O-GlcNAcylation is essential for therapeutic mitochondrial transplantation

• Published in: Communications medicine 2023-11, Vol.3 (1), p.169-12, Article 169
• Main Authors: Park, Ji Hyun, Tanaka, Masayoshi, Nakano, Takafumi, Licastro, Ester, Nakamura, Yoshihiko, Li, Wenlu, Esposito, Elga, Mandeville, Emiri T., Chou, Sherry Hsiang-Yi, Ning, MingMing, Lo, Eng H., Hayakawa, Kazuhide
• Format: Article
• Language: English
• Subjects: 101/58; 13; 13/1; 13/51; 13/89; 38; 38/1; 42; 42/70; 45; 59; 631/337/458; 631/80/642/333; 692/699/375/534; Experiments; Glucose; Ischemia; Laboratory animals; Medicine; Medicine & Public Health; Metabolism; Mitochondria; Patients; Sodium; Temperature; Transplants & implants
• ISSN: 2730-664X; 2730-664X
• DOI: 10.1038/s43856-023-00402-w

Background Transplantation of mitochondria is increasingly explored as a novel therapy in central nervous system (CNS) injury and disease. However, there are limitations in safety and efficacy because mitochondria are vulnerable in extracellular environments and damaged mitochondria can induce unfavorable danger signals. Methods Mitochondrial O-GlcNAc-modification was amplified by recombinant O-GlcNAc transferase (OGT) and UDP-GlcNAc. O-GlcNAcylated mitochondrial proteins were identified by mass spectrometry and the antiglycation ability of O-GlcNAcylated DJ1 was determined by loss-of-function via mutagenesis. Therapeutic efficacy of O-GlcNAcylated mitochondria was assessed in a mouse model of transient focal cerebral ischemia-reperfusion. To explore translational potential, we evaluated O-GlcNAcylated DJ1 in CSF collected from patients with subarachnoid hemorrhagic stroke (SAH). Results We show that isolated mitochondria are susceptible to advanced glycation end product (AGE) modification, and these glycated mitochondria induce the receptor for advanced glycation end product (RAGE)-mediated autophagy and oxidative stress when transferred into neurons. However, modifying mitochondria with O-GlcNAcylation counteracts glycation, diminishes RAGE-mediated effects, and improves viability of mitochondria recipient neurons. In a mouse model of stroke, treatment with extracellular mitochondria modified by O-GlcNAcylation reduces neuronal injury and improves neurologic deficits. In cerebrospinal fluid (CSF) samples from SAH patients, levels of O-GlcNAcylation in extracellular mitochondria correlate with better clinical outcomes. Conclusions These findings suggest that AGE-modification in extracellular mitochondria may induce danger signals, but O-GlcNAcylation can prevent glycation and improve the therapeutic efficacy of transplanted mitochondria in the CNS. Plain language summary Mitochondria are the part of a cell that generate most of its energy to perform its functions. In injury or disease, mitochondrial function can become disrupted. Transplantation of healthy mitochondria is being explored as a potential therapy to replace damaged mitochondria and restore normal cellular function. However, this approach is difficult to perform because mitochondria are not able to maintain their healthy state outside of cells. Here, we show that one of the reasons for this is due to a molecular process called advanced glycation end product modification. We show that simple mo