Mesenchymal Stem Cell Therapy for Corneal Cystinosis

间充质干细胞治疗角膜胱氨酸病

基本信息

批准号：
8655888
负责人：
Jennifer Lynne Simpson
金额：
$ 37.49万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8655888
关键词：
Address Affect Age Age-Months Animal Model Biochemical Blepharospasm Bone Marrow Cell Density Cells Cicatrix Clinical Collaborations Confocal Microscopy Cornea Corneal Diseases Corneal Injury Corneal Stroma Crystal Formation Cysteamine Cystine Cystinosis Data Defect Deposition Deterioration Development Disease Disease Progression Drug usage Engraftment Epithelial Eyedrops Foundations Funding Goals Hereditary Disease Hour Human Infiltration Inflammatory Injection of therapeutic agent Injury Kidney Diseases Knockout Mice Knowledge Laser Scanning Confocal Microscopy Lysosomal Storage Diseases Lysosomes Measurement Measures Mesenchymal Stem Cell Transplantation Mesenchymal Stem Cells Methods Molecular Biology Multiple Organ Failure Mus Natural regeneration Oral Organ Orphan Patients Phenotype Photophobia Population Recurrence Refractory Renal function Reporting Research Research Personnel Staging Systemic Therapy Therapeutic Therapeutic Effect Time Tissues Transplantation Umbilical cord structure Universities Visual Wound Healing aged base body system immunocytochemistry in vivo loss of function mouse model novel research study response response to injury stem cell therapy

项目摘要

Project Summary The long term goal of this proposal is to develop a successful therapeutic strategy for treating corneal cystinosis. Cystinosis is a rare lysosomal storage disease caused by a defect in the cystine lysosomal transporter cystinosin (CTNS) that leads to cystine crystal formation in various tissues and organs resulting in multiple organ failure at an early age. In the cornea, progressive visual deterioration due to photophobia, blepharospasm and recurrent corneal erosions are associated with increasing concentrations of corneal crystals and are a major long-term burden for patients with cystinosis despite successful treatment of other organ systems. Recently, a cystinosin knockout mouse (Ctns-/-) has been generated that develops cystine crystals in tissues and shows progression of disease similar to that of cystinosis patients, including decreased kidney function and formation of corneal cystine crystals. In collaboration with Drs. Stephanie Cherqui at Scripps and Winston Kao at University of Cincinnati, we have conducted preliminary experiments to evaluate potential novel therapies for treating corneal cystinosis. These preliminary studies suggest that intra- corneal injection of human umbilical cord mesenchymal stem cells (hUCMSC) inhibits progression and scarring of Ctns-/- mouse corneas, and reduces the presence of intrastromal corneal crystals, suggesting that hUCMSC can replace dead or dying corneal keratocytes and restore corneal function. Based on these findings we propose the HYPOTHESIS: That hUCMSC intrastromal transplantation can rescue or block the development of corneal cystinosis in the Ctns-/- mouse. This hypothesis prompts the following important QUESTIONS that need to be addressed. What percent of the host keratocyte population needs to be replaced by hUCMSC to inhibit the development of corneal cystinosis (QUESTION 1)? Does stage of disease affect engraftment and differentiation of hUCMSC (QUESTION 2)? And, does engraftment of hUCMSC affect normal corneal responses to injury (QUESTION 3)? To address these questions have proposed the following specific aims: 1. Determine the effect of intrastromal injection of hUCMSC on the development of corneal cystinosis by injecting different numbers of hUCMSC into young (3 month old) mice and measuring in vivo cystine crystal volume, hUCMSC keratocyte differentiation and cell density over time. (QUESTION 1) 2. Assess the effect of stage of corneal cystinosis on hUCMSC engraftment by injecting the optimal number of hUCMSC in different aged mice and measuring in vivo cystine crystal volume, hUCMSC keratocyte differentiation and cell density over time, (QUESTION 2) 3. Characterize the wound healing response of hUCMSC engrafted corneas by performing epithelial scrape injuries in optimally engrafted mouse corneas and measure epithelial and keratocyte regeneration and return to keratocyte phenotype. (QUESTION 3)

项目概要该提案的长期目标是开发一种成功的治疗角膜的治疗策略胱氨酸病。胱氨酸贮积症是一种罕见的溶酶体贮积病，由胱氨酸溶酶体缺陷引起胱氨酸转运蛋白（CTNS），导致各种组织和器官中胱氨酸晶体形成，从而导致早年多器官衰竭。在角膜中，由于畏光而导致视力逐渐恶化，眼睑痉挛和复发性角膜糜烂与角膜浓度增加有关尽管其他药物治疗成功，但晶体仍是胱氨酸病患者的主要长期负担器官系统。最近，已经产生了一种胱氨酸敲除小鼠（Ctns-/-），它可以产生胱氨酸组织中出现晶体，并且显示出与胱氨酸病患者相似的疾病进展，包括减少肾功能和角膜胱氨酸晶体的形成。与博士合作。斯蒂芬妮·切尔基辛辛那提大学的 Scripps 和 Winston Kao，我们进行了初步实验来评估治疗角膜胱氨酸病的潜在新疗法。这些初步研究表明，内部角膜注射人脐带间充质干细胞（hUCMSC）可抑制进展并 Ctns-/-小鼠角膜的疤痕，并减少基质内角膜晶体的存在，表明hUCMSC可以替代死亡或垂死的角膜细胞并恢复角膜功能。基于这些发现，我们提出假设：hUCMSC 基质内移植可以挽救或阻止 Ctns-/- 小鼠角膜胱氨酸病的发展。这一假设提出了以下需要解决的重要问题。占百分之几宿主角膜细胞群需要被hUCMSC替代以抑制角膜的发育胱氨酸中毒（问题 1）？疾病阶段是否影响 hUCMSC 的植入和分化（问题 2）？并且，hUCMSC 的植入是否会影响正常角膜对损伤的反应（问题 3）？针对这些问题提出了以下具体目标： 1. 确定基质内注射 hUCMSC 对角膜胱氨酸沉着症发展的影响将不同数量的 hUCMSC 注射到幼年（3 个月大）小鼠体内并测量体内胱氨酸晶体体积、hUCMSC 角膜细胞分化和细胞密度随时间的变化。（问题1） 2. 通过注射最佳的角膜胱氨酸沉积症阶段对 hUCMSC 植入的影响不同年龄小鼠hUCMSC数量及体内胱氨酸晶体体积测定，hUCMSC 角膜细胞分化和细胞密度随时间的变化（问题 2） 3. 通过进行上皮细胞移植来表征 hUCMSC 移植角膜的伤口愈合反应刮擦最佳移植小鼠角膜的损伤并测量上皮细胞和角膜细胞再生并返回角膜细胞表型。（问题3）