Structural Dynamics of the Iron-Sulfur Cluster Assembly Machinery in Patient-Derived FRDA Cells Using Super-Resolution Microscopy

Dr. Javier Santos’s project will investigate how the balance between frataxin (FXN) and its partner protein FDX2 influences the organization, dynamics, and function of the iron-sulfur (Fe-S) cluster assembly machinery in mitochondria. Using control and FA patient-derived human cellular models, the team will precisely manipulate the FXN/FDX2 ratio to mimic disease conditions or rescue cellular function. To study these effects at high resolution, they will employ advanced imaging techniques, including the Proximity Ligation Assay and DNA-PAINT super-resolution microscopy. Nanobodies specific to FXN, NFS1, and ISCU2, combined with anti-FDX2 monoclonal antibodies, will allow multiplexed measurements of protein interactions and supercomplex dynamics. By mapping how changes in the Fe-S cluster machinery affect mitochondrial structure and metabolism within cells, this research aims to provide the first detailed picture of these processes in FA. The insights gained could help identify new therapeutic strategies to restore mitochondrial function in affected tissues.