Nanoparticle-based synthetic transcription factor to induce stem cell myogenesis

基于纳米颗粒的合成转录因子诱导干细胞肌发生

• 批准号: 9461879
• 负责人: Kibum Lee
• 金额: $ 16.82万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-26 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9461879
• 关键词: Achievement; Address; Adipose tissue; Advanced Development; Animals; Area; Biology; Cell Lineage; Cell Therapy; Cell physiology; Cells; Chemicals; Clinical; Consensus; DNA Binding Domain; Data; Degenerative Disorder; Development; Disease; Elements; Epigenetic Process; Future; Gene Expression; Gene Expression Regulation; Gene Targeting; Generations; Genes; Genetic Transcription; Goals; Human; Libraries; Mediating; Mesenchymal Stem Cells; Methods; Muscle; Muscle Cells; Muscular Dystrophies; Musculoskeletal Diseases; Myogenic Regulatory Factors; Myogenin; Natural regeneration; Pathway interactions; Patients; Plasmids; Property; Proteins; Replacement Therapy; Research; Research Personnel; Safety; Signal Pathway; Skeletal Muscle; Source; Stem cells; Structure; Testing; Transfection; Translations; Virus; Work; base; clinical application; design; expectation; in vivo; innovation; innovative technologies; interdisciplinary approach; myogenesis; nano; nanomedicine; nanoparticle; novel strategies; precision medicine; protein expression; small molecule; stem; stem cell biology; stem cell differentiation; stem cell fate; therapeutic development; tool; transcription factor

PROJECT SUMMARY Recent advances in stem cell biology hold great potential in developing new approaches for the treatment of many devastating diseases, including musculoskeletal disorders (MSDs). Stem cell-based therapies for regenerating functional muscle cells and restoring muscular functions to damaged skeletal muscles can be critical for the development of therapeutic advances in musculoskeletal disease and disorder. Such approaches, however, require the generation of engraftable cell sources of functional myogenic cells and better control of stem cell myogenic differentiation in an effective, selective, and safe manner. To this end, the main goal of this proposal is to develop a bio-inspired platform that can replicate the structure and function on endogenous proteins called transfection factors (TFs; MyoD and Myogenin), which are specific for muscle-specific genes and responsible for orchestrating overall stem differentiation into muscle cells. Our bio-inspired platform called NanoScript, is a nanoparticle-based transcription factor that behaves and function just like natural TF proteins. This NanoScript platform is designed to be gene-specific and can effectively activate targeted gene expressions (e.g. MyoD, Myogenin and the related endogenous genes) in a non-toxic and non- viral manner. Out central hypothesis, based upon recent achievement and preliminary data, is that our proposed NanoScript platform can effectively generate functional muscle cells from human patient-derived adipose- derived mesenchymal stem cells (AMDSCs), which are an abundant source of stem cells, with patient-specific stem cells treatment possibilities. We propose to test our central hypothesis and achieve our objectives by addressing the following specific aims: Aim1: Design and synthesize muscle cell-specific TFs (MRF) and epigenetic modulators for the construction of enhanced muscle cell-specific NanoScripts [NanoScripts-MRF]. Aim2: Utilize NanoScript to activate muscle-specific genes in ADMSCs for generating muscle cells. The proposed research is innovative, as this concept of developing a TF emulator by integrating two multidisciplinary approaches (chemical biology and nanomedicine) onto a single nano-platform for non-viral gene regulation in stem cells has not been developed. The proposed research is significant, since we will develop an innovative technology platform and our NanoScript is an easily tunable and robust platform, it can be further developed to combine with epigenetic modulators or other synergists for effective and selective induction of functional muscle cells. Collectively, upon successful completion of the proposed study, our expectations are that NanoScript-MRF will activate transcription of the muscular-specific genes containing their cognate TF consensus DNA binding domain, which will lead to an enhanced stem differentiation into muscle cells. Because NanoScript is non-toxic and non-viral, the generated muscle cells will be considered for translation into in vivo animal studies in future studies.

项目摘要 干细胞生物学的最新进展在开发新方法的治疗方面具有很大的潜力 许多毁灭性疾病，包括肌肉骨骼疾病（MSDS）。基于干细胞的疗法 再生功能性肌肉细胞并恢复肌肉功能为受损的骨骼肌可能是 对于发展肌肉骨骼疾病和疾病的治疗进展至关重要。这样的方法， 但是，需要产生功能性肌源性细胞的植入细胞来源，并更好地控制 干细胞肌生成的分化，有效，选择性和安全的方式。 为此，该提案的主要目标是开发一个可以复制结构的生物启发的平台 并在称为转染因子（TFS; Myod和Myogenin）的内源性蛋白上的功能，这是特定的 用于肌肉特异性基因，并负责策划整体茎分化为肌肉细胞。我们的 生物启发的平台称为纳米文字，是一种基于纳米颗粒的转录因子，其行为和功能 就像天然TF蛋白一样。该纳米录制平台设计为基因特异性，可以有效地激活 靶向基因表达（例如myod，肌生成素和相关内源基因）在无毒和非毒性中 病毒方式。基于最近的成就和初步数据，中心假设是我们提出的 Nanoscript平台可以有效地从人类患者衍生的脂肪中产生功能性肌肉细胞 衍生的间充质干细胞（AMDSC），它是干细胞的丰富来源，患者特异性 干细胞治疗可能性。 我们建议通过解决以下特定的特定来检验我们的中心假设并实现目标 目标：AIM1：设计和合成肌肉细胞特异性TF（MRF）和表观遗传调节剂 构建增强的肌肉细胞特异性纳米物品[nanoscripts-MRF]。 AIM2：利用纳米字样 激活ADMSC中的肌肉特异性基因以产生肌肉细胞。 拟议的研究具有创新性，因为这是通过整合两个来开发TF模拟器的概念 多学科方法（化学生物学和纳米医学）在单个纳米平台上用于非病毒基因 干细胞中的调节尚未开发。拟议的研究很重要，因为我们将开发 创新的技术平台和我们的纳米文字是一个易于调谐且健壮的平台，它可以进一步 开发以与表观遗传调节剂或其他协同作用相结合，以有效和选择性地诱导 功能性肌肉细胞。总的来说，在成功完成拟议的研究后，我们的期望是 该纳米文字MRF将激活包含其同源TF的肌肉特异性基因的转录 共识DNA结合结构域，这将导致茎分化增强对肌肉细胞。因为 纳米文本是无毒的，无毒的，生成的肌肉细胞将被考虑转化为体内 未来研究的动物研究。