Lysosome Dynamics Shape Ferroptosis Sensitivity [Nov. 17, 2025] DOJINDO LABORATORIES

Ferroptosis is an iron-dependent form of cell death shaped by lipid peroxidation as well as intracellular factors such as iron localization and lysosomal acidity. Lysosomes function as central sites for iron storage and mobilization, and emerging evidence suggests that iron activation and lipid peroxidation within lysosomes contribute to ferroptosis induction. Recent studies show that BDH2-dependent iron transfer between mitochondria and lysosomes in melanoma prevents Fe(II) sequestration, whose loss increases ferroptosis vulnerability. Another study demonstrates that lysosomal alkalinization during senescence causes abnormal Fe(II) retention, forming a major basis of ferroptosis resistance. These findings highlight lysosome-centered iron regulation as a key determinant of ferroptosis sensitivity.

1. BDH2-driven lysosome-to-mitochondria iron transfer shapes ferroptosis vulnerability of the melanoma cell states (Nature Metabolism, 2025)
Summary: Melanoma cells exist in a melanocytic (MEL) state and a drug-tolerant mesenchymal-like (MES) state. BDH2 mediates Fe²⁺ transfer at mitochondria–lysosome contacts in MEL cells and maintains lysosomal and mitochondrial homeostasis, and its loss in MES cells causes lysosomal iron sequestration and ferroptosis sensitivity, which can be reversed by restoring BDH2 or adding 2,5-DHBA, also increasing metastatic ability.

Highlighted technique: In this study, the authors aimed to determine how Fe²⁺ is distributed among intracellular organelles by performing live-cell imaging–based visualization and quantification of iron. They combined fluorescent probes to visualize Fe(II) within lysosomal regions and compare iron levels between MEL and MES cells, and further quantified mitochondrial iron using a mitochondria-specific probe to analyze iron compartmentalization across organelles.

Related technique Intracellular Fe²⁺ Detection, Mitochondrial Fe²⁺ Detection

2. Senescence-associated lysosomal dysfunction impairs cystine deprivation-induced lipid peroxidation and ferroptosis (Nature Communications, 2025)
Summary: This study showed that senescence impairs lysosomal acidification, leading to the abnormal retention of Fe²⁺ within lysosomes and thereby inhibiting cystine-deprivation-induced lipid peroxidation and ferroptosis. The authors also demonstrated that restoring lysosomal acidity through V-ATPase activation increases ferroptosis sensitivity in senescent cells.

Highlighted technique: This study aimed to clarify how lysosomal dysfunction in senescent cells alters intracellular Fe²⁺ distribution and ferroptosis sensitivity. To address this question, the authors used live-cell imaging with fluorescent probes to detect lysosomal Fe²⁺ and measure lysosomal pH, allowing them to evaluate iron sequestration and changes in intracellular iron distribution.

Related technique Lysosomal Fe²⁺ Detection, Lysosome function Analysis

Application Note
> When Lysosomes Go Neutral: Iron Loss Unveiled

We investigated the transition of cellular metabolisms in A549 cells treated with erastin, a known ferroptosis inducer. Our results revealed the following.

Results
- The inhibition of cystine uptake by erastin led to a depletion of cysteine, which in turn increased the compensatory uptake of other amino acids.
- Glucose uptake, which typically promotes ferroptosis*, was found to decrease upon erastin treatment, suggesting a potential cellular self-defense mechanism.
- The depletion of cysteine resulted in a decrease in glutathione levels and an increase in Fe²⁺, ROS, and lipid peroxides, all of which are recognized markers of ferroptosis.

Cell Line: A549
Incubation Conditions: 100 μmol/l Erastin/MEM, 37℃, 3h
*Reference: Xinxin Song, et al., Cell Reports, (2021)

Products in Use
① Amino Acid Uptake: Amino Acid Uptake Assay Kit
② Glucose Uptake: Glucose Uptake Assay Kit-Green
③ Cystine Uptake: Cystine Uptake Assay Kit
④ Intracellular glutathione: GSSG/GSH Quantification Kit
⑤ Intracellular labile Fe: FerroOrange
⑥ Intracellular total ROS: ROS Assay Kit -Highly Sensitive DCFH-DA-
⑦ Lipid Peroxides: Liperfluo

Ferroptosis Indicators

Target	Kit & Probes
Lysosomal ferrous ion (Fe²⁺) detection	Lyso-FerroRed
Ferroptosis Indicator: ferrous ion (Fe²⁺)	FerroOrange(intracellular), Mito-FerroGreen(mitochondria)
Ferroptosis Indicator: lipid peroxidation	Liperfluo(intracellular), MitoPeDPP(mitochondria)
Lipid Peroxidation Assay	Lipid Peroxidation Probe -BDP 581/591 C11-
Malondialdehyde Detection	MDA Assay Kit
Mitochondrial superoxide detection	MitoBright ROS Deep Red - Mitochondrial Superoxide Detection
Total ROS detection	Highly sensitive DCFH-DA or Photo-oxidation Resistant DCFH-DA
Glutathione Quantification	GSSG/GSH Quantification Kit
Cystine Uptake detection	Cystine Uptake Assay Kit