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: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=587898
• 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.

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