OPTIMIZING SYSTEMIC STEM/PROGENITOR CELL THERAPY FOR AMD

优化 AMD 的系统干细胞/祖细胞治疗

• 批准号: 9507559
• 负责人: Michael Edwin Boulton
• 金额: $ 54.62万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9507559
• 关键词: Acute; Address; Age; Age related macular degeneration; American; Back; Blindness; Blood Circulation; Bone Marrow Transplantation; Cell Differentiation process; Cell Therapy; Cell Transplantation; Cell Transplants; Cells; Cessation of life; Chronic; Circadian Rhythms; Clinical; Dependence; Development; Disease; Elderly; Electroretinography; Engraftment; Ensure; Environment; Epithelial; Equilibrium; Ethics; Eye; Failure; Functional disorder; Genes; Health Care Costs; Hematopoietic stem cells; Histology; Home environment; Human; Immunohistochemistry; Inflammatory; Injectable; Injection of therapeutic agent; Injury; Knowledge; Measures; Mesenchymal Stem Cells; Microglia; Modeling; Mus; Natural regeneration; Nonexudative age-related macular degeneration; Outcome; Pathology; Patients; Pattern; Peripheral; Phagocytosis; Phase; Phenotype; Photoreceptors; Proteins; Quality of life; RPE65 protein; Recruitment Activity; Regulation; Retina; Retinal; Retinal Diseases; Retinal Pigments; Route; SCID Mice; SOD2 gene; Stem cells; Structure of retinal pigment epithelium; Subfamily lentivirinae; System; Therapeutic; Therapeutic Intervention; Time; Translating; Transplantation; Trauma; Vision; base; cell type; chemokine receptor; genetic approach; geographic atrophy; human stem cells; improved; injured; knock-down; monocyte; monolayer; mouse model; preconditioning; public health relevance; reconstitution; repaired; social; stem

DESCRIPTION (provided by applicant): Age-related macular degeneration (AMD) is the leading cause of visual loss in the elderly. Despite knowledge of the cell types involved, therapeutic intervention has been limited and there is currently no treatment for "nonexudative AMD". While RPE cell transplantation into the subretinal space of patients offered a promising therapeutic approach, outcomes to date have been limited due to: 1) transplantation in late stage disease, 2) the invasive route of administration, and 3) incomplete differentiation status of the transplanted cells. To address these limitations, we have made a number of exciting discoveries: 1) forced expression of the RPE65 gene allows mouse hematopoietic stem cells (mHSC), when injected back into the circulation, to home to the retina and renew the RPE monolayer in both acute and chronic mouse models of RPE loss and re-establish visual function; 2) the circadian pattern to endogenous HSC release impacts reconstitution following bone marrow transplantation; 3) microglial activation in AMD will require "modulation" to ensure efficient RPE regeneration by HSCs; 4) a highly effective non-viral protein delivery machinery (T3SS) is able to deliver target proteins into host HSCs and promote their differentiation; 5) human hematopoietic progenitor cells (hHPCs), when programmed with RPE65, express RPE cell markers. Based on these observations we hypothesize that: "Successful therapeutic utilization of human or murine HSC requires their programming prior to injection into the systemic circulation, their injection at the time of optimal engraftment potential and preconditioning of the retina by either suppression of resident microglia activation and/or restoring the balance of peripheral pro-inflammatory and homeostatic monocytes." The hypothesis is addressed in three Aims. In Aim 1 we will determine the dependence of recruitment and incorporation of programmed mHSC into the injured RPE in the SOD2 KD mouse model upon the time of day of injection and the age of the donor HSC as well as the age of the recipient. Aim 2 will investigate the importance of manipulating the retinal environment, by controlling either the activation state of the resident microglia or the influx of peripheral monocytes on the efficiency of systemically administered programmed mHSC to repair the RPE layer in the SOD2 KD model. Aim 3 will translate our mouse findings into hHPCs. We will express RPE65, in human CD133+, CD 34- , CD38- cells to differentiate these cells toward RPE cells and allow RPE regeneration in SCID mice undergoing the SOD2 KD model. Co-injection of mesenchymal stem cells will be utilized to reduce activation of resident microglia. Our approach overcomes many of the current limitations of human stem cell therapies for AMD.

描述（由申请人提供）：年龄相关性黄斑变性（AMD）是老年人视力丧失的主要原因。尽管了解所涉及的细胞类型，但治疗干预仍然有限，目前还没有针对“非渗出性 AMD”的治疗方法。虽然将 RPE 细胞移植到患者视网膜下腔提供了一种有前途的治疗方法，但迄今为止的结果有限，原因如下：1) 晚期疾病移植，2) 侵入性给药途径，以及 3) RPE 细胞的不完全分化状态。 移植的细胞。为了解决这些限制，我们取得了许多令人兴奋的发现：1) RPE65 基因的强制表达使小鼠造血干细胞 (mHSC) 在注射回循环系统时能够回归视网膜并更新视网膜中的 RPE 单层。 RPE 丧失和重建视功能的急性和慢性小鼠模型； 2) 内源性 HSC 释放的昼夜节律模式影响骨髓移植后的重建； 3) AMD 中的小胶质细胞激活需要“调节”以确保 HSC 有效再生 RPE； 4）高效的非病毒蛋白递送机制（T3SS）能够将靶蛋白递送至宿主HSC并促进其分化； 5) 当用 RPE65 编程时，人类造血祖细胞 (hHPC) 表达 RPE 细胞标记。基于这些观察，我们假设：“人类或小鼠 HSC 的成功治疗利用需要在注射到体循环之前对其进行编程，在最佳植入潜力时进行注射，并通过抑制驻留小胶质细胞激活和预处理视网膜来预处理它们。” /或恢复外周促炎单核细胞和稳态单核细胞的平衡。”该假设在三个目标中得到解决。在目标 1 中，我们将确定在 SOD2 KD 小鼠模型中，程序化的 mHSC 募集和掺入到受损 RPE 中的依赖性取决于注射时间、供体 HSC 的年龄以及受体的年龄。目标 2 将研究操纵视网膜环境的重要性，方法是 控制常驻小胶质细胞的激活状态或外周单核细胞的流入，以影响系统施用程序化 mHSC 修复 SOD2 KD 模型中 RPE 层的效率。目标 3 将把我们的小鼠发现转化为 hHPC。我们将在人 CD133+、CD 34-、CD38- 细胞中表达 RPE65，以将这些细胞分化为 RPE 细胞，并允许接受 SOD2 KD 模型的 SCID 小鼠中 RPE 再生。间充质干细胞的共注射将用于减少驻留小胶质细胞的激活。我们的方法克服了目前人类干细胞疗法治疗 AMD 的许多局限性。