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

项目概要/摘要 受损颅面组织的再生是极其困难的，并且由于不充分的组织再生而常常不能令人满意。 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/或肿瘤支持特性使得目前可获得的多能和多能细胞 不适合组织再生，尤其是颅面组织。因此，生产更安全，更容易 用于功能性颅面组织再生的可用细胞来源仍然是再生的主要挑战。 医学，特别是颅面重建。为了回应这一需求，我们建立了一个新的 其中真皮成纤维细胞通过连续暴露于细胞外 基质（ECM）蛋白聚糖、纤调蛋白（FMOD）。重要的是，尽管两者 FMOD重编程（FReP）细胞和诱导多能干细胞（iPSC）表达类似的多能干细胞分化。 标志物并保持三胚层分化潜能，它们各自的致瘤潜能完全不同。 不同的，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iPSC。与通过癌基因激活产生的iPSC不同，FReP细胞是 通过不需要基因组整合的简单ECM刺激程序重编程，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（颅面疾病和创伤重建的重要组成部分）， 肌肉隔室的高度血管化的微环境可特别有利于肿瘤的发生 阵实际上，当检测细胞毒性时，肌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， 获得足够的功能性组织再生。然而，与间充质干细胞相比， 尽管FReP细胞是间充质干细胞（MSC）和诱导多能干细胞（iPSC）的主要来源，但FReP细胞的研究仍处于起步阶段。公开访问的进一步比较 FReP细胞和iPSC之间的转录组学数据将为FMOD重编程提供更多的理解 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 进行深入的转录组学分析，以确认非致瘤性或FReP细胞不依赖于 其亲本成纤维细胞（AIM 1）的性别和年龄，并跟踪生物能量相关的转录组学改变。 FMOD重编程过程中的基因，以深入了解推进这项技术的新策略（AIM 2）。 完成这些AIM将丰富我们对细胞命运决定和重编程的知识， 肿瘤发生，从而为R 01阶段的未来研究奠定基础。因此目前 这项研究将直接有利于开发基于FReP细胞的颅面或 肌肉骨骼重建和再生医学。因此，这项建议是严格 与当前NIDCR R 03赠款的使命一致。

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.