Fluorescent Probe Development for Detection of Oxidative Stress-Induced Carbonylation in Cells

There is an urgent need for precise and robust diagnostic tools for monitoring disease progression in biological systems for healthcare and biomedical research. The development of a non-invasive fluorescence probe that is capable of detecting oxidative stress-induced carbonylation in cells is vital to advance our knowledge of oxidative stress-related diseases. Oxidative stress occurs when there is an excessive amount of reactive oxygen species (ROS) in a biological system that can damage crucial components of the cell and increase carbonylation in the cell environment. This chemical process can be associated with several health complications such as cancer, neurodegenerative disorders, and diabetes. The detection of oxidative stress-induced carbonylation with novel fluorophores can therefore be very helpful for monitoring different cell line systems in vitro. In order to introduce fluorophores into the cellular systems, the design of new probes is essential. Probes should have particular chemical and photophysical characteristics to achieve successful imaging: high stability, less toxicity, fast kinetics, good spectral properties such as large Stokes shifts, reasonable quantum yields, drastic changes on absorption and emission spectra. In this work, we focused on synthesizing a few probe systems for targeting carbonyl moieties of biomolecules in living cells using a click reaction to produce a fluorescent product so that we can be able to visualize the carbonylation process in various cancer cell lines. In the near future, the development of non-invasive fluorescent probes that are capable of detecting oxidative stress-induced carbonylation in cells will be pivotal in understanding the mechanisms that govern disease. In addition, we anticipate that it will help immensely improve diagnostics and drug development in biomedical sciences.

Hello, everyone! My name is Amira Anwar and this is my OSCAR URSP Spring 2024 project. My research focuses on the development of fluorescent probes for the detection of oxidative stress-induced carbonylation in cells. I would like to express my sincere gratitude to my mentor Dr. Dilek, for their invaluable guidance and support throughout this research project. I want to extend my thanks to Eva-Maria Rudler for her wonderful support as well. There is a critical need for precise diagnostic tools to accurately assess disease progression in biological systems for healthcare and biomedical research. These tools are important for early disease detection, guiding treatment decisions, and understanding the mechanisms that govern health and disease. Below are some different types of probes. Oxidative stress occurs due to an imbalance between the production of reactive oxygen species and the biological system’s ability to detoxify them. When there is an excessive amount, reactive oxygen species overwhelm the biological system and can cause chaos in important cellular components including lipids, proteins, and nucleic acids. Oxidative stress can further exacerbate this by increasing carbonylation levels within the cell environment potentially triggering apoptosis or programmed cell death. This type of chemical process is linked to various health complications including cancer, metabolic disorders like diabetes, and neurodegenerative disorders like Alzheimer’s or Parkinson’s disease. Here is a diagram of a normal cell undergoing oxidative stress and then resulting in apoptosis. The development of a non-invasive fluorescence probe capable of detecting oxidative stress-induced carbonylation in cells is crucial for advancing our understanding of oxidative stress-related diseases. These probes can provide a means to directly visualize live cells in real-time and offer incredible insights into the molecular mechanisms. This is a picture of a fluorescent probe that is detecting mitochondrial oxidative stress in cells of mice. Using novel fluorophores to detect oxidative stress-induced carbonylation can be highly advantageous for monitoring various cell line systems in vitro. The design of new fluorescent probes is key in order to introduce them into cellular systems. They need very particular chemical and photophysical properties for successful imaging like high stability to ensure their integrity and reliability in harsh cellular environments. It must have low toxicity levels to allow accurate detection while promising the safety of the biological system. Fast kinetics to enable rapid binding and response to target molecules. There also needs to be good spectral properties like significant Stokes shift, favorable quantum yield, and notable alterations in the absorption and emission spectrums. I focused on synthesizing probe systems to target carbonyl groups in biomolecules found in live cells, utilizing click chemistry reactions to produce fluorescent products. This was done to enable the visualization of the carbonylation process across various cell lines. Here is an example of a click chemistry reaction: this click reaction is referred to as the copper-catalyzed alkyne”“azide cycloaddition. The development of non-invasive fluorescent probes capable of detecting OSIC within cells will play a pivotal role in understanding the mechanisms that govern health and disease. I anticipate this development will advance diagnostics and facilitate drug development in biomedical sciences. Looking ahead I plan to continue my research with OSCAR and focus on several key areas to advance in such as cellular fluorescence imaging techniques, applications of cell culture labeling, potentially investigating additional molecules for metal sensing, and computational calculations. Finally, I would like to say thank you to everyone watching and thank you to OSCAR for giving me this opportunity.