Bioengineering Cornea with Autologous Stem Cells

自体干细胞生物工程角膜

基本信息

批准号：
10367567
负责人：
Yiqin Du
金额：
$ 57.31万
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/10367567
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 Therapy Cells Clinic Clinical Collagen Cornea Corneal Endothelium Corneal Opacity Corneal Stroma Corneal edema Cues 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 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 injured innovation limbal mechanical load monolayer 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 项目结合了两个具有不同专业领域的研究小组的努力，以解决开发生物工程角膜基质和内皮组织以提供治疗的长期目标这些组织将由成体干细胞进行生物工程改造。由于干细胞是从待处理的自体细胞或同种异体细胞储存的个体中获得的在过去的 5 年里，我们已经证明这些细胞可以组织成组织。可以通过由天然细胞外基质蛋白构建的支架来引导，该支架是使用仿生材料制造的，匹兹堡大学杜实验室将从角膜缘获得干细胞。他们对这些角膜基质干细胞（CSSC）的广泛研究表明。它们分化为基质角膜细胞和角膜内皮细胞（负责大多数角膜的组织）我们已经证明可以从活检样本中获得 CSSC，这为我们提供了机会。卡内基梅隆大学 Feinberg 实验室生成患者特异性自体生物工程组织。开发了组装天然细胞外基质蛋白以产生组织样蛋白的新方法具有定义的 3D 架构的脚手架将建立在我们之前的工作之上，这表明我们可以做到。设计支架提供的空间和生化线索，以从 CSSC 生成基质样组织可以堆叠形成类似于角膜基质的多层 3D 组织。在拟议的实验中产生的将受到生物力学负载以模拟角膜开发并进一步提高生物工程基质的机械和光学特性。板层角膜移植术将在体内兔子模型中进行评估，Aim 2 将建立在我们区分的工作的基础上。 CSSC 进入内皮细胞并在工程基底膜蛋白支架上生长以形成表达角膜内皮典型基因的多边形单层细胞之前，我们证明了这种能力。创造相当于后弹力层的膜，对整张内皮细胞进行生物工程改造，适合板层角膜移植术，但我们还开发了另一种方法来设计小块内皮细胞并通过简单的注射来输送这些在拟议的研究中产生的生物工程内皮细胞。我们新开发的“收缩包装”技术可创建微型角膜内皮补片，被注入前房以实现无损伤植入的功能。在兔子体内模型中，重点是提高细胞密度以改善泵功能治疗内皮疾病的板层角膜移植术的替代方案该项目将建立在创新实验的基础上。重要的是，我们在该项目的前 5 年开发了组织并进行了验证。在这项拟议的研究中，体内有可能进入临床相关研究，以提供解决角膜失明这一古老问题的新治疗方法。