Proj 4: Chemical Chaperone Therapy of Batten Disease

项目 4：Batten 病的化学伴侣疗法

基本信息

批准号：
8150188
负责人：
Glyn Dawson
金额：
$ 20.23万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8150188
关键词：
Address Adolescent Age Amino Acids Biological Biological Models Blood - brain barrier anatomy Blood capillaries Borates Brain CLN1 gene CLN2 gene Cells Central cord canal structure Chemicals Chickens Child Cholesterol Collaborations Complex Cultured Cells Defect Disease Drug Delivery Systems Embryo Endosomes Enzymes Fibroblasts Glass Glutamine Hereditary Disease Heterozygote Hippocampus (Brain)Histidine Human Hydrolase Hydrolysis Individual Infantile neuronal ceroid lipofuscinosis Injection of therapeutic agent Instruction Label Ligands Lysosomes Mediating Membrane Membrane Microdomains Mental Retardation Methods Methyl Green Missense Mutation Modeling Molecular Chaperones Monoglycerides Mutation Neonatal Nerve Degeneration Neuroglia Neurons Patients Peptide Hydrolases Peptides Pharmaceutical Preparations Point Mutation Proline Protein Region Proteins Psyche structure Quantum Dots Rat-1 Rattus Residual state Rhodamine Seizures Signal Transduction Slice Sphingolipids Spielmeyer-Vogt Disease Spinal Cord Structure Sulfhydryl Compounds Surface System Testing Therapeutic Tissues Toxic effect Work base capillary design disease-causing mutation enzyme activity esterase feeding fluorophore in vitro activity inhibitor/antagonist inorganic phosphate link protein lymphoblast lysyl-aspartyl-glutamyl-leucine novel novel strategies null mutation palmitoyl-protein hydrolase postnatal protein aminoacid sequence protein misfolding rabies virus glycoprotein G research study thioester trafficking tripeptidyl aminopeptidase tripeptidyl-peptidase I uptake

项目摘要

PROJECT SUMMARY (See instructions): Pharmacological chaperones (eg: AcGDap(Palm)VKIKK)can be internalized by cells and re-fold misfolded proteins to an active configuration, but need to cross the blood brain barrier, enter neurons and escape endosomes. We have shown that the palmitoylated peptide motif is uniquely able to allow drugs to escape endosomes and that such chaperones can reactivate misfolded proteins such as palmitoyhprotein thioesterase (PPTl). We now propose to design sequences to permit the chaperones to cross the blood brain barrier by either attaching a fluorophore and target motif (eg: rabies virus glycoprotein coat peptide (RVG)) or coating on 6-1 Onm quantum dots. We will test short sequences of proline and histidine, with a glutamine spacer to attach to the surface of 635 nm red QDs capped with the 4-thiol PEG ligand. We will test this in cultured postnatal neurons, and lymphoblasts from patients with defined point mutations in PPTl origin, and then use either the E3 embryonic chick spinal cord injection system, in collaboration with Project I, or the rat hippocampal slice system through collaboration with Project II. The three model systems represent embryonic brain, neonatal brain and postnatal brain and the collaborations will allow us to better assess the toxicity (if any) of these drugs, their efficacy and their ultimate cellular distribution by adding our chaperones into their experimental systems. We will also test the idea that the palmitoylated peptide motif works by specifically localizing to lipid rafts, microdomains in membranes which are greatly enriched in cholesterol and sphingolipids and appear to be used to assemble signaling complexes. Chaperones can only treat 20-50% of the mutations so for the remaining 50-80% of INCL patients we propose that hydrophobic thiols such as thiocholesterol could chemically facilitate hydrolysis of the storage material itself. Finally we will extend our approach to the most common form of Batten disease caused by mutations in the CLN2 gene (tripepfidyl-peptidase-1). Compound heterozygote patients with milder disease should benefit from chaperone therapy based on borate complexes of the inhibitor (AAFX) delivered to the CNS with our unique peptide sequences either directly or coated on quantum dots.

项目摘要（请参阅说明）：药理学伴侣（例如：ACGDAP（PALM）VKIKK）可以被细胞内部化，并将错误折叠的蛋白质重折成主动构型，但需要越过血脑屏障，进入神经元并逃脱内体。我们已经表明，棕榈酰化的肽基序具有独特的能力允许药物逃脱内体，并且这种伴侣可以重新激活错误折叠的蛋白质，例如棕榈酰蛋白酶硫酯酶（PPTL）。现在，我们建议设计序列，以允许伴侣通过连接荧光团和靶基序（例如：狂犬病病毒糖蛋白涂料肽（RVG））或在6-1 onM量子点上跨越血脑屏障。我们将用A的短序列测试脯氨酸和组氨酸的短序列谷氨酰胺间隔物连接到用4-硫醇PEG配体盖的635 nm红色QD的表面。我们将在培养的产后神经元中测试这一点，以及来自PPTL起源定义点突变患者的淋巴细胞，然后与Project I或通过与Project II合作的Rat Rat Hippocampal Slice System合作使用E3胚胎鸡脊髓注射系统。三个模型系统代表胚胎大脑，新生儿大脑和产后大脑，这些合作将使我们能够通过将伴侣伴侣添加到实验系统中来更好地评估这些药物的毒性（如果有的话），它们的功效和最终细胞分布。我们还将测试以下想法：棕榈酰化的肽基序是通过专门定位于脂质筏，在膜中极大地富集胆固醇和鞘脂的膜中的微域而起作用的，并且似乎用于组装信号传导络合物。伴侣只能治疗20-50％的突变，因此对于其余50-80％的含患者，我们建议疏水性硫醇（例如硫胆固醇）可以化学促进储存材料本身的水解。最后，我们将我们的方法扩展到由CLN2基因突变（Tripepfidyl-肽酶-1）引起的最常见形式的淡淡疾病。患有温和疾病的复合杂合子患者应受益于基于抑制剂（AAFX）的伴侣疗法（AAFX），并直接使用我们独特的肽序列或涂有量子点涂有我们独特的肽序列。