Autophagy Toolbox
Autophagy (“self-eating”) is an evolutionarily conserved survival mechanism in living organisms during which intracellular components (‘cargo’) are identified and delivered to lysosomes for degradation. Basal autophagy plays a vital role in maintaining cellular homeostasis; dysregulation of autophagy has been found to associate with diseases ranging from neurodegeneration, early stage of cancers, cardiovascular disease to infectious disease, with different stages of autophagy being impaired. Research efforts have identified multiple molecular targets to rectify autophagy with the promise for therapeutic intervention. As autophagy is a multi-step process, it is important to identify and assess which stage of autophagy is affected by a modulator, to quantify the extent of the regulation and to draw a clear mechanistic picture. However, tools that allow real-time monitoring of the dynamics of autophagy, especially with quantitative readout, are still scarce and highly desirable. Our lab endeavours to develop next generation autophagy probes based on small molecules that are highly specific to autophagy, which can be used in live cells without the need of permeability or genetic modifications. These probes will be used to track the dynamic process of autophagy and measure its activity in cells and in vivo, with applications ranging from fundamental mechanistic studies, drug evaluation and screening, to disease diagnostics.
Proteostasis Sensors
The proper folding of polypeptide chains into their native structure is crucial for protein functionality. In eukaryotic systems, a network of approximately 800 proteins, including molecular chaperones, protein disaggregases, and proteasomes, maintains proteostasis, ensuring the foldedness of the proteome. This highly conserved quality control system plays a pivotal role in all organisms. Disruption of proteostasis, caused by external stress or aberrant proteins, results in accumulation of improperly folded or aggregated proteins, leading to increased susceptibility to damage—a common characteristic of aging and diseases like cancer, diabetes, and neurodegeneration.
Our laboratory has successfully developed a series of fluorogenic probes for quantifying and imaging intracellular unfolded protein loads, providing a measure of proteostasis capacity in cells. Furthermore, we have created fluorescent probes that target amyloid protein aggregates in vitro, enabling the detection of prefibrillar species during early stages of protein aggregation. These probes offer valuable insights into how cells maintain functional proteomes and respond to stress, holding promise for biomarker discovery in disease diagnosis and the development of effective treatment strategies.
Bioanalytical Tools
Leveraging our expertise in designing and synthesizing fluorescent probes, we are dedicated to developing cutting-edge bioanalytical tools. Our research focuses on creating novel fluorescent probes for the detection of urinary proteins, enabling effective monitoring of chronic kidney disease. Additionally, we aim to advance enzymatic activity detection in urine, cells, and soil samples through the development of innovative probes.
In order to analyse the intracellular environment, we have successfully developed a range of photostable organelle imaging agents. These agents facilitate precise tracking of cellular components such as mitochondria, lysosomes, endoplasmic reticulum (ER), nucleus, and the plasma membrane. Moreover, we have combined these agents with advanced fluorescence techniques to analyse critical intracellular parameters, including pH, viscosity, macromolecular crowding, and polarity. Our approaches allow for in-situ quantification of diverse physical parameters within live cells, enabling us to investigate their pivotal roles in driving fundamental biomolecular interactions, such as liquid-liquid phase separation and protein aggregation.