Lacrimal Gland Repair Using Progenitor Cells

泪腺(LG)是一种外分泌的小管腺，分泌泪膜的水层。 LG产生的泪液在数量和/或质量上的任何改变都会导致缺水干燥 眼病(ADDE)。ADDE是一种慢性疾病，影响着数百万美国人，其症状包括 从干痒到视力模糊，并伴随着眼睛感染风险的增加。再生性 而针对LG修复的干细胞疗法现在正在崭露头角，然而，我们对LG的理解 干细胞和祖细胞生物学仍然不完整。我们之前在祖细胞上的实验 移植标记保留细胞(LRC)分析和克隆分析的细胞谱系追踪提示成年 LG有储备的或极具可塑性的祖细胞，特别是在Sox10细胞谱系中，LG 祖细胞可能在ADDE中起治疗作用。我们最近的研究表明，肌上皮细胞 (MECs)保持高度可塑性，在LG损伤后能够分化为腺泡细胞 移植。此外，体内MECs的基因消除表明它们是LG所必需的 祖细胞和腺泡细胞功能。我们还表明LG炎症可以通过 PAnnexin-1(Panx1)膜通道糖蛋白--炎症体组装的关键调节因子。 炎性小体是一种大的细胞内多蛋白复合体，可以激活促炎细胞因子 对感染和组织损伤或慢性炎症的反应。我们的研究表明，上皮细胞 细胞感知损伤/炎症，并通过产生促炎症反应来促进炎症反应 炎性细胞因子IL-1β(IL-1β)和IL-18。此外，我们还发现，阻断PAnnexin-1 LG中的Panx1或Caspase 4减少促炎细胞因子的释放并改善移植细胞 嫁接。在一项新的计划中，我们将研究LG前体细胞的分子和细胞性质 在健康和慢性炎症的LG周围有分化的细胞。首先，我们将评估Sox10 (MEC和ACENA)在健康和疾病的LGs中建立谱系并研究Sox10的作用 MEC谱系建立过程中Krt5表达祖细胞的研究第二，我们将调查 炎性小体通路在SS中LG祖细胞及其他上皮细胞功能和LG修复中的作用 小鼠模型第三，我们将使用抗炎Panx1通路联合阻断疗法 以及用于LG修复的细胞移植。我们希望找出LG功能障碍的关键机制。 我们对炎症体途径的分析可能有助于开发全新的药物治疗 加德。我们的研究也将为LG祖细胞在细胞中的应用提供重要的使能信息 无法有效治疗或治愈的干眼病的替代疗法。

Lacrimal gland (LG) is an exocrine tubuloacinar gland that secretes the aqueous layer of the tear film. Any alteration in the quantity and/or quality of tears produced by LG can result in aqueous deficiency dry eye disease (ADDE). ADDE is a chronic condition affecting millions of Americans, with symptoms ranging from a dry itchiness to blurred vision and accompanied by an increased risk of eye infections. Regenerative and stem cell therapies that target LG repair are now coming to the fore, however, our understanding of LG stem and progenitor cell biology is still incomplete. Our previous experiments on progenitor cell transplantation label retaining cell (LRC) analysis and cell lineage tracing with clonal analyses suggest adult LG has reserve or extremely plastic progenitor cells especially in the Sox10+ cell lineage and that LG progenitor may have a therapeutic role in ADDE. Our recent studies demonstrate that myoepithelial cells (MECs) retained high level of plasticity and are able to differentiate into acinar cells upon LG injury or transplantation. Moreover, genetic elimination of MECs in vivo showed that they are required for LG progenitor and acinar cell function. We also showed that LG inflammation could be mediated by the Pannexin-1 (Panx1) membrane channel glycoprotein - a key regulator of inflammasome assembly. Inflammasomes are large intracellular multiprotein complexes that activate proinflammatory cytokines in response to infection and tissue damage or chronic inflammation. Our study suggests that the epithelial cells sense damage/inflammation and contribute to the inflammatory response by producing the pro- inflammatory cytokines interleukin-1 Beta(IL-1Beta) and IL-18. Moreover, we showed that blocking pannexin-1 (Panx1) or Caspase 4 in LG reduces pro-inflammatory cytokine release and improves transplanted cell engraftment. In a new proposal we will study the molecular and cellular nature of LG progenitor and surrounding differentiated cells in healthy and chronically inflamed LG. First we will evaluate the Sox10+ (MEC and acinar) lineage establishment in healthy and diseased LGs and investigate the role Sox10 expression in Krt5 expressing progenitors in establishment of the MEC lineage. Second, we will investigate the role of inflammasome pathways in LG progenitor and other epithelial cell function and LG repair in SS mouse models Third, we will use a combination of anti-inflammatory Panx1 pathways blocking therapies and cell transplantation for LG repair. We expect to identify key mechanisms responsible for LG dysfunction. Our analysis of inflammasome pathways may facilitate development of entirely new drug treatments for ADDE. Our studies will also provide important enabling information for use of LG progenitor cells in cell replacement therapy for dry eye diseases that have no effective treatment or cure.