Optimizing systemic stem/progenitor cell therapy for AMD

优化 AMD 的全身干/祖细胞治疗

• 批准号: 8561485
• 负责人: Michael Edwin Boulton
• 金额: $ 58.96万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 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 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8561485
• 关键词: Accounting; Acute; Address; Age; Age related macular degeneration; American; Back; Blindness; Blood Circulation; Bone Marrow Transplantation; Cell Therapy; Cell Transplantation; Cell Transplants; Cells; Cessation of life; Chronic; Circadian Rhythms; Clinical; Controlled Environment; Dependence; Development; Disease; Elderly; Electroretinography; Engraftment; Ensure; Epithelial; Equilibrium; Ethics; Eye; Failure; Functional disorder; Genes; Genetic; Health Care Costs; Hematopoietic stem cells; Histology; Home environment; Human; Immunohistochemistry; Inflammatory; Injection of therapeutic agent; Injury; Knowledge; Lentivirus; 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; Regulation; Retina; Retinal; Retinal Pigments; Route; SCID Mice; Staging; Structure of retinal pigment epithelium; System; Therapeutic; Therapeutic Intervention; Time; Translating; Transplantation; Trauma; Vision; base; cell type; chemokine receptor; geographic atrophy; human SOD2 protein; human stem cells; improved; injured; monocyte; monolayer; mouse model; preconditioning; programs; public health relevance; reconstitution; repaired; social; stem; stem cell therapy; stem cells

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.

视网膜相关性黄斑变性（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，但治疗干预是有限的， 目前没有治疗“非渗出性AMD”的方法。将RPE细胞移植到视网膜下， 患者的空间提供了一种有前途的治疗方法，但迄今为止的结果有限 由于：1）晚期疾病中的移植，2）侵入性给药途径，以及3） 移植细胞的不完全分化状态。为了解决这些局限性，我们 有了一些令人兴奋的发现：1）RPE 65基因的强制表达使小鼠 造血干细胞（mHSC），当注射回循环，回家视网膜， 在急性和慢性RPE丧失小鼠模型中更新RPE单层， 视觉功能; 2）内源性HSC释放的昼夜节律模式影响重建 3）AMD中的小胶质细胞活化将需要“调节”， 确保HSC的高效RPE再生; 4）高效的非病毒蛋白递送 T3 SS能够将靶蛋白递送到宿主HSC中并促进其分化; 5)当用RPE 65编程时，人造血祖细胞（hHPC）表达RPE 细胞标记物。基于这些观察，我们假设：“成功的治疗 利用人或鼠HSC需要在注射入HSC之前对其进行编程。 体循环，在最佳植入潜力时注射， 通过抑制驻留的小胶质细胞活化和/或 恢复外周促炎和稳态单核细胞的平衡。“那 假设在三个目标中得到解决。在目标1中，我们将确定招聘的依赖性 以及在SOD 2 KD小鼠模型中程序化mHSC掺入受损RPE中， 注射的时间和供体HSC的年龄以及受体的年龄。目的2 将研究操纵视网膜环境的重要性，通过控制 常驻小胶质细胞的激活状态或外周单核细胞的流入对 全身施用程序化mHSC以修复SOD 2 KD模型中的RPE层。目的 3将我们的小鼠发现转化为hHPC。我们将在人CD 133+、CD 34- CD 38-细胞使这些细胞向RPE细胞分化并允许SCID中的RPE再生 进行S 0 D2 KD模型的小鼠。间充质干细胞的共注射将用于 减少常驻小胶质细胞的激活。我们的方法克服了许多目前的限制， 人类干细胞治疗AMD