Development of an Effective Strategy to Block Nab Activity for AAV Brain Transduction

开发一种有效策略来阻断 AAV 脑转导的 Nab 活性

• 批准号: 10600969
• 负责人: Charles H Askew
• 金额: $ 26.96万
• 依托单位: NABGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10600969
• 关键词: Address; Adoptive Transfer; Affect; Affinity; Animals; Antibodies; Antibody titer measurement; Antigen-Antibody Complex; Antigens; B-Lymphocytes; Binding; Binding Proteins; Biological; Blindness; Blood Coagulation Disorders; Blood Component Removal; Brain; Brain Diseases; Capsid; Cellular Tropism; Central Nervous System; Central Nervous System Diseases; Chemicals; Chromosomes; Clinical; Clinical Research; Clinical Trials; Control Groups; DNA Sequence Alteration; Data; Dependence; Dependovirus; Development; Disease; Dose; Engineering; FDA approved; Family suidae; Gene Delivery; Genetic Diseases; High Prevalence; Human; Immunization; Immunize; Immunoglobulins; Immunologics; In Vitro; Individual; Injections; Intravenous Immunoglobulins; Intraventricular; Intraventricular Injections; Light; Marketing; Mediating; Medicine; Mendelian disorder; Modification; Mus; Muscular Atrophy; Mycoplasma; Nature; Orphan Drugs; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Phase; Plasma; Population; Preparation; Production; Proteins; Quality of life; Rare Diseases; Route; Safety; Serotyping; Serum; Spinal; Structure; Technology; Therapeutic; Vertebral column; Work; adeno-associated viral vector; clinical development; comparison control; efficacy testing; gene therapy; improved outcome; in vitro activity; intravenous injection; mortality; multiple myeloma M Protein; mutant; nervous system disorder; neutralizing antibody; novel; prevent; protective effect; rare genetic disorder; side effect; success; targeted treatment; technology development; tool; transduction efficiency; transgene expression; vector; virus tropism

Adeno-associated virus (AAV) vectors have been successfully employed in patients with rare neurological diseases. Recently, two AAV based gene therapy drugs have been approved by the FDA, Luxturna been valued at $850,000 with a one-time application for blindness and Zolgensma at $2,100,000 for spinal muscle atrophy. AAV vector mediated gene therapy has shown to be a potentially huge market. Although successful in clinical studies for neurological disorders, one of the major concerns for effective AAV brain application is high prevalence of neutralizing antibody (Nab) in humans. In the general human population, over 95 % of individuals are infected by AAV and, on average, 50 % of them develop Nabs. The inhibition effect of Nabs on AAV brain transduction has been well documented regardless of delivery routes (direct intra-brain injection or systemic administration). Several approaches have been exploited to escape AAV Nabs, including chemical modification, use of different AAV vector serotypes, AAV capsid engineering, and biological depletion of Nab titer (empty capsid utilization, B cell depletion, plasma-apheresis, and Ig proteases). Generally, these approaches have low efficiency, unwanted side effects, or AAV tropism change. Recently, Nabgen has developed a vector independent protein-based strategy to universally block Nabs using a unique mycoplasma derived protein, termed Protein-M. Protein-M is able to interact with immunoglobulin from any species without antigen dependence by binding to variable regions on the antibody light and heavy chains. Using human IVIG and serum from AAV immunized mice, we have found that Protein-M reduced AAV vector neutralization over 100 fold when compared to control group without Protein-M in vitro. Most importantly, we have observed that Protein-M was able to retain AAV transduction over 1000 fold in mice with 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 patients with AAV brain targeted therapy, it is imperative to address the efficacy of Protein-M to protect AAV from Nabs for brain transduction in subjects with Nabs. In this proposal, we will first study the effect of Protein- M co-administered with AAV vectors on AAV Nab blockage via direct injection into the brain in mice with pre- immunization of AAV (Aim 1). Next, we will study the effect of Protein-M via systemic injection on brain transduction after direct delivery of AAV vectors (Aim 2). If successful, this novel and effective technology will extend the benefits of AAV targeted gene therapy to every patient with brain disorders and AAV Nabs.

腺相关病毒（AAV）载体已成功用于罕见神经系统疾病患者， 疾病最近，两种基于AAV的基因治疗药物已被FDA批准，Luxturna被 价值850 000美元，一次性用于失明，Zolgensma价值2 100 000美元用于脊髓肌肉 萎缩AAV载体介导的基因治疗已显示出潜在的巨大市场。尽管成功地 在神经系统疾病的临床研究中，有效的AAV脑应用的主要关注点之一是高的 中和抗体（Nab）在人类中的流行率。在普通人群中，超过95%的 个体被AAV感染，平均50%的个体发展成Nab。Nabs的抑制作用 AAV脑转导已被充分记录，无论递送途径如何（直接脑内注射或直接脑内注射）。 全身给药）。已经利用了几种方法来逃避AAV Nabs，包括化学方法。 修饰、不同AAV载体血清型的使用、AAV衣壳工程改造和Nab的生物学消耗 滴度（空衣壳利用率、B细胞耗竭、血浆单采术和IG蛋白酶）。通常这些 这些方法具有低效率、不希望的副作用或AAV向性改变。最近，Nabgen 开发了一种载体独立的基于蛋白质的策略，使用独特的支原体 衍生的蛋白质，称为蛋白质-M。蛋白-M能够与来自任何物种的免疫球蛋白相互作用， 通过与抗体轻链和重链上的可变区结合而产生抗原依赖性。使用人免疫球蛋白 和来自AAV免疫的小鼠的血清，我们发现蛋白-M降低了AAV载体中和， 100倍，与无蛋白-M的对照组相比。最重要的是，我们观察到， 蛋白-M能够在具有Nab阳性的过继转移的小鼠中保持超过1000倍的AAV转导。 血清的到目前为止，这是逃避AAV Nabs的最有效策略。探讨蛋白M在免疫治疗中的应用 对于AAV脑靶向治疗的患者，解决Protein-M保护AAV的功效势在必行 用于Nabs受试者的脑转导。在这个建议中，我们将首先研究蛋白质的作用- M与AAV载体共同施用，通过直接注射到具有预处理的小鼠的脑中进行AAV Nab阻断。 AAV免疫（目的1）。接下来，我们将研究蛋白-M通过全身注射对脑的影响， 在直接递送AAV载体后的转导（Aim 2）。如果成功，这种新颖有效的技术将 将AAV靶向基因治疗的益处扩展到每一位患有脑部疾病和AAV Nab的患者。