Global Transcriptional Analyses of Fibromodulin Reprogrammed Cells - A New Multipotent Cell Source for Craniofacial Tissue Reconstruction

项目摘要/摘要 受损的头面部组织的再生极其困难，而且往往由于不足而不能令人满意。 内源性前体细胞以及相应的有限再生能力。此外，固有的 致瘤性和/或支持肿瘤的特性使目前可用的多能和多能细胞 不适合组织再生，尤其是头面部组织。因此，生产更安全、更容易 可用于功能性颅面组织再生的细胞来源仍然是再生的主要挑战 医学，特别是颅面重建术。为了回应这一需求，我们建立了一部小说 真皮成纤维细胞通过持续暴露在细胞外获得多能性的技术平台 基质(ECM)蛋白多糖，纤维调素(FMOD)，在无血清条件下。重要的是，尽管两者 FMOD重编程(FReP)细胞和诱导多能干细胞(IPSCs)表达相似的多能性 标记，并具有三倍母细胞分化潜能，它们各自的致瘤潜能完全是 不同，这可能与重新编程真皮的根本不同的方法有关 成纤维细胞转化为FReP细胞或IPSCs。与通过癌基因激活产生的IPSCs不同，FReP细胞是 通过简单的ECM刺激程序重新编程，不需要基因组整合或 癌基因激活。值得注意的是，低致瘤性是使用潜在细胞来源的先决条件 骨骼肌再生(颅面紊乱和创伤重建的重要组成部分) 肌肉间高度血管化的微环境特别有利于肿瘤的发生。 队形。事实上，当细胞检测时，肌肉内植入是畸胎瘤形成的常见途径。 多能性。因此，FReP细胞似乎是一种潜在的安全和有效的细胞来源，可以 获得足够的功能组织再生。然而，与间充质干细胞相比， 对于MSCs和IPSCs，FReP细胞的研究还处于初级阶段。进一步比较可公开访问的 FReP细胞和IPSCs之间的转录数据将提供对FMOD重新编程的更多了解 以及协调细胞命运决定和肿瘤发生的细胞信号通路。在这项研究中，我们将 执行深入的转录图谱以确认非致瘤性质或FReP细胞独立于 其亲代成纤维细胞的性别和年龄(AIM 1)，并追踪与生物能量相关的转录变化 FMOD重新编程期间的基因，以深入了解推动这项技术的新战略(AIM 2)。 完成这些目标将丰富我们对细胞命运决定和重新编程的知识，以及 肿瘤的发生，从而为R01阶段的未来研究奠定了基础。因此，当前 这项研究将直接有利于FReP细胞治疗颅面部创伤的发展 肌肉骨骼重建和再生医学。因此，这项提议是严格的 与当前NIDCRR03赠款的使命相一致。

PROJECT SUMMARY / ABSTRACT Regeneration of damaged craniofacial tissue is extremely difficult and often unsatisfactory due to insufficient endogenous precursor cells as well as a corresponding finite regenerative capability. Furthermore, inherent tumorigenicity and/or tumor supporting properties render the currently available pluripotent and multipotent cells unsuitable for tissue regeneration, especially for craniofacial tissues. Thus, the production of safer and readily available cell sources for functional craniofacial tissue regeneration remains a major challenge for regenerative medicine, and particularly for craniofacial reconstruction. In response to this demand, we established a novel technology platform in which dermal fibroblasts acquire multipotency by continuous exposure to an extracellular matrix (ECM) proteoglycan, fibromodulin (FMOD), under serum-free conditions. Importantly, although both FMOD reprogrammed (FReP) cells and induced pluripotent stem cells (iPSCs) express similar pluripotent markers and hold the triploblastic differentiation potentials, their respective tumorigenic potencies are entirely different, which could relate back to the fundamentally disparate methodologies for reprogramming dermal fibroblasts into FReP cells or iPSCs. Unlike iPSCs generated through oncogene activation, FReP cells are reprogrammed through a simple ECM stimulation procedure that does not require genome integration or oncogene activation. It is worth noting that low tumorigenicity is a prerequisite for using a potential cell source in skeletal muscle regeneration (an essential component of craniofacial disorder and trauma reconstruction) since the highly vascularized microenvironment of the muscle compartment can be especially conducive to tumor formation. Indeed, intramuscular implantation is a common route for teratoma formation when testing for cellular pluripotency. Therefore, FReP cells appear to be a potentially safe and effective cell source that can be adequately obtained for functional tissue regeneration. However, compared with mesenchymal stem cells (MSCs) and iPSCs, FReP cells’ investigation is still in its infancy. A further comparison of the publicly accessible transcriptomic data between FReP cells and iPSCs will provide more understanding into FMOD reprogramming and the cell signal pathways that orchestrate the cell fate determination and tumorigenesis. In this study, we will perform an in-depth transcriptomic profile to confirm the non-tumorigenic nature or FReP cells is independent of the sex and age of their parental fibroblasts (AIM 1) and track the transcriptomic alteration of bioenergetic-related genes during FMOD reprogramming to gain insight into a novel strategy to advance this technology (AIM 2). Completing these AIMs will enrich our knowledge about cell fate determination and reprogramming, and tumorigenesis, thus building the fundamentals for future investigations in the R01 stage. Therefore, the current study will directly benefit the development of FReP cell-based therapies for the craniofacial or musculoskeletal reconstruction and regenerative medicine in general. As such, this proposal is tightly aligned with the mission of the current NIDCR R03 grant.