Nanoprobes to reveal signature biomarkers

纳米探针在细胞适应中的功能生物标志物******为什么要研究细胞适应？细胞适应不同有害刺激的能力是推动进化的主要力量之一。细胞适应的机制尚不完全清楚。溶酶体是一种动态细胞器，长期以来被认为是“垃圾焚烧炉”，最近被认为是细胞适应的重要参与者。溶酶体的功能在转录和转录后水平上受到调节，这些功能对细胞外和细胞内的信号作出反应。转录因子EB （TFEB）是溶酶体功能的主要调节因子。它的状态取决于上游激酶的活性。我们的工作假设是细胞对创伤（机械）和化学损伤的适应是通过溶酶体适应发生的。我们的长期研究将集中在与溶酶体适应有关的机制和开发新的（简单和敏感的）纳米工具，这些纳米工具将广泛应用于细胞对损伤的适应过程。***我们的短期目标是：开发基于纳米结构的酶功能分析，如胞质激酶Erk2（细胞外调节激酶）和GSK3 β（糖原合成酶激酶），它们是TFEB的调节因子，以及溶酶体蛋白酶（组织蛋白酶）。2. 生成纳米探针，识别选定的质膜生物标志物，并以比现有工具更高的灵敏度测量其表达。3. 生成“纳米脚本”（人造的、基于纳米颗粒的转录因子），旨在模拟TFEB的结构和功能。我们将利用半导体纳米晶体（量子点，QDs）和金纳米颗粒（AuNP）独特的物理和化学性质。相对简单的测量，廉价的仪器，稳健性和潜在的高通量分析，使提出的分析具有吸引力。***预期结果和效益。我们的纳米工具用于选定蛋白质和功能酶分析的例子将广泛适用于酶的功能评估，而不仅仅是细胞损伤和适应。例如，培养积极的生活方式，特别是与孩子一起，会带来许多伤害。从长远来看，了解细胞在不同组织中的适应机制将为促进“细胞适应性”提供手段。细胞越健康，适应性和功能完整性就越好，从而防止长期慢性残疾，大大减轻经济负担。这些研究将使本科生、研究生和博士后水平的受训者获得跨越化学、物理和生物的多学科研究经验

Nanoprobes for functional biomarkers in cell adaptation******Why study cell adaptation? The ability of cells to adapt to different injurious stimuli is one of the major forces driving evolution. The mechanisms underlying cell adaptation are not fully understood. Lysosomes, dynamic cellular organelles which were considered for a long time only "garbage incinerators" have been recently proposed as essential players in cellular adaptation. Lysosomal functions are regulated at transcriptional and posttranscriptional levels and these functions respond to extracellular and intracellular cues. Transcription factor EB (TFEB) is a master regulator of lysosomal functions. Its status depends on upstream kinase activities. ***Our working hypothesis is that cell adaptation to traumatic (mechanical) and chemical injury occurs through lysosomal adaptation. Our long-term studies will focus on the mechanisms implicated in lysosomal adaptation and the development of new (simple and sensitive) nanotools which will be widely applicable beyond processes in cell adaptation to insults. ***Our short-term objectives are: 1. To develop nanostructure-based functional assays for enzymes, such as cytoplasmic kinases Erk2 (extracellular regulated kinase) and GSK3 beta (glycogen synthase kinase), which are regulators of TFEB, and for lysosomal proteases (cathepsins). 2. To generate nanoprobes recognizing selected plasma membrane biomarkers and to measure their expression with superior sensitivity compared to currently available tools. 3. To generate "NanoScript" (artificial, nanoparticle-based transcription factor), which is designed to mimic the structure and function of TFEB. We will exploit unique physical and chemical properties of semiconductor nanocrystals (quantum dots, QDs) and gold nanoparticles (AuNP). The relative simplicity of the measurement, the inexpensive instrumentation, robustness and potential for high-throughput assaying, makes the proposed assays attractive.***Anticipated outcomes and benefits. Our examples of nanotools for selected proteins and functional enzymatic assays will be widely applicable for functional assessments of enzymes beyond cell injury and adaptation. For instance, fostering active life style, especially with children, brings about numerous injuries. Understanding the mechanisms of cell adaptation in different tissues will in the long run provide the means of promoting "cell fitness". The fitter the cell the better adaptability and functional integrity will be achieved thereby preventing long-term chronic disabilities and considerably reducing the economic burden. The studies will allow trainees at undergraduate, graduate and post-doctoral level to gain experience in multidisciplinary research bridging chemistry, physics and biology.**