Multipotent ABCB5-positive cell therapeutics for corneal disease

用于治疗角膜疾病的多能 ABCB5 阳性细胞疗法

• 批准号: 9884771
• 负责人: NATASHA Y FRANK
• 金额: $ 117.63万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9884771
• 关键词: Affect; Allogenic; Allografting; Apoptotic; Applications Grants; Autologous; Bilateral; Binding; Blindness; Boston; Cell Maintenance; Cell Surface Proteins; Cell Therapy; Cell Transplantation; Cells; Chronic; Clinical; Cornea; Corneal Diseases; Corneal Stroma; Data; Dermal; Disease; Ear; Epithelial Cells; Eye; Genes; Genetic; Goals; Hospitals; Human; Immune system; Immunity; In Vitro; Inflammation; Intestines; Laboratories; Laboratory Research; Light; Malignant Neoplasms; Methods; Monoclonal Antibodies; Multipotent Stem Cells; Mus; Natural regeneration; Nature; Ophthalmology; Patients; Pediatric Hospitals; Procedures; Production; Publishing; Reporter; Research; Research Personnel; Retina; Role; Skin; Source; Specimen; Stem cell transplant; Therapeutic; Therapeutic Uses; Tissue Transplantation; Tissues; Transgenic Organisms; Translating; Translations; Transplantation; Vision; Woman; biobank; blind; clinically relevant; corneal epithelium; experimental study; human model; immunodeficient mouse model; improved; limbal; member; mouse model; neovascularization; novel strategies; patient population; prospective; real time monitoring; regenerative; restoration; stem cell niche; stem cell population; stem cell therapy; stem cells; success; transplant model

PROJECT SUMMARY 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. However, these treatments remain technically challenging. Moreover, the larger population of patients with bilateral LSCD has no source of autologous LSC and much less success was observed with allogeneic limbal tissue transplants, indicating new approaches are needed for successful treatment of bilateral LSCD patients. Our collaborative research group has now discovered two new sources of purified donor ABCB5+ stem cells for patients with bilateral LSCD (i) allogeneic immunosuppressive ABCB5+ LSC, and (ii) autologous multipotent dermal-skin-derived ABCB5+ DSC (Dermal Stem Cells). We recently discovered that the ABCB5 gene, a new member of the ATP-binding cassette (ABC) superfamily of active transporters, identifies multipotent stem cells in the limbus (LSC) and the skin (DSC) in mice and humans. Importantly, ABCB5 is a cell surface protein and specific monoclonal antibodies developed by our laboratories are capable of prospectively isolating pure ABCB5-positive stem cells from the limbus and skin. Recently published proof-of-principle experiments demonstrated that purified human ABCB5+ (but not ABCB5 negative) LSC were capable of long-term restoration of the corneal epithelium in an immunodeficient mouse model of LSCD, indicating that this purified LSC population has the potential to significantly improve therapy for corneal disease associated with LSCD. The goal of the current grant application is to translate our laboratory research findings into new ABCB5+ stem cell-based therapies to treat unilateral and bilateral LSCD patients. Our overall hypothesis is that ABCB5+ stem cells from the limbus or skin can be isolated and expanded in vitro as a source of stem cells to regenerate the corneal epithelium when transplanted to recipients with either a unilateral or bilateral LSCD. The three Modules of this project will: Validate the regenerative potential of purified in vitro-expanded ABCB5+ LSC (Module 1) and purified in vitro-expanded ABCB5+ DSC (Module 2) for unilateral and bilateral LSCD patients; and (Module 3) create an LSC Biobank for clinical transplantation and conduct studies resulting in FDA IND approval.

项目概要 角膜缘含有一小部分罕见的角膜缘干细胞 (LSC)，它们不断地重新填充角膜缘。 角膜上皮。角膜缘干细胞缺乏症（LSCD）患者无法再生角膜 上皮细胞，导致角膜基质“结膜化”，从而引发新血管形成，慢性 炎症，并最终因不可逆的不透明角膜而失明。几种方法有 已用于通过移植角膜缘组织或离体扩增的角膜缘细胞来替代LSC。这些程序 取得了一些成功，主要使用单侧 LSCD 患者的自体角膜缘组织。 然而，这些治疗方法在技术上仍然具有挑战性。此外，更多的患者群体 双侧 LSCD 没有自体 LSC 来源，同种异体的成功率要低得多 角膜缘组织移植，表明需要新的方法来成功治疗双侧 LSCD 患者。我们的合作研究小组现已发现纯化供体 ABCB5+ 的两种新来源 双侧 LSCD 患者的干细胞 (i) 同种异体免疫抑制 ABCB5+ LSC，以及 (ii) 自体多能真皮衍生的 ABCB5+ DSC（真皮干细胞）。我们最近发现 ABCB5 基因是主动转运蛋白 ATP 结合盒 (ABC) 超家族的新成员， 鉴定小鼠和人类的角膜缘 (LSC) 和皮肤 (DSC) 中的多能干细胞。重要的是， ABCB5是一种细胞表面蛋白，我们实验室开发的特异性单克隆抗体是 能够前瞻性地从角膜缘和皮肤中分离纯 ABCB5 阳性干细胞。最近 已发表的原理验证实验表明，纯化的人类 ABCB5+（但不是 ABCB5 阴性）LSC 能够长期恢复免疫缺陷小鼠的角膜上皮 LSCD 模型，表明这种纯化的 LSC 群体有可能显着改善治疗 用于与 LSCD 相关的角膜疾病。当前拨款申请的目标是将我们的 新的 ABCB5+ 干细胞疗法的实验室研究结果可治疗单侧和双侧 LSCD 患者。我们的总体假设是可以分离来自角膜缘或皮肤的 ABCB5+ 干细胞 并在体外扩增作为干细胞来源，移植到角膜上皮后再生 患有单侧或双侧 LSCD 的接受者。该项目的三个模块将： 验证 纯化的体外扩增的 ABCB5+ LSC（模块 1）和纯化的体外扩增的再生潜力 ABCB5+ DSC（模块 2）适用于单侧和双侧 LSCD 患者；以及（模块 3）创建 LSC 生物库 用于临床移植并进行研究，从而获得 FDA IND 批准。