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.

项目概要 导致光感受器 (PR) 细胞死亡的外视网膜退行性疾病 (RDD) 是导致光感受器 (PR) 细胞死亡的主要原因 全世界范围内都有视力障碍，但对许多患者来说，挽救或恢复视力的选择是有限的。 人类多能干细胞 (hPSC) 衍生的 PR 移植作为一种治疗方法越来越多地被研究 针对这些患者的策略，视网膜内的神经再生最近被确定为一个领域 国家眼科研究所 (NEI) 的战略重点。多项临床前研究表明，在一定程度上 在动物模型中通过推注 PR 移植进行视力恢复，以及研究安全性的临床试验 推注胎儿源性视网膜前体细胞对严重视网膜变性患者的疗效 目前正在进行中。尽管最近取得了这些成功，但该领域在之前仍面临几个关键障碍 大多数RDD患者都可以实现临床PR替代治疗。当前视网膜下推注策略 分离的 PR 的传递无法准确地重建外视网膜的复杂组织，并且它们 通常伴随着混乱、不可预测的剂量和细胞总数低 由于回流到玻璃体腔而导致注射。此外，目前还不清楚视觉反应是否普遍存在 归因于移植供体 PR 的原因实际上是由于解剖整合和功能性突触形成 宿主视网膜内发生退化。事实上，PR 移植后突触形成的效率，以及 从头突触发生和视觉功能测量之间的关系尚未经过测试 日期。 在这里，我们寻求使用最先进的生物材料和 PR 支架以及严格的突触追踪 在严重光感受器退化的大鼠模型中解决这些挑战的方法。在目标 1 中，我们 将使用一种新型微图案、可生物降解支架进行靶向 hPSC-PR 移植，以评估 推注递送和支架递送的 PR 在体内的保留、存活和成熟。在目标 2 中，我们将 定义退化视网膜外植体和活宿主退化视网膜中 hPSC-PR 的突触连接 组织与创新的单突触逆行追踪测定。威斯康星大学麦迪逊分校培养 成功实现这些目标的理想科学和智力环境，具有强大的、 跨眼科、生物医学工程和 再生医学。这里详述的研究提案和奖学金培训计划旨在解决 解决 PR​​ 替代领域当前的障碍，同时还提供必要的技能来解决 视网膜再生这一新兴领域面临着下一代挑战。