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: 10210547
• 负责人: Daniel Kacy Cullen
• 金额: $ 42.88万
• 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/10210547
• 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; Nude Rats; Out-Migrations; Parkinson Disease; Pathway interactions; Patients; 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 由于创伤性脑损伤（TBI）造成的慢性残疾影响总人口的2%，神经元损失是 通常被认为是永久性的，这是由于成年哺乳动物大脑的神经再生能力有限。 目前还没有被批准的用于改善TBI后恢复的治疗方法，并且没有创新的方法来改善TBI后的恢复。 增强神经再生是迫切需要的。有趣的是，新的神经元产生于 脑室下区（SVZ），然后通过吻侧皮层引导至嗅球/嗅束（可能还有纹状体）。 迁移流（RMS），用于集成到现有电路中。最近的出版物表明， 成神经细胞可以被重定向到病变中，分化成区域特异性神经元细胞类型，整合到 电路，并改善成年啮齿动物的功能恢复，但一个翻译策略，以指导和增强 成神经细胞迁移到损伤中还有待建立。为了应对这一挑战，我们组织了一个 由干细胞专家、神经生物学家、临床医生和组织工程师组成的多学科团队， 第一个解剖学启发的微组织，设计用于在结构上和功能上模仿神经胶质管的神经胶质管， RMS。在一个令人兴奋的突破中，我们的团队开发了新的微组织工程技术， 星形胶质细胞自我组装成纵向排列的纤维束，以重现神经胶质细胞的组织结构 RMS管。到目前为止，我们已经生物制造了这种组织工程吻迁移流（TE-RMS） 使用啮齿动物来源的星形胶质细胞以及人类干细胞来源的星形胶质细胞，重要的是，我们 表明TE-RMS直接促进体外和体内未成熟神经元的排列和迁移， vivo.在目前的建议中，我们将首先验证TE-RMS作为体外试验床，以阐明机制 神经元祖细胞迁移和细胞命运决定（目的1）。然后我们将测试TE-RMS的能力 在体内转移内源性神经元祖细胞并修复实验性脑损伤后的大脑皮层， 大鼠TBI（目标2）。在这个目的中，TE-RMS将在急性损伤期后立体定位显微注射， 从SVZ到受损组织的跨越，以及迁移神经元的重定向以重新填充皮层区域， 将评估与残余回路的功能整合以及行为恢复的促进作用。最后作为 作为临床转化的第一步，我们将进行体外和体内研究，以验证构建的TE-RMS 使用从成年人牙龈收集的干细胞衍生的星形胶质细胞来开发 最终从容易获得的细胞来源产生自体的、患者衍生的植入物（目标3）。的 TE-RMS概括了大脑自己的新神经元的传递和整合方法。因此，执行 这些目标将大大推进翻译生物工程的方法，能够提供有针对性的， 以及神经创伤和/或变性后的持续细胞替换。我们的团队具有独特的优势， 提供了一种可行的，但高度创新的神经再生方法，可以产生重大影响， 患有其他难以治愈的TBI后果的患者。