Single domain antibodies for diagnosis and treatment of synucleinopathies

用于诊断和治疗突触核蛋白病的单域抗体

基本信息

项目摘要

Abstract Immunotherapies targeting aggregates of amyloid-β (Aβ), tau and α-synuclein (αsyn) are the most common experimental approach for diseases characterized by such depositions. Most of these potential therapies are whole antibodies. Antibody fragments have certain advantages over antibodies but their diagnostic and therapeutic potential has not been well explored. Five anti-αsyn IgG antibodies and one αsyn vaccine are in Phase 1-2 clinical trials, and one Aβ antibody recently showed clinical benefit in a Phase 3 trial, which strongly supports this therapeutic approach. Regarding antibody fragments, several anti-αsyn single chain variable fragments (scFvs) are effective in culture and in vivo, and two anti-αsyn single domain antibodies (sdAbs) have been characterized in a binding assay but their therapeutic efficacy or diagnostic imaging potential have not been reported. SdAbs have several advantages over scFvs, including higher affinity, binding to cryptic epitopes and simpler engineering. They are also more suitable for gene therapy because of uncomplicated expression and predictable protein folding, given their single domain, resulting in high solubility. We have developed 58 anti-αsyn sdAb clones with unique binding regions from a screen of phage display libraries generated from B-cells of a llama immunized with αsyn. Three aims are proposed to clarify their potential to detect and clear αsyn in vivo. Aim 1 will determine the in vivo imaging potential of sdAbs targeting αsyn. Aim 2 will clarify αsyn clearance potential and mechanism of action of sdAbs targeting αsyn, and Aim 3 will explore the feasibility of sdAb gene therapy targeting αsyn. Preliminary findings indicate that the sdAbs: 1) recognize various forms of αsyn with high affinity; 2) clear αsyn and prevent its toxicity in culture models; 3) clear αsyn from brain interstitial fluid in αsyn mice; 4) bind to αsyn in the brain of αsyn mice after intravenous injection with in vivo brain signal that correlates well with αsyn brain burden; 5) clear αsyn from the brain after intravenous injection of sdAb with improved clearance by its PROTAC derivative, and; 6) clear αsyn from the brain after intravenous sdAb gene therapy. It is hypothesized that the small size of the sdAbs will provide diagnostic and therapeutic benefits over whole antibodies, primarily because of greater access into the brain and to the target, proper folding for gene therapy, and to some extent due to their binding to novel epitopes that the larger antibodies cannot access. Overall, the proposed studies are likely to aid in clarifying αsyn pathogenesis, decipher the mechanism of action of the sdAbs, and identify promising diagnostic and therapeutic sdAbs for clinical trials on synucleinopathies such as Lewy Body Dementia, Parkinson's disease, Alzheimer's disease with Lewy Bodies, and Multiple System Atrophy.
摘要 针对淀粉样蛋白β(Aβ)、tau和α突触核蛋白(αsyn)聚集体的免疫疗法最多 以这种沉积为特征的疾病的常见实验方法。其中大部分潜力 治疗都是完全的抗体。抗体片段比抗体有一定的优势,但它们的 诊断和治疗的潜力还没有得到很好的开发。5例抗αSYN抗体和1例αSYN 疫苗正处于1-2期临床试验,一种Aβ抗体最近在3期试验中显示出临床益处, 这有力地支持了这种治疗方法。关于抗体片段,几种抗α同源单抗 单链可变区(ScFv)在培养和体内都是有效的,两个抗α同源结构域 抗体(SdAbs)的特征是结合分析,但它们的治疗效果或诊断 成像潜力尚未见报道。与单链抗体相比,SdAbs有几个优点,包括更高的 亲和力、与神秘表位的结合以及更简单的工程设计。它们也更适合于基因治疗。 由于其简单的表达和可预测的蛋白质折叠,给定其单一的结构域,导致 高溶解度。 我们从噬菌体的筛选中获得了58个具有独特结合区的抗αsyn sdAb克隆 展示由αSYN免疫的骆驼B细胞生成的文库。提出了三个目标来澄清 它们在体内检测和清除αSYN的潜力。目标1将确定sdAbs的体内成像潜力 目标是αSYN。目的2将阐明针对αSYN的单抗清除αSYN的潜力和作用机制, 目的3探讨以αSYN为靶点的双链抗体基因治疗的可行性。 初步研究结果表明,sdAbs:1)识别多种形式的高亲和力的α-syn; 清除αSYN并预防其在培养模型中的毒性;3)清除αSYN小鼠脑间质液中的SYN;4) 静脉注射后与α同种异体小鼠脑内α同源蛋白结合的体内脑信号良好相关 α-SYN脑负荷;5)静脉注射改良的α-SYN单抗后清除脑内 6)静脉注射α基因治疗后,清除脑组织中的SdAb。它 假设sdAbs的小尺寸将提供整体的诊断和治疗益处 抗体,主要是因为更容易进入大脑和靶标,适当折叠以进行基因治疗, 在某种程度上,这是因为它们与较大的抗体无法获得的新表位结合。 总体而言,拟议的研究可能有助于阐明αSYN的发病机制,破译其机制。 研究sdAbs的作用,并确定有希望的诊断和治疗sdAbs用于临床试验 路易小体痴呆、帕金森氏病、阿尔茨海默病合并路易小体、 和多系统萎缩。

