Modulated Thermal Stress to Manipulate Cell Protein Expression

CBET-0828131 Diller概述。分子生物学的进展表明，细胞可能通过交替的途径对分级热应激做出反应，导致细胞坏死或增加热休克蛋白(HSP)的产生，从而提高存活率。在治疗癌症等应用中，当有合理的基础来预测发生坏死和存活的领域时，热疗是最有效的设计。当特定细胞类型的构成行为被定量地表征为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，就有可能设计一种热方案来在靶细胞和组织中引发特定的治疗或预防反应。这项建议旨在开发一套生物工程数据和工具，用于操纵HSP在正常和前列腺癌细胞中的表达。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体系，用荧光蛋白克隆显示热休克蛋白的表达和细胞坏死。特别感兴趣的是根据施加的温度和时间分布来量化表达过程的动力学。根据施加的暴露温度和时间，细胞将受到受控的热应激方案，并通过热休克蛋白27、60和70的表达、细胞凋亡和坏死来测量反应。由于已有大量关于热损伤效应的知识，本研究将集中于热休克蛋白表达的动力学特征。本构性能数据将以可用于工艺设计的数学公式表示。分析和建模方法将转化为基于最先进的学习科学技术的生物运输的新学习工具。这项拟议的研究是围绕我们公认的测量、表征和应用通过受控热应激在选定细胞类型中操纵HSP表达的能力而设计的。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加热方案，以平衡靶向细胞破坏和提高存活率，这可以应用于患者的特定治疗。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构成数据，我们预计这将是该研究的开创性贡献。它将通过控制组织自我修复的能力来提高控制对组织施加的治疗性损伤的能力，并应用于预防和治疗的热疗程序的设计。更广泛的影响。这项研究的特点是研究和教育方面的紧密结合。教育部分以人们如何学习的科学(HPL)为基础，专注于开发和应用学习材料，帮助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挑战问题，这些问题涉及现实的工程问题，并基于现有的研究结果，如将在拟议的研究中开发的。这些学习工具可以很容易地出口，供其他成熟的教育合作伙伴使用，包括德克萨斯大学泛美分校(该校的西语裔入学率是美国大陆所有院校中最高的)和德克萨斯州埃尔金高中(一个农村少数民族学区，PI在该学区获得了目前的德克萨斯州STEM奖)。这些教育材料还将在档案学、科学和工程教育文献中传播，在这些文献中，非政府组织定期出版。研究结果将通过国际癌症研究所与德克萨斯大学MD安德森癌症中心的同事建立的翻译伙伴关系，在癌症治疗方面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科学生的新教育工具中。将开发两种新类型的学习挑战问题，用于HPL教育框架。其中一个重点是确定热休克蛋白表达的热动力学作为施加温度应力模式的函数。第二个将涉及反问题的定义和求解，以设计所施加的热应力以在目标组织中产生所需的热休克蛋白和细胞损伤模式。

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.