Experimental Confirmation of Progerin–Small Peptide Interaction Using NanoLuc Binary Technology Assay in Human Cell Lines to Validate in Silico–Designed Progerin Interactors

Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare genetic disorder characterized by accelerated aging, primarily caused by the accumulation of a mutant form of lamin A known as progerin. One of the hallmarks of HGPS is increased oxidative stress, often associated with elevated levels of reactive oxygen species (ROS). In this study, we employed flow cytometry to quantitatively assess ROS levels in three different human fibroblast cell lines: MRC5 cells, MRC5 cells expressing progerin, and primary fibroblasts derived from HGPS patients. ROS detection was performed using the Total ROS Assay Kit (Thermo Fisher Scientific), which enables the measurement of total intracellular ROS with high sensitivity. This approach allowed us to compare the oxidative stress profiles among the different cell types and investigate the contribution of progerin expression to cellular redox imbalance. The measurements we took revealed a decrease in ROS levels following treatment, supporting the effectiveness of our therapeutic approach in counteracting oxidative stress associated with Progeria.