Tissue Engineered Rostral Migratory Stream for Directed Neuronal Replacement

用于定向神经元替换的组织工程嘴侧迁移流

• 批准号: 10373065
• 负责人: Daniel Kacy Cullen
• 金额: $ 53.69万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10373065
• 关键词: Acute; Address; Adult; Affect; Aging; Alzheimer' s Disease; Anatomy; Area; Astrocytes; Autologous; Beds; Behavioral; Biomedical Engineering; Brain; Brain Injuries; Cells; Cerebral cortex; Chronic; Chronic Brain Injury; Clinical; Cognitive; Corpus striatum structure; Cues; Dependence; Development; Engineering; Exhibits; Future; Gingiva; Harvest; Human; Implant; In Vitro; Injury; Lateral; Lesion; Location; Mesenchymal Stem Cells; Methods; Morphology; Natural regeneration; Nerve Degeneration; Nerve Regeneration; Nervous System Trauma; Neurodegenerative Disorders; Neurons; Neurosphere; Nude Rats; Out-Migrations; Parkinson Disease; Pathway interactions; Patients; Persons; Population; Positioning Attribute; Pre-Clinical Model; Prosencephalon; Proteins; Publications; Punch Biopsy; Rattus; Recovery; Recovery of Function; Regenerative Medicine; Residual state; Rodent; Signal Transduction; Site; Source; Specialist; Stream; Structure; Target Populations; Techniques; Testing; Tissue Engineering; Tissues; Traumatic Brain Injury; Traumatic Brain Injury recovery; Tube; base; biofabrication; cell type; clinical translation; controlled cortical impact; dentate gyrus; design; disability; efficacy evaluation; efficacy testing; human stem cells; improved; in vitro testing; in vivo; injury recovery; innovation; insight; migration; minimally invasive; motor deficit; multidisciplinary; nerve stem cell; neural circuit; neuroblast; neurogenesis; neuron loss; neuronal replacement; newborn neuron; non-genetic; novel; olfactory bulb; prevent; protein expression; repaired; self assembly; stem cell migration; stem cells; subventricular zone; tissue stem cells; translational approach

PROJECT SUMMARY Chronic disability due to traumatic brain injury (TBI) affects 2% of the total population, and neuronal loss is generally considered permanent, owing to limited capacity for neuroregeneration in the adult mammalian brain. There are currently no approved treatments for improving recovery after TBI, and innovative approaches to enhance neuroregeneration are desperately needed. Intriguingly, new neurons are generated in the subventricular zone (SVZ) and then guided to the olfactory bulb/tract (and possibly striatum) via the rostral migratory stream (RMS) for integration into existing circuitry. Recent publications have demonstrated that SVZ neuroblasts can be redirected into lesions, differentiate into region-specific neuronal cell types, integrate into circuitry, and improve functional recovery in adult rodents, but a translational strategy to direct and enhance neuroblast migration into lesions has yet to be established. To address this challenge, we have assembled a multi-disciplinary team of stem cell specialists, neurobiologists, clinicians, and tissue engineers to develop the first anatomically-inspired microtissue designed to structurally and functionally emulate the glial tube of the RMS. In an exciting breakthrough, our team developed novel microtissue engineering techniques that promote the self-assembly of astrocytes into longitudinally aligned bundles that recapitulate the organization of the glial tube of the RMS. To date, we have biofabricated this Tissue Engineered Rostral Migratory Stream (TE-RMS) using rodent derived astrocytes as well as human stem cell derived astrocytes and, importantly, we have shown that the TE-RMS directly facilitates the alignment and migration of immature neurons in vitro and in vivo. In the current proposal, we will first validate the TE-RMS as an in vitro test bed to elucidate mechanisms of neuronal progenitor migration and cell fate determination (Aim 1). We will then test the ability of the TE-RMS to divert endogenous neuronal progenitors in vivo and repair damaged cerebral cortex following experimental TBI in rats (Aim 2). In this Aim, the TE-RMS will be stereotaxically microinjected after the acute injury period to span from the SVZ into lesioned tissue, and the redirection of migrating neurons to repopulate cortical areas, functional integration with residual circuitry, and facilitation of behavioral recovery will be assessed. Finally, as a first step towards clinical translation, we will perform in vitro and in vivo studies to validate the TE-RMS built using astrocytes derived from stem cells harvested from adult human gingiva to develop methods for the eventual creation of autologous, patient-derived implants from an easily accessible cell source (Aim 3). The TE-RMS recapitulates the brain's own method for delivery and integration of new neurons. Thus, the execution of these Aims will significantly advance a translational bioengineering approach capable of providing targeted and sustained cell replacement following neurotrauma and/or degeneration. Our team is uniquely positioned to provide a feasible, yet highly innovative neuroregenerative approach that can have a significant impact on patients suffering from the otherwise intractable consequences of TBI.

项目摘要 由于脑外伤引起的慢性残疾（TBI）影响总人口的2％，而神经元丧失为 由于成人哺乳动物大脑中神经发生的能力有限，通常被认为是永久性的。 目前尚无批准的治疗方法来改善TBI之后的恢复，以及创新的方法 迫切需要增强神经创成。有趣的是，在 脑室下区（SVZ），然后引导到嗅球/小块（以及可能的纹状体）， 迁移流（RMS）集成到现有电路中。最近的出版物表明SVZ 神经细胞可以重定向到病变中，分化为特定区域的神经元细胞类型，集成到 电路，并改善成年啮齿动物的功能恢复，但是指导和增强的转化策略 神经细胞迁移到病变尚未确定。为了应对这一挑战，我们组装了 干细胞专家，神经生物学家，临床医生和组织工程师的多学科团队 首先是在解剖学启发的微动物 RMS。在一个令人兴奋的突破中，我们的团队开发了促进的小型微动工工程技术 星形胶质细胞的自组装成纵向对齐的束，概括了神经胶质的组织 RMS的管。迄今为止，我们已经生物制作了该组织工程的鼻迁移流（TE-RMS） 使用啮齿动物衍生的星形胶质细胞以及人类干细胞衍生的星形胶质细胞，重要的是，我们有 表明TE-RM直接促进了未成熟神经元在体外和IN的对齐和迁移 体内。在当前的提案中，我们将首先验证TE-RMS作为体外测试床以阐明机制 神经元祖细胞迁移和细胞命运的确定（目标1）。然后，我们将测试TE-RMS的能力 在实验后，在体内转移内源性神经元祖细胞和修复受损的脑皮质 大鼠的TBI（AIM 2）。在此目的中，TE-RMS将在急性受伤期之后进行立体定位显微注射到 从SVZ到病变的组织，并将迁移神经元重定向以重新填充皮质区域， 将评估与残余电路的功能整合，并评估行为恢复的促进。最后，如 迈向临床翻译的第一步，我们将在体外和体内进行研究以验证构建的TE-RMS 使用源自从成年人类牙龈收获的干细胞的星形胶质细胞来开发用于 最终从易于访问的细胞来源创建自体源性植入物（AIM 3）。这 TE-RMS概括了大脑自身传递和整合新神经元的方法。因此，执行 这些目标将大大提高一种能够提供目标的转化生物工程方法 神经瘤和/或变性后持续的细胞置换。我们的团队独特地定位 提供一种可行但高度创新的神经增合方法，可以对 患有原本棘手的TBI后果的患者。