Manipulating N-acetyl-L-aspartate transport to treat Canavan leukodystrophy

操纵 N-乙酰基-L-天冬氨酸转运来治疗 Canavan 脑白质营养不良

基本信息

批准号：
10406711
负责人：
David E. Pleasure
金额：
$ 23.55万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10406711
关键词：
Ablation Acetyltransferase Affect Amino Acids Anticonvulsants Antisense Oligonucleotides Applications Grants Arachnoid mater Aspartoacylase Astrocytes Ataxia Autopsy Blindness Brain Brain Stem Canavan Disease Cell membrane Cells Cerebellum Cerebrospinal Fluid Child Citrates Cognitive deficits Coupled Data Development Drainage procedure Enzymes Epithelial Excretory function Genes Infant Kidney Knock-out Knockout Mice Lesion Ligase Lithium Lymphatic Endothelium Mediating Megalencephaly Meningeal Meninges Motor Mus Mutation N-acetylaspartate Neurodegenerative Disorders Neurologic Deficit Neurons Oligodendroglia Prosencephalon Reporting Route Seizures Sensory Sodium Specific qualifier value Testing Tubular formation Vacuole base brain magnetic resonance imaging cytotoxic dicarboxylate dicarboxylate-binding protein dietary manipulation effective therapy gene therapy human subject immunoreactivity knock-down leukodystrophy loss of function lymphatic vessel motor deficit mouse model prevent targeted treatment therapeutic target transcription factor urinary young adult

项目摘要

Canavan disease is a vacuolar (“spongiform”) leukodystrophy caused by ASPA mutations that diminish brain aspartoacylase activity. Current treatments for this neurodegenerative disorder, which include dietary manipulations, administration of lithium citrate and anticonvulsants, and AAV-mediated brain parenchymal ASPA gene therapy, do not reverse or prevent progression of neurological deficits in affected infants and children. Because aspartoacylase is necessary for cleavage of the abundant brain amino acid N-acetyl-L- aspartate (NAA), brain NAA is markedly elevated in Canavan disease. We were the first to report that knocking out or knocking down Nat8l, the gene that encodes the NAA-synthesizing enzyme N-acetytransferase 8-like, prevented development of motor deficits and brain vacuolation by aspartoacylase-deficient Canavan disease (“CD”) mice. Later, we showed that intrathecal administration of an Nat8l-inhibiting antisense oligonucleotide to young adult CD mice reversed their established brain NAA elevation, motor deficits and brain vacuolation. Together, those results strongly support the hypothesis that motor deficits and brain vacuolation in Canavan disease are consequences of elevated brain NAA. Our most recent studies, presented in this grant application, indicate that constitutive deletion of Slc13a3, which encodes NaDC3, a plasma membrane sodium-coupled transporter for NAA and other dicarboxylates, normalizes brain NAA content and prevents ataxia and brain vacuolation in CD mice. Also, we have confirmed prior reports that astroglia and renal proximal tubular epithelium express Slc13a3 and NaDC3, and have localized meningeal NaDC3 expression to arachnoid mater-associated Prox1+ cells. We now plan conditional deletion studies to determine whether selective ablation of NaDC3 from astroglia (Specific Aim 1), arachnoid (Specific Aim 2), or renal proximal tubular epithelium (Specific Aim 3) is sufficient to normalize brain NAA content and reverse motor deficits and brain vacuolation in symptomatic CD mice. Results of these studies will serve to guide development of selective Slc13a3 and NaDC3 therapeutic targeting strategies for infants and children with Canavan disease.

Canavan病是由ASPA突变引起的一种真空（“海绵状”）白细胞营养不良的减少大脑的天际囊酶活性。目前对这种神经退行性疾病的治疗其中包括饮食中的操纵，柠檬酸锂和抗惊厥药的给药以及 AAV介导的脑实质ASPA基因治疗，请勿逆转或防止受影响的婴儿和儿童的神经缺陷。因为天冬氨酸酶对于裂解丰富的脑氨基酸N-乙酰基-L-裂解是必需的天冬氨酸（NAA），脑NAA在Canavan病中明显升高。我们是第一个举报的人敲除或击倒NAT8L的那是编码NAA合成酶的基因 n-环控制酶8样，防止运动的发展定义和大脑真空化天冬氨酸酶缺陷式的Canavan病（“ CD”）小鼠。后来，我们显示了鞘内的施用NAT8L抑制反义寡核苷酸对年轻成人CD小鼠的逆转他们已建立的大脑NAA升高，运动定义和大脑真空化。在一起，那些结果强烈支持马达定义和脑真空吸尘的假设疾病是脑NAA升高的后果。我们最近在此赠款应用中提出的最新研究表明，构成删除 SLC13A3，编码NADC3，Naa和NaA的质膜钠耦合转运蛋白其他二羧酸盐，使脑NAA含量归一化，并防止共济失调和大脑吸尘 CD小鼠。另外，我们已经证实了先前的报告说星形胶质细胞和肾近端管状上皮Express SLC13A3和NADC3，并具有局部脑膜NADC3表达蛛网膜相关的prox1+细胞。现在，我们计划有条件的删除研究，以确定是否从星形胶质细胞（特定目标1），蛛网膜（特定AIM 2）或肾近端管状上皮（特定目标3）足以使大脑NAA含量正常化，并且反向运动定义和大脑有症状的CD小鼠的真空化。这些研究的结果将有助于指导发展选择性SLC13A3和NADC3治疗靶向策略 Canavan病。