Van Hove Johan L K, Josefsberg Sagi, Freehauf Cynthia, Thomas Janet A, Thuy Le Phuc, Barshop Bruce A, Woontner Michael, Mock Donald M, Chiang Pei-Wen, Spector Elaine, Meneses-Morales Iván, Cervantes-Roldán Rafael, León-Del-Río Alfonso
Department of Pediatrics, University of Colorado at Denver, CO, USA.
Mol Genet Metab. 2008 Dec;95(4):201-5. doi: 10.1016/j.ymgme.2008.09.006. Epub 2008 Oct 29.
We investigated in a patient with holocarboxylase synthetase deficiency, the relation between the biochemical and genetic factors of the mutant protein with the pharmacokinetic factors of successful biotin treatment. A girl exhibited abnormal skin at birth, and developed in the first days of life neonatal respiratory distress syndrome and metabolic abnormalities diagnostic of multiple carboxylase deficiency. Enzyme assays showed low carboxylase activities. Fibroblast analysis showed poor incorporation of biotin into the carboxylases, and low transfer of biotin by the holocarboxylase synthetase enzyme. Kinetic studies identified an increased Km but a preserved Vmax. Mutation analysis showed the child to be a compound heterozygote for a new nonsense mutation Q379X and for a novel missense mutation Y663H. This mutation affects a conserved amino acid, which is located the most 3' of all recorded missense mutations thus far described, and extends the region of functional biotin interaction. Treatment with biotin 100mg/day gradually improved the biochemical abnormalities in blood and in cerebrospinal fluid (CSF), corrected the carboxylase enzyme activities, and provided clinical stability and a normal neurodevelopmental outcome. Plasma concentrations of biotin were increased to more than 500 nM, thus exceeding the increased Km of the mutant enzyme. At these pharmacological concentrations, the CSF biotin concentration was half the concentration in blood. Measuring these pharmacokinetic variables can aid in optimizing treatment, as individual tailoring of dosing to the needs of the mutation may be required.
我们在一名全羧化酶合成酶缺乏症患者中,研究了突变蛋白的生化和遗传因素与生物素成功治疗的药代动力学因素之间的关系。一名女孩出生时皮肤异常,并在出生后的头几天出现新生儿呼吸窘迫综合征和多种羧化酶缺乏症的代谢异常诊断结果。酶分析显示羧化酶活性较低。成纤维细胞分析显示生物素掺入羧化酶的能力较差,全羧化酶合成酶对生物素的转运能力较低。动力学研究确定Km增加但Vmax保持不变。突变分析显示该儿童是新的无义突变Q379X和新的错义突变Y663H的复合杂合子。这种突变影响一个保守氨基酸,该氨基酸位于迄今为止描述的所有错义突变的最3'端,并扩展了功能性生物素相互作用的区域。每天100mg生物素治疗逐渐改善了血液和脑脊液(CSF)中的生化异常,纠正了羧化酶活性,并提供了临床稳定性和正常的神经发育结果。生物素的血浆浓度增加到超过500 nM,从而超过了突变酶增加的Km。在这些药理浓度下,脑脊液中的生物素浓度是血液中浓度的一半。测量这些药代动力学变量有助于优化治疗,因为可能需要根据突变的需求对剂量进行个体化调整。