BIOMOLECULAR ANALYSIS USING LIQUID CRYSTALS

使用液晶进行生物分子分析

基本信息

批准号：
8520195
负责人：
NICHOLAS L ABBOTT
金额：
$ 35.19万
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 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8520195
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 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 screening 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合作伙伴（UW-BRP）正在开发新的分子分析工具来识别和验证生物终点，从而可以更准确、更快速地评估新型抗癌药物的活性。分子机制和临床相关性。这项工作的重点是表皮生长因子受体（EGFR），因为它的过度表达和突变与一些最无法治愈的癌症密切相关。然而，这些工具的通用性足以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它们对受体或其他信号分子富集制剂的体外分子分析的应用，但 UW-BRP 寻求建立也可应用于分析培养细胞样品的工具原理，以及来自异种移植和自发肿瘤组织的活检。这种能力最终将实现一种跨越分子、细胞和组织水平的基本方法，并将用于基础研究以及动物和人类临床试验。在过去的 4 年里，UW-BRP 定义了基于液晶的分子分析工具的关键分析特征，并展示了 EGFR 表达和激活（磷酸化）以及小分子对 EGFR 酪氨酸激酶活性的抑制的报告（在复杂的样品，例如膜提取物和细胞裂解物）。本提案寻求继续支持在化学和生物工程、化学和生物化学以及生物分子和生物医学领域具有不同专业知识的多学科研究人员团队，通过整合以下领域的进展进一步开发这种广泛适用的生物分析方法：a) 识别和优化均匀固定的单链抗体，识别细胞裂解物中关键 EGFR 突变体的表位，b) 整合基于液晶的分析方法和样品制备使用小样本（少数细胞）对野生型和突变型 EGFR 的表达、激活和激酶抑制进行定量，以及 c) 研究参与癌发生相关过程的关键细胞信号传导蛋白。具体来说，UW-BRP 分子分析工具将在一项研究中与传统方法进行比较，该研究将 a) 快速、灵敏地评估生物样品中野生型和突变型人类 EGFR 的水平和活性，b) 测试野生型和突变型人类 EGFR 的假设EGFR 类型和致癌形式将表现出不同的抑制剂特异性，并且 c) 评估在体外有效抑制 EGF 介导事件的药物是否也表现出拮抗细胞培养物中 EGFR 表达和/或活性的能力。从长远来看，这些新工具应该有助于评估抗癌药物的分子机制和后果，从而促进从基础生物学到临床疗效评估的研究。民众

项目成果

期刊论文数量（57）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The use of native chemical functional groups presented by wound beds for the covalent attachment of polymeric microcarriers of bioactive factors.

DOI：
10.1016/j.biomaterials.2012.09.055
发表时间：
2013-01
期刊：
Biomaterials
影响因子：
14
作者：
Jain R;Agarwal A;Kierski PR;Schurr MJ;Murphy CJ;McAnulty JF;Abbott NL
通讯作者：
Abbott NL

Principles for manipulation of the lateral organization of aqueous-soluble surface-active molecules at the liquid crystal-aqueous interface.

DOI：
10.1021/la803475c
发表时间：
2009-02-17
期刊：
Langmuir : the ACS journal of surfaces and colloids
影响因子：
0
作者：
Gupta JK;Abbott NL
通讯作者：
Abbott NL

Interfacial ordering of thermotropic liquid crystals triggered by the secondary structures of oligopeptides.

DOI：
10.1039/c5cc06996c
发表时间：
2015-11
期刊：
Chemical communications
影响因子：
4.9
作者：
Xiaoguang Wang;Pei Yang;Frédéric Mondiot;Yaoxin Li;Daniel S. Miller;Zhan Chen;N. Abbott
通讯作者：
Xiaoguang Wang;Pei Yang;Frédéric Mondiot;Yaoxin Li;Daniel S. Miller;Zhan Chen;N. Abbott

Enantiomeric interactions between liquid crystals and organized monolayers of tyrosine-containing dipeptides.