TY - UNPB
T1 - Tagless LysoIP method for molecular profiling of lysosomal content in clinical samples
AU - Saarela, Daniel
AU - Lis, Paweł
AU - Gomes, Sara
AU - Nirujogi, Raja
AU - Dong, Wentao
AU - Rawat, Eshaan S.
AU - Glendinning, Sophie
AU - Zeneviciute, Karolina
AU - Bagnoli, Enrico
AU - Fasimoye, Rotimi
AU - Lin, Cindy
AU - Nyame, Kwamina
AU - Boros, Fanni A.
AU - Zunke, Friederike
AU - Lamoliatte, Fred
AU - Elshani, Sadik
AU - Jaconelli, Matthew
AU - Jans, Judith J. M. Jans
AU - Huisman, Margriet A.
AU - Posern, Christian
AU - Westermann, Lena M. Westermann
AU - Schulz, Angela
AU - van Hasselt, Peter M.
AU - Alessi, Dario
AU - Abu-Remaileh, Monther
AU - Sammler, Esther
N1 - The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
PY - 2024/5/27
Y1 - 2024/5/27
N2 - 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 allowed major discoveries in the field, however studying lysosomal dysfunction in human patients remains challenging. Here, we report the development of the “tagless LysoIP method” to enable 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 (iPSCs) derived neurons). Isolated lysosomes are intact and suitable for subsequent multimodal omics analyses. To validate our approach, we employed the tagless LysoIP to enrich lysosomes from peripheral blood mononuclear cells (PBMCs) derived from fresh blood from patients with CLN3 disease, a neurodegenerative LSD. Metabolic profiling of isolated lysosomes showed 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 novel biomarkers and discover human-relevant disease mechanisms.
AB - 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 allowed major discoveries in the field, however studying lysosomal dysfunction in human patients remains challenging. Here, we report the development of the “tagless LysoIP method” to enable 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 (iPSCs) derived neurons). Isolated lysosomes are intact and suitable for subsequent multimodal omics analyses. To validate our approach, we employed the tagless LysoIP to enrich lysosomes from peripheral blood mononuclear cells (PBMCs) derived from fresh blood from patients with CLN3 disease, a neurodegenerative LSD. Metabolic profiling of isolated lysosomes showed 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 novel biomarkers and discover human-relevant disease mechanisms.
U2 - 10.1101/2024.05.17.594681
DO - 10.1101/2024.05.17.594681
M3 - Preprint
BT - Tagless LysoIP method for molecular profiling of lysosomal content in clinical samples
PB - BioRxiv
ER -