Tagless LysoIP for immunoaffinity enrichment of native lysosomes from clinical samples
Lysosomes are implicated in a wide spectrum of human diseases, including monogenic lysosomal storage disorders (LSDs), age-associated neurodegeneration, and cancer. Profiling lysosomal content using tag-based lysosomal immunoprecipitation (LysoTagIP) in cell and animal models has substantially moved the field forward, but studying lysosomal dysfunction in patients remains challenging. Here, we report the development of the 'tagless LysoIP' method, designed to enable the rapid enrichment of lysosomes, via immunoprecipitation, using the endogenous integral lysosomal membrane protein TMEM192, directly from clinical samples and human cell lines (e.g., induced pluripotent stem cell-derived neurons). Isolated lysosomes were intact and suitable for subsequent multimodal omics analyses. To validate our approach, we applied the tagless LysoIP to enrich lysosomes from peripheral blood mononuclear cells derived from fresh blood of healthy donors and patients with CLN3 disease, an autosomal recessive neurodegenerative LSD. Metabolic profiling of isolated lysosomes revealed massive accumulation of glycerophosphodiesters (GPDs) in patients' lysosomes. Interestingly, a patient with a milder phenotype and genotype displayed lower accumulation of lysosomal GPDs, consistent with their potential role as disease biomarkers. Altogether, the tagless LysoIP provides a framework to study native lysosomes from patient samples, identify disease biomarkers, and discover human-relevant disease mechanisms.
Authors
- Fanni Annamária Boros (ORCID: https://orcid.org/0000-0003-2922-4537)
- Raja Sekhar Nirujogi (ORCID: https://orcid.org/0000-0002-3177-8566)
- Enrico Bagnoli (ORCID: https://orcid.org/0000-0002-0664-9981)
- Eshaan S. Rawat (ORCID: https://orcid.org/0000-0002-2595-7605)
- Lena Marie Westermann
- Paweł Lis (ORCID: https://orcid.org/0000-0002-4978-7671)
- Wentao Dong (ORCID: https://orcid.org/0000-0001-5490-4265)
- Kwamina Nyame (ORCID: https://orcid.org/0000-0001-8898-4689)
- Monther Abu-Remaileh (ORCID: https://orcid.org/0000-0003-3772-0477)
- Dario R. Alessi (ORCID: https://orcid.org/0000-0002-2140-9185)
- Rotimi Fasimoye (ORCID: https://orcid.org/0000-0001-8057-0879)
- Sara Gomes (ORCID: https://orcid.org/0000-0002-9086-2303)
- Friederike Zunke (ORCID: https://orcid.org/0000-0002-0408-6388)
- Cindy Lin (ORCID: https://orcid.org/0000-0003-4555-195X)
- Esther Sammler (ORCID: https://orcid.org/0000-0003-3218-7116)
- Peter M. van Hasselt (ORCID: https://orcid.org/0000-0001-9821-8398)
- Frédéric Lamoliatte (ORCID: https://orcid.org/0000-0002-0549-6514)
- Judith Jans (ORCID: https://orcid.org/0000-0003-0960-6263)
- Sadik Elshani
- Daniel Saarela (ORCID: https://orcid.org/0000-0001-8854-7438)
- Christian Posern
- Karolina Zeneviciute (ORCID: https://orcid.org/0000-0002-9628-3783)
- Matthew Jaconelli
- Angela Schulz (ORCID: https://orcid.org/0000-0001-9156-4861)
- S. G. Glendinning (ORCID: https://orcid.org/0009-0001-2969-198X)
- Margriet Huisman
Institutions
- Universität Hamburg (DE)
- University of Dundee (GB)
- Friedrich-Alexander-Universität Erlangen-Nürnberg (DE)
- Neurosciences Institute (US)
- Utrecht University (NL)
- University Medical Center Utrecht (NL)
- Universitätsklinikum Erlangen (DE)
- Stanford Health Care (US)
- University Medical Center Hamburg-Eppendorf (DE)
- MRC Protein Phosphorylation and Ubiquitylation Unit (GB)
- German Centre for Cardiovascular Research (DE)
- Research Network (United States) (US)
- Wilhelmina Children's Hospital (NL)
- Medical Research Council (GB)
Publication Details
- Journal
- Journal of Clinical Investigation
- Published
- 2024-12-26
- DOI
- https://doi.org/10.1172/jci183592
- Citations
- 11
- Primary Topic
- Lysosomal Storage Disorders Research
- Type
- article
- Field-Weighted Citation Impact
- 3.59
Funders
- Michael J. Fox Foundation for Parkinson's Research
- GlaxoSmithKline
- University of Dundee
- Merck KGaA
- Wellcome Trust
- Medical Research Council