Development of Novel Cell Free Therapies to Prevent Age-related Motor Neuron loss

开发新型无细胞疗法来预防与年龄相关的运动神经元损失

• 批准号: BB/I015787/1
• 金额: $ 11.71万
• 依托单位: University of Reading
• 依托单位国家: 英国
• 项目类别: Training Grant
• 财政年份: 2011
• 资助国家: 英国
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI015787%2F1
• 关键词: Development; Novel; Cell; Free; Therapies

Addressing the adverse effects of ageing is a priority research area not only of the BBSRC but also of the Industrial sector. This project aims to develop new and novel cell free reagents to prevent age-induced motor neuron loss by forging an alliance between academia (University of Reading) and industry (NATURAL BIOSCIENCES LTD) to exploit a new finding made at the University of Reading. An extensive body of literature exists showing that as age increases beyond 60 years, human muscle undergoes denervation, due to a loss of MN that leads to a poor quality of life and often death. Permanently denervated muscle is ultimately lost and replaced by fat and fibrous tissue. This project will exploit our timely finding that deletion of the gene Myostatin in the mouse results in an unprecedented survival of MN, which are normally eliminated by programmed cell death during (Elashry et al 2010 Journal of Anatomy, submitted). Since the myostatin mutation only affects skeletal muscle, we have developed the central hypothesis 'that the skeletal muscle produces diffusible trophic factors that act as neuro-protective agents that prevent age related motor neuron death'. The hypothesis will be tested by the student. This study will take advantage of a strategic alliance between the groups of Professor Patel, who at the University of Reading have made this key discovery and Dr Steve Ray of NATURAL BIOSCIENCES LTD who brings to the research arena extensive (including expertise development of cell free therapies to promote neural regeneration) and research reagents (mouse lines displaying premature MN death, including SODG93A mouse line. Project aims: (1) To determine using in-vitro platforms whether MN that develop in the absence of myostatin are resistant to neuro-toxins. (2) To determine in-vivo whether the ageing resistant MN that form in the absence of myostatin display the survival characteristics of young MN. (3) To conduct in-vitro experiments to determine whether cell free reagents (conditioned media, micro vesicles, RNA) generated from myostatin null tissue harbour neuro-protective properties. (4) To determine whether cell free reagents developed in aim (3) offer neuro-protective properties in an in-vivo context. Timeline: 0-9 months. Isolation and culturing foetal MN from wild type and myostatin null mice by standard methods (Kume et al (2005) Neuroscience Letters 383:199-202). Training: cell culture, working with transgenic animals. Venue: University of Reading 10-18 months. Neuroprotective properties of MN from myostatin nulls to exposure to glutamate or nitric oxide agonists will be established. Training: cell culture, statistical analysis of data. Venue: NATURAL BIOSCIENCES LTD 12-18 months. Establish myostatin/SODG93A line. Training: animal handling, molecular biology. Venue: University of Reading 19-30 months. Histological examination of motor nerves in myostatin/ SODG93A line. Training: con-focal microscopy, electron microscopy. Venue: University of Reading 28-33 months. Isolation of cell free biologically active reagents (conditioned media, micro vesicles, RNA) from myostatin null muscle. Training: molecular biology, biochemistry. Venue: NATURAL BIOSCIENCES LTD 28-36 months. Confirm the neuroprotective properties of cell free reagents derived from myostatin null mice to prevent the toxic effects of glutamate or nitric oxide agonists. Training: toxicology, statistics. Venue: NATURAL BIOSCIENCES LTD 36-48 months. Quantify evidence of neuroprotection of cell free reagents from myostatin nulls in SODG93A mice. Training: histology. Venue: University of Reading The student will spend about each 24 months in the laboratory of Professor Patel and NATURAL BIOSCIENCES LTD. Natural Biosciences SA will provide £15,000 per year for animal, tissue culture, reagent and travel costs. Outputs from this project will be used for a future project grant application- details in later sections.

应对老龄化的不利影响不仅是BBSRC的优先研究领域，也是工业部门的优先研究领域。该项目旨在开发新型无细胞试剂，通过建立学术界（阅读大学）和工业界（NATURAL BIOSCIENCES LTD）之间的联盟，利用大学的新发现来预防年龄引起的运动神经元丢失。阅读。大量文献表明，随着年龄的增长，超过60岁，人体肌肉会因MN的丧失而经历去神经支配，从而导致生活质量下降，甚至死亡。永久性失神经支配的肌肉最终会丢失，并被脂肪和纤维组织取代。该项目将利用我们及时发现的小鼠中基因肌生长抑制素的缺失导致MN的前所未有的存活，MN通常通过过程中的程序性细胞死亡消除（Elashry等人，2010 Journal of Anatomy，提交）。由于肌生长抑制素突变只影响骨骼肌，我们提出了一个中心假设，即骨骼肌产生可扩散的营养因子，这些营养因子作为神经保护剂，防止与年龄相关的运动神经元死亡。这个假设将由学生来检验。本研究将利用Patel教授和NATURAL BIOSCIENCES LTD的Steve Ray博士之间的战略联盟，Patel教授在阅读大学做出了这一关键发现，Steve Ray博士为研究竞技场带来了广泛的（包括无细胞疗法的专业知识开发，以促进神经再生）和研究试剂（显示过早MN死亡的小鼠系，包括SODG 93 A小鼠系）。项目目的：（1）使用体外平台确定在没有肌肉生长抑制素的情况下发育的MN是否对神经毒素具有抗性。(2)为了确定在不存在肌生长抑制素的情况下形成的抗老化MN是否显示年轻MN的存活特征。(3)进行体外实验，以确定由肌生长抑制素缺失组织生成的无细胞试剂（条件培养基、微泡、RNA）是否具有神经保护特性。(4)确定目标（3）中开发的无细胞试剂是否在体内环境中提供神经保护特性。时间轴：0-9个月。通过标准方法从野生型和肌生长抑制素缺失小鼠中分离和培养胎儿MN（Kume等人（2005）Neuroscience Letters 383：199-202）。培训：细胞培养，转基因动物工作。地点：阅读大学10-18个月。将建立从肌生长抑制素无效到暴露于谷氨酸或一氧化氮激动剂的MN的神经保护特性。培训：细胞培养，数据统计分析。地点：自然生物科学有限公司12-18个月。建立肌生成抑制素/SODG 93 A细胞系。培训：动物处理，分子生物学。地点：阅读大学19-30个月。肌生成抑制素/SODG 93 A系中运动神经的组织学检查。培训：共聚焦显微镜，电子显微镜。地点：阅读大学28-33个月。从无肌生长抑制素的肌肉中分离无细胞生物活性试剂（条件培养基、微泡、RNA）。培训：分子生物学，生物化学。地点：自然生物科学有限公司28-36个月。确认来自肌生长抑制素缺失小鼠的无细胞试剂的神经保护特性，以防止谷氨酸或一氧化氮激动剂的毒性作用。培训：毒理学、统计学。地点：自然生物科学有限公司36-48个月。在SODG 93 A小鼠中定量无细胞试剂对肌生长抑制素无效的神经保护的证据。培训：组织学。会议地点：阅读大学学生将在帕特尔教授和自然生物科学有限公司的实验室度过大约每24个月。自然生物科学公司每年将提供15,000英镑的动物，组织培养，试剂和旅行费用。本项目的成果将用于未来的项目资助申请-详细信息在后面的章节中。