Lacrimal gland repair using progenitor cells

使用祖细胞修复泪腺

• 批准号: 9199215
• 负责人: Helen P. Makarenkova
• 金额: $ 61.45万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9199215
• 关键词: Acinar Cell; Adult; Affect; Age; Alpha Cell; American; Autoimmune Process; Biological; Birth; Blindness; CD34 gene; Cell Lineage; Cell Transplantation; Cell Transplants; Cell physiology; Cells; Chronic; Clinical; Clone Cells; Data; Development; Disease; Disease Progression; Duct (organ) structure; Ductal; Ductal Epithelial Cell; Endothelial Cells; Epithelial; Epithelial Cells; Eye diseases; Female; Functional disorder; Gene Expression Profiling; Genetic Recombination; Gland; Goals; Human; Image; Infiltration; Inflammation; Inflammatory; Label; Lacrimal gland structure; Life Style; Maintenance; Molecular; Morphogenesis; Morphology; Mouse Strains; Mus; Myoepithelial; Myoepithelial cell; Neurons; PTPRC gene; Pathogenesis; Patients; Population; Proto-Oncogene Protein c-kit; Replacement Therapy; Reporter; Signal Transduction; Sjogren' s Syndrome; Smooth Muscle Actin Staining Method; Spatial Distribution; Stem cells; System; TACSTD1 gene; Tamoxifen; Testing; Therapeutic; Thrombospondin 1; Tissues; Transplantation; Visual Acuity; Work; aqueous; effective therapy; eye dryness; functional restoration; in vivo; injured; irritation; loss of function; monolayer; mouse model; novel; novel therapeutics; ocular surface; pluripotency; progenitor; public health relevance; repaired; stem; three dimensional cell culture; tool

 DESCRIPTION (provided by applicant): Aqueous deficiency dry eye (ADDE) is characterized by a lack of tear secretion from the lacrimal glands (LG). Currently there is no cure and no satisfactory treatment for ADDE. One of the challenges of understanding the mechanism of human ADDE pathogenesis is the inability to perform the biological and molecular studies prior to obvious clinical signs. As a result, it remains difficult to understand the precise steps of disease development. The goal of this proposal is to track stem and progenitor cell fate during tissue maintenance and disease and to identify the optimum progenitor cells to repair the "diseased" LG. We propose to use the recently developed TSP-1-/- mice that fully mimic the chronic development of autoimmune dry eye disease with loss of function preceding cellular inflammatory cell infiltration. We propose the novel idea that the LG has two functionally distinct progenitor cell populations: EPCP and MECP of the adult LG contains common (or bipotent) stem/progenitor cells able to give rise to both EPC and MEC cell lineages. We will follow each cell lineage in vivo and determine whether the MECP's or EPCP's have a common stem/progenitor cell producing acinar, ductal, and MEC's or a lineage specific progenitor. We will also investigate the fate of MECP's and EPCP's cells in `diseased' LG's and test the ability of normal MECP's and EPCPs to restore the function of `diseased' LG's. Our recent data shows that among c-kit-positive epithelial cell (EPC) populations sorted from mouse LG the c-kit+dim/EpCAM+/Sca1-/CD34-/CD45- cells are a putative epithelial (acinar and ductal) cell progenitor (EPCP) population. Isolated EPCP's express pluripotency factors and markers of the epithelial cell lineage Runx1 and EpCAM and form branches when grown in reaggregated 3D cultures. When transplanted into injured or diseased LG's, they restore the functional acinar and ductal epithelial component of the LG. We also found that myoepithelial cells (MEC's) have a high level of plasticity and are able to differentiate into neuronal, endothelial, and MEC lineages Availability of new tamoxifen- inducible epithelial and myoepithelial lineage specific-reporter mice allows us to directly isolate MEC's/MECP's and EPC's/EPCP's from the healthy and diseased LG's for morphological, functional and gene expression analyses, and also for progenitor cell purification, and transplantation. Our study will provide proof of concept for use f LG progenitor cells in cell replacement therapy for dry eye diseases that have no effective treatment or cure. Our work would also open up new therapeutic possibilities to treat ADDE.

 描述（由申请人提供）：水液缺乏性干眼症（ADDE）的特征是泪腺（LG）缺乏泪液分泌。目前ADDE尚无治愈方法，也没有令人满意的治疗方法。了解人类 ADDE 发病机制的挑战之一是无法在明显的临床症状之前进行生物学和分子研究。因此，仍然很难理解疾病发展的精确步骤。该提案的目标是追踪组织维持和疾病期间干细胞和祖细胞的命运，并确定修复“患病”LG 的最佳祖细胞。我们建议使用最近开发的 TSP-1-/- 小鼠，该小鼠完全模拟自身免疫性干眼病的慢性发展，并在细胞炎症细胞浸润之前丧失功能。我们提出了一个新颖的想法，即 LG 有两个功能不同的 祖细胞群：成体 LG 的 EPCP 和 MECP 含有共同（或双能）干/祖细胞，能够产生 EPC 和 MEC 细胞谱系。我们将跟踪体内的每个细胞谱系，并确定 MECP 或 EPCP 是否具有产生腺泡、导管和 MEC 的共同干/祖细胞或谱系特异性祖细胞。我们还将研究“患病”LG 中 MECP 和 EPCP 细胞的命运，并测试正常 MECP 和 EPCP 恢复“患病”LG 功能的能力。我们最近的数据显示，在从小鼠 LG 分选的 c-kit 阳性上皮细胞 (EPC) 群体中，c-kit+dim/EpCAM+/Sca1-/CD34-/CD45- 细胞是假定的上皮（腺泡和导管）细胞祖细胞 (EPCP) 群体。分离的 EPCP 表达上皮细胞谱系 Runx1 和 EpCAM 的多能因子和标记，并在重新聚集的 3D 培养物中生长时形成分支。当移植到受伤或患病的 LG 中时，它们可以恢复 LG 的功能性腺泡和导管上皮成分。我们还发现肌上皮细胞 (MEC) 具有高水平的可塑性，能够分化为神经元、内皮和 MEC 谱系。新的他莫昔芬诱导上皮和肌上皮谱系特异性报告小鼠的出现使我们能够直接从健康和患病 LG 中分离出 MEC's/MECP's 和 EPC's/EPCP's，以进行形态学、 功能和基因表达分析，也用于祖细胞纯化和移植。我们的研究将为使用 LG 祖细胞进行细胞替代疗法治疗没有有效治疗或治愈的干眼病提供概念证明。我们的工作还将为治疗 ADDE 开辟新的治疗可能性。