Anti-inflammatory and hMSC combination therapy for traumatic brain injury

抗炎与hMSC联合治疗创伤性脑损伤

• 批准号: 10486391
• 负责人: SUBHRA MOHAPATRA
• 金额: --
• 依托单位: JAMES A. HALEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2026-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10486391
• 关键词: Acute; Address; Anti-Inflammatory Agents; Anxiety; Astrocytes; Attenuated; Behavioral; Brain; Brain Injuries; Brain-Derived Neurotrophic Factor; CCL20 gene; CCR6 gene; Cause of Death; Cells; Clinical; Closed head injuries; Cognitive deficits; Collaborations; Combined Modality Therapy; Communication; Consensus; Data; Dendrimers; Dose; Drug Targeting; Encephalitis; Engraftment; FDA approved; Functional disorder; Funding; Future; Genes; Gliosis; Goals; Health; Human; Human Resources; Immune; Impaired cognition; Infiltration; Inflammasome; Inflammation; Inflammatory; Injury; Integral Membrane Protein; Interferons; Interleukin-1 beta; Intravenous; Lateral; Lesion; Ligands; Liquid substance; Long-Term Effects; Memory Loss; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Methods; Microglia; Military Personnel; Modeling; Mus; Nerve Degeneration; Nerve Regeneration; Nervous System Trauma; Neurogenic Inflammation; Neurologic; Neurons; Outcome; Pathogenesis; Pathology; Patients; Percussion; Peripheral; Pharmaceutical Preparations; Phase; Pioglitazone; Plasmids; Play; Population; Prevalence; Proteins; Rattus; Reagent; Regimen; Reporting; Research; Role; Severities; Signal Transduction; Spleen; Stimulus; Surrogate Markers; TBI treatment; Testing; Therapeutic; Therapeutic Agents; Translational Research; Traumatic Brain Injury; Uncertainty; Work; adult stem cell; anxiety reduction; anxiety-like behavior; astrogliosis; behavioral outcome; chemokine; chemokine therapy; clinical translation; combat; combinatorial; depressive behavior; disability; effective therapy; evidence base; glial activation; improved; inhibitor; innovation; learned behavior; motor behavior; mouse model; nano; nanoformulation; nanoparticle; neurogenesis; neuropathology; novel; novel therapeutics; pre-clinical research; programs; rational design; receptor; response; small hairpin RNA; stem cell therapy; systemic inflammatory response; targeted treatment; therapy outcome

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. The prevalence of TBI from all causes is estimated at ~2.5 to 3.7 million cases in the USA alone. Currently, there are no FDA- approved drugs available to treat TBI. While TBI illness ranges from mild to severe, moderate TBI is clinically most relevant, and If not treated immediately the secondary injury in these patients could spread quickly and may lead to long-term consequences including cognitive and memory loss. Mesenchymal stem cells (MSCs) transplantation has emerged as one of the major approaches to treat TBI. However, the viability of sole MSC therapy is being debated due to the uncertainty of variable engraftment in inflammatory state. Hence, there is a genuine unmet need to discover and develop new therapeutic agents for all forms of TBI, especially moderate TBI. It has been reported that adding an anti-inflammatory agent prior to MSC treatment increased the latter’s efficacy. During the last funding period, we have established an experimental mouse model to enable characterization of the consequences of repeated TBI (rTBI) in closed head injury, which shows extensive neuropathologic changes associated with the cognitive deficit, consistent with human TBI. We made the striking discovery that the expression of chemokine (C-C motif) ligand 20 (CCL20) (MIP3α) is pivotal to inflammation in TBI, implicating CCL20 as a potential TBI drug target. We have demonstrated that CCL20 inhibitor, pioglitazone (PG), played a key role in reducing astrogliosis, microglial activation and inflammasome activation in TBI. A combination of PG and hMSC treatment not only reduced inflammation but also improved behavioral outcomes by increasing the expression of brain-derived neurotrophic factor (BDNF). Because PG is not a specific CCL20 inhibitor and may have off-target effects, we have established and validated a nanoformulation of short hairpin RNA (shRNA) plasmids encoding CCL20 and its receptor CCR6 using dendrimer. A combination treatment involving shCCL20/shCCR6 (shCombo) and human MSC (hMSC) showed a significant reduction in inflammation, a concomitant increase in BDNF and neurogenesis in rTBI mouse model. Based on our preliminary data, we hypothesize that the shCombo dendriplex (DPX) or/and hMSC combination regimen will attenuate both acute brain- and systemic-inflammation by decreasing activation of glial cells and astrocytes in the brain, and secreting trophic factors over a prolonged period, which in turn promotes neuronal remodeling in the damaged brain and provides sustained relief from TBI-induced pathology. Three specific aims are proposed to test this hypothesis. In Aim #1, we will determine the optimal dose of pshCombo-DPX and the therapeutic window of treatment in rTBI mice. In Aim #2 we will examine the short-term and long-term effects of shCombo-DPX or/and hMSC therapy on pathophysiology and behavioral deficits in rTBI mice. In Aim #3, we will determine the mechanism of action of CCL20 in rTBI-induced pathogenesis and behavioral deficits. This highly innovative proposal aims to investigate whether shCombo-DPX as a single agent or in combination with hMSC will reduce acute brain- and systemic-inflammation and behavioral deficits, and plans to address the mechanism of action and surrogate markers of efficacy based on genes induced during acute inflammation. The proposed translational research based on a rationally designed better anti-inflammatory therapy is expected to increase our understanding of the long-term outcome of the therapy for mild to moderate TBI and pave the way for future clinical translation.

