Exploiting cellular discrimination through GLUTs with small-molecule GLUT-targeting probes

摘要 果糖摄取的变化与癌症的发展有关，并向更具侵略性的转变。 表型已经进行了许多努力来使用GLUT作为治疗靶点以区分癌细胞，以用于治疗。 疾病诊断和治疗。然而，特异性靶向疾病相关的GLUT是一个未满足的需求。我们 假设靶向疾病相关GLUT的组成和活性将能够快速准确地 代谢改变的鉴定以及疾病诊断和治疗。然而，一个重要的限制是， 阻碍了基于GLUT的小区识别。那就是缺乏分子工具来区分 葡萄糖。因此，我们的目标是能够特异性靶向不同的GLUT，以有效区分 正常细胞和癌细胞以及癌症亚型之间的关系，具体目标如下： 目标1。探索果糖GLUT与荧光团的货物容量。我们建议进行一项 两种主要果糖转运蛋白- GLUTs 2和5 -的货物偏好的系统探索 荧光团作为货物和摄取的同时实时报告。了解差异， GLUT之间的存在将直接促进GLUT特定分析和生物医学工具的发展。 目标二。探索糖构象对果糖转运蛋白底物选择的影响。 细胞中的果糖转运由GLUT 5和非特异性GLUT 2、7、9、11和12促进。虽然显著 文献通过GLUT 5解决底物选择，其他转运蛋白的底物摄取关系， 它们与癌症的相关性还没有被很好地理解。在此目标下，GLUT 2和GLUT 5区分 糖构象之间的关系将通过集中的结构-摄取关系研究进行评估。的 获得的知识将为设计针对果糖特异性与果糖的分子探针奠定基础， 非特异性GLUT。 目标3。在癌细胞鉴别中使用基于小分子的GLUT分析。的重大 正常细胞和癌细胞之间果糖摄取的差异支持果糖摄取的定量分析 作为致癌作用的衡量标准。本研究的重点是探讨是否有针对性地分析GLUT 5和细胞 代谢的检测可以提供鉴定代谢受损细胞如癌症的有效方法。的 方法是基于使用GLUT特异性分子探针。 本文所产生的知识、探测和分析方法将为 开发诊断剂和可能的药用剂作为糖摄取和细胞增殖的调节剂， 新陈代谢.概述的研究计划也是为了促进下一代科学家的教育 因为该计划大量涉及本科生，研究生和博士后水平的研究人员。拟议 研究项目直接有助于实现NIH区域奖的使命，并加强研究和 密歇根理工大学健康相关领域的教育环境。

Abstract Changes in fructose uptake have been implicated in cancers development, and transition into more aggressive phenotypes. Many efforts have been directed to use GLUTs as therapeutic targets to distinguish cancer cells for disease diagnosis and treatment. However, specific targeting of disease-relevant GLUTs is an unmet need. We hypothesize that targeting the composition and activity of disease-relevant GLUTs will enable rapid and accurate identification of metabolic alterations and disease diagnosis and therapy. However, a significant limitation hampers the GLUT-based cell identification. That is the absence of molecular tools to discriminate between GLUTs. Thus, it is our goal to enable specific targeting of different GLUTs for effective discrimination between normal and cancer cells and between cancer subtypes through the following specific aims: Aim 1. Exploring the cargo capacity of fructose GLUTs with fluorophores. We propose to carry out a systematic exploration of cargo preferences for two major fructose transporters – GLUTs 2 and 5 - using fluorophores as cargoes and simultaneous real-time reporters of the uptake. Understanding the differences that exist between GLUTs will directly promote the development of GLUT-specific analytical and biomedical tools. Aim 2. Exploring the impact of sugar conformation on substrate selection by fructose transporters. Fructose transport in cells is facilitated by GLUT5 and non-specific GLUTs 2, 7, 9, 11 and 12. While significant literature addresses substrate selection by GLUT5, the substrate-uptake relationship for other transporters and their relevance to cancer is not well understood. Within this aim, the ability of GLUT2 and GLUT5 to discriminate between sugar conformations will be assessed through focused structure-uptake relationship studies. The acquired knowledge will lay the foundation for designing molecular probes to target fructose-specific vs. fructose- nonspecific GLUTs. Aim 3. Employing small molecule-based GLUT analysis in cancer cell discrimination. The significant difference in fructose uptake between normal and cancer cells supports quantitative analysis of fructose uptake as a measure of carcinogenesis. The focus of this aim is to explore whether targeted analysis of GLUT5 and cell metabolism can provide effective approach to identify metabolically-compromised cells such as cancer. The approach is based on using GLUT-specific molecular probes. The knowledge, probes, and analytical approaches produced herein will lay foundation for the development of diagnostic agents and, possibly, medicinal agents as regulators of sugar uptake and cellular metabolism. The outlined research plan is also geared to promote the education of next generations of scientists as the program heavily involves researchers at undergraduate, graduate, and postdoctoral levels. The proposed research project directly contributes to fulfilling the mission of NIH AREA award and strengthen the research and an educational environment in health-related fields at Michigan Tech.