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：
8272241
负责人：
Jennifer Lynne Simpson
金额：
$ 38.25万
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/8272241
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

DESCRIPTION (provided by applicant): 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 failur 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) PUBLIC HEALTH RELEVANCE: This study will evaluate the potential therapeutic effects of human umbilical cord mesenchymal stem cells (hUCMSC) transplantation into to the corneal stroma to treat a mouse model of corneal cystinosis. Progression of corneal cystinosis and the formation of intrastromal cystine crystals will be evaluated by quantitative in vivo confocal microscopy. Laser scanning confocal microscopy, immunocytochemistry and molecular biology will be used to evaluate differentiation of hUCMSC to mouse corneal keratocytes and their response to corneal injury.

描述（由申请人提供）：该提案的长期目标是开发一种成功的治疗角膜胱氨酸病的治疗策略。胱氨酸病是一种罕见的溶酶体贮积病，由胱氨酸溶酶体转运蛋白胱氨酸（CTNS）缺陷引起，导致多种组织和器官中胱氨酸晶体形成，导致早期多器官衰竭。在角膜中，由于畏光、眼睑痉挛和复发性角膜糜烂导致的进行性视力恶化与角膜晶体浓度的增加有关，尽管其他器官系统得到了成功的治疗，但对于胱氨酸中毒患者来说，这仍然是一个主要的长期负担。最近，已经产生了一种胱氨酸基因敲除小鼠（Ctns-/-），它在组织中产生胱氨酸晶体，并显示出与胱氨酸中毒患者相似的疾病进展，包括肾功能下降和角膜胱氨酸晶体形成。与博士合作。斯克里普斯大学的 Stephanie Cherqui 和辛辛那提大学的 Winston Kao 进行了初步实验，以评估治疗角膜胱氨酸病的潜在新疗法。这些初步研究表明，角膜内注射人脐带间充质干细胞 (hUCMSC) 抑制 Ctns-/- 小鼠角膜的进展和疤痕形成，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。需要用 hUCMSC 替代百分之几的宿主角膜细胞群才能抑制角膜胱氨酸沉着症的发展（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）？为了解决这些问题，提出了以下具体目标： 1. 通过向年轻（3个月大）小鼠注射不同数量的hUCMSC并测量体内胱氨酸晶体体积，确定基质内注射hUCMSC对角膜胱氨酸病发生的影响角膜细胞分化和细胞密度随时间的变化。（问题 1） 2. 通过在不同年龄的小鼠中注射最佳数量的 hUCMSC 并测量体内胱氨酸晶体体积、hUCMSC 角膜细胞分化和随时间变化的细胞密度，评估角膜胱氨酸沉着症阶段对 hUCMSC 植入的影响，（问题 2）3通过在最佳移植的小鼠角膜中进行上皮刮擦损伤并测量上皮，表征 hUCMSC 移植角膜的伤口愈合反应。以及角膜细胞再生并返回角膜细胞表型。（问题3）公共健康相关性：本研究将评估将人脐带间充质干细胞 (hUCMSC) 移植到角膜基质中治疗小鼠角膜胱氨酸病模型的潜在治疗效果。角膜胱氨酸病的进展和基质内胱氨酸晶体的形成将通过体内定量共聚焦显微镜进行评估。激光扫描共聚焦显微镜、免疫细胞化学和分子生物学将用于评估 hUCMSC 向小鼠角膜角膜细胞的分化及其对角膜损伤的反应。