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.

项目摘要 边缘包含稀有膜层干细胞（LSC）的少量亚群，该细胞不断地重新流行 角膜上皮。水缘干细胞缺乏症（LSCD）的患者无法再生角膜 上皮，导致角膜基质的“结膜化”，从而触发新血管形成，慢性 炎症，最终由于不可逆的不透明的角膜而导致失明。有几种方法 用于通过移植边缘组织或离体扩展的膜膜细胞来替代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。 但是，这些治疗在技术上仍然具有挑战性。此外，较大的患者人口 双侧LSCD没有自体LSC来源，并且同种异体的成功率要少得多 边缘组织移植，表明成功治疗双侧LSCD需要新的方法 患者。我们的合作研究小组现在发现了两个纯化的供体ABCB5+的新来源 双侧LSCD患者（I）同种异体免疫抑制ABCB5+ LSC的干细胞，（II） 自体多能皮肤皮肤衍生的ABCB5+ DSC（真皮干细胞）。我们最近发现 ABCB5基因是ATP结合盒（ABC）的新成员 在小鼠和人类中鉴定小鼠（LSC）中的多能干细胞和皮肤（DSC）。重要的是， ABCB5是一种细胞表面蛋白，我们的实验室开发的特定单克隆抗体是 能够前瞻性地将纯ABCB5阳性干细胞从边缘和皮肤中分离出来。最近 发表的原理证明实验表明，纯化的人ABCB5+（但不是ABCB5 LANG）LSC能够长期恢复免疫缺陷小鼠中角膜上皮 LSCD的模型，表明这种纯化的LSC种群有可能显着改善治疗 与LSCD相关的角膜疾病。当前赠款申请的目标是翻译我们 实验室研究结果对新的ABCB5+干细胞基疗法治疗单侧和双侧治疗 LSCD患者。我们的总体假设是，可以分离来自边缘或皮肤的ABCB5+干细胞 并在体外扩展作为干细胞来源，以重新生成角膜上皮时 具有单侧或双侧LSCD的接受者。该项目的三个模块将：验证 纯化的体外扩展ABCB5+ LSC（模块1）的再生潜力，并在体外膨胀 单侧和双侧LSCD患者的ABCB5+ DSC（模块2）； （模块3）创建LSC生物库 用于临床移植和进行研究，导致FDA IND批准。