项目成果

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Einar M Sigurdsson其他文献

Einar M Sigurdsson的其他文献

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{{ truncateString('Einar M Sigurdsson', 18)}}的其他基金

Efficacy and Mechanism of Action of Heavy-Chain α-Synuclein Antibody Fragments Derived from Llama
美洲驼重链α-突触核蛋白抗体片段的功效和作用机制
  • 批准号:
    9762785
  • 财政年份:
    2018
  • 资助金额:
    $ 71.75万
  • 项目类别:
Clearance and In Vivo Detection of Tau Pathology
Tau 病理学的清除和体内检测
  • 批准号:
    8673411
  • 财政年份:
    2013
  • 资助金额:
    $ 71.75万
  • 项目类别:
Epitope-Specific Targeting of Tau Aggregates.
Tau 聚集体的表位特异性靶向。
  • 批准号:
    8673382
  • 财政年份:
    2013
  • 资助金额:
    $ 71.75万
  • 项目类别:
Immune Therapy and Imaging in Mouse and Primate Models of Alzheimer's Disease.
阿尔茨海默病小鼠和灵长类动物模型的免疫治疗和成像。
  • 批准号:
    8676081
  • 财政年份:
    2013
  • 资助金额:
    $ 71.75万
  • 项目类别:
Epitope-Specific Targeting of Tau Aggregates
Tau 聚集体的表位特异性靶向
  • 批准号:
    10594553
  • 财政年份:
    2011
  • 资助金额:
    $ 71.75万
  • 项目类别:
Epitope-Specific Targeting of Tau Aggregates.
Tau 聚集体的表位特异性靶向。
  • 批准号:
    8230884
  • 财政年份:
    2011
  • 资助金额:
    $ 71.75万
  • 项目类别:
Epitope-Specific Targeting of Tau Aggregates.
Tau 聚集体的表位特异性靶向。
  • 批准号:
    8464819
  • 财政年份:
    2011
  • 资助金额:
    $ 71.75万
  • 项目类别:
Epitope-Specific Targeting of Tau Aggregates
Tau 聚集体的表位特异性靶向
  • 批准号:
    10467481
  • 财政年份:
    2011
  • 资助金额:
    $ 71.75万
  • 项目类别:
Epitope-Specific Targeting of Tau Aggregates.
Tau 聚集体的表位特异性靶向。
  • 批准号:
    8320099
  • 财政年份:
    2011
  • 资助金额:
    $ 71.75万
  • 项目类别:
Epitope-Specific Targeting of Tau Aggregates.
Tau 聚集体的表位特异性靶向。
  • 批准号:
    10187658
  • 财政年份:
    2011
  • 资助金额:
    $ 71.75万
  • 项目类别:
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