Nanoprobes to reveal signature biomarkers

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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.**