) integration of liquid crystal-based analytic methodologies and sample preparation for quantification of expression, activation and kinase inhibition of wild-type and mutant EGFRs using small samples (a few cells), and c) the investigation of key cell signaling proteins that participate in processes associated with carcinogenesis. Specifically, the UW-BRP molecular analysis tools will be compared to conventional methods in a study that will a) rapidly and sensitively assess the levels and activity of wild-type and mutant human EGFRs in biological samples, b) test the hypothesis that wild-type and oncogenic forms of the EGFR will exhibit differential inhibitor specificity, and c) assess if agents that potently inhibit EGF-mediated events in vitro will also exhibit a capacity to antagonize EGFR expression and/or activity in cell culture. In the long term, these new tools should be useful for the assessment of the molecular mechanisms and consequences of anti-cancer agents, thereby facilitating their research from basic biology through to clinical assessment of efficacy. PUBLIC

描述（由申请人提供）：威斯康星大学生物工程研究伙伴关系（UW-BRP）正在开发新的分子分析工具，以识别和验证生物学终点，从而可以对新型抗癌药物的活性进行更准确，更快地评估其分子机制和临床相关性。鉴于其过表达和突变与某些最无法治愈的癌症密切相关，因此该工作集中在表皮生长因子受体（EGFR）上。但是，这些工具足够多功能，适用于其他关键信号分子。尽管许多现有筛选的基础限制了其应用于富集受体或其他信号分子的体外分子分析，但UW-BRP试图为工具建立原理，这些工具也可以用于分析培养细胞的样品，以及Xenography和Xenography and Xenography and tumor tumor tumor tumor组织的样品。这种能力最终将实现一种基本方法，该方法将跨越分子，细胞和组织水平，并将用于基础研究以及动物和人类临床试验。在过去的四年中，UW-BRP定义了基于液晶的分子分析工具的关键分析特性，并证明了EGFR表达和激活（磷酸化）的报告以及抑制小分子的EGFR酪氨酸激酶活性（在像膜提取物等复杂样品中）。本提案寻求对化学和生物工程，化学和生物化学方面具有多样专业知识的多学科研究人员团队的持续支持，以及生物分子和生物医学科学的生物分子和生物医学科学，以进一步开发这种广泛的生物分析方法，通过在以下领域中识别和优化单一的链条，a），通过识别单一的群体，a），a）单一的群体，a）。细胞裂解物中的突变体，b）基于液晶的分析方法和样品制备，用于定量表达，激活和激酶抑制野生型和突变体EGFR，并使用小样品（几个细胞）（几个细胞）以及c）研究，研究的关键细胞信号蛋白参与与交往的过程相关的过程。具体而言，将在一项研究中将UW-BRP分子分析工具与常规方法进行比较，该研究将a）快速和敏感地评估生物样品中野生型和突变体EGFR的水平和活动的水平和活动，b）测试以下假说，即野生型和过对EGFR的野生型和过对egfr的egfied-fied if fimied if ventied formied formied formied formied formied fornied formied formied formied formied formied formied formied formied contied formied contied formied contied formied contied formied contied formied contied。还将表现出在细胞培养中拮抗EGFR表达和/或活性的能力。从长远来看，这些新工具应有助于评估抗癌剂的分子机制和后果，从而促进其从基本生物学到疗效临床评估的研究。民众