Bone marrow derived stem cells for therapy of eye disease

骨髓干细胞用于治疗眼部疾病

• 批准号: 7322015
• 负责人: Audra J Johnson
• 金额: $ 3.1万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7322015
• 关键词: Adult; Affinity; Age related macular degeneration; Animal Model; Antibodies; Binding; Biological; Biological Assay; Blindness; Blood Vessels; Bone Marrow; CD44 gene; Cell Fraction; Cell Lineage; Cell Separation; Cells; Chemicals; Classification; Clinical; Degenerative Disorder; Development; Diabetic Retinopathy; Eye; Eye diseases; Generations; Growth; Hematopoietic; Hematopoietic stem cells; Hyaluronic Acid; Incubated; Libraries; Magnetic Bead Technology; Methodology; Modification; Mononuclear; Mus; Nerve Degeneration; Neurons; Pharmaceutical Preparations; Phase; Plan B; Population; Procedures; Process; Property; Reagent; Reporting; Research; Research Training; Retina; Retinal; Retinal Degeneration; Retinal Diseases; Screening procedure; Side; Site; Solutions; Sorting - Cell Movement; Stem cells; Testing; Therapeutic; Tissues; Vascular Diseases; Vascularization; base; inhibiting antibody; laser photocoagulation; malformation; mouse model; neovascularization; receptor; small molecule; small molecule libraries; stem cell therapy

DESCRIPTION (provided by applicant): Bone marrow-derived hematopoietic stem cells (HSCs) have been shown to target sites of neovascularization in the retina. These HSCs have vasculo- and neurotrophic rescue in mouse models of retinal degeneration, and have enormous potential for clinical use as therapeutics for treating retinal degenerative diseases. However, the mechanism by which these cells exert their trophic activity remains unknown. Therefore, in an effort to characterize the activity of these cells, significant improvements need to be made in their extraction/identification. This research will develop a cell-based positive selection assay utilizing small molecules. We will accomplish this through three specific aims Specific Aim 1: Develop a high through-put screening assay to test for small molecules that preferentially bind to CD44HI-expressing cells. Using a pre-competitive cell based ELISA-like assay, we will screen a non-peptidic library for small molecules that bind efficiently to CD44HI-expressing cells. A colorimetric assay will be used to assay the small molecules ability to inhibit binding to bone marrow derived cells by an antibody to CD44. Specific Aim 2: Optimize the binding efficiency of the small molecule through chemical modifications. Enlisting the solution phase methodology used for the library synthesis, secondary libraries will be prepared based on systematic modifications of the validated leads identified in the high through-put assay and optimized for binding affinity inhibiting antibody binding with CD44Hi-expressing cells. Specific Aim 3: Utilize identified small molecules to positively sort out CD44Hi-expressing cells and assess trophic/ biological activity of these cells in animal models of retinal neuronal/vascular degeneration. By incubating whole bone marrow with the identified small molecule(s) and then sorting for cells bound to the small molecule, we can analyze these cells to see if they are CD44Hi-expressing and/or possess trophic activity. This will be accomplished by injecting this subpopulation of cells intravitreally into mice with retinal degeneration and observing rescue of vascular and neuronal degeneration as previously described. In retinal diseases such as diabetic retinopathy, and age-related macular degeneration, the leading causes of vision loss in adults, abnormal blood vessel growth occurs. This research will contribute to the development of therapeutic drugs aimed at treating retinal diseases caused by blood vessel malformation that can result in blindness.

描述（由申请人提供）：骨髓来源的造血干细胞（HSC）已显示靶向视网膜中的新生血管形成部位。这些HSC在视网膜变性的小鼠模型中具有血管和神经营养性拯救，并且具有作为治疗视网膜变性疾病的治疗剂的临床应用的巨大潜力。然而，这些细胞发挥其营养活性的机制仍然未知。因此，为了表征这些细胞的活性，需要在它们的提取/鉴定中进行显著的改进。本研究将开发一种利用小分子的基于细胞的阳性选择试验。我们将通过三个具体目标来实现这一目标具体目标1：开发一种高通量筛选试验，以检测优先与表达CD 44 HI的细胞结合的小分子。使用基于竞争前细胞的ELISA样测定，我们将筛选非肽文库中有效结合CD 44 HI表达细胞的小分子。将使用比色测定法测定小分子抑制CD 44抗体与骨髓衍生细胞结合的能力。 具体目标2：通过化学修饰优化小分子的结合效率。采用用于文库合成的溶液相方法，将基于高通量试验中鉴定的经验证的先导化合物的系统性修饰制备二级文库，并针对抑制抗体与CD 44 Hi表达细胞结合的结合亲和力进行优化。 具体目标3：在视网膜神经元/血管变性的动物模型中，利用鉴定的小分子阳性分选出表达CD 44 Hi的细胞，并评估这些细胞的营养/生物活性。通过将整个骨髓与鉴定的小分子一起孵育，然后分选与小分子结合的细胞，我们可以分析这些细胞，看看它们是否表达CD 44 Hi和/或具有营养活性。这将通过将该细胞亚群玻璃体内注射到患有视网膜变性的小鼠中并观察如前所述的血管和神经元变性的挽救来实现。在视网膜疾病如糖尿病视网膜病变和年龄相关性黄斑变性中，发生异常血管生长，这是成年人视力丧失的主要原因。这项研究将有助于开发治疗药物，旨在治疗血管畸形引起的视网膜疾病，可导致失明。