Global Cellular Proteo-Lipidomic Profiling of LSD Mutants (nMOST)

This repository contains R scripts, ImageJ macros, CellProfiler pipelines, and supporting data files used in the publication:

Global cellular proteo-lipidomic profiling of diverse lysosomal storage disease mutants using nMOST
Science Advances (doi: 10.1126/sciadv.adu5787)

📄 Science Advances, 2025
📄 bioRxiv Preprint
📋 Protocols.io Collection

Abstract

Lysosomal storage diseases (LSDs) encompass ~50 monogenic disorders characterized by lysosomal accumulation. We developed a nanoflow-based multiomic single-shot technology (nMOST) to quantify proteomes and lipidomes from >24 LSD mutants in HeLa cells. Cross-correlation of lipid–protein data revealed autophagic trafficking defects, iron–lipid metabolism disruptions, and mitochondrial dysfunction—particularly in cholesterol-accumulating NPC1/2 mutants. These defects were partially rescued via transferrin-mediated iron delivery. This study demonstrates the power of nMOST for high-resolution phenotyping across LSDs.

Contents

• CellProfiler pipelines:
  • sf3d_4KO-FilipinEval.cppipe
  • sf5ab_Filipin_FTH1_DNAsyp555_HA_eval_v1.cppipe
• ImageJ macros:
  • f6c_SR_Cristae_v1.ijm
  • sf3c_LC3-Lyso_Count.ijm
  • sf6c_MTDR-TMRE_RatioMIP_v2.ijm

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