Novel strategy to block AAV neutralizing anitbody activity

阻止 AAV 中和抗体活性的新策略

基本信息

批准号：
10080225
负责人：
Chengwen Li
金额：
$ 30万
依托单位：
NABGEN, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10080225
关键词：
Address Adoptive Transfer Affect Affinity Animal Model Antibodies Antibody titer measurement Antigen-Antibody Complex Antigens B-Lymphocytes Binding Binding Proteins Biological Blindness Blocking Antibodies Blood Blood Coagulation Disorders Blood Component Removal Canis familiaris Capsid Cell Nucleus Cells Cellular Tropism Chemicals Chromosomes Clinic Clinical Clinical Research Clinical Trials Control Groups DNA Sequence Alteration Data Dependence Dependovirus Development Disease Dose FDA approved Family suidae Gene Delivery Genetic Diseases Hemophilia A Hemorrhage Hereditary Disease High Prevalence Human Immunization Immunize Immunoglobulin Variable Region Immunoglobulins Immunologics In Situ In Vitro Individual Injections Intramuscular Intramuscular Injections Intravenous Immunoglobulins Light Luciferases Mediating Medicine Mendelian disorder Modification Mus Muscle Muscular Atrophy Mutagenesis Mycoplasma Myopathy Nature Neurologic Orphan Drugs Patients Pharmaceutical Preparations Pharmacological Treatment Phase Plasma Population Preparation Production Property Proteins Quality of life Route Safety Serotyping Serum Spinal Structure Surface Testing Therapeutic Time Tissues Transgenes Treatment Efficacy Virion Work adeno-associated viral vector analog base clinical development combinatorial design dosage efficacy testing gene therapy improved improved outcome in vitro activity in vivo mortality multiple myeloma M Protein mutant neutralizing antibody novel strategies pre-clinical prevent protective effect side effect success technology development tissue tropism tool trafficking transduction efficiency transgene expression vector virus tropism

项目摘要

Adeno-associated virus (AAV) vectors have been successfully applied in clinical trials in patients with blindness, neurological/muscular disorders and bleeding diseases. Two AAV based gene therapy drugs have been recently approved by the FDA, Luxturna has been valued at $850,000 for a one-time treatment for a rare form of blindness and Zolgensma at $2,100,000 for spinal muscle atrophy. Gene therapy with AAV vectors has shown a potentially huge market. Although successful in clinical studies, one of major concerns for broader AAV vector application for patients is high prevalence of neutralizing antibody (Nab). AAV Nabs are able to bind to the surface of virions and interfere with AAV binding to target cells or intracellular trafficking or uncoating in the nucleus, and then block effective AAV transduction. In the general human population, over 95% of individuals have been infected by AAV and, on average, 50% of them have Nabs. Several approaches have been explored to evade AAV Nabs, including chemical modification, different serotype of AAV vector, rational design and combinatorial mutagenesis of the capsid in situ, as well as biological depletion of Nab titer (empty capsid utilization, B cell depletion and plasma-apheresis). Generally, these approaches have low efficiency, troubling side effects, or alter beneficial AAV properties like infectivity and production yield. Therefore, it is imperative to develop ideal strategies to evade Nabs, but without a changing tissue tropism from modification of capsids or negative side-effects of pharmacological treatment. Recently, we have developed a vector independent protein-based strategy to universally block Nabs. We have pioneered the use of a unique mycoplasma derived protein and its analogues, termed Protein-M, to enable successful gene delivery by preventing AAV neutralization of Nabs. Protein-M is able to interact with immunoglobulins from any species without antigen dependence by universally binding to variable regions on the antibody light and heavy chains. We have validated Protein-M mediated Nab escape in vitro using human IVIG and serum from AAV immunized mice, and found that Protein-M protected AAV vector neutralization over 100-fold when compared to control group without Protein-M. Most importantly, we found that 1000-fold protection of AAV transduction could be achieved in vivo when using Protein-M in mice after adoptive transfer of Nab positive serum. So far, this is the most effective strategy to evade AAV Nabs. To explore the application of Protein-M in clinical trials, it is imperative to address the efficacy of Protein-M for Nab blockage in subjects with pre-immunization. In this proposal, we will first study the effect of Protein-M on AAV Nabs blockage for systemic AAV gene delivery (Aim 1). Preliminary results have also shown that circulating Nabs block transduction during local administration of AAV vectors. Next, we will study the effect of Protein-M administered via different routes on muscle transduction after direct muscular injection of AAV vectors (Aim 2). If successful in preclinical animal models, these AAV mutants can be immediately transitioned to the clinic.

腺相关病毒（AAV）载体已成功地用于患有临床试验中失明，神经/肌肉疾病和出血疾病。两种基于AAV的基因治疗药物具有卢克斯特纳（Luxturna 脊柱肌肉萎缩的失明和Zolgensma的形式为$ 2,100,000。用AAV载体的基因疗法具有显示了一个潜在的巨大市场。尽管在临床研究方面取得了成功，但对更广泛的主要关注点之一患者的AAV载体应用是中和抗体（NAB）的高患病率。 aav nabs能够与病毒体表面结合并干扰与靶细胞或细胞内运输或在细胞核中取消涂覆，然后阻止有效的AAV转导。在一般人口中 95％的个体已被AAV感染，平均有50％的人患有NAB。几种方法已经探索以逃避AAV NAB，包括化学修饰，AAV载体的不同血清型， Capsid原位的合理设计和组合诱变，以及NAB滴度的生物学耗竭（空的衣壳利用率，B细胞耗竭和等离子体代理）。通常，这些方法很低效率，令人不安的副作用或改变有益的AAV特性，例如感染性和生产产量。因此，必须制定理想的策略来逃避NAB，但没有变化的组织偏向主义来自药理学治疗的衣壳的修饰或负面副作用。最近，我们有开发了一种基于媒介的蛋白质策略来普遍阻止NAB。我们已经开创了使用独特的支原体衍生的蛋白质及其类似物称为蛋白-M，以实现成功的基因通过防止NAB的AAV中和。蛋白质M能够与任何来自任何的免疫球蛋白相互作用通过普遍结合抗体光和重的可变区域，没有抗原依赖性的物种链。我们使用人IVIG和AAV的血清验证了蛋白质M介导的NAB逃脱的体外逃脱免疫性小鼠，发现蛋白质M比较了100倍以上的AAV载体中和对照组无蛋白质-M。最重要的是，我们发现对AAV转导的1000倍保护 Nab阳性血清的过继转移后，在小鼠中使用蛋白质M时可以在体内实现。迄今为止，这是逃避AAV NABS的最有效策略。为了探索蛋白质M在临床试验中的应用，必须解决蛋白质M对NAB阻塞的功效，对具有预免疫的受试者。在这个提案，我们将首先研究蛋白质-M对全身AAV基因递送的AAV NABS阻塞的影响（目标1）。初步结果还表明，循环NABS阻止局部的转导 AAV向量的管理。接下来，我们将研究通过不同途径给予蛋白质的影响直接肌肉注射AAV载体后的肌肉转导（AIM 2）。如果在临床前动物成功模型，这些AAV突变体可以立即转移到诊所。