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（由适用提供）：缺乏水性干眼（ADDE）的特征是缺乏泪液烤架（LG）的撕裂分泌。目前尚无治愈方法，也没有补充的满意度。理解人类添加发病机理机制的挑战之一是在明显的临床体征之前无法进行生物学和分子研究。结果，很难理解疾病发展的确切步骤。该提案的目的是在组织维持和疾病期间跟踪茎和祖细胞命运，并确定最佳祖细胞以修复“患病” LG。我们建议使用最近开发的TSP-1 - / - 小鼠，以完全模仿自身免疫性眼病的慢性发展，并且在细胞炎性细胞浸润之前的功能丧失，功能丧失。我们提出了一个新颖的想法，即LG具有两个功能上不同的想法 祖细胞群体：成年LG的EPCP和MECP包含常见（或二强）茎/祖细胞，可以引起EPC和MEC细胞谱系。我们将在体内遵循每个细胞谱系，并确定MECP或EPCP是否具有共同的茎/祖细胞产生腺泡，导管和MEC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“疾病” LG功能的能力。我们最近的数据表明，在C-KIT阳性上皮细胞（EPC）中，用小鼠LG排序的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）的种群。分离的EPCP的EPCP表达多能因素和上皮细胞谱系Runx1和Epcam的标记并在重新培养的3D培养物中生长时形成分支。当移植到受伤或解散的LG中时，它们会恢复LG的功能性腺泡和导管上皮成分。 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 allow us to directly isolate MEC's/MECP's and EPC's/EPCP's from the healthy and disseased LG's for morphological,功能和基因表达分析，还用于祖细胞纯化和移植。我们的研究将提供概念证明，用于使用F LG祖细胞在细胞替代疗法中用于没有有效治疗或治愈的干眼疾病。我们的工作还将为治疗加法提供新的治疗可能性。