Combined Nano and Cell therapy for the Treatment of TBI

 描述（由申请人提供）： 我们研究计划的长期目标是为创伤性脑损伤（TBI）开发安全有效的治疗方法，这是我们战斗中军事人员的主要健康问题，估计有150，000 - 300，000名来自伊拉克自由行动和持久自由行动的军事人员患有某种程度的TBI。据估计，仅在美国，所有原因引起的TBI的患病率就为250万至370万。目前，没有药物可用于治疗TBI。虽然TBI疾病的范围从轻度到重度，但中度TBI在临床上是最相关的，并且如果不进行治疗，其通常进展为难以治疗的重度TBI。因此，确实需要发现用于中度TBI的新治疗剂。包括间充质干细胞（MSC）在内的成体干细胞疗法正在研究中，然而，由于使用不同的模型和技术，递送途径的不确定性，可变的植入和潜在的不良事件，因此对这种MSC疗法的可行性仍然缺乏共识。因此，发现和开发新的治疗靶点仍然是一个主要的未满足的需求。我们发现趋化因子（C-C基序）配体20（CCL 20）（MIP 3 α）的表达在啮齿类动物LFP-TBI（一种侧向液压冲击（LFP）损伤模型）后的炎症中起关键作用，暗示CCL 20是潜在的TBI药物靶点。我们的数据表明，TBI的急性期涉及来自外周的CCL 20驱动的炎性细胞的流入，这些炎性细胞似乎在随后的神经炎症和神经变性中发挥作用。为了开发基于CCL 20的用于TBI的纳米基因药物，我们合成了多功能壳聚糖-聚乙烯亚胺（PEI）磁性胶束（CPI-PEG），以将药物和基因递送到受损的大脑。此外，我们已经发现鼻内递送的MSC在TBI后容易迁移到脑和脾，并减少受损脑中的反应性星形胶质细胞增生。这些结果使我们假设，包括在TBI后1-2天鼻内/静脉内施用CPI 40以递送针对CCL 20的小干扰RNA（siRNA），随后在TBI后3-10天MSC疗法的组合纳米细胞疗法将分别改善急性炎症和继发性神经元损伤，从而为中度TBI提供有效治疗。提出了三个具体目标来检验这一假设。在目标#1中，我们将合成和表征靶向CCL 20信号传导的TRIC（靶向的、活性氧物质诱导的CPMM）纳米颗粒，并检查CCL 20抑制对LFP诱导的TBI病理的影响。在目标#2中，我们将评估为MSC递送提供更好的治疗窗口的递送途径以及MSC递送在修复LFP诱导的中度TBI后大鼠脑中的神经功能缺陷中的功效。在目标#3中，我们计划评估两步纳米细胞治疗方法对中度TBI的治疗效果。我们将比较靶向CCL 20/CCR 6的TRIC NP随后MSC治疗在改善中度TBI后的缺陷方面的功效，并确定作用机制。这种涉及携带CCL 20信号传导抑制剂的NP和用于治疗LFP诱导的中度TBI的hMSC疗法的两步疗法的高度跨学科和创新方法是高度创新的。我们预测，拟议的研究将增加我们对TRIC-siCCL 20治疗TBI的潜力和作用的了解。所有必要的试剂、方法和合作都已到位，我们的调查团队已准备好进行拟议的研究。

 DESCRIPTION (provided by applicant): The long-term goal of our research program is to develop safe and effective therapies for traumatic brain injury (TBI), which is a major health issue with our military personnel in combat, with estimates of 150,000-300,000 military personnel from Operation Iraqi Freedom and Operation Enduring Freedom suffering some level of TBI. The prevalence of TBI from all causes has been estimated to be 2.5 to 3.7 millions in the USA alone. Currently, there are no drugs available to treat TBI. While TBI illness ranges from mild to severe, moderate TBI is clinically most relevant, and if left untreated it often progresses to severe TBI, which is difficult to treat Hence, there is a genuine need to discover new therapeutic agents for moderate TBI. Adult stem cell therapy including mesenchymal stem cells (MSCs), is under investigation, however, there remains a lack of consensus about the viability of this MSC therapy due to use of different models and techniques, uncertainty about routes of delivery, variable engraftment and the potential for adverse events. Thus, discovering and developing new therapeutic targets remains a major unmet need. We made the striking discovery that expression of chemokine (C-C motif) ligand 20 (CCL20) (MIP3α) is pivotal to inflammation following LFP-TBI (a lateral fluid percussion (LFP) injury model) in rodents, implicating CCL20 as a potential TBI drug target. Our data demonstrates that the acute phase of TBI involves an influx of CCL20-driven inflammatory cells from the periphery that appear to play a role in subsequent neuroinflammation and neurodegeneration. Toward developing a CCL20-based nanogene drug for TBI, we have synthesized multifunctional chitosan- polyethyleneimine (PEI) magnetic micelles (CPMMs) to deliver drugs and genes to the injured brain. In addition, we have found that intranasally delivered MSCs readily migrate to brain and spleen post TBI and reduce reactive astrogliosis in the injured brain. These results have led us to hypothesize that a combinatorial nano-cell therapy comprising intranasal/intravenous administration of CPMMs 1-2 days after TBI to deliver small interfering RNAs (siRNAs) against CCL20 followed by MSC therapy 3-10 days after TBI would respectively, ameliorate acute inflammation and secondary neuronal injury thus providing an effective treatment for moderate TBI. Three specific aims are proposed to test this hypothesis. In Aim #1, we will synthesize and characterize TRIC (targeted, reactive oxygen species-inducible CPMM) nanoparticles targeting CCL20 signaling and examine the effects of CCL20 inhibition on LFP-induced TBI pathology. In Aim #2, we will evaluate the route of delivery that provides a better therapeutic window for MSC delivery and the efficacy of MSC delivery in repairing neurological function deficits in the rat brain after LFP-induced moderate TBI. In Aim #3, we plan to evaluate therapeutic efficacy of a two-step nano-cell treatment approach for moderate TBI. We will compare the efficacy of TRIC NPs targeting CCL20/CCR6 followed by MSC therapy in ameliorating deficits following moderate TBI and determine mechanisms of action. This highly interdisciplinary and innovative approach of a two-step therapy involving NPs carrying the inhibitor(s) of CCL20 signaling and hMSC therapy for treatment of LFP-induced moderate TBI is highly innovative. We predict that the proposed research will increase our understanding of the potential and role of TRIC-siCCL20 for TBI. All the necessary reagents, methods and collaborations are in place and our investigative team is uniquely poised to conduct the proposed studies.