Domestic animal stem cell biology for understanding tissue development, maintenance and therapeutic interventions

用于了解组织发育、维护和治疗干预的家畜干细胞生物学

• 批准号: RGPIN-2014-04587
• 负责人: Koch, Thomas
• 金额: $ 2.55万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=655787
• 关键词: Domestic; animal; stem; cell; biology

Introduction*Horses are multi-use livestock that contribute more than $19 billion annually to the Canadian economy. Injuries to tendon and joints are among the most common causes of lost training days or premature retirement in equine athletes. Stem cells hold the promise of novel therapeutic approaches to these difficult-to-treat injuries. I am an Early Career Researcher (ECR) and the overall goal of my research program is to use domestic animal stem cells as potent experimental systems to investigate and potentially treat common, economically important livestock and companion animal diseases while simultaneously enhancing our basic understanding of tissue regeneration biology. Students in my laboratory will gain fundamental knowledge in the regulation of various stem cell populations, develop unique skills in the experimental investigation of disease processes, and in the use of these cells as innovative bioengineered therapies for specific orthopedic conditions in domestic animals. * This research program will focus on understanding the role of microRNAs (miRNAs) in stem cell homeostasis, stem cell cartilage differentiation and stem cell immunomodulatory functions. miRNAs are a class of endogenous non-coding RNAs thought to be involved in regulating one third of all mammalian genes. This year we reported that miR-140 is upregulated during chondrogenesis of equine cord blood-derived mesenchymal stromal cells (eCB-MSC). miRNAs in vivo stability and the availability of synthetic mimics and inhibitors makes them ideal candidates for diagnostic, prognostic and therapeutic approaches. I will, in the next five years, interrogate specific miRNAs with regard to their ability to modulate eCB-MSC chondrogenic capacity, enhance biomechanical properties of formed neocartilage, and modulate host animal immune responses in vitro. miRNA and mRNA transcriptome signatures will be determined using next generation sequencing (NGS) to discover novel miRNAs that regulate these processes and correlate miRNA expression to possible mRNA targets. Development of MSC-based therapies, strategic use of selected miRNA mimics or inhibitors for immediate in vivo testing for cartilage repair and osteoarthritic treatments as well as novel diagnostic and prognostic joint biomarkers is expected from the projected knowledge gain. **Background*I was the first to report the isolation of eCB-MSC. eCB-MSC is better at forming neocartilage than MSC from bone marrow and adipose tissue. The mechanisms are unknown. Neocartilage generated by chemical induction of MSC has inferior mechanical properties compared to normal hyaline cartilage. Mechanical MSC stimulation is increasingly used to replicate the load-bearing environment of hyaline cartilage. Inflammation is a central component of osteoarthritis. The ability of eCB-MSC to suppress lymphocyte proliferation, as a proxy for anti-inflammatory cell properties, was recently confirmed in my lab. **Objectives*During this first 5-year funding period, my primary aim is to complement previous descriptive studies on eCB-MSCs with fundamental cell mechanistic investigation. Based on my program goals, the specific hypothesis for the proposed studies is: miRNAs determine eCB-MSC chondrogenic and lymphocyte suppressive potency. The 3 key elements of this proposal are: 1) To identify miRNA signatures associated with high chondrogenic potency of MSC and examine the roles of selected miRNAs in the regulation of important target mRNAs. 2) To characterize the roles of specific mechano-responsive miRNAs in chondrogenic differentiation. 3) To identify and characterize miRNAs associated with immune-modulatory processes in eCB-MSCs.

简介*马是多种用途的牲畜，每年为加拿大经济贡献超过190亿美元。肌腱和关节受伤是马术运动员失去训练天数或提前退役的最常见原因之一。干细胞有望为这些难以治疗的损伤提供新的治疗方法。我是一名早期职业研究人员(ECR)，我的研究计划的总体目标是将家畜干细胞作为强大的实验系统来研究和潜在地治疗常见的、经济上重要的家畜和伴随的动物疾病，同时增强我们对组织再生生物学的基本理解。我实验室的学生将获得各种干细胞种群调控的基本知识，在疾病过程的实验研究中发展独特的技能，并将这些细胞用作家畜特定整形外科条件下的创新生物工程疗法。*这项研究计划将重点了解microRNAs(MiRNAs)在干细胞稳态、干细胞软骨分化和干细胞免疫调节功能中的作用。MiRNAs是一类内源性非编码RNA，被认为参与调控三分之一的哺乳动物基因。今年我们报道了miR-140在马脐带血来源的间充质基质细胞(ECB-MSC)软骨形成过程中表达上调。MiRNAs在体内的稳定性以及合成模拟物和抑制剂的可用性使它们成为诊断、预后和治疗方法的理想候选者。在接下来的五年里，我将询问特定的miRNAs是否有能力调节ECB-MSC的软骨形成能力，增强形成的新生软骨的生物力学特性，以及在体外调节宿主动物的免疫反应。将使用下一代测序(NGS)来确定miRNA和mRNA转录组的特征，以发现调节这些过程的新miRNAs，并将miRNA表达与可能的mRNA靶标关联起来。基于MSC的治疗方法的开发，选定的miRNA模拟物或抑制剂的战略性使用，用于软骨修复和骨关节炎治疗的即时体内测试，以及新的诊断和预后关节生物标记物的预期知识增长。**背景*我是第一个报道ECB-MSC分离的人。ECB-MSC在骨髓和脂肪组织中形成新生组织的能力优于MSC。其机制尚不清楚。与正常透明软骨相比，MSC化学诱导产生的新生软骨具有较差的力学性能。机械刺激MSC越来越多地被用来复制透明软骨的承载环境。炎症是骨性关节炎的中心组成部分。ECB-MSC抑制淋巴细胞增殖的能力，作为抗炎细胞特性的替代品，最近在我的实验室得到证实。**目标*在这第一个5年的资助期内，我的主要目标是用基础细胞机制研究来补充之前对ECB-MSCs的描述性研究。根据我的计划目标，建议研究的具体假设是：miRNAs决定ECB-MSC的软骨生成和淋巴细胞抑制能力。这项建议的3个关键要素是：1)鉴定与MSC的高软骨形成能力相关的miRNA信号，并研究选定的miRNAs在调节重要的靶向mRNAs中的作用。2)研究特异性机械反应miRNAs在软骨细胞分化中的作用。3)鉴定和鉴定ECB-MSCs中与免疫调节相关的miRNAs。