Project #4- Therapeutic approaches to CoQ10 deficiencies.

项目摘要 辅酶Q10（CoQ 10）是一种由苯醌环和辅酶A环组成的亲脂分子。 疏水性癸烯基尾。在线粒体呼吸链中，辅酶Q10转运电子 从复合物I和II到复合物III。它也是一种抗氧化剂，辅助因子嘧啶 生物合成和细胞凋亡的调节剂。人类辅酶Q10缺乏症是一种遗传和 临床异质性疾病。对治疗的反应是可变的。我们的研究 来自携带PDSS 2、COQ 2、ADCK 3和COQ 9突变的患者的成纤维细胞， 显示辅酶Q10水平和氧化应激之间的相关性; <15% and >60%的残留 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。 伴随氧化应激、线粒体超极化，随后是去极化和细胞死亡。 在具有原发性CoQ 10缺陷的细胞系中，将CoQ 10缺陷的成纤维细胞孵育1周 与5 mM辅酶Q10（但不是短尾泛醌类似物）改善的生物能量学表明， CoQ 10的药代动力学限制和剂量可能限制CoQ 10缺乏患者的疗效。 基于我们的研究，我们假设：1）基因型异质性有助于 辅酶Q10缺乏的表型变异; 2）氧化应激水平，呼吸链 辅酶Q10缺乏的严重程度不同，细胞凋亡也不同，3）辅酶Q10缺乏 其类似物在辅酶Q10缺乏症中具有不同的功效。为了验证这些假设，我们 提出以下两个具体目标：目标1）测试遗传和药物治疗， 通过评估ADCK 3过表达和4-羟色胺类似物的体外CoQ 10缺乏症 羟基苯甲酸，作为拯救内源性辅酶Q10生物合成缺陷的方法;目的2） 在两种CoQ 10缺乏症小鼠模型中，通过比较口服和 CoQ 10和艾地苯醌的技术内给药预防或延迟 在Pdss 2 kd/kd和Coq 9 X/X突变小鼠中的疾病。

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.