Global cellular proteo-lipidomic profiling of diverse lysosomal storage disease mutants using nMOST.

Kraus, Felix; He, Yuchen; Swarup, Sharan; Overmyer, Katherine A; Jiang, Yizhi; Brenner, Johann; Capitanio, Cristina; Bieber, Anna et al. · Sci Adv · 2025

basic_science · Level V

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Abstract

Lysosomal storage diseases (LSDs) comprise ~50 monogenic disorders marked by the buildup of cellular material in lysosomes, yet systematic global molecular phenotyping of proteins and lipids is lacking. We present a nanoflow-based multiomic single-shot technology (nMOST) workflow that quantifies HeLa cell proteomes and lipidomes from over two dozen LSD mutants. Global cross-correlation analysis between lipids and proteins identified autophagy defects, notably the accumulation of ferritinophagy substrates and receptors, especially in <i>NPC1</i><sup>-/-</sup> and <i>NPC2</i><sup>-/-</sup> mutants, where lysosomes accumulate cholesterol. Autophagic and endocytic cargo delivery failures correlated with elevated lysophosphatidylcholine species and multilamellar structures visualized by cryo-electron tomography. Loss of mitochondrial cristae, MICOS complex components, and OXPHOS components rich in iron-sulfur cluster proteins in <i>NPC2</i><sup>-/-</sup> cells was largely alleviated when iron was provided through the transferrin system. This study reveals how lysosomal dysfunction affects mitochondrial homeostasis and underscores nMOST as a valuable discovery tool for identifying molecular phenotypes across LSDs.

Medical subject headings