Magnetic Steering and Longitudinal Visualization of Stem Cells for Trabecular Meshwork Therapy in Glaucoma

用于青光眼小梁网治疗的干细胞磁控和纵向可视化

• 批准号: 10653277
• 负责人: STANISLAV Y EMELIANOV
• 金额: $ 7.78万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10653277
• 关键词: Abbreviations; Anterior; Apoptosis; Area; Blindness; CASP3 gene; Cell Culture Techniques; Cell Survival; Cells; Cellularity; Data; Disease; Dyes; Effectiveness; Eye; Eye diseases; Family suidae; Glaucoma; Glossary; Goals; Histologic; Homeostasis; Human; Hydrogen Peroxide; Image; In Situ; Inflammatory; Label; Location; Magnetic nanoparticles; Magnetism; Mesenchymal Stem Cells; Modeling; Monitor; Mus; Mutate; Nanotechnology; Ocular Hypertension; Organ Culture Techniques; Patients; Peptides; Photosensitivity; Physiologic Intraocular Pressure; Primary Open Angle Glaucoma; Property; RNA; Reporter; Research; Risk; Risk Factors; Spatial Distribution; Specificity; System; Technology; Testing; Time; Tissues; Trabecular meshwork structure; Transgenic Mice; Vesicle; Visualization; Work; anterior chamber; cellular imaging; comparative efficacy; cytokine; delivery vehicle; design; experimental study; imaging platform; imaging system; improved; in vivo; induced pluripotent stem cell; innovation; instrument; interest; myocilin; nanoparticle; novel; outcome prediction; photoacoustic imaging; pressure; serial imaging; stem cell delivery; stem cell technology; stem cell therapy; stem cells; superparamagnetism; ultrasound; virtual

Project Summary/Abstract Glaucoma is a major cause of blindness and current treatments are insufficient. A major risk factor for glaucoma, and the only treatable risk factor, is elevated intraocular pressure (IOP). Current IOP-lowering therapies fail too often, and thus there continues to be great interest in novel IOP control strategies. The trabecular meshwork (TM), the key tissue determining IOP, has reduced cellularity in glaucoma, which has led a number of groups to study stem cell-based therapies for the TM. An obstacle to such therapies is cell delivery: current approaches have low cell delivery efficiency and specificity for the TM, and cannot deliver cells to all parts of the TM. Here a novel technological solution for “steering” stem cells to the TM is proposed, which can be additionally used to visualize stem cell delivery and to monitor stem cell apoptosis. This approach will be tested in several models of ocular hypertension. The key technology is superparamagnetic and photosensitive nanoparticles. Their superparamagnetic properties mean that cells which have taken up these nanoparticles can be rapidly steered to the TM by a magnet placed at the limbus. Their photosensitivity means that cells can be visualized by ultrasound/photoacoustic imaging in the living eye. The functionality of these nanoparticles will be further enhanced by using a photosensitive marker of active caspase-3 to monitor stem cell apoptosis. Our overall objective is to validate these technologies as a safe and effective approach for monitoring and steering of stem cells to the TM, thereby restoring intraocular pressure (IOP) homeostasis in glaucoma patients. Three specific aims towards this long-term goal are proposed, building on our significant preliminary data. In aim 1, a novel caspase-3-sensitive reporter for monitoring apoptosis will be synthesized and characterized, and magnets for steering stem cells to the TM will be optimized. In aim 2, an instrument capable of imaging of labeled stem cells in whole eyes, including longitudinal monitoring of stem cell distribution and apoptosis, will be developed. In aim 3, stem cells will be delivered to the TM in two glaucoma models, and their ability to restore IOP homeostasis will be evaluated. The ability of ultrasound/photoacoustic imaging to monitor stem cell delivery to the TM and stem cell apoptosis will also be validated; and mesenchymal stem cells (MSCs) will be compared to differentiated induced pluripotent stem cells (iPSC-TMs) for their efficacy in restoring IOP homeostasis. This project is highly innovative: it the first study to steer and visualize stem cells as part of a treatment for ocular hypertension. It is also the first to compare the efficacy of MSCs vs. iPSC-TMs for treating ocular hypertension. We expect, as suggested by our strong preliminary data, to discover that stem cells can be efficiently and selectively steered to the TM by a simple magnet placed at the limbus for as little as 15 minutes; and that it will be possible to accurately monitor the location of stem cells in the eye and stem cell apoptosis over time. Further, we expect that TM function will be improved by stem cells steered in this way, as tested in 2 glaucoma models.

项目摘要/摘要 青光眼是失明的主要原因，目前的治疗不足。青光眼的主要危险因素， 唯一可治疗的危险因素是眼内压（IOP）升高。当前降低IOP的疗法也失败了 通常，因此，对新型IOP控制策略的兴趣仍然很大。小梁网 （TM）是确定IOP的关键组织，在青光眼中降低了细胞，这导致许多组达到 研究TM的研究干细胞疗法。这种疗法的障碍是细胞输送：当前方法 TM的细胞输送效率和特异性低，并且无法将细胞传递到TM的所有部位。 这里提出了一种新型的“转向”干细胞向TM的技术解决方案，可以另外 用于可视化干细胞输送并监测干细胞凋亡。这种方法将在几个 眼高血压模型。关键技术是超磁和光敏纳米颗粒。 它们的超副磁特性意味着吸收这些纳米颗粒的细胞可以迅速 用位于边缘的磁铁转向TM。它们的光敏性意味着可以通过 超声/光声成像在活眼中。这些纳米颗粒的功能将进一步 通过使用活性caspase-3的光敏标记来监测干细胞凋亡。我们的整体 目的是验证这些技术是一种安全有效的方法来监视和转向STEM 细胞到TM，从而恢复青光眼患者的眼内压（IOP）稳态。 基于我们重要的初步数据，提出了针对这个长期目标的三个具体目标。目标 1，将合成和表征用于监测凋亡的新型caspase-3敏感记者，并且 转向干细胞到TM的磁铁将被优化。在AIM 2中，能够成像标记的仪器 整个眼睛的干细胞，包括干细胞分布和凋亡的纵向监测，将是 发达。在AIM 3中，干细胞将在两个青光眼模型中传递到TM，并将其恢复能力传递 IOP稳态将进行评估。超声/光声成像监测干细胞输送的能力 TM和干细胞凋亡也将得到验证；将比较间充质干细胞（MSC） 分化的诱导多能干细胞（IPSC-TMS）在恢复IOP稳态方面的效率。这 项目具有很高的创新性：这是第一个研究和可视化干细胞作为眼部治疗的一部分的研究 高血压。它也是第一个比较MSC与IPSC-TMS治疗眼高血压的效率的方法。 正如我们强大的初步数据所建议的那样，我们期望干细胞可以有效，并且 通过在边缘放置的简单磁铁将其选择性地蒸至TM，短短15分钟；而且它将 随着时间的流逝，可以准确监测干细胞在眼睛和干细胞凋亡中的位置。更远， 我们希望以这种方式蒸的干细胞将改善TM功能，如在2种青光眼模型中测试。