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）载体已成功地用于罕见神经系统的患者 疾病。最近，FDA批准了两种基于AAV的基因疗法药物，Luxturna已成为 价值850,000美元，一次性申请盲目申请，Zolgensma的脊柱肌肉为$ 2,100,000 萎缩。 AAV媒介介导的基因疗法已证明是一个潜在的巨大市场。虽然成功 神经系统疾病的临床研究，有效AAV脑应用的主要问题之一是高 人类中和抗体（NAB）的患病率。在普通人口中，超过95％ 个体被AAV感染，平均有50％的人患有NAB。 NABS对 无论输送路线如何（直接脑内注入或 系统管理）。已经利用了几种方法来逃避AAV NABS，包括化学 修改，使用不同的AAV矢量血清型，AAV CAPSID工程以及NAB的生物学耗竭 滴度（空的衣壳利用率，B细胞耗竭，等离子体代表和Ig蛋白酶）。通常，这些 方法的效率低，不需要的副作用或AAV的朝向主义改变。最近，纳本（Nabgen）有 开发了一种基于媒介的蛋白质策略，以使用独特的支原体来普遍阻止NABS 衍生的蛋白质，称为蛋白质。蛋白质M能够与任何物种的免疫球蛋白相互作用 抗原依赖性通过与抗体光和重链上的可变区结合。使用人类IVIG 和来自AAV免疫小鼠的血清，我们发现蛋白质M降低了AAV载体中和 与没有蛋白质M的对照组相比，100倍。最重要的是，我们观察到 蛋白质M能够在小鼠中保留超过1000倍的AAV转导，而NAB阳性的产物转移 血清。到目前为止，这是逃避AAV NABS的最有效策略。探索蛋白质M在 患有AAV脑靶向治疗的患者，必须解决蛋白质M的疗效以保护AAV 从NABS的NABS进行NABS转导。在此提案中，我们将首先研究蛋白质的影响 m通过直接注入小鼠的大脑，与AAV媒介共同辅助AAV向量 AAV的免疫（AIM 1）。接下来，我们将研究蛋白质M通过全身注射对脑的影响 直接传递AAV向量后的转导（AIM 2）。如果成功，这项小说有效的技术将 将AAV靶向基因治疗的好处扩展到每个患有脑疾病和AAV NABS的患者。