Microparticle Therapy for Cerebral Amyloid Angiopathy

项目摘要 脑淀粉样血管病（CAA），一种微血管病变，其中β-淀粉样蛋白（A β）积聚在壁中 与血管脆性和继发于血管壁的出血有关 崩溃CAA对血管平滑肌细胞（VSMC）尤其有害。CAA存在于70 - 90%的 阿尔茨海默病（AD）病例，增加出血性中风的风险，并加剧了活性淀粉样蛋白 免疫疗法，从而损害这种有前途的AD治疗剂。CAA目前尚无有效的治疗方法。 尽管CAA在AD中普遍存在，并且AD和CAA是不同的疾病，但CAA通常是 在AD研究中被忽视，因为A β被广泛认为是导致血管壁VSMC丢失的原因。 A β-装载血管。CAA中VSMC的丢失是由于补体系统的细胞溶解膜的形成 攻击复合物（MAC）的图尼卡中膜的A β负载CAA血管。根据我们的发现， 科学团队，我们的目标是开发一种基于抑制MAC形成的CAA治疗方法， 我们假设这将防止CAA诱导的血管脆性。我们计划以抑制MAC形成为目标， 与全身性MAC抑制相反，由于MAC对于 对微生物感染的免疫保护。我们提出了一种鼻脑纳米粒子疗法， 用二乙基乙胺（DEAE）修饰的去热原壳聚糖，其递送包封的CD59质粒 其表达废除了MAC的形成。我们已经开发出一种技术， 这使得能够进行内部放置，因为市售的壳聚糖被内毒素污染。这些 污染物干扰质粒转染和基因表达。我们的壳聚糖去热原技术 是基于氮等离子体的应用-相同的技术，用于净化潜在的 911后寄给国会的炭疽信件我们完成了第一阶段SBIR， 用含壳聚糖基因的纳米颗粒转染原代人VSMCs，诱导表面表达 通过纳米颗粒转染的VSMCs中的CD59，并建立了CD59转染细胞的成功保护 由MAC引发的细胞溶解在第二阶段SBIR中，我们打算（1）提高均匀、可重复的 DEAE壳聚糖纳米颗粒随时间推移而稳定，从而在体内实现一致的、可预测的 转染脑微血管并提供实质保护，（2）表征治疗性 我们的鼻-脑CS-CD59纳米颗粒CAA疗法在CAA的相关转基因小鼠模型中的功效， 和（3）确定增强的CAA转基因小鼠模型中的组织学、生理学和认知效应。 CD59表达。该项目的成功完成将有助于使我们的DEAE的知情翻译 壳聚糖纳米粒治疗CAA进入临床。

PROJECT SUMMARY Cerebral Amyloid Angiopathy (CAA), a microvasculopathy in which beta-amyloid (Aβ) accumulates in the walls of cerebral blood vessels, is associated with vascular fragility and bleeding secondary to blood vessel wall breakdown. CAA is especially deleterious to vascular smooth muscle cells (VSMC). CAA is found in 70-90% of Alzheimer's disease (AD) cases, increases hemorrhagic stroke risk, and is exacerbated by active amyloid immunotherapy thereby compromising this promising AD therapeutic. There is no effective therapy for CAA. Despite the prevalence of CAA in AD and the fact that AD and CAA are different diseases, CAA is often overlooked in AD studies as Aβ has been widely presumed to be responsible for the VSMC loss in the walls of Aβ-laden vessels. VSMC loss in CAA occurs due to formation of the complement system's cytolytic membrane attack complex (MAC) in the tunica media of Aβ-laden CAA blood vessels. Based on this discovery by our scientific team, our goal is to develop a first-ever therapeutic for CAA based on inhibition of MAC formation which we hypothesize will prevent CAA-induced vascular fragility. We plan to target inhibition of MAC formation in the walls of Aβ-laden CAA blood vessels, as opposed to systemic MAC inhibition, due to the importance of MAC for immune protection against microbial infection. We propose a nose-to-brain nanoparticle therapy made of depyrogenated chitosan modified with diethylethylamine (DEAE) that delivers an encapsulated CD59 plasmid whose expression abrogates the formation of MAC. We have developed a technique to depyrogenate chitosan that enables internal placement since commercially available chitosans are contaminated with endotoxins. These contaminants interfere with plasmid transfection and gene expression. Our chitosan depyrogenation technique is based on the application of nitrogen plasma – the same technology employed to decontaminate potential anthrax letters sent to Congress after 9/11. We completed a Phase I SBIR in which we established successful transfection of primary human VSMCs with chitosan gene-containing nanoparticles, induced surface expression of CD59 in VSMCs via nanoparticle transfection, and established successful protection of CD59 transfected cells from MAC-initiated cell lysis. In this Phase II SBIR, we intend to (1) improve production of uniform, reproducible DEAE chitosan nanoparticles that are stable over time so as to achieve consistent, predictable in vivo transfection of brain microvasculature and provide parenchymal protection, (2) characterize the therapeutic efficacy of our nose-to-brain CS-CD59 nanoparticle CAA therapy in a relevant transgenic mouse model of CAA, and (3) determine histologic, physiologic, and cognitive effects in the transgenic CAA mouse model of enhanced CD59 expression. Successful completion of this project will help enable informed translation of our DEAE chitosan nanoparticle therapy for CAA into the clinic.