Targeted gene transfer across specific synapse types

跨特定突触类型的靶向基因转移

基本信息

批准号：
9212208
负责人：
ALFRED I. GELLER
金额：
$ 31.94万
依托单位：
LSU HEALTH SCIENCES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2020-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9212208
关键词：
Alzheimer&apos s Disease Amyloid beta-Protein Amyotrophic Lateral Sclerosis Antibodies Area Behavior Binding Biological Assay Biological Neural Networks Brain Cells Complex Dense Core Vesicle Disease Disease Progression Enhancers Enzymes Epilepsy Etiology Exhibits Gene Targeting Gene Transfer Genes Genus staphylococcus Glutamates Goals HSV vector Human Huntington Disease Immunoglobulin Variable Region Learning Mediating Methods Mus N-Methylaspartate NR1 gene Neocortex Neurodegenerative Disorders Neurology Neurons Neurosciences Neurotransmitter Receptor Neurotransmitters Parkinson Disease Peptides Physiological Physiology Procedures Prosencephalon Proteins Rattus Reagent Receptor Protein-Tyrosine Kinases Role Schizophrenia Simplexvirus Sorting - Cell Movement Specificity Stroke Structure Surface Synapses Synaptic plasticity System Tauopathies Technology Virion base density entorhinal cortex experimental study gene therapy genetic analysis mouse model neocortical nervous system disorder neural circuit neuronal circuitry neurotrophic factor novel particle postsynaptic neurons presynaptic neurons prototype public health relevance receptor receptor binding synucleinopathy theories vector

项目摘要

DESCRIPTION (provided by applicant): Developing gene therapies for specific neurological diseases, elucidating the etiology of these diseases, and genetic analyses of neuronal circuits, behaviors, and learning will all benefit from targeted gene transfer that can deliver different genes into presynaptic neurons and a selected subset of their postsynaptic neurons, based on both projection area and synapse type. Diseases that exhibit deficits in specific circuits include the major neurodegenerative diseases, and other major neurological disorders. Moreover, synaptic plasticity and neural network theories hypothesize that specific behaviors are encoded in specific circuits. Thus, targeted gene transfer across specific synapses will have broad applications to understanding and treating neurological diseases, and to basic neuroscience. To target gene transfer to specific neuron types, we modified a protein on the surface of Herpes Simplex Virus (HSV-1) vector particles; we replaced a panspecific cell binding domain with a neuron type-specific binding domain. In the initial study, we added specific neurotrophic factors to the surface of HSV-1 particles, targeting gene transfer to neurons that contain the cognate receptors. Next, we developed a general method to target gene transfer to specific neuron types, antibody-mediated targeting. We added the Staphylococcus A protein antibody binding domain to a vector particle protein. Complexes of these vector particles and specific antibodies supported targeted gene transfer. We developed targeted gene transfer to deliver different genes into presynaptic neurons and a selected subset of their postsynaptic neurons, based on both projection area and synapse type. The initial study targeted gene transfer across glutamatergic synapses. The first gene transfer, into the presynaptic neurons, uses standard procedures. The vector expresses an artificial peptide neurotransmitter that contains a dense core vesicle sorting domain, a neurotransmitter receptor binding domain (for NMDA NR1 subunits), and the His tag. Upon release, this peptide neurotransmitter binds to the cognate receptors on the postsynaptic neurons. Antibody-mediated targeting to these postsynaptic neurons uses a His tag antibody, as the peptide neurotransmitter contains the His tag. This targeting supported an ~10-fold increase in specificity. The long-term goal of this project is to develop targeted gene transfer across specific synapse types, for elucidating disease etiologies, developing gene therapies, and basic neuroscience. The proposed experiments will optimize targeted gene transfer across synapses, develop targeting across specific synapse types critical in neurology and neuroscience, and apply these advances to a problem in the etiology of Alzheimer's disease, thereby developing a novel gene therapy. Of note, these targeting strategies and reagents can be used in any gene transfer system.

描述（由申请人提供）：开发针对特定神经系统疾病的基因疗法，阐明这些疾病的病因，以及神经元回路、行为和学习的遗传分析都将受益于靶向基因转移，该基因转移可以将不同的基因传递到突触前神经元和神经元中。根据投影面积和突触类型选择突触后神经元的子集。特定回路出现缺陷的疾病包括主要的神经退行性疾病和其他主要的神经系统疾病。此外，突触可塑性和神经网络理论假设特定行为被编码在特定电路中。因此，跨特定突触的靶向基因转移将在理解和治疗神经系统疾病以及基础神经科学方面具有广泛的应用。为了将基因转移到特定的神经元类型，我们修饰了单纯疱疹病毒（HSV-1）载体颗粒表面的蛋白质；我们用神经元类型特异性结合域替换了泛特异性细胞结合域。在最初的研究中，我们在 HSV-1 颗粒的表面添加了特定的神经营养因子，将基因转移到含有同源受体的神经元。接下来，我们开发了一种将基因转移到特定神经元类型的通用方法，即抗体介导的靶向。我们将葡萄球菌 A 蛋白抗体结合域添加到载体颗粒蛋白中。这些载体颗粒和特异性抗体的复合物支持靶向基因转移。我们开发了靶向基因转移，根据投影面积和突触类型将不同的基因传递到突触前神经元及其选定的突触后神经元子集。最初的研究针对跨谷氨酸突触的基因转移。第一次基因转移进入突触前神经元，使用标准程序。该载体表达人工肽神经递质，其中包含致密核心囊泡分选结构域、神经递质受体结合结构域（针对 NMDA NR1 亚基）和 His 标签。释放后，这种肽神经递质与突触后神经元上的同源受体结合。抗体介导的针对这些突触后神经元的靶向使用 His 标签抗体，因为肽神经递质包含 His 标签。这种靶向支持特异性提高约 10 倍。该项目的长期目标是开发跨特定突触类型的靶向基因转移，以阐明疾病病因、开发基因疗法和基础神经科学。拟议的实验将优化跨突触的靶向基因转移，开发对神经病学和神经科学至关重要的特定突触类型的靶向，并将这些进展应用于阿尔茨海默病的病因学问题，从而开发出一种新型基因疗法。值得注意的是，这些靶向策略和试剂可用于任何基因转移系统。