FTO-targeted siRNA delivery by MSC-derived exosomes synergistically alleviates dopaminergic neuronal death in Parkinson's disease via m6A-dependent regulation of ATM mRNA.

BACKGROUND: Parkinson's disease (PD), characterized by the progressive loss of dopaminergic neurons in the substantia nigra and striatum of brain, seriously threatens human health, and is still lack of effective treatment. Dysregulation of N6-methyladenosine (m6A) modification has been implicated in PD pathogenesis. However, how m6A modification regulates dopaminergic neuronal death in PD remains elusive. Mesenchymal stem cell-derived exosomes (MSC-Exo) have been shown to be effective for treating central nervous disorders. We thus propose that the m6A demethylase FTO-targeted siRNAs (si-FTO) may be encapsulated in MSC-Exo (Exo-siFTO) as a synergistic therapy against dopaminergic neuronal death in PD. METHODS: In this study, the effect of m6A demethylase FTO on dopaminergic neuronal death was evaluated both in vivo and in vitro using a MPTP-treated mice model and a MPP + -induced MN9D cellular model, respectively. The mechanism through which FTO influences dopaminergic neuronal death in PD was investigated with qRT-PCR, western blot, immumohistochemical staining, immunofluorescent staining and flow cytometry. The therapeutic roles of MSC-Exo containing si-FTO were examined in PD models in vivo and in vitro. RESULTS: The total m6A level was significantly decreased and FTO expression was increased in PD models in vivo and in vitro. FTO was found to promote the expression of cellular death-related factor ataxia telangiectasia mutated (ATM) via m6A-dependent stabilization of ATM mRNA in dopaminergic neurons. Knockdown of FTO by si-FTO concomitantly suppressed upregulation of α-Synuclein (α-Syn) and downregulation of tyrosine hydroxylase (TH), and alleviated neuronal death in PD models. Moreover, MSC-Exo were utilized to successfully deliver si-FTO to the striatum of animal brain, resulting in the significant suppression of α-Syn expression and dopaminergic neuronal death, and recovery of TH expression in the brain of PD mice. CONCLUSIONS: MSC-Exo delivery of si-FTO synergistically alleviates dopaminergic neuronal death in PD via m6A-dependent regulation of ATM mRNA.

DANTE-SPACE: a new technical tool for DVT on 1.5T MRI.

The aim of the study was to compare CE-MRV with DANTE-SPACE on a 1.5T MRI system for the diagnosis of DVT. The patients were diagnosed with deep venous thrombosis of the lower extremities based on swelling, pain, and superficial varicose veins of the lower extremities. MRI examination confirmed the diagnosis. DANTE-SPACE images were obtained before the conventional contrast-enhanced MRV, which uses gadolinium. The scanning field started from the end of the inferior vena cava to the end of the ankle, divided into five observation segments, namely, the common iliac vein, external iliac vein, femoral vein, popliteal vein, and calf vein. The DANTE-SPACE and CE-MRV results were used for a consistency analysis. For the DANTE-SPACE and CE-MRV images, the signal intensity ratios of the thrombus/cavity and thrombus/muscle were calculated, and the ratio difference was compared using the paired t test. Twenty-six patients completed the examination; one of the patients underwent a right lower limb amputation, yielding a total of 255 lower limb vascular segments. The analysis of the DANTE-SPACE images showed that there were 14 iliac vein thromboses, 18 external iliac vein thromboses, 23 femoral vein thrombi, 21 popliteal vein thromboses, and 18 calf vein thromboses; these findings were consistent with the diagnostic results of CE-MRV. The ratio of the thrombus/cavity signal intensity measured in the DANTE-SPACE and CE-MRV images were as follows: 20.51 ± 12.96 vs. 0.51 ± 0.46; P < 0.05, n = 51; the difference was statistically significant. The ratio of the thrombus/muscle signal intensity measured on the DANTE-SPACE and CE-MRV images were as follows: 1.74 ± 0.57 vs. 0.99 ± 0.53; P < 0.05, n = 51; the difference was statistically significant. Compared with CE-MRV, DANTE-SPACE showed no significant difference in the ability to detect deep venous thrombosis of the lower extremities. DANTE-SPACE did not use contrast-enhancing agents and showed no evidence of inflammatory enhancement, and the display effect of small diameter veins was slightly poor. However, deep venous thrombosis of the lower extremities presents different levels of high signal in the DANTE-SPACE images, making it easy to identify and diagnose. It can also indicate the different components and age of the thrombus and help with the selection of a more accurate clinical treatment plan. MRI DANTE-SPACE is the preferred imaging modality for patients with deep venous thrombosis who are unable or unwilling to use gadolinium contrast agents.