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: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=685486
• 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.

马是多用途牲畜，每年为加拿大经济贡献超过190亿加元。肌腱和关节损伤是马运动员失去训练日或过早退役的最常见原因之一。干细胞为这些难以治疗的损伤提供了新的治疗方法。我是一名早期职业研究员（ECR），我的研究计划的总体目标是使用家畜干细胞作为有效的实验系统来研究和治疗常见的，经济上重要的牲畜和伴侣动物疾病，同时提高我们对组织再生生物学的基本理解。在我的实验室的学生将获得各种干细胞群体的调节的基础知识，发展疾病过程的实验研究的独特技能，并在使用这些细胞作为创新的生物工程疗法在家畜特定的骨科条件。* 该研究计划将重点关注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，被认为参与调节所有哺乳动物基因的三分之一。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）的软骨形成过程中上调。miRNA在体内的稳定性以及合成模拟物和抑制剂的可用性使其成为诊断、预后和治疗方法的理想候选者。在接下来的五年里，我将研究特定的miRNAs在体外调节eCB-MSC软骨形成能力、增强形成的新软骨的生物力学特性和调节宿主动物免疫反应的能力。将使用下一代测序（NGS）来确定miRNA和mRNA转录组特征，以发现调节这些过程并将miRNA表达与可能的mRNA靶点相关联的新型miRNA。基于MSC的疗法的开发，选择的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志物，预计将从预计的知识获得。** 背景 * 我是第一个报道分离eCB-MSC的人。eCB-MSC在形成新软骨方面优于来自骨髓和脂肪组织的MSC。其机制尚不清楚。与正常透明软骨相比，MSC化学诱导产生的新软骨具有较差的机械性能。机械MSC刺激越来越多地用于复制透明软骨的承重环境。炎症是骨关节炎的核心组成部分。eCB-MSC抑制淋巴细胞增殖的能力，作为抗炎细胞特性的代表，最近在我的实验室得到了证实。** 目标 * 在第一个5年的资助期内，我的主要目标是通过基本的细胞机制研究来补充以前对eCB-MSC的描述性研究。基于我的项目目标，所提出的研究的具体假设是：miRNAs决定eCB-MSC软骨形成和淋巴细胞抑制效力。该方案的3个关键要素是：1）鉴定与MSC的高软骨形成潜能相关的miRNA特征，并检查所选miRNA在重要靶mRNA的调节中的作用。2)研究特异性机械应答miRNAs在软骨细胞分化中的作用。3)鉴定和表征eCB-MSC中与免疫调节过程相关的miRNA。