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：
8449080
负责人：
Jennifer Lynne Simpson
金额：
$ 36.34万
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/8449080
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)

项目摘要该建议的长期目标是制定成功治疗角膜的治疗策略囊肿。囊肿性是由胱氨酸溶酶体缺陷引起的一种罕见的溶酶体储存疾病转运蛋白塞司霉素（CTN），导致各种组织和器官的胱氨酸晶体形成，导致从小就多器官衰竭。在角膜中，由于恐惧症引起的渐进视觉恶化，骨痉挛和复发性角膜侵蚀与角膜浓度的增加有关尽管成功治疗其他治疗器官系统。最近，已经产生了一种cystinosin基因敲除小鼠（CTN-/ - ）组织中的晶体，显示出类似于囊肿患者的疾病的进展，包括降低角膜胱氨酸晶体的肾功能和形成。与Drs合作。斯蒂芬妮·切尔奎（Stephanie Cherqui）辛辛那提大学的Scripps和Winston Kao，我们进行了初步实验以评估治疗角膜囊肿病的潜在新疗法。这些初步研究表明 - 角膜注射人脐带间充质干细胞（HUCMSC）抑制进展和 ctns - / - 小鼠角膜的疤痕，并减少了基质内角膜晶体的存在提示HUCMSC可以取代死亡或垂死的角膜角膜细胞并恢复角膜功能。基于这些发现，我们提出了一个假设：HUCMSC内部移植可以营救或阻止CTNS-/ - 小鼠中角膜囊肿的发展。该假设促使以下重要问题需要解决。百分之几宿主角膜细胞种群需要用HUCMSC取代，以抑制角膜的发展囊肿病（问题1）？疾病的阶段会影响植入和HUCMSC的分化（问题2）？并且，HUCMSC的植入会影响正常的角膜对损伤的反应（问题 3）？解决这些问题已提出以下具体目的： 1。确定基质内注射HUCMSC对通过将不同数量的HUCMSC注射到年轻（3个月大）的小鼠中，并在体内测量Cystine 晶体体积，HUCMSC角化细胞分化和细胞密度随时间的时间。（问题1） 2。通过注入最佳的角膜囊肿阶段对HUCMSC植入的影响不同老年小鼠中的HUCMSC数量，并在体内胱氨酸晶体体积，HUCMSC中测量随着时间的流逝，角膜细胞分化和细胞密度，（问题2） 3。表征HUCMSC植入角膜的伤口愈合反应，通过执行上皮在最佳植入的小鼠角膜中刮擦伤害，并测量上皮和角膜细胞再生并返回角膜细胞表型。（问题3）