Bioengineering Cornea with Autologous Stem Cells

自体干细胞生物工程角膜

基本信息

批准号：
10584599
负责人：
Yiqin Du
金额：
$ 49.08万
依托单位：
CARNEGIE-MELLON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10584599
关键词：
3-Dimensional Address Alkalies Allogenic Anterior Architecture Area Autologous Basement membrane Biochemical Biomechanics Biomedical Engineering Biomimetics Biopsy Specimen Bioreactors Blindness Cadaver Cell Density Cell Separation Cell Therapy Cells Clinic Clinical Collagen Cornea Corneal Endothelium Corneal Opacity Corneal Stroma Corneal edema Cues Cytoskeleton Data Descemet&apos s membrane Development Disease Disease model Endothelial Cells Endothelium Engineering Engraftment Evaluation Excision Extracellular Matrix Extracellular Matrix Proteins Eye Failure Generations Genes Goals Graft Rejection Healthcare Human Hydration status Immune In Vitro Individual Injections Injury Integrins Keratoplasty Lamellar Keratoplasty Mechanics Medical Device Melons Modeling Natural regeneration Optics Oryctolagus cuniculus Patients Penetrating Keratoplasty Persons Procedures Process Productivity Property Proteoglycan Pump Qualifying Research Scaffolding Protein Structure Surface Technology Thick Tight Junctions Tissue Donors Tissue Engineering Tissue Transplantation Tissues Translating Translations Transplantation Universities Validation Vision Visually Impaired Persons Work adult stem cell anterior chamber clinical translation clinically relevant corneal repair cost density disability efficacy validation experimental study functional restoration high risk human old age (65+)human stem cells improved in vivo in vivo Model indexing innovation limbal mechanical load monolayer nano nanofabrication novel strategies novel therapeutic intervention repaired scaffold sight restoration stem cell differentiation stem cells technology development

项目摘要

This Multi-PI project combines the efforts of two research groups with different areas of expertise to address the long-term goal of developing bioengineered corneal stroma and endothelial tissues to provide therapy for individuals with corneal blindness. These tissues will be bioengineered from adult stem cells, which can be obtained from the individuals to be treated as autologous or from allogeneic cell storage since the stem cells are immunosuppressive. Over the past 5 years we have demonstrated that organization of these cells into tissues can be guided by scaffolds constructed of native extracellular matrix proteins, fabricated using a biomimetic, surface-induced assembly process. The Du Lab at the University of Pittsburgh will obtain stem cells from limbal stroma of donated human corneas. Their extensive work with these corneal stromal stem cells (CSSC) shows that they differentiate to stromal keratocytes and to corneal endothelial cells, tissues responsible for most corneal opacity. We have demonstrated that CSSC can be obtained from biopsy samples, presenting the opportunity to generate patient-specific, autologous bioengineered tissues. The Feinberg Lab at Carnegie Melon University has developed novel approach of assembling native extracellular matrix proteins to produce tissue-like scaffolding with defined 3-D architecture. Aim 1 will build on our previous work, which has showed that we can engineer spatial and biochemical cues provided by the scaffolding to generate stroma-like tissue from CSSCs that can be stacked to form multilamellar 3-D tissue similar to that of the corneal stroma. Bioengineered stroma produced in the proposed experiments will be subjected to biomechanical loading to simulate cornea development and further improve mechanical and optical properties. Function of the bioengineered stroma in lamellar keratoplasty will be evaluated in an in vivo rabbit model. Aim 2 will build on our work differentiating CSSC into endothelial cells and growing these on engineered basement membrane protein scaffolds to form polygonal monolayers that express genes typical of corneal endothelium. Previously, we demonstrated the ability to create the equivalent of Descemet's membrane to bioengineer an entire sheet of endothelium suitable for lamellar keratoplasty, but we also developed an alternative approach to engineer small patches of endothelium and deliver these via simple injection. Bioengineered endothelium produced in the proposed studies will use our newly developed “shrink-wrapping” technology to create microscale patches of corneal endothelium that can be injected into the anterior chamber for injury-free engraftment. Functionality of the constructs will be demonstrated in rabbit models in vivo, focused on boosting cell-density to improve pump function as an alternative to lamellar keratoplasty for endothelial disease. This project will build on the innovative experimental approaches we developed during the first 5 years of this project. Importantly, tissues developed and validated in vivo during this proposed study hold the potential of being advanced into clinically relevant studies to provide a novel therapeutic approach to the age-old problem of corneal blindness.

这个多PI的项目结合了两个研究小组与不同专家领域的努力，以解决开发生物工程的角膜基质和内皮组织的长期目标，以提供治疗角膜失明的人。这些时间将从成年干细胞中进行生物工程，这可以是从个体获得以自体或同种异体细胞存储而获得的，因为干细胞是免疫抑制。在过去的5年中，我们证明了这些细胞将这些细胞组织到组织可以由由天然细胞外基质蛋白构成的支架来指导，该蛋白使用仿生型制成表面诱导的组装过程。匹兹堡大学的DU实验室将从边缘获得干细胞捐赠人角膜的基础。他们在这些角膜基质干细胞（CSSC）中的广泛工作显示它们分化为基质角膜细胞和角膜内皮细胞，该组织负责大多数角膜不透明度。我们已经证明，可以从活检样本中获得CSSC，并提供了机会产生患者特异性的自体生物工程组织。卡内基瓜大学的Feinberg实验室已经开发了组装天然细胞外基质蛋白以产生组织样的新方法用定义的3-D体系结构进行脚手架。 AIM 1将以我们以前的工作为基础，这表明我们可以脚手架提供的工程师空间和生化提示，以产生CSSC的基质样组织可以堆叠以形成类似于角膜基质的多层3-D组织。生物工程基质在拟议的实验中产生的将经过生物力学负载以模拟角膜开发并进一步改善了机械和光学特性。生物工程基质的功能层状角膜成形术将在体内兔模型中进行评估。 AIM 2将基于我们的工作与众不同 CSSC进入内皮细胞，并在工程的地下膜蛋白支架上种植它们以形成表达典型的角膜植物基因的多边形单层。以前，我们证明了能力为了创建相当于Descemet膜的物质，以生物工程师一整片适合的内皮层状角膜造成术，但我们还开发了一种替代方法来设计内皮的小斑块并通过简单的注射传递。拟议的研究中生产的生物工程的内皮将使用我们新开发的“收缩包装”技术，以创建角膜内皮微观斑块，可以被注入前室以进行无损伤的植入。结构的功能将是在体内兔模型中证明，重点是提高细胞密度以提高泵的功能替代内皮疾病的层状角膜膜成形术。该项目将基于创新的实验我们在该项目的前5年中开发的方法。重要的是，组织在在这项拟议的研究中，体内有可能进入临床相关研究以提供新型的热方法解决了古老的角膜失明问题。