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三维视网膜组织移植到视网膜下空间 为2 在持续长达12个月的纵向研究中，使用具有光感受器（PR）变性的动物模型。我们将 使用具有Crx突变的早发性RD的大眼动物模型（Rdy阿比西尼亚猫）和免疫缺陷 SD-Foxn 1 Tg（S334 ter）3Lav（RD裸）PR变性大鼠。当前的目标是证明 存活、分层、成熟以及移植物与受体视网膜的结构和突触整合， 的 和 视力改善6-12个月， 并制定标准操作程序来改造此类设备 技术用于治疗由PR死亡引起的失明。总体目标是评估这一点 在视网膜色素变性（RP）患者中进行的先导性临床试验中使用的技术，RP是FDA快速通道的孤儿疾病 批准 .失明是美国人的一个主要健康问题，影响生活质量， 和 沉重的情感 负担两种主要的视网膜疾病与 感光体 变性的 RP和干性年龄相关性黄斑变性（AMD）。 我们迫切需要找到新的治疗方法 RP和AMD的光感受器替代。 近15-20年的研究表明， 一块退化的哺乳动物视网膜可以用健康的胎儿视网膜组织代替，这可以改善 视野研究表明 接枝 胎儿视网膜将完成分化，在受体视网膜上形成突触 神经节细胞神经元，并重新建立与视觉皮层的连接。人胚胎视网膜组织 供应非常有限，其临床治疗用途在伦理上是不可接受的。BioTime推测， 6-8- 周龄 人胚胎干细胞衍生三维视网膜组织 将 在结构上和突触上整合到 退化的受体视网膜， 改善视力 动物中 高级RD .我们预测， hESC-3D视网膜组织移植在“大眼睛”动物模型中的积极治疗作用， 研发部将 使 我们搬 这 在RP患者中进行临床试验的技术。我们已经开发了hESC-3D视网膜 组织，并证明它携带一层RPE， PRS 二级神经元和神经节细胞， 我们还报道说， 改善RP和失明大鼠动物模型的视力，并激活上级丘。我们建议 在具有失明的RD（Rdy猫）的“大眼睛”动物模型中以及在SD-Foxn 1的大队列中的体内测试 Tg（S334 ter）3Lav（RD裸）大鼠与RD进行统计学评价，以评估该疗法在RP患者中的可行性。在 目标1（BioTime），我们将扩大从cGMP级hESC生产hESC-3D视网膜组织，并评估 使用FDA标准的几个批次。在目标2（UCI）中，我们将对盲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，我们将对Rdy/+猫进行视网膜下移植， 还评估视力改善和移植物-宿主连接。结果和程序将纳入 临床方案，使临床试验的失明所造成的PR退化 轴突发生，突触发生，并逐渐变得电活跃。 .

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. .