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

项目详情：干细胞可促进人体内众多组织的再生。然而，支持其修复能力的机制仍有待阐明。越来越多的证据表明，干细胞不会结合到新形成的组织中，而是通过分泌的分子指示驻留细胞激活修复机制。因此，一种范式的转变已经接受了干细胞产生的因子而不是干细胞是控制组织修复的有效实体。这一概念代表着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，这些化合物不仅被定义，而且还被单独分析，从而限制了它们在不涉及细胞的情况下促进组织再生的安全使用。两个主要障碍阻碍了系统地识别分泌干细胞因子的仿生方法。第一个问题是缺乏一个强大的平台来对干细胞衍生因子做出生物学反应。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，首先是我们发现人类脂肪来源的间充质干细胞(ADMSC)分泌促进细胞增殖、迁移和防止应激诱导的衰老的因子(SF)。第二个发展是在各种细胞中对FDA批准的1300多种化合物进行图谱分析，以开发大型数据集，允许对每种药物进行图谱分析，而不是根据其生物效应，而是根据基因转录(连接性图谱，C-Map)。关键的创新是利用生物信息学的方法，将ADMSC衍生因子在调控增殖、迁移和衰老方面的转录变化与安全化合物库中公布的那些相匹配，从而识别调控组织再生的干细胞衍生因子的仿生学。目的：1.通过ADMSC-SF的活性，在细胞分裂、细胞迁移和细胞衰老分析中发展转录谱变化。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细胞分析转录组特征与FDA化合物数据库[C-Map]进行比较，以鉴定、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实验中考察了10个最强化合物的仿生特性。5.比较ADMSC SF和3种仿生剂在动物组织再生模型中的应用。在加速衰老模型中比较ADMSC SF和3种仿生药物。

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.