Unpuzzle - Unravelling molecular motifs from dietary fibers responsible for interaction with recognition receptors in human cells

Dietary fibres (DF) can induce beneficial health effects by interacting with receptors at the surface of intestinal epithelial cells and immune system cells. Although the knowledge of the structure-function relationship between DF and their biological effects in human cells is highly desirable for successful translational applications, the molecular motifs within the often-complex DF structure responsible for binding to human cell receptors remain poorly understood. In Unpuzzle, we propose a new methodological framework to explore the interaction between DF and Toll-like receptors (TLR) in human cells. We will reduce the structural complexity of selected DF by generating oligomers through non-enzymatic hydrolysis. The produced oligomers (structural motifs) will be further fractionated and evaluated for interaction with TLR using reporter cells. The structural motifs that can interact with TLR will be characterized through high-resolution tandem mass spectrometry techniques and explored for biological effects in human macrophages through metabolomics-based approaches and cytokine array. Finally, network analysis will link structural data of molecular motifs with effects in human cells. Results will clarify how DF interacts with human cells and the potential consequences of such interaction on our gut health. Findings will also provide information for the identification and tailored production of DF with desired biological effects.