我们研究计划的长期目标是开发安全有效的脑外伤疗法 创伤性脑损伤（TBI），这是我军在战斗中面临的一个主要健康问题。 据估计，仅在美国就有约 2.5 至 370 万例此类病例，目前 FDA 还没有相关规定。 批准用于治疗 TBI 的药物虽然 TBI 疾病范围从轻度到重度，但中度 TBI 是 临床上最相关，如果不立即治疗，这些患者的继发性损伤可能会扩散 快速并可能导致长期后果，包括认知和记忆丧失。 干细胞（MSC）移植已成为治疗 TBI 的主要方法之一。 由于不同细胞植入的不确定性，单一 MSC 疗法的可行性正在受到争论。 因此，发现和开发新疗法确实存在未满足的需求。 适用于所有形式的 TBI，尤其是中度 TBI。 据报道，在 MSC 治疗前添加抗炎剂可增加 在最后的资助期间，我们建立了一个实验小鼠模型来研究后者的功效。 能够描述闭合性颅脑损伤中重复 TBI (rTBI) 的后果，这表明 与认知缺陷相关的广泛神经病理学变化，与人类 TBI 一致。 做出了惊人的发现，趋化因子（C-C基序）配体20（CCL20）（MIP3α）的表达 是 TBI 中的关键炎症，表明 CCL20 是潜在的 TBI 药物靶标。 证明 CCL20 抑制剂吡格列酮 (PG) 在减少星形胶质细胞增生中发挥关键作用， TBI 中的小胶质细胞激活和炎症小体激活 PG 和 hMSC 联合治疗。 不仅可以减少炎症，还可以通过增加表达来改善行为结果 因为 PG 不是特定的 CCL20 抑制剂，并且可能是脑源性神经营养因子 (BDNF) 的抑制剂。 存在脱靶效应，我们建立并验证了短发夹 RNA 的纳米制剂 使用树枝状聚合物 A 组合处理编码 CCL20 及其受体 CCR6 的 (shRNA) 质粒。 涉及 shCCL20/shCCR6 (shCombo) 和人 MSC (hMSC) 的结果显示， rTBI 小鼠模型中炎症、BDNF 和神经发生的伴随增加。 根据我们的初步数据，我们捕获了 shCombo dendriplex (DPX) 或/和 hMSC 联合方案将通过以下方式减轻急性脑部和全身炎症： 减少大脑中神经胶质细胞和星形胶质细胞的活化，并在一段时间内分泌营养因子 长时间，这反过来又促进受损大脑中的神经元重塑，并提供 提出了三个具体目标来检验这一假设。 目标#1，我们将确定 pshCombo-DPX 的最佳剂量和治疗窗口 在 rTBI 小鼠中，我们将检查 shCombo-DPX 或/和的短期和长期影响。 hMSC 疗法对 rTBI 小鼠病理生理学和行为缺陷的影响 在目标#3 中，我们将确定。 CCL20 在 rTBI 诱导的发病机制和行为缺陷中的作用机制。 这项高度创新的提案旨在研究 shCombo-DPX 是否作为单一药物或 与 hMSC 结合将减少急性脑部和全身炎症以及行为缺陷， 并计划根据诱导的基因来解决作用机制和功效替代标记 所提出的基于合理设计的更好的转化研究。 抗炎治疗有望增加我们对长期结果的了解 治疗轻至中度 TBI 并为未来的临床转化铺平道路。