Phenotypic screening reveals coordinated multi-organelle remodeling driven by impaired mitochondrial surveillance

Mitochondria are essential organelles involved in energy production and multiple cellular processes, relying on dynamic quality control mechanisms to maintain cellular homeostasis. However, the extent to which mitochondrial surveillance gene disruptions affect the morphology of mitochondria and other organelles remains unclear. We performed a high-content phenotypic screening of human HeLa cells treated with siRNA knockdowns targeting 103 mitochondrial surveillance genes, and assessed mitochondrial mass, membrane potential, and motility, as well as morphological changes across nine cytoplasmic compartments. We discovered a coordinated multi-organelle remodeling linked to mitochondrial network alterations, with several Parkinson’s disease-associated genes among the top modulators. Validation of PINK1 , NIPSNAP1 , HTRA2 , and MICOS13 knockdowns further revealed distinct mechanisms influencing mitochondrial and endomembrane system homeostasis. Our results provide a comprehensive view of organelle interplay following mitochondrial surveillance perturbations and offer a novel framework for investigating molecular pathways underlying human diseases.