Global cellular proteo-lipidomic profiling of diverse lysosomal storage disease mutants using nMOST
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.
Authors
- Florian Wilfling (ORCID: https://orcid.org/0000-0002-6559-7261)
- Benton J. Anderson (ORCID: https://orcid.org/0000-0001-6156-342X)
- Katherine A. Overmyer (ORCID: https://orcid.org/0000-0002-1929-1229)
- Brenda Arlene Schulman (ORCID: https://orcid.org/0000-0002-3083-1126)
- João A. Paulo (ORCID: https://orcid.org/0000-0002-4291-413X)
- Johann Brenner (ORCID: https://orcid.org/0000-0003-1306-3564)
- Ian R. Smith (ORCID: https://orcid.org/0000-0002-4966-9333)
- Sharan Swarup (ORCID: https://orcid.org/0000-0003-0226-5582)
- Anna Bieber (ORCID: https://orcid.org/0000-0001-5014-6620)
- Nicole M. Nightingale (ORCID: https://orcid.org/0009-0004-6917-2406)
- Cristina Capitanio (ORCID: https://orcid.org/0000-0002-5297-9156)
- J. Wade Harper (ORCID: https://orcid.org/0000-0002-6944-7236)
- Yizhi Jiang (ORCID: https://orcid.org/0000-0001-9016-1104)
- Felix Kraus (ORCID: https://orcid.org/0000-0002-3757-541X)
- Annie Jen (ORCID: https://orcid.org/0000-0001-5569-8387)
- Steven P. Gygi (ORCID: https://orcid.org/0000-0001-7626-0034)
- Jürgen M. Plitzko (ORCID: https://orcid.org/0000-0002-6402-8315)
- Joshua J. Coon (ORCID: https://orcid.org/0000-0002-0004-8253)
- Yuchen He (ORCID: https://orcid.org/0000-0002-6096-4915)
- Chan Lee (ORCID: https://orcid.org/0000-0002-5512-8146)
Institutions
- University of Wisconsin–Madison (US)
- Harvard University (US)
- Morgridge Institute for Research (US)
- Boston VA Research Institute (US)
- Max Planck Institute of Biophysics (DE)
- Max Planck Institute of Psychiatry (DE)
- Max Planck Institute of Biochemistry (DE)
- Research Network (United States) (US)
Publication Details
- Journal
- Science Advances
- Published
- 2025-01-22
- DOI
- https://doi.org/10.1126/sciadv.adu5787
- Citations
- 3
- Primary Topic
- Mosquito-borne diseases and control
- Type
- article
- Field-Weighted Citation Impact
- 5.64