Transcriptome-based approach to identify biomimetics for stem cell-derived factors regulating tissue regeneration

项目说明：干细胞促进人体众多组织的再生。然而，支撑其恢复能力的机制仍有待阐明。越来越多的证据表明，干细胞不会融入新形成的组织，而是通过分泌分子来指导驻留细胞激活修复机制。因此，一种范式的转变是接受由干细胞而不是干细胞产生的因子是控制组织修复的有效实体。这个概念代表了一个前所未有的机会，可以在工业规模上识别和开发化合物，不仅可以定义，还可以单独分析，从而加快它们的安全使用，在没有细胞参与的情况下促进组织再生。两个主要障碍阻碍了系统的方法来识别分泌干细胞因子的仿生学。首先是缺乏一个对干细胞衍生因子产生生物反应的强大平台。第二个是在发现平台的另一端；缺乏大量能引起细胞表型变化的生物安全化合物。这两个障碍最近都被克服了，首先是通过我们发现人类脂肪源性间充质干细胞（ADMSC）分泌因子（SF），促进细胞增殖、迁移和防止应激诱导的衰老。第二个发展是对各种细胞中超过1300种fda批准的化合物进行分析，以开发大型数据集，使每种药物不仅可以根据其生物效应进行分析，还可以根据基因转录进行分析（连接图，C-Map）。关键创新是利用生物信息学方法，将admsc衍生因子在调节增殖、迁移和衰老能力方面引发的转录变化与安全化合物文库中公布的转录变化相匹配，从而鉴定出干细胞衍生因子调节组织再生的仿生学。通过ADMSC-SF的活性，在细胞分裂、细胞迁移和细胞衰老试验中发展转录谱变化。2. 对转录组数据进行生物信息学分析，识别激活的信号通路。3. 将ADMSC SF细胞检测转录组特征与FDA化合物数据库[C-Map]进行比较，以识别、评分和排名相关的仿生学。4. 在细胞分裂、细胞迁移和细胞衰老试验中研究10种最强化合物的仿生特性。5. 比较ADMSC SF与3种仿生材料在动物组织再生模型中的应用。比较ADMSC SF与3种仿生学在加速衰老模型中的作用。

Project Details: Stem cells promote the regeneration of numerous tissues in the human body. However the mechanisms underpinning their restorative ability remain to be elucidated. An increasing body of evidence demonstrates that stem cells do not incorporate into newly formed tissue but rather instruct resident cells, via secreted molecules, to activate repair mechanisms. Thus a paradigm shift has been to accept that factors produced by stem cells rather than the stem cells are the efficacious entities controlling tissue repair. This concept represents an unprecedented opportunity to identify and develop compounds, on an industrial scale, that are not only defined but also individually analysed, thus expiditing their safe use to promote tissue regeneration without the involvement of cells.Two major impediments have prevented a systematic approach to identifying biomimetics for secreted stem cells factors. The first was the absence of a robust platform that gives a biological response to stem cells-derived factors. The second was at the other end of the discovery platform; the lack of large number of biologically safe compounds that evoke a change in cell phenotype. Both these hurdles have been recently overcome, firstly through our discovery that human adipose-derived mesenchymal stem cells (ADMSC) secrete factors (SF) that promote cell proliferation, migration and protect against stress-induced senescence. The second development has been the profiling of over 1300 FDA-approved compounds in a variety of cells to develop large data sets that allows each drug to be profiled not on its biological effect but in terms of gene transcription (The Connectivity Map, C-Map). The key innovation is to use bioinformatic approaches to match the changes in transcription evoked by ADMSC-derived factors in their ability to regulate proliferation, migration and senescence to those promulgated by the library of safe compounds, leading to the identification of biomimetics for stem cell-derived factors regulating tissue regeneration.Aims:1. Develop transcriptional profile changes in cell division, cell migration and cell senescence assays through the activity of ADMSC-SF. 2. Perform bioinformatic analysis of transcriptome data and identify activated signalling pathways. 3. Compare the ADMSC SF cell assay transcriptome signatures with the database of FDA compounds [C-Map] to identify, score and rank correlating biomimetics. 4. Investigate biomimetic properties of the 10 strongest compounds in cell division, cell migration and cell senescence assays. 5. Compare ADMSC SF to 3 biomimetics in animal tissue regeneration models.6. Compare ADMSC SF to 3 biomimetics in accelerated aging models.