Molecular probes for lysosomal accumulation, release and environmental sensing.

该项目将在EPSRC的医疗保健技术研究主题，技术触及生活的优先研究领域，以及化学生物学，分析科学和合成有机化学领域运作。溶酶体是一种膜结合的细胞器，其特征是低pH值，其中含有水解酶，作为细胞循环系统的一部分。与这些细胞器相关的低pH值为使用小分子进行积累、释放和传感提供了独特的机会，从而允许了解这种化学环境及其与疾病的关系。我们将设计、合成和监测小分子，这些小分子能够使用生物信息学来处理和解释所生成的成像数据。该项目将在三个不同的领域进行，这些领域将反馈以实现探针开发。领域1：合成化学。设计和合成将在细胞的溶酶体区域积累的小分子。探针将含有三个不同的区域（i）响应于细胞的pH或氧化还原环境的环境敏感基团。(ii)由共轭双芳基丁二炔部分组成的拉曼报告物。(iii)溶酶体靶向基团，其最初将包含仲胺，或将在溶酶体中暴露的受保护仲胺。将确定化合物作为比率探针报告pH或氧化还原电位的能力。如果成功，将建立探针与溶酶体积聚的能力。在识别适当的探针时，我们将建立化学生物学工作流程，以时空方式监测外源分子对溶酶体环境的影响，识别屏幕以监测疾病模型中的表型读数。

The project will operate in the EPSRC research theme of Healthcare Technologies, in the priority research area of Technology Touching Life, and the areas of Chemical Biology, Analytical Science, and Synthetic Organic Chemistry.Lysosomes are membrane bound organelles characterised by a low pH, which contain hydrolytic enzymes that function as part of the cellular recycling system. The low pH associated with these organelles provides a unique opportunity for accumulation, release, and sensing using small molecules, permitting an understanding of this chemical environment and its relationship to disease. We will design, synthesise, and monitor small molecules that are able to examine each of these areas using bioinformatics to process and interpret the imaging data generated.The Project will be conducted in three distinct areas that will feedback to enable probe development.Area 1: Synthetic chemistry. The design and synthesis of small molecules that will accumulate in the lysosomal region of a cell. The probes will contain three distinct regions (i) an environmentally sensitive group that is responsive to either the pH or redox environment of a cell. (ii) a Raman reporter consisting of a conjugated bisarylbutadiyne moiety. (iii) an lysosome targeting group that will initially comprise of a secondary amine, or protected secondary amine that will be unmasked in the lysosome.Area 2: Analytical Chemistry. The ability of the compounds to report back on either pH or redox potential as ratiometric probes will be determined. If successful, the ability of probes to accumulate with lysosomes will be established.Area 3. On identification of an appropriate probe, we will establish chemical biology workflows to monitor the effect of exogenous molecules on lysosome environment in a spatiotemporal manner, identifying screens to monitor phenotypic readouts in models of disease.