Optimizing systemic stem/progenitor cell therapy for AMD

优化 AMD 的全身干/祖细胞治疗

基本信息

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
项目类别：
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

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

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项目摘要

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.

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，但治疗干预仍有限，并且有目前尚未治疗“非X级AMD”。而RPE细胞移植到视网膜下患者的空间提供了一种有希望的治疗方法，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）移植细胞的不完全分化状态。为了解决这些限制，我们有做出了许多令人兴奋的发现：1）RPE65基因的强制表达允许鼠标造血干细胞（MHSC），当注射回到循环中时，回到了视网膜的家中在RPE损耗的急性和慢性小鼠模型中续订RPE单层和重新建立视觉功能； 2）内源性HSC释放的昼夜模式会影响重组骨髓移植后； 3）AMD中的小胶质细胞激活将需要“调制”到确保HSC有效的RPE再生； 4）高效的非病毒蛋白递送机械（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：“成功的治疗性人类或鼠类HSC的利用需要在注射到全身循环，在最佳植入潜力时注射和通过抑制居民小胶质细胞激活和/或恢复外围促炎和稳态单核细胞的平衡。在三个目标中解决了假设。在AIM 1中，我们将确定招聘的依赖性并将编程的MHSC掺入SOD2 KD鼠标模型中的受伤RPE中一天中的注射时间和捐助者HSC的年龄以及接受者的年龄。目标2 将通过控制要么驻留小胶质细胞的激活状态或外周单核细胞在效率上的涌入系统地管理编程的MHSC，以修复SOD2 KD模型中的RPE层。目的 3将我们的鼠标发现转化为HHPC。我们将在人CD133+，CD 34-中表达RPE65- ，CD38-细胞将这些细胞区分为RPE细胞，并允许RPE再生经历了SOD2 KD模型的小鼠。间充质干细胞的共同注射将用于减少居民小胶质细胞的激活。我们的方法克服了许多当前的局限 AMD的人类干细胞疗法。