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)载体已成功地应用于罕见的神经系统疾病患者 疾病。最近，两种基于AAV的基因治疗药物已经获得FDA的批准，Luxturna被批准 价值850,000美元，一次性申请失明，Zolgensma，脊柱肌肉，2,100,000美元 萎缩。AAV载体介导的基因治疗已被证明是一个潜在的巨大市场。虽然在 神经疾病的临床研究，有效应用AAV脑的主要关注点之一是高 中和抗体(NAB)在人类中的流行率。在普通人口中，超过95%的人 个人感染甲型流感病毒，平均有50%的人患上新城疫。NaBS对小鼠肺泡巨噬细胞的抑制作用 AAV脑转导已经被很好地记录下来，而不考虑传递途径(直接脑内注射或 系统管理)。已经利用了几种方法来逃避AAV Nabs，包括化学 不同AAV载体血清型的修饰、使用、AAV衣壳工程和NAB的生物耗竭 效价(空衣壳利用率、B细胞耗竭、血浆分离和Ig蛋白水解酶)。一般而言，这些 治疗方法效率低、副作用大或AAV取向改变。最近，纳布根已经 开发了一种基于载体的非依赖蛋白质的策略，利用一种独特的支原体普遍阻断NAB 衍生蛋白质，称为蛋白质-M。蛋白-M能够与任何物种的免疫球蛋白相互作用，而不需要 通过结合抗体轻链和重链上的可变区来依赖抗原。使用人类静脉注射免疫球蛋白 和AAV免疫小鼠的血清，我们发现Protein-M降低了AAV载体的中和能力。 与不加蛋白-M的对照组相比，在体外是对照组的100倍。最重要的是，我们观察到 在过继转移NAB阳性的小鼠中，Protein-M能够保留1000倍以上的AAV转导 血清。到目前为止，这是规避AAV NAB最有效的策略。探讨蛋白质-M在生物医学中的应用 AAV患者脑内靶向治疗，解决蛋白-M对AAV的保护作用势在必行 用于NABS受试者的脑传导。在这项提案中，我们将首先研究蛋白质的影响- 小鼠脑内直接注射M与AAV载体联合应用阻断AAV NAB的实验研究 甲型肝炎病毒免疫(目标1)。接下来，我们将研究蛋白-M通过全身注射对大脑的影响 AAV载体直接传递后的转导(目标2)。如果成功，这项新颖而有效的技术将 将AAV靶向基因治疗的好处扩大到每一位患有脑部疾病和AAV NAB的患者。