New Lysosome Restoration Pathways for Disease and Aging [Aug. 27, 2025] DOJINDO LABORATORIES

Discovery of New Transcriptional Programs Restoring Lysosomes Suggests New Therapeutic Opportunities
Lysosomes are essential organelles for maintaining cellular homeostasis, and their dysfunction is closely associated with a variety of diseases, including neurodegeneration, making it critical to understand how cells restore lysosomal function. Recent work in C. elegans showed that the GATA transcription factor ELT-2 activates a lysosomal surveillance response (LySR), thereby enhancing lysosomal activity and extending lifespan¹. In addition, studies in mouse models of lysosomal storage disorders identified STING, best known as a DNA sensor in innate immunity, as a lysosomal repair sensor that activates TFEB, the master regulator of lysosomal and autophagy genes, uncovering new pathways of lysosomal quality control and suggesting strategies to counter proteotoxicity and neurodegenerative disease².

1. A lysosomal surveillance response to stress extends healthspan (Nature Cell Biology, 2025)

Summary: Researchers found that silencing vacuolar H+-ATPase subunits in the intestine of C. elegans creates a stress that activates the lysosomal surveillance response (LySR), governed by the GATA transcription factor ELT-2, to boost lysosomal activity and clear protein aggregates. This mechanism enhances lysosomal function, extends lifespan, and suggests that activating the newly identified LySR pathway could be a strategy to treat lysosome-related diseases and promote healthy aging.

Highlighted technique: To examine how silencing v-ATPase subunits affects lysosomal function, the authors used two approaches in living C. elegans. They applied a fluorescent probe that labels lysosomes and a pH-sensitive lysosomal reporter to monitor changes in acidification, and these assays demonstrated that LySR activation boosts lysosomal activity.

Related technique Lysosome Detection, Lysosomal Acidic pH Detection

2. STING mediates lysosomal quality control and recovery through its proton channel function and TFEB activation in lysosomal storage disorders (Molecular Cell, 2025)

Summary: STING, long recognized as a cytosolic DNA sensor that initiates immune responses, is now shown to have a distinct role in maintaining lysosomal homeostasis. This study reveals that lysosomal dysfunction activates a STING–TFEB pathway, in which STING’s proton channel function triggers TFEB-driven expression of lysosomal and autophagy genes, thereby facilitating lysosomal repair and highlighting new therapeutic avenues for lysosomal storage disorders and neurodegenerative diseases.

Highlighted technique: To evaluate lysosomal damage and repair, researchers used LLOME to rupture lysosomal membranes. When lysosomes were damaged, cytosolic Galectin-3 bound to the exposed glycans and was visualized as fluorescent puncta with antibodies, allowing the appearance and disappearance of these puncta over time to reveal the repair process.

Related technique Lysosomal Function, Autophagic Flux Assay

All Related Techniques (click to open/close)

Target	Kit & Probes
Lysosomal function	Lysosomal Acidic pH Detection Kit -Green/Red and Green/Deep Red
First-time autophagy research	Autophagic Flux Assay Kit
Mitophagy detection	Mitophagy Detection Kit
Endocytosis Detection	ECGreen-Endocytosis Detection
Cellular senescence detection	SPiDER-βGal for live-cell imaging or flow cytometry / microplate reader / tissue samples Blue cellular senescence detection dye for fixed cells, SPiDER Blue
Mitochondrial superoxide detection	MitoBright ROS Deep Red - Mitochondrial Superoxide Detection
Total ROS detection	Highly sensitive DCFH-DA or Photo-oxidation Resistant DCFH-DA
Apoptosis detection in multiple samples	Annexin V Apoptosis Plate Assay Kit
Cell proliferation/ cytotoxicity assay	Cell Counting Kit-8 and Cytotoxicity LDH Assay Kit-WST

Application Note (click to open/close)
> Accurate Measurement for Lysosomal pH changes

With existing reagents, it was difficult to determine whether lysosomal mass or their function (pH) fluctuated because the discussion was based on changes in the fluorescence brightness of a single dye. This kit contains pHLys Green, which is highly specific to lysosomes and shows pH-dependent changes in fluorescence, and pH-resistant LysoPrime Deep Red. Using these two dyes, lysosomal pH and volume of the same sample can be measured for a detailed analysis of lysosomal function.
Existing lysosomal pH detection reagents have issues with dye localization, pH sensitivity, and retention. pHLys Green is a dye that solves these issues. The improved dye retention and localization enable detection of normal lysosomes, and the improved pH sensitivity enables detection of slight pH changes.
1. High sensitive pH detection Comparison of pH response of cells treated with low concentrations of lysosomal acidification inhibitor Bafilomycin A1	2. High specificity for lysosomes Comparison of specificity for lysosomes using lysosomal marker protein LAMP1-GFP expressing cells	3. High retention in lysosomes Comparison of intracellular retention

Product in Use: - Lysosomal Acidic pH Detection Kit-Green/Deep Red Related Product: - Lysosomal Acidic pH Detection Kit-Green/Red - LysoPrime Deep Red - High Specificity and pH Resistance - pHLys Red - Lysosomal Acidic pH Detection