Targeted gene transfer across specific synapse types

跨特定突触类型的靶向基因转移

• 批准号: 9004657
• 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
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9004657
• 关键词: Alzheimer' 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 Transfer; Genes; Genus staphylococcus; Glutamates; Goals; Health; Herpesvirus 1; 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; Staging; Stroke; Structure; Surface; Synapses; Synaptic plasticity; System; Tauopathies; Technology; base; density; entorhinal cortex; gene therapy; genetic analysis; learned behavior; mouse model; neocortical; nervous system disorder; neural circuit; neuronal circuitry; neurotrophic factor; novel; particle; postsynaptic neurons; presynaptic neurons; prototype; receptor; receptor binding; research study; synucleinopathy; theories; vector

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.

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.在特定电路中表现出缺陷的疾病包括主要的神经退行性疾病和其他主要神经系统疾病。此外，突触可塑性和神经网络理论假设特定行为是在特定电路中编码的。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本神经科学方面具有广泛的应用。 为了靶向基因转移到特定的神经元类型，我们在单纯疱疹病毒（HSV-1）载体颗粒表面修饰了蛋白质。我们用神经元类型特异性结合结构域代替了Panspific细胞结合结构域。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，将基因转移到包含同源受体的神经元中。接下来，我们开发了一种通用方法，将基因转移靶向特定的神经元类型，即抗体介导的靶向。我们将葡萄球菌A蛋白抗体结合结构域与载体颗粒蛋白添加在一起。这些载体颗粒和特定抗体的复合物支持靶向基因转移。 我们基于投影区域和突触类型开发了靶向基因转移，以将不同的基因传递到突触前神经元及其突触后神经元的选定子集中。最初的研究针对跨谷氨酸能突触的基因转移。第一个基因转移到突触前神经元中使用标准程序。该载体表达一个人工肽神经递质，其中包含一个密集的核囊泡排序结构域，神经递质受体结合结构域（对于NMDA NR1亚基）以及他的标签。释放后，该肽神经递质与突触后神经元上的同源受体结合。这些突触后神经元的抗体介导的靶向使用他的标记抗体，因为肽神经递质包含他的标签。该目标支持特异性增加约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，发展基因疗法和基本神经科学。提出的实验将优化跨突触的靶向基因转移，在神经病学和神经科学中至关重要的特定突触类型中发展靶向，并将这些进步应用于阿尔茨海默氏病病因的问题，从而发展出一种新型的基因疗法。 值得注意的是，这些靶向策略和试剂均可在任何基因转移系统中使用。