Collection Citation: mai.nguyen , Jack Collier, Olesia Ignatenko, Sidong Huang, Michel Desjardins, heidi.mcbride 2025. Protocols for "MAPL regulates gasdermin-mediated release of mtDNA from lysosomes to drive pyroptotic cell death". protocols.io https://dx.doi.org/10.17504/protocols.io.8epv5x266g1b/v2Version created by Lilia Rodriguez
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this collection and it's working
Created: September 19, 2025
Last Modified: September 21, 2025
Collection Integer ID: 227743
Keywords: ASAPCRN, pyroptosis, mitochondrial dna, mitochondria, inflammatory pathway, mapl, lysosome, mtdna, immune signalling, mitochondrial control of cell death, pyroptosis through an inflammatory pathway, induced pyroptosi, pyroptosi, mitochondrial control, mitochondrial dna trafficking, lysosome pathway at the nexus, lysosome pathway, outer mitochondrial membrane, mitochondria, mapl overexpression, mechanisms from cancer, placing mapl, inflammatory cell death, depletion of mapl, inflammatory pathway, protein ligase, mapl, involving mitochondria, lysosome, multiple parkinson, pyroptotic cell, release of mtdna, cell death, inflammatory cell death in primary macrophage, mtdna, neurodegeneration, cell survival, wide functional genetic screening
Funders Acknowledgements:
Aligning Science Across Parkinson’s (ASAP)
Grant ID: ASAP 000525
Abstract
Mitochondrial control of cell death is of central importance to disease mechanisms from cancer to neurodegeneration. Mitochondrial anchored protein ligase (MAPL) is an outer mitochondrial membrane small ubiquitin-like modifier ligase that is a key determinant of cell survival, yet how MAPL controls the fate of this process remains unclear. Combining genome-wide functional genetic screening and cell biological approaches, we found that MAPL induces pyroptosis through an inflammatory pathway involving mitochondria and lysosomes. MAPL overexpression promotes mitochondrial DNA trafficking in mitochondrial-derived vesicles to lysosomes, which are permeabilized in a process requiring gasdermin pores. This triggers the release of mtDNA into the cytosol, activating the DNA sensor cGAS, required for cell death. Additionally, multiple Parkinson’s disease-related genes, including VPS35 and LRRK2, also regulate MAPL-induced pyroptosis. Notably, depletion of MAPL, LRRK2 or VPS35 inhibited inflammatory cell death in primary macrophages, placing MAPL and the mitochondria–lysosome pathway at the nexus of immune signalling and cell death.
Files
Culturing of cell lines.docx
Generation of Bone Marrow derived macrophages.docx
Transfection and generation of stable cell lines.docx
