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Extracellular Vesicles for CNS Delivery of Therapeutic Enzymes to Treat Lysosomal Storage Disorders

细胞外囊泡用于中枢神经系统递送治疗酶以治疗溶酶体贮积症

基本信息

批准号：
9768769
负责人：
ELENA BATRAKOVA
金额：
$ 32.26万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9768769
关键词：
Address Adhesives Adverse effects Affect Antiinflammatory Effect Astrocytes Binding Biodistribution Biological Biological Availability Blood - brain barrier anatomy Brain C-Type Lectins CD47 gene Carrier Proteins Cell Communication Cells Cellular Membrane Central Nervous System Diseases Child Data Development Dichloromethylene Diphosphonate Disadvantaged Disease Disease Progression Dose Drug Delivery Systems Eating Electrophoretic Mobility Shift Assay Enzymes Evaluation FDA approved Fingerprint Formulation Freezing Genes Genetic Materials Goals Histologic Home environment Homing Human Image Imaging Techniques Immune system In Vitro Inflammation Injections Integrins Intraventricular Infusion Jansky-Bielschowsky Disease Label Lead Ligands Liposomes Longevity Lysosomal Storage Diseases Magnetic Resonance Imaging Measures Mediating Membrane Methods Microglia Modality Modeling Modification Mus Mutation NF-kappa B Nature Nerve Degeneration Nervous System Physiology Neuraxis Neurodegenerative Disorders Neuronal Ceroid-Lipofuscinosis Neurons Nuclear Extract Organ Patients Peptide Hydrolases Phagocytes Pharmaceutical Preparations Procedures Proteins Proteomics Recombinants Research Retina Risk Role Route Schedule Side Signal Transduction Spielmeyer-Vogt Disease Structure Surface Techniques Testing Therapeutic Therapeutic Index Tissues Toxicology Transfection Transgenic Mice Treatment Efficacy Validation Vesicle Vision Western Blotting base behavioral study bioimaging biomaterial compatibility brain cell brain endothelial cell brain tissue cytotoxicity effective therapy enzyme replacement therapy experimental study extracellular vesicles high risk immunogenicity improved in vitro Model in vivo in vivo imaging infrared spectroscopy innovation insight macromolecule macrophage magnetic resonance spectroscopic imaging mouse model nanocarrier nanosized neuroimaging neuroprotection neurovascular unit novel novel therapeutics optical imaging pediatric patients plasmid DNA preservation protein transport real-time images receptor relating to nervous system safety study symptomatic improvement syncytin targeted treatment therapeutic enzyme therapeutic protein therapy development tripeptidyl aminopeptidase uptake vector vesicular release

项目摘要

The neuronal ceroid lipofuscinoses or Batten disease (BD) is one of a broad class of severe neurodegenerative diseases that are known as lysosomal storage disorders (LSDs). BD is characterized by the intracellular accumulation of storage material in neural tissues and progressive loss of neurological functions and vision that primarily affect children. CLN2 is a specific type of BD that results from mutations in the TPP1 gene causing an insufficiency or complete lack of a soluble lysosomal enzyme tripeptidyl peptidase-1 (TPP1). Last year, the first FDA-approved treatment, intraventricular infusions of a recombinant form of human TPP1, was shown to provide symptomatic improvements in pediatric patients. Unfortunately, this invasive procedure carries a high risk of adverse effects. The less invasive systemic administration provides no therapeutic benefits, because the blood brain barrier (BBB) severely restricts transport of macromolecules - including TPP1- to the brain. To circumvent this problem, we propose using extracellular vesicles (EVs) released by macrophages as biocompatible nanocarriers for systemic delivery of TPP1. Comprised of cellular membranes with multiple adhesive proteins on their surface, EVs are known to specialize in cell-cell communications facilitating transport of proteins and genetic material to target cells. Our research has previously demonstrated that therapeutic proteins, including TPP1, can be efficiently incorporated into EVs without losing their biological activity. In particular, TPP1 was loaded into EVs using two methods: (i) transfection of parental EV-producing macrophages with TPP1-encoding plasmid DNA (pDNA), or (ii) loading therapeutic protein TPP1 into naive empty EVs. The resulting EVs carrier ensemble was shown to readily migrate into the brains of late-infantile neuronal ceroid lipofuscinosis (LINCL) mice upon systemic administration. Importantly, multiple lines of evidence for therapeutic efficacy were observed in LINCL mice, including significant neuroprotection and improved life span. Noteworthy, we developed different methods for EVs isolation, purification, characterization, and storage in sufficient quantities for therapeutic application. In the current proposal, we will utilize two EV-based formulations of TPP1 to demonstrate the proof of concept using a BD mouse model, LINCL mice. Planned studies include: (SA1) elucidation the nature of selective fingerprinting of macrophage-derived EVs, and mechanism of EVs interactions and TPP1 facilitated uptake in cells of neurovascular unit; (SA2) evaluation of brain bioavailability for EV-TPP1 with MRI and optical imaging in vivo, and (SA3) validation the therapeutic potential of this novel drug delivery system by measuring its neuroprotective and anti-inflammatory effects in LINCL mice. The proposed research addresses a critical problem in the effective delivery of therapeutic proteins to the central nervous system (CNS), and will provide fundamental insights into, how EVs communicate with target brain cells, and selectively deliver their cargo.

神经性脑蜡样脂褐变病或称巴顿病（BD）是一类严重的神经退行性疾病