ABCB5-positive stem cells for LSCD therapy

用于 LSCD 治疗的 ABCB5 阳性干细胞

• 批准号: 9106905
• 负责人: NATASHA Y FRANK
• 金额: $ 53.42万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9106905
• 关键词: Address; Alkalies; Allogenic; Aniridia; Autologous; Bilateral; Binding; Blindness; Boston; Burn injury; Cell Maintenance; Cell Surface Proteins; Cell Survival; Cell Therapy; Cells; Chronic; Clinical; Colon; Conjunctival Epithelium; Cornea; Corneal Diseases; Corneal Injury; Corneal Stroma; Development; Disease; Disease model; Ear; Epithelial; Eye; Genes; Genetic; Hospitals; Human; Immunodeficient Mouse; In Vitro; Inflammation; Injury; Intestines; Knockout Mice; Laboratories; Lysosomal Storage Diseases; Monoclonal Antibodies; Mus; Natural regeneration; Nature; Ophthalmology; Organ; Patients; Pediatric Hospitals; Procedures; Regenerative Medicine; Research; Role; Skin; Source; Stem cell transplant; Stem cells; Stomach; Stratified Squamous Epithelium; Surface; Therapeutic; Time; Tissues; Transplantation; Transplanted tissue; Vision; Woman; adult stem cell; clinically relevant; corneal epithelium; effective therapy; human tissue; improved; limbal; member; neovascularization; prevent; public health relevance; regenerative; repaired; self-renewal; stem cell population; stem cell therapy; success

 DESCRIPTION (provided by applicant): Many mammalian organs (skin, stomach, intestines, colon, and eye) possess a source of adult stem cells that continually replenishes their rapidly self-renewing epithelial surface. One important challenge in regenerative medicine is replacing these stem cells when they are eliminated following an injury or disease. The eye contains two highly specialized stratified squamous epithelia- the conjunctival epithelium and the corneal epithelium. A healthy corneal epithelium is essential for maintaining a clear cornea and normal vision. The limbus contains a small subpopulation of rare limbal stem cells (LSC) that continually repopulates the corneal epithelium. Patients with limbal stem cell deficiency (LSCD) are unable to regenerate the corneal epithelium, resulting in "conjunctivalization" of the corneal stroma that triggers neovascularization, chronic inflammation, and ultimately blindness due to an irreversibly opaque cornea. Several approaches have been used to replace LSC by transplanting limbal tissue or ex vivo expanded limbal cells. These procedures have obtained some success, mainly using autologous limbal tissue from patients with unilateral LSCD. Patients with bilateral LSCD have no source of autologous LSC and much less success was observed with allogeneic limbal tissue transplants. However, success of all these procedures was severely limited by the inability to prospectively identify and purify LSC. This problem was recently addressed by a new collaborative research group from the departments of: Ophthalmology, Genetics, and Transplantation Research (Mass Eye & Ear, Brigham & Women's Hospital and Boston Children's Hospital) that comprises the three PIs of this proposal, who discovered that the ABCB5 gene, a new member of the ATP-binding cassette (ABC) superfamily of active transporters, is expressed by stem cells of the limbus in both mouse and human tissues. Normal function of ABCB5+ LSC is required for corneal development and repair, through critical roles in stem cell maintenance and survival, and knockout mice that lack ABCB5 do not develop a fully differentiated mature corneal epithelium. Importantly, ABCB5 is a cell surface protein and specific monoclonal antibodies developed by the laboratories of Co-PIs Drs. M. Frank and N. Frank are capable of isolating pure ABCB5-positive cells from the limbus. Transplantation of purified human ABCB5+ (but not ABCB5-) LSC onto the corneal stroma of immunodeficient mice with induced LSCD restored the corneal epithelium, indicating that this purified LSC population has the potential to significantly improve therapy for corneal disease associated with LSCD. The current application builds upon these results to address the important challenges that prevent successful stem cell therapy for patients with unilateral or bilateral LSCD. Our overall hypothesis is that ABCB5+ stem cells from the limbus can be isolated and expanded ex vivo as a source of stem cells to regenerate the corneal epithelium when transplanted to recipients with either a unilateral or bilateral LSCD.

 描述（由适用提供）：许多哺乳动物器官（皮肤，摊位，肠，结肠和眼睛）具有成年干细胞的来源，这些干细胞不断地复制其快速自我更新的上皮表面。再生医学中的一个重要挑战是在受伤或疾病后消除这些干细胞时取代这些干细胞。眼睛包含两个高度专业的鳞状上皮 - 结膜上皮和角膜上皮。健康的角膜上皮对于保持清晰的角膜和正常视力至关重要。边缘含有稀有膜膜干细胞（LSC）的小亚群，该细胞不断地重新占据角膜上皮。水缘干细胞缺乏症（LSCD）的患者无法再生角膜上皮，从而导致角膜基质的“结膜”，从而触发了新生血管性，慢性炎症，最终由于使用多种方法来代替LSC，从而通过移植边缘组织或Ex ex ex viv膨胀的毛极细胞来代替LSC。这些程序已经取得了一些成功，主要使用单侧LSCD患者的自体膜层组织。双侧LSCD的患者没有自体LSC来源，并且在同种异体边缘组织移植中观察到的成功率要少得多。但是，所有这些程序的成功受到了前瞻性识别和净化LSC的严重限制。 This problem was recently addressed by a new collaborative research group from the departments of: Ophthalmology, Genetics, and Transplantation Research (Mass Eye & Ear, Brigham & Women's Hospital and Boston Children's Hospital) that comprises the three PIs of this proposal, who discovered that the ABCB5 gene, a new member of the ATP-binding cassette (ABC) superfamily of active transporters, is expressed by stem cells of the小鼠和人体组织中的边缘。 ABCB5+ LSC的正常功能是角膜发育和修复所必需的，通过在干细胞维持和存活中的关键作用，而缺乏ABCB5的敲除小鼠不会形成完全分化的成熟角膜上皮。重要的是，ABCB5是一种细胞表面蛋白，是由Co-Pis DRS实验室开发的特定单克隆抗体。 M. Frank和N. Frank能够从边缘分离纯ABCB5阳性细胞。用诱导的LSCD将纯化的人ABCB5+（但未ABCB5-）LSC移植到免疫缺陷小鼠的角膜基质中，恢复了角膜上皮，这表明这种纯化的LSC群体有可能显着改善与LSCD相关的角膜疾病的治疗。当前的应用基于这些结果，以应对为单侧或双侧LSCD患者提供成功的干细胞疗法的重要挑战。我们的总体假设是，可以分离出来自边缘的ABCB5+干细胞，并在体内扩展，作为干细胞的来源，当移植到具有单侧或双侧LSCD的受体时，可以再生角膜上皮。