Project #4- Therapeutic approaches to CoQ10 deficiencies.

项目摘要 辅酶Q10(CoQ10)是由苯醌环和辅酶Q10组成的脂多糖分子。 疏水十碳烯基尾巴。在线粒体呼吸链中，辅酶Q10运输电子 从络合物I和II到络合物III。它也是一种抗氧化剂，是嘧啶的辅助因子 生物合成，以及细胞凋亡的调节器。人类辅酶Q10缺乏症是一种遗传和 临床异质性疾病。对治疗的反应是不同的。我们对培养的研究 携带PDSS2、COQ2、ADCK3和COQ9突变的患者的成纤维细胞 辅酶Q10水平与氧化应激、15%和60%残留量的相关性 辅酶Q10与氧化应激无关，而30-50%的残留辅酶Q10与氧化应激有关 在氧化应激下，线粒体超极化，随后是去极化和细胞死亡。 在原代辅酶Q10缺陷细胞系中，辅酶Q10缺陷成纤维细胞孵育1周 使用5 mM辅酶Q10(但不是短尾泛醌类似物)改善了生物能量学，表明 辅酶Q10的药代动力学限制和剂量可能限制辅酶Q10缺陷患者的疗效。 根据我们的研究，我们假设：1)基因异质性对 辅酶Q10缺乏症的表型变异性；2)氧化应激水平、呼吸链 CoQ10缺乏的严重程度不同，以及3)CoQ10缺乏的严重程度不同 其类似物对辅酶Q10缺陷有不同的疗效。为了检验这些假设，我们 提出以下两个具体目标：目标1)测试遗传和药物疗法 评估ADCK3及其类似物在体外的辅酶Q10缺乏症 羟基苯甲酸，作为修复内源性辅酶Q10生物合成缺陷的方法；目标2) 比较两种辅酶Q10缺乏症小鼠模型的治疗效果 辅酶Q10和艾地苯酮预防或延缓癫痫发作的临床研究 Pdss 2kd/kd和Coq9X/X突变小鼠的疾病。

Project Summary Coenzyme Q10 (CoQ10) is a lipophillic molecule composed of a benzoquinone ring and a hydrophobic decaprenyl tail. In the mitochondrial respiratory chain, CoQ10 transports electrons from complexes I and II to complex III. It is also an antioxidant, co-factor for pyrimidine biosynthesis, and modulator of apoptosis. Human CoQ10 deficiency is a genetically and clinically heterogeneous disease. Response to therapy is variable. Our studies of cultured fibroblasts from patients carrying mutations in PDSS2, COQ2, ADCK3, and COQ9, have revealed correlations between level of CoQ10 and oxidative stress; <15% and >60% residual CoQ10 are not associated with oxidative stress, whereas 30-50% residual CoQ10 is associated with oxidative stress, mitochondrial hyperpolarization followed by depolarization and cell death. In cell lines with primary CoQ10 deficiency, incubation of CoQ10 deficient fibroblasts for 1 week with 5mM CoQ10 (but not short-tail ubiquinone analogs) improved bioenergetics indicating pharmacokinetic constraints and dose of CoQ10 may limit efficacy in CoQ10 deficient patients. Based on our studies, we hypothesize that: 1) genotypic heterogenity contributes to the phenotypic variability of CoQ10-deficiency; 2) levels of oxidative stress, respiratory chain deficiency, and apoptosis differ depending on the severity CoQ10 deficiency, and 3) CoQ10 and its analogs have variable efficacy in CoQ10 deficiencies. To test these hypotheses, we propose the following two specific aims: Aim 1) To test genetic and pharmacologic therapies for CoQ10 deficiency in vitro by assessing ADCK3 overexpression and analogs of 4- hydroxybenzoic acid, as approaches to rescue endogenous CoQ10 biosynthesis defects; Aim 2) To test therapies in two mouse model of CoQ10 deficiency by comparing efficacies of oral and intratechal administration of CoQ10 and idebenone in preventing or delaying the onset of the disease in the Pdss2 kd/kd and Coq9X/X mutant mice.