Retinal tissue derived from human pluripotent stem cells for vision restoration

摘要 该项目将测试人类胚胎干细胞衍生的3-D视网膜组织移植到视网膜下空间 两个中的一个 在一项长达12个月的纵向研究中，建立了光感受器(PR)变性的动物模型。我们会 使用具有CRX突变和免疫缺陷的早发性RD的大眼睛动物模型(Rdy阿比西尼亚猫) PR变性大鼠SD-Foxn1 TG(S334ter)3Lav(RD裸鼠)。眼下的目标是证明 移植物与受体视网膜的存活、分层、成熟以及结构和突触的整合， 这个 和 视力改善6-12个月， 并生成标准操作程序，以将这种 将技术转化为治疗公关死亡所致失明的方法。首要目标是评估这一点 对视网膜色素变性(RP)患者进行试点临床试验的技术，这是FDA的快车道孤儿疾病 批核 。失明是美国人的一个主要健康问题，影响着生活质量，经济负担也很高 和 沉重的情绪 负担。与以下疾病相关的两种主要视网膜疾病 光感受器 退变是 RP和干性老年黄斑变性(AMD)。 迫切需要找到新的治疗方法。 基于光感受器替代的RP和AMD。 过去15-20年的研究表明， 退化的哺乳动物视网膜可以被健康的胎儿视网膜组织取代，这可以改善 幻象。研究表明， 嫁接 胎儿视网膜将完成分化，在受体视网膜上突触 神经节细胞神经元，并重新建立与视觉皮质的连接。人胎视网膜组织 药物的供应非常有限，从伦理上讲，它在治疗中的临床应用是不可接受的。BioTime假设 6-8- 周岁 人胚胎干细胞来源的三维视网膜组织 将要 在结构上和语法上整合到 变性的受者视网膜和 改善视力 在动物身上 高级研发 。我们预测，示威活动 HESC-3D视网膜组织移植对“大眼”动物模型的积极治疗作用 RD将会 启用 美国将搬家 这 在RP患者中进行临床试验的技术。我们已经开发出hESC-3D视网膜 组织，并证明它携带一层RPE， 无线市话 ，二级神经元和神经节细胞，能够 我们还报道了它 改善RP致盲大鼠动物模型的视力，并激活上丘。我们提议去做 在SD-Foxn1大眼盲动物模型中的活体测试 Tg(S334ter)3Lav(RD裸鼠)对RD大鼠进行治疗，以统计评价该疗法治疗RP患者的可行性。在……里面 目标1(BioTime)，我们将从cGMP级hESCs中放大生产hESC-3D视网膜组织并评估 几个批次使用了FDA的标准。在目标2(UCI)中，我们将对失明大鼠进行视网膜下移植，并评估视力 改进和移植物-宿主连接。在目标3(MSU)中，我们将为Rdy/+猫进行视网膜下移植，并 还要评估视力改善和移植物与宿主的连通性。结果和程序将整合到 实现PR变性致盲的临床试验的临床方案 轴突发生，突触发生，并逐渐变得活跃的电。 。

ABSTRACT This project will test human embryonic stem cell-derived 3-D retinal tissue transplanted in subretinal space of 2 animal models with photoreceptor (PR) degeneration in a longitudinal study lasting for up to 12 months. We will use a large-eye animal model of early-onset RD with a Crx mutation (Rdy Abyssinian cats) and immunodeficient rats with PR degeneration SD-Foxn1 Tg(S334ter)3Lav (RD nude). The immediate goal is to demonstrate survival, lamination, maturation, and structural and synaptic integration of grafts with the recipient retina, the and improvement in vision by 6-12 months, and to generate standard operating procedures for transforming such technology into therapy to treat blindness caused by PR death. The overarching goal is to evaluate this technology in pilot clinical trials in patients with retinitis pigmentosa (RP), an orphan disease with fast track FDA approval . Blindness is a major health concern among Americans, affecting quality of life and with a high financial and heavy emotional burden. The two major retinal diseases associated with photoreceptor degeneration are RP and dry age-related macular degeneration (AMD). There is a critical and urgent need to find new treatments of RP and AMD based on photoreceptor replacement. The research in the past 15-20 years indicates that a piece of degenerating mammalian retina can be replaced with healthy fetal retinal tissue, which can improve vision. The research shows that grafted fetal retina will complete differentiation, synapse on the recipient retinal ganglion cell neurons, and re-establish connectivity with the visual cortex. Human fetal-derived retinal tissue supply is very limited, and its clinical use in therapy is ethically not acceptable. BioTime hypothesizes that 6-8- week old human embryonic stem cell-derived 3-D retinal tissue will integrate structurally and synaptically into the degenerating recipient retina and improve vision in animals with advanced RD . We predict that demonstrating positive therapeutic impact of hESC-3D retinal tissue grafting in a “large eye” animal model with RD will enable us to move this technology to pilot clinical trials in RP patients. We have already developed hESC-3D retinal tissue and demonstrated that it carries a layer of RPE, PRs , second order neurons and ganglion cells, is capable We also reported that it improves vision in a rat animal model with RP and blindness and activates superior colliculus . We propose to do in vivo testing in a “large eye” animal model of RD (Rdy cats) with blindness, and in a large cohort of SD-Foxn1 Tg(S334ter)3Lav (RD nude) rats with RD to statistically evaluate the feasibility of this therapy in RP patients. In Aim 1 (BioTime), we will scale-up production of hESC-3D retinal tissue from cGMP-grade hESCs and evaluate several lots using FDA criteria. In Aim 2 (UCI), we will do subretinal grafting into blind rats and evaluate vision improvements and graft-host connectivity. In Aim 3 (MSU), we will do subretinal grafting into Rdy/+ cats and also evaluate vision improvements and graft-host connectivity. The results and procedures will be integrated into clinical protocols to enable clinical trials of blindness caused by PR degeneration of axonogenesis, synaptogenesis and becomes progressively electrically active. .