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制定成功治疗角膜囊肿的治疗策略。囊肿性是一种罕见的溶酶体储存疾病，由胱氨酸溶酶体转运蛋白转运蛋白（CTN）的缺陷引起，可导致各种组织和器官的胱氨酸晶体形成，从而在早期导致多个器官故障。在角膜中，由于恐惧症，孔隙痉挛和复发性角膜侵蚀引起的进行性视觉恶化与角膜晶体浓度的增加有关，尽管成功治疗了其他器官系统，但对于囊肿性患者来说，这是重大的长期负担。最近，已经产生了一种cystinosin基因敲除小鼠（CTN-/ - ），该小鼠在组织中发展了胱氨酸晶体，并显示出类似于囊肿性患者的疾病的进展，包括肾功能降低和角膜胱氨酸晶体的形成。与Drs合作。辛辛那提大学斯克里普斯（Scripps）的斯蒂芬妮·切尔奎（Stephanie Cherqui）和辛辛那提大学的温斯顿·考尔（Winston Kao）进行了初步实验，以评估潜在的新型治疗角膜囊肿性疗法。这些初步研究表明，角膜内注射人脐带间充质干细胞（HUCMSC）抑制了CTN的进展和疤痕 - / - 小鼠角膜，并减少了基质内角膜晶体的存在，这表明HUCMSC可以取代死亡或垂死的角膜角膜细胞和恢复角膜角膜功能。基于这些发现，我们提出了一个假设：HUCMSC基质内移植可以挽救或阻止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。 HUCMSC抑制角膜囊肿的发展（问题1），需要将多少％的宿主角化细胞种群取代？疾病的阶段会影响HUCMSC的植入和分化（问题2）吗？并且，HUCMSC的植入是否会影响正常的角膜对损伤的反应（问题3）？为了解决这些问题，提出了以下具体目的：1。通过将不同数量的HUCMSC注射到年轻（3个月大）的小鼠中，并测量体内胱氨酸晶体体积，HUCMSC角膜差异和细胞差异，通过将不同数量的HUCMSC注射到年轻（3个月大）的小鼠中，确定HUCMSC内注入HUCMSC对角膜囊肿的发展的影响。（问题1）2。通过在不同老年小鼠中注入最佳的HUCMSC数量来评估角膜囊肿的阶段对HUCMSC植入的影响，并测量体内胱氨酸晶体体积，HUCMSC角化细胞分化和细胞密度的hucmsc ceratocyte分化和细胞密度（问题2）3。植入的小鼠角膜，测量上皮和角膜细胞再生，并返回角膜细胞表型。（问题3）公共卫生相关性：本研究将评估人脐带间充质干细胞（HUCMSC）移植到角膜基质中的潜在治疗作用，以治疗角膜囊肿的小鼠模型。角膜膀胱变性的进展和基质内胱氨酸晶体的形成将通过体内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与小鼠角膜角膜细胞的分化及其对角膜损伤的反应。