Lrig1-Expressing Colonic Stem Cells in Homeostasis and Repair

表达 Lrig1 的结肠干细胞在稳态和修复中的作用

• 批准号: 9341258
• 负责人: Anne E Zemper
• 金额: $ 16.8万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2018-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9341258
• 关键词: Acute; Affect; Age; Biochemical; Biological Assay; Cell Differentiation process; Cell Proliferation; Cell physiology; Cells; Cessation of life; Chronic; Colon; Colorectal Cancer; Complement; Coupling; Data; Development Plans; Disease; Environment; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Epithelial; Epithelium; Evolution; Extramural Activities; Fluorescence Microscopy; Funding; Genetic Transcription; Goals; Growth; Growth Factor; Health; Histologic; Homeostasis; Human; Image; Immunoglobulin Domain; Impaired wound healing; Inflammation; Injury; Internal Ribosome Entry Site; Intestines; Knockout Mice; Knowledge; LGR5 gene; Leucine-Rich Repeat; Ligation; Link; Literature; Maintenance; Malignant - descriptor; Mediating; Mentored Research Scientist Development Award; Mentors; Mesenchyme; Microscopic; Microscopy; Modeling; Molecular; Mucous Membrane; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Natural regeneration; Phase; Play; Population; Process; Production; Property; Proteins; Recovery; Regenerative response; Regulation; Reporter; Repression; Research; Research Proposals; Resolution; Role; Running; Signal Transduction; Small Intestines; Sodium Dextran Sulfate; Stem cells; Tamoxifen; Testing; Tissues; Training; Transcript; Work; Wound Healing; Wounds and Injuries; adult stem cell; career development; cell growth; cohort; colonic crypt; design; human tissue; imaging study; improved; in vivo; injury and repair; insight; migration; mouse model; progenitor; programs; public health relevance; regenerative; repaired; response; response to injury; skills; stem; stem cell population; tissue regeneration; tissue repair; tool

DESCRIPTION (provided by applicant): This NIDDK Mentored Research Scientist Development Award application describes a three-year training plan designed to allow me to acquire additional skill and knowledge so that I can transition into directing my own independent and productive research program. In carrying out the proposed research, and by following a carefully mentored career development plan in a collegial research environment, I will add to my scientific repertoire by acquiring expertise in intestinal epithelial injury models and as well as live-cell and super-resolution microscopy. Using this newly acquired expertise, I will establish a scientific niche that will set me apart from my mentors and pave the way to a robust, extramurally funded research program. The research proposed in this application will focus on understanding how colonic stem cells function in repair after epithelial injury. It is known that Egfr signaling is activated after injury and this contributes to repair; however, this activation ad subsequent growth and proliferation, must be tightly regulated to avoid aberrant overgrowth. I have found that an Egfr inhibitor, Lrig1, marks a population of largely quiescent colonic stem cells, unlike the well-characterized, proliferative stem cell population marked by Lgr5. My proposal seeks to clarify the role of both Lrig1+ stem cells, and the role of Lrig1 as a negative regulator of Egfr, in colonic tissue stem cell homeostasis and wound healing. The requirement of stem cell populations in repair after injury has not yet been tested; this is critical to our understanding of the colonic epithelial crypt niche requirements for promoting proper regeneration. My Specific Aims are designed to test my over-arching hypothesis that Lrig1 is expressed in regenerative progenitor cells, in response to an active growth factor signal, such as activated Egfr signaling. In Aim 1, I will eliminate both Lrig1- and Lgr5-expressing stem cells before and after wound healing, to test their requirement in injury repair. I will evaluate initial injury and end-stage wound-repair by histological, molecular, and super-resolution analysis. In Aim 2, to examine the influence of Lrig1 on Egfr in wound healing, I will examine the effects of the absence of Lrig1 in wound healing, using Lrig1 null mice, and use advanced microscopy to examine the dynamics of Egfr activation. With the evolution of tools to manipulate stem cell populations in vivo, only now are the studies proposed here possible. Moreover, Vanderbilt's super-resolution microscopic capabilities will enable me to examine colon cell dynamics in tissue repair after injury at a resolution not previously possible. By coupling sophisticated mouse modeling with super-resolution imaging, the studies proposed here directly test the relationship between Lrig1- and Lgr5-expressing stem cells in injury repair, as well as examine the role of Lrig1 as a negative regulator of Egfr activity in this wound-healing context. The goal of this Mentored Research Scientist Development Award is to develop the expertise that will allow me to run an independent, funded research program that will contribute to improving human health and disease.

该NIDDK指导研究科学家发展奖申请描述了一个为期三年的培训计划，旨在让我获得额外的技能和知识，以便我可以过渡到指导我自己的独立和富有成效的研究计划。在开展拟议的研究，并遵循一个精心指导的职业发展计划，在大学的研究环境，我将通过获得肠上皮损伤模型的专业知识，以及活细胞和超分辨率显微镜添加到我的科学剧目。利用这一新获得的专业知识，我将建立一个科学的利基，这将使我从我的导师，并铺平道路，一个强大的，校外资助的研究计划。 这项申请中提出的研究将集中在了解结肠干细胞在上皮损伤后的修复中如何发挥作用。已知Egfr信号传导在损伤后被激活，这有助于修复;然而，必须严格调节这种激活和随后的生长和增殖，以避免异常过度生长。我发现Egfr抑制剂Lrig 1标记了一群基本上静止的结肠干细胞，与Lgr 5标记的特征良好的增殖干细胞群不同。我的建议旨在阐明Lrig 1+干细胞的作用，以及Lrig 1作为Egfr的负调节因子在结肠组织干细胞稳态和伤口愈合中的作用。损伤后修复中干细胞群的需求尚未得到测试;这对我们理解结肠上皮隐窝生态位促进适当再生的需求至关重要。我的特定目标旨在测试我的过度假设，即Lrig 1在再生祖细胞中表达，以响应活性生长因子信号，如激活的Egfr信号。在目标1中，我将在伤口愈合前后消除表达Lrig 1和Lgr 5的干细胞，以测试它们在损伤修复中的需求。我将评估初始 通过组织学、分子学和超分辨率分析研究损伤和终末期创伤修复。在目标2中，为了研究Lrig 1对Egfr在伤口愈合中的影响，我将使用Lrig 1缺失小鼠来研究Lrig 1缺失对伤口愈合的影响，并使用先进的显微镜来研究Egfr激活的动力学。 随着体内操纵干细胞群的工具的发展，只有现在才有可能进行这里提出的研究。此外，范德比尔特的超分辨率显微镜能力将使我能够检查结肠细胞动力学在组织修复损伤后在一个决议以前不可能。通过耦合复杂的鼠标 通过使用超分辨率成像建模，本文提出的研究直接测试了表达Lrig 1和Lgr 5的干细胞在损伤修复中的关系，并检查了Lrig 1在这种伤口愈合背景下作为Egfr活性的负调节剂的作用。这个指导研究科学家发展奖的目标是发展专业知识，使我能够运行一个独立的，资助的研究计划，这将有助于改善人类健康和疾病。