Modulated Thermal Stress to Manipulate Cell Protein Expression

调节热应激来操纵细胞蛋白表达

• 批准号: 0828131
• 负责人: Kenneth Diller
• 金额: $ 29.96万
• 依托单位: University of Texas at Austin
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0828131&HistoricalAwards=false
• 关键词: Modulated; Thermal; Stress; Manipulate; Cell

CBET-0828131DillerOverview. Advances in molecular biology have demonstrated that cells may respond to graded thermal stress by alternate pathways leading to either cell necrosis or enhanced production of heat shock proteins (HSP) which will enhance survival. In applications such as treating cancer, thermal therapies are designed most effectively when there is a rational basis for predicting the domains for which necrosis and survival occur. When the constitutive behavior of a specific cell type is characterized quantitatively as coupling the time course and magnitude of hsp expression enhancement with the time and temperature dimensions of an applied thermal stress, it then becomes possible to design a thermal protocol to elicit a specific therapeutic or prophylactic response in target cells and tissues. This proposal is directed toward developing a set of bioengineering data and tools for manipulating hsp expression in normal and cancerous cells of the prostate.Approach. The experimental system is two and three dimensional cell cultures established from canine prostate normal and cancerous tissues, cloned with fluorescent proteins to display hsp expression and cell necrosis. Of particular interest is quantification of the kinetics of the expression process in terms of the applied temperature and time profile. Cells will be subjected to controlled thermal stress regimens as defined by the imposed exposure temperature and time, and the response measured in terms of hsp 27, 60 and 70 expression, apoptosis and necrosis. Since there already exists a significant body of knowledge about thermal injury effects, this study will focus on characterizing the kinetics of hsp expression. The constitutive property data will be expressed in mathematical formulations that can be used for process design. The analysis and modeling methods will be transformed into new learning tools for biotransport based on state-of-the-art learning science techniques.Intellectual Merit. The proposed study is designed around our well established capability to measure, characterize and apply the manipulation of hsp expression in selected cells types via controlled thermal stress. A strength of this work is the development of a quantitative method for designing thermal protocols to apply to living systems that balances targeted cell destruction and enhanced survival which can be applied for patient specific therapies. A critical component of the success of this approach is to identify constitutive data for the kinetics of temperature driven processes in specific cell species, which we anticipate will be a pioneering contribution of the study. It will lead to an increased ability to control therapeutic injury imposed on tissue by manipulating the ability of tissue to repair itself, with applications for the design of thermal procedures for prophylaxis and therapy. Broader Impacts. This study features a close integration of research and education aspects. The educational component is based on the science of How People Learn (HPL) and is focused on developing and applying learning materials that facilitate students acquiring adaptive expertise for problem solving. A key component is the use of open ended challenge problems that deal with realistic engineering issues and that are based on present research findings, such as will be developed in proposed study. These learning tools are readily exported for use by other established educational partners including the University of Texas - Pan American (which has the highest Hispanic enrollment of any continental institution in the US) and the Elgin, TX High School (a rural minority school district with which the PI has obtained a current Texas STEM award). The educational materials also will be disseminated in the archival learning science and engineering education literature in which the PI publishes regularly. The research results will be of direct benefit to society in terms of medical advances in cancer treatment via established translational partnerships of the PI with colleagues at UT MD Anderson Cancer Center.Integration of Research and Education. Knowledge gained from the research results will be incorporated into new educational tools for engineering students. Two new types of learning challenge problems will be developed for use in the HPL educational framework. One will focus on determination of the thermal kinetics of hsp expression as a function of an applied temperature stress pattern. A second will involve definition and solution of an inverse problem to design an applied thermal stress to produce a desired hsp and cell injury pattern in a target tissue.

CBET-0828131 Diller概述。 分子生物学的进展已经证明，细胞可以通过交替途径对分级的热应激作出反应，导致细胞坏死或增加热休克蛋白（HSP）的产生，这将提高存活率。 在诸如治疗癌症的应用中，当有合理的基础来预测坏死和存活发生的区域时，热疗法被设计得最有效。 当特定细胞类型的组成性行为被定量表征为将hsp表达增强的时间过程和幅度与所施加的热应力的时间和温度维度相耦合时，则可以设计热方案以在靶细胞和组织中引起特定的治疗或预防反应。 该建议是针对开发一套生物工程数据和工具，用于操纵热休克蛋白在前列腺的正常和癌细胞中的表达。 实验系统是从犬前列腺正常组织和癌组织建立的二维和三维细胞培养物，用荧光蛋白克隆以显示热休克蛋白表达和细胞坏死。 特别令人感兴趣的是定量的表达过程中的动力学的应用温度和时间曲线。 细胞将经受受控的热应激方案，如通过施加的暴露温度和时间所定义的，并且根据hsp 27、60和70表达、凋亡和坏死来测量响应。 由于已经存在一个显着的机构的知识，热损伤的影响，这项研究将集中在表征热休克蛋白表达的动力学。 本构性质数据将以可用于工艺设计的数学公式表示。 基于最先进的学习科学技术，分析和建模方法将转化为生物运输的新学习工具。智力优点。 拟议的研究是围绕我们建立良好的能力，测量，表征和应用在选定的细胞类型，通过控制热应力热休克蛋白表达的操纵。 这项工作的一个优势是开发了一种定量方法，用于设计适用于生命系统的热协议，平衡靶向细胞破坏和提高生存率，可用于患者特异性治疗。 这种方法成功的一个关键组成部分是确定特定细胞物种中温度驱动过程动力学的组成数据，我们预计这将是这项研究的开创性贡献。 它将导致通过操纵组织自身修复的能力来控制施加在组织上的治疗性损伤的能力增加，并应用于预防和治疗的热程序的设计。更广泛的影响。 这项研究的特点是研究和教育方面的密切结合。 教育部分是基于人们如何学习（HPL）的科学，并专注于开发和应用学习材料，促进学生获得解决问题的适应性专业知识。 一个关键组成部分是使用开放式的挑战问题，处理现实的工程问题，并根据目前的研究结果，如将在拟议的研究开发。 这些学习工具很容易出口到其他已建立的教育合作伙伴使用，包括德克萨斯大学泛美分校（在美国任何大陆机构中拥有最高的西班牙裔入学率）和德克萨斯州埃尔金高中（一个农村少数民族学区，PI已获得当前德克萨斯州STEM奖）。 还将在PI定期出版的科学和工程教育文献档案中传播教育材料。 通过PI与UT MD安德森癌症中心的同事建立的翻译伙伴关系，研究结果将在癌症治疗的医学进步方面直接造福社会。 从研究成果中获得的知识将被纳入工程专业学生的新教育工具。 两种新类型的学习挑战问题将被开发用于HPL教育框架。 一个将集中在确定热休克蛋白表达的热动力学作为一个应用的温度应力模式的函数。 第二个将涉及反问题的定义和解决方案，以设计所施加的热应力，从而在靶组织中产生所需的热休克蛋白和细胞损伤模式。