Chronic Versus Acute Transplantation of Neural Tissues for TBI-Induced Cortical Injuries

慢性与急性神经组织移植治疗 TBI 引起的皮质损伤

• 批准号: 10657622
• 负责人: HAN-CHIAO ISAAC CHEN
• 金额: $ 40.56万
• 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: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657622
• 关键词: Acute; Acute Brain Injuries; Affect; Anatomy; Animals; Architecture; Brain; Cell Line; Cells; Cerebral cortex; Chronic; Chronic Phase; Cicatrix; Clinical; Cognitive deficits; Data; Development; Excision; Fluorescent Protein Tracings; Fostering; Future; Gliosis; Goals; Green Fluorescent Proteins; Health; Histologic; Histology; Human; In Vitro; Injury; Knowledge; Maintenance; Mission; National Institute of Neurological Disorders and Stroke; Nature; Neurologic Deficit; Neurons; Organoids; Outcome; Outcome Measure; Patients; Phase; Photic Stimulation; Process; Public Health; Rabies virus; Rattus; Recovery; Research; Sensory; Severities; Stroke; Structure; Synapses; System; Techniques; Testing; Therapeutic; Thinness; Tissue Transplantation; Tissues; Translations; Transplantation; Traumatic Brain Injury; Visual; Visual Cortex; aspirate; brain circuitry; brain repair; cell replacement therapy; cohort; controlled cortical impact; disability; efficacy evaluation; extracellular; functional outcomes; functional restoration; improved; improved outcome; in vivo; induced pluripotent stem cell; injured; innovation; nervous system disorder; neural; novel strategies; optogenetics; organoid transplantation; personalized medicine; post-transplant; repair strategy; repaired; response; stem cells; tissue repair; translational approach; visual stimulus

PROJECT SUMMARY/ABSTRACT Injury to the cerebral cortex occurs frequently across the spectrum of severity in traumatic brain injury (TBI). No therapies exist to counter the neurological and cognitive deficits caused by these injuries, which are responsible for substantial disability after TBI. A promising strategy for restoring brain function after injury is cell replacement. Neural tissues that connect with host cortex locally and function as supplementary cortical processing modules are especially intriguing candidates for this approach. Currently available tissue substrates that are suitable for translation, including human brain organoids derived from patient-matched stem cell lines, do not fully recapitulate the architecture or micro-circuitry of cortex. However, they can still be used to investigate outstanding questions regarding neural tissue integration with the host brain. One essential issue that has not been examined systemically is the optimal timing of cell replacement after TBI. The overall objective of the current proposal is to evaluate how the interplay between the timing of neural tissue transplantation after TBI and the state of the cortical microenvironment affects anatomic and functional outcomes. Our central hypothesis is that acute neural tissue transplantation after TBI and removal of the injury perimeter will improve outcomes as a result of enhanced integration of graft neurons with host brain networks and maintenance of host cortex integrity. To test this hypothesis, we will transplant human cortical organoids into rat visual cortex in the chronic or acute setting after a controlled cortical impact injury and assess anatomic and functional outcome measures. In Aim 1, organoids will be transplanted directly into a chronic injury cavity or after resection of the glial scar at the border of the cavity. In Aim 2, organoid grafts will be inserted directly into an acute injury cavity or after the injury margin as been removed. In both of these Aims, organoid health and cell composition as well as host cortex integrity will be assessed histologically. The extent of formation of graft efferents (green fluorescent protein tracing) and afferents (modified rabies virus system for retrograde trans-synaptic tracing) also will be determined. Functional integration of organoid grafts with the host cortex will be investigated using in vivo techniques for recording extracellular neural activity and visual stimulation of the host animal. In Aim 3, we will examine how modulating the activity of organoids using optogenetic stimulation impacts their connectivity and integration with the chronically or acutely injured brain. The proposed research is innovative in its use of human brain organoids as structured neural tissues for cortical repair after TBI and because it explicitly assesses how the timing of transplantation affects outcomes. We expect that the proposed studies will elucidate conditions that result in improved outcomes after organoid transplantation while also identifying the limitations of currently available neural tissue substrates. These expected outcomes will advance the field of cortical repair after TBI by reinvigorating the concept of cell replacement therapy and inspiring novel strategies for modulating graft integration with the brain to achieve specific therapeutic goals.

项目总结/摘要 在创伤性脑损伤（TBI）的严重程度范围内，大脑皮层损伤经常发生。没有 存在对抗由这些损伤引起的神经和认知缺陷的疗法， 造成严重残疾。一种有希望的恢复脑损伤后功能的策略是 细胞置换与宿主皮层局部连接并起辅助皮层作用的神经组织 处理模块是这种方法的特别有趣的候选者。目前可用的组织基质 适用于翻译的细胞，包括源自患者匹配干细胞系的人脑类器官， 并不能完全重现大脑皮层的结构或微电路。但是，它们仍然可以用于 研究有关神经组织与宿主大脑整合的悬而未决的问题。一个基本问题是 TBI后细胞置换的最佳时机是什么？整体 目前的建议的目的是评估如何之间的相互作用神经组织的时间 TBI后移植和皮质微环境的状态影响解剖和功能 结果。我们的中心假设是，TBI后急性神经组织移植和损伤的去除， 由于移植神经元与宿主脑网络的整合增强，周长将改善结果 和维持宿主皮层的完整性。为了验证这一假设，我们将移植人类皮质类器官 在慢性或急性情况下，在受控的皮质撞击损伤后， 和功能结果测量。在目标1中，类器官将直接移植到慢性损伤腔中 或在切除腔边缘处的胶质瘢痕之后。在目标2中，将直接插入类器官移植物 进入急性损伤腔或在损伤边缘被移除之后。在这两个目标中，类器官健康 并从组织学上评估细胞组成以及宿主皮质完整性。的形成程度 移植物传出（绿色荧光蛋白示踪）和传入（用于逆行的修饰的狂犬病病毒系统 跨突触追踪）也将被确定。类器官移植物与宿主皮质的功能整合将 使用记录细胞外神经活动和视觉刺激的体内技术进行研究。 宿主动物在目标3中，我们将研究如何使用光遗传学刺激来调节类器官的活性。 影响它们与慢性或急性损伤的大脑的连接和整合。拟议的研究是 创新地使用人脑类器官作为结构化神经组织，用于TBI后的皮质修复， 因为它明确评估了移植时机对结果的影响。我们预计， 研究将阐明导致类器官移植后结果改善的条件， 确定目前可用的神经组织基质的局限性。这些预期成果将 通过重振细胞替代疗法的概念，推进TBI后皮质修复领域， 激发了调节移植物与脑整合以实现特定治疗目标的新策略。

项目成果

Protocol for human brain organoid transplantation into a rat visual cortex to model neural repair.

• DOI: 10.1016/j.xpro.2023.102470
• 发表时间: 2023-09-15
• 期刊: STAR PROTOCOLS
• 作者: Jgamadze, Dennis;Harary, Paul M.;Castellanos, Mackenzie;Blue, Rachel;Song, Hongjun;Ming, Guo-li;Isaac, H.
• 通讯作者: Isaac, H.