Toward enhancing organization and defining synaptic connectivity of transplanted human pluripotent stem cell-derived photoreceptor grafts

旨在增强移植的人多能干细胞衍生光感受器移植物的组织和定义突触连接

• 批准号: 9911500
• 负责人: Allison Lyn Ludwig
• 金额: $ 4.17万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9911500
• 关键词: 3-Dimensional; Address; Affect; Age related macular degeneration; Anatomy; Animal Model; Animals; Area; Behavior; Biocompatible Materials; Biological Assay; Biomedical Engineering; Blindness; Bolus Infusion; Cell Count; Cell Death; Cells; Clinic; Clinical; Clinical Trials; Clinical Trials Design; Coculture Techniques; Collaborations; Communities; Complex; Custom; Development; Disease; Disease model; Doctor of Philosophy; Donor person; Dose; Environment; Face; Fellowship; Fostering; Foundations; Future; Goals; Home environment; In Vitro; Inherited; Injections; Institution; Leadership; Measurement; Mentorship; Methodology; Methods; Modeling; National Eye Institute; Natural regeneration; Nerve Regeneration; Ophthalmology; Organoids; Patients; Pattern; Photoreceptors; Physiological; Positioning Attribute; Production; Rattus; Reflux; Regenerative Medicine; Replacement Therapy; Reproducibility; Research; Research Personnel; Research Proposals; Research Training; Retina; Retinal Degeneration; Retinal Diseases; Retinal Dystrophy; Safety; Scientist; Source; Synapses; Technology; Testing; Therapeutic; Thinking; Tissues; Training; Training Support; Transplantation; Universities; Vision; Visual; Visual impairment; Wisconsin; Xenograft procedure; bench to bedside; career; design; fetal; human pluripotent stem cell; improved; in vivo; innovation; migration; multidisciplinary; next generation; novel; novel therapeutics; photoreceptor degeneration; polarized cell; polyglycerol; pre-clinical; preclinical study; reconstitution; response; restoration; retinal regeneration; safety study; scaffold; skills; stem cell biology; stem cell technology; success; synaptic function; synaptogenesis; targeted delivery; vision science

PROJECT SUMMARY Outer retinal degenerative diseases (RDDs) resulting in photoreceptor (PR) cell death are a leading cause of visual impairment worldwide, but options for rescuing or restoring vision in many of these patients are limited. Human pluripotent stem cell (hPSC)-derived PR transplantation is increasingly being studied as a therapeutic strategy for these patients, and neural regeneration within the retina has recently been identified as an area of strategic focus by the National Eye Institute (NEI). Several preclinical studies have shown some degree of visual restoration with bolus-delivered PR transplants in animal models, and clinical trials studying the safety and efficacy of bolus-delivered fetal-derived retinal precursors in patients with severe retinal degeneration are currently underway. Despite these recent successes, the field still faces several critical roadblocks before clinical PR replacement therapy can be realized for most RDD patients. Current strategies for bolus subretinal delivery of dissociated PRs fail to accurately reconstitute the complex organization of the outer retina, and they are often accompanied by disorganization, unpredictable dosing, and overall low cell counts immediately after injection due to reflux into the vitreous cavity. Further, it remains unclear whether visual responses commonly attributed to transplanted donor PRs are actually due to anatomic integration and functional synapse formation within the host degenerate retina. Indeed, the efficiency of synapse formation following PR transplantation, and the relationship between de novo synaptogenesis and measurements of visual function has not been tested to date. Here, we seek to use state-of-the-art biomaterials and PR scaffolds along with rigorous synaptic tracing methodologies to address these challenges in a rat model of severe photoreceptor degeneration. In Aim 1, we will use a novel micro-patterned, biodegradable scaffold for targeted hPSC-PR transplantation to assess the retention, survival, and maturation of bolus-delivered and scaffold-delivered PRs in vivo. In Aim 2, we will define the synaptic connectivity of hPSC-PRs in degenerate retinal explants and live host degenerate retinal tissue with an innovative monosynaptic retrograde tracing assay. The University of Wisconsin-Madison fosters the ideal scientific and intellectual environment for successful completion of these aims with strong, collaborative research communities spanning the fields of ophthalmology, biomedical engineering, and regenerative medicine. The research proposal and fellowship training plans detailed here seek to address current roadblocks within the field of PR replacement while also providing the necessary skillset to address the next generation of challenges facing the burgeoning field of retinal regeneration.

项目摘要 导致光感受器（PR）细胞死亡的视网膜外变性疾病（RDD）是主要原因 全球视觉障碍，但是许多患者的拯救或恢复视力的选择受到限制。 人类多能干细胞（HPSC）衍生的PR移植被越来越多地研究为治疗性 这些患者的策略以及视网膜内的神经再生最近被确定为 国家眼科研究所（NEI）的战略重点。一些临床前研究表明 在动物模型中使用推注的PR移植的视觉恢复，以及研究安全性的临床试验 和推注胎儿衍生的视网膜前体在严重视网膜变性患者中的功效是 目前正在进行中。尽管最近取得了这些成功，但该领域仍然面临着几个关键的障碍 对于大多数RDD患者，可以实现临床PR替代疗法。注网下注网策略 解散的PR的交付无法准确重建外部视网膜的复杂组织，他们 通常伴随着混乱，不可预测的给药和整体低细胞计数。 注射导致玻璃体腔。此外，尚不清楚视觉响应是否常见 归因于移植的供体PRS实际上是由于解剖整合和功能突触的形成引起的 在主机退化视网膜内。实际上，PR移植后突触形成的效率，以及 从头突触发生与视觉功能的测量之间的关系尚未测试 日期。 在这里，我们寻求使用最先进的生物材料和公关支架以及严格的突触跟踪 在严重感光器变性的大鼠模型中解决这些挑战的方法。在AIM 1中，我们 将使用一种新型的微观图案，可生物降解的支架进行靶向HPSC-PR移植来评估 在体内保留和脚手架的PR的保留，生存和成熟。在AIM 2中，我们将 定义退化视网膜外植体中HPSC-PR的突触连通性和现场宿主退化视网膜 具有创新的单突触逆行跟踪测定法的组织。威斯康星大学麦迪逊分校养育者 成功完成这些目标的理想科学和知识环境， 跨越眼科，生物医学工程和 再生医学。这里详细介绍的研究建议和奖学金培训计划寻求解决 当前的PR替换领域内的障碍，同时还提供了必要的技能来解决 下一代视网膜再生领域面临的挑战。