Roles of LTF and HAMP in Neuronal Ferroptosis in Alzheimer's Disease Cell Model

Abstract

Dysregulation of iron homeostasis and ferroptosis is increasingly implicated in Alzheimer's disease (AD) pathology. Lactotransferrin (LTF) and hepcidin (HAMP) are key regulators of neuronal iron metabolism, but their role in AD-related ferroptosis remains unclear. Bioinformatics analysis of hippocampal RNA-sequencing data from AD patients and healthy controls identified differentially expressed genes (DEGs) associated with iron metabolism. SH-SY5Y cells were treated with Aβ oligomers to establish AD cell models and further subjected to siRNA-mediated knockdown of LTF or HAMP. Ferroptosis was evaluated using Fe²⁺ accumulation, lipid reactive oxygen species (ROS), mitochondrial morphology, MDA/GSH levels, and the expression of core ferroptosis regulators (GPX4, SLC7A11, ACSL4). Cell viability, LDH release, and apoptosis were also assessed. Bioinformatics analysis revealed LTF and HAMP as key DEGs related to iron homeostasis, potentially linked to ferroptosis in AD. In vitro, Aβ treatment decreased LTF and HAMP expression, induced Fe²⁺ accumulation, oxidative stress, mitochondrial damage, and ferroptotic cell death. Knockdown of LTF or HAMP further exacerbated these effects, while treatment with the ferroptosis inhibitor Fer-1 partially restored cellular homeostasis and viability. The data suggest a regulatory role of LTF and HAMP in neuronal ferroptosis. Notably, hyper-expression of LTF and HAMP observed in post-mortem AD hippocampal samples likely reflects compensatory negative feedback mechanisms in late-stage disease. LTF and HAMP act as protective modulators against neuronal ferroptosis in AD. Their early downregulation promotes iron overload and oxidative stress, whereas compensatory upregulation in vivo delays ferroptosis. Targeting LTF and HAMP-mediated pathways may provide novel therapeutic strategies for mitigating AD progression.