Targeted Nano-therapeutics for Neural Regeneration

项目主任/首席调查员(最后、第一、中间)：Vyavahare Narendra R. 项目摘要 创伤性脑损伤(TBI)引发复杂的生理反应，涉及两个进行性组织 修复过程的损伤和激活，包括神经发生、血管生成和可塑性 轴突萌发和突触重组。成人大脑重塑的有限能力是 可归因于与年龄相关的外源性神经微环境的变化，如髓鞘堆积- 相关抑制物(MAI)和硫酸软骨素蛋白多糖(CSPGs)，以及 神经元生化，如环磷酸腺苷(CAMP)水平。几个组织已经开始调查 使用抗MAIS、CSPG-1的单抗克服这些障碍的治疗策略 降解酶和调节cAMP水平的药物。尽管结果令人鼓舞，但临床试验 这些方法的翻译受到侵入性递送方法、移植 异种细胞，以及细菌衍生酶的使用。这个项目的目标是开发新的 用于药物和小干扰RNA(SiRNA)联合递送的神经元特异性纳米疗法 针对神经可塑性的外在和内在障碍。这些纳米疗法将由聚合体 (lactide-co-glycolide)-graft-polyethyleneimine(PGP)共聚胶束负载罗利普兰， 磷酸二酯酶抑制剂在疏水核心稳定神经元cAMP水平，2)siRNA结合 靶向RhoA的阳离子外壳，RhoA是一种由多个神经元生长激活的细胞内信号分子 抑制剂，以及3)抗Nogo受体的单抗(MNgR1)，它与神经元的MAI结合 靶向和抑制MAI/受体结合。具体目标是：1.合成和评价PGP- MNgR1纳米粒作为药物和siRNA载体，2.评价PgP-mNgR1纳米治疗的能力 负载罗利普兰和RhoA siRNA抑制RhoA表达，升高cAMP并刺激轴突 在抑制底物上生长，以及3.评估神经可塑性和功能恢复对 在脑外伤大鼠皮质直接撞击模型中纳米治疗药物的输送。这些研究将依赖于 与细胞、组织和分子分析核心以及生物工程和生物成像核心合作 材料表征、细胞反应分析、动物成像以及PI的专业知识 在基础和临床神经科学方面指导马克·金迪博士和迈克尔·林恩博士。通过这些研究，我们将 开发PGP作为一种靶向、组合的药物输送系统，能够解决复杂的病理 TBI的。由于它们的模块化设计，这些NPs可以被修饰以用于各种药物/siRNA和靶向 配体，为各种病理提供了广泛的适用性。 OMB编号0925-0001/0002(08/12版批准至2015年8月31日)页面续格式页面

Program Director/Principal Investigator (Last, First, Middle): Vyavahare Narendra R. Project Summary Traumatic brain injury (TBI) initiates a complex physiological response involving both progressive tissue damage and activation of reparative processes including neurogenesis, angiogenesis, and plasticity through axonal sprouting and synaptic reorganization. The limited capacity for remodeling in the adult brain is attributable to age-related changes in the extrinsic neuronal microenvironment such as accumulation of myelin- associated inhibitors (MAIs), and chondroitin sulfate proteoglycans (CSPGs), as well as intrinsic changes in neuronal biochemistry such as cyclic AMP (cAMP) levels. Several groups have begun to investigate therapeutic strategies to overcome these barriers using monoclonal antibodies against MAIs, CSPG- degradative enzymes, and drugs that modulate cAMP levels. Despite encouraging results, the clinical translation of these approaches is limited by the need for invasive delivery methods, transplantation of xenogenic cells, and use of bacterially-derived enzymes. The objective of this project is to develop novel neuron-specific nanotherapeutics for combinatorial delivery of drug and small interfering RNA (siRNA) targeting both extrinsic and intrinsic barriers to neuroplasticity. These nanotherapeutics will consist of poly (lactide-co-glycolide)-graft-polyethyleneimine (PgP) copolymer micelles loaded with 1) rolipram, phosphodiesterase inhibitor in the hydrophobic core to stabilize neuronal cAMP levels, 2) siRNA bound to the cationic shell targeting RhoA, an intracellular signaling molecule activated by multiple neuronal growth inhibitors, and 3) a monoclonal antibody against the Nogo receptor (mNgR1) that binds MAIs for neuronal targeting and inhibition of MAI/receptor binding. The Specific Aims are: 1. to synthesize and evaluate PgP- mNgR1 nanoparticles as a drug and siRNA carrier, 2. to evaluate the ability of PgP-mNgR1 nanotherapeutics loaded with rolipram and RhoA siRNA to inhibit RhoA expression, elevate cAMP, and stimulate neurite outgrowth on inhibitory substrates, and 3. to evaluate neuroplasticity and functional recovery in response to delivery of nanotherapeutics in a rat direct cortical impact model of TBI. These studies will rely upon close collaboration with the Cell, Tissue, and Molecular Analysis Core and Bioengineering and Bioimaging Core for material characterization, analysis of cell response, and animal imaging and on the expertise of the PI's mentors Drs. Mark Kindy and Michael Lynn in basic and clinical neuroscience. Through these studies, we will develop PgP as a targeted, combinatorial drug-delivery system capable of addressing the complex pathology of TBI. Due to their modular design, these NPs can be modified for use with various drugs/siRNA and targeting ligands, providing broad applicability to a diverse range of pathologies. OMB No. 0925-0001/0002 (Rev. 08/12 Approved Through 8/31/2015) Page Continuation Format Page