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）：与年龄相关的黄斑变性（AMD）是老年人视觉丧失的主要原因。尽管了解涉及的细胞类型，但治疗性干预措施受到限制，目前尚无治疗“非量AMD”的治疗方法。尽管RPE细胞移植到患者的视网膜下空间提供了有前途的治疗方法，但迄今为止的结果受到限制，由于：1）在晚期疾病中移植，2）侵入性给药途径，3）3）） 移植的细胞。为了解决这些局限性，我们做出了许多令人兴奋的发现：1）将RPE65基因强迫表达允许小鼠造血干细胞（MHSC）（MHSC），当注入循环中时，将RPE单层恢复到视网膜的家中，并在急性和慢性小鼠损失和重复型构想中恢复RPE单层。 2）内源性HSC释放的昼夜节律影响骨髓移植后的重建； 3）AMD中的小胶质细胞激活将需要“调制”以确保HSC有效再生； 4）高效的非病毒蛋白输送机制（T3SS）能够将靶蛋白传递到宿主HSC中并促进其分化； 5）人类造血祖细胞（HHPC）用RPE65进行编程时，表示RPE细胞标记。基于这些观察结果，我们假设：“成功的治疗性利用人类或鼠类HSC需要在注入全身循环之前进行编程，在最佳植入潜力和预处理预处理时，它们在居住的小胶质细胞激活和/或恢复围绕围harm的平衡和/或恢复围位概念和主持人的平衡和/或恢复。该假设在三个目标中解决。在AIM 1中，我们将在注射日和供体HSC的年龄以及接收者的年龄的情况下确定在SOD2 KD小鼠模型中募集和掺入程序的MHSC中受伤的RPE。 AIM 2将调查操纵视网膜环境的重要性， 控制驻留小胶质细胞的激活状态或周围单核细胞在系统管理的程序MHSC效率上涌入以修复SOD2 KD模型中的RPE层。 AIM 3将我们的鼠标发现转化为HHPC。我们将在人CD133+，CD 34-，CD38-细胞中表达RPE65，以将这些细胞区分为RPE细胞，并允许在经历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居民小胶质细胞的激活。我们的方法克服了人类干细胞疗法对AMD的许多当前局限性。