Shedding light on cellular function: Combining kinetic, mechanistic modelling approaches with label free microspectroscopic imaging

Cellular physiology involves complex and dynamic processes that are not well represented by static models based on limited biomarkers. CYTOMECHSPEC introduces an innovative approach by combining several advanced techniques:

- Raman microspectroscopic subcellular analysis: A label-free spectroscopic method to study cells and their function.

- Advanced chemometric algorithms: Used for analysing complex data and particular the signatures of how the cells are changing with time.

- Kinetic, mechanistic modeling: Helps to describe and visualise cellular function.

- Metabolic Pathway Analysis: Provides a comprehensive view of cellular events and response pathways.

This project aims to validate this combined approach as a routine, label-free bio-analytical method. It seeks to enhance traditional labeled and omics approaches for fundamental research and establish a High Content Spectroscopic Analysis protocol for in-vitro cellular analysis.

The improved understanding of cellular function at a metabolic level is relevant for understanding disease mechanisms, drug toxicity, and has potential applications in preclinical pharmacological development, toxicology screening, and personalized medicine.

CYTOMECHSPEC is based on established expertise at TU-Dublin in spectroscopic techniques and numerical modeling of cellular pathways, with support from collaborations in cellular biology (TU-Dublin) and biotechnology (DCU).

Although much is known about how human cells function, current understanding is largely based on a static picture of networks of organelles, and biological molecules (e.g. proteins, lipids and DNA). The mechanisms by which they interact and evolve with time is less well explored and understood. This
project proposes a combination of established and emerging, complementary analytical techniques to better describe cellular responses in terms of dynamic pathways. Examples of model cellular metabolic pathways are chosen to demonstrate the validity of the approach, and the benefit of the approaches to
basic biological research, with potential applications in pharmacology and healthcare.

The project has established open source numerical models for commercial assays of glycolysis processes, and extended their applicability to glutaminolysis. It has spectroscopically profiled the influence of the glycolysis and glutaminolysis processes on the extracellular environment.