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

纤维调节蛋白重编程细胞的整体转录分析 - 用于颅面组织重建的新的多能细胞来源

基本信息

批准号：
10438868
负责人：
Zhong Zheng
金额：
$ 15.6万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-06-30
项目状态：
已结题

项目摘要

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.

项目概要/摘要受损颅面组织的再生极其困难，而且往往由于再生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。致瘤性和/或肿瘤支持特性使得目前可用的多能和专能细胞不适合组织再生，特别是颅面组织，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满足这一需求，我们建立了一种新颖的方法。真皮成纤维细胞通过持续暴露于细胞外获得多能性的技术平台重要的是，在无血清条件下，基质（ECM）蛋白聚糖、纤维调节蛋白（FMOD）。 FMOD 重编程 (FReP) 细胞和诱导多能干细胞 (iPSC) 表达相似的多能性标记物并具有三叶细胞分化潜能，它们各自的致瘤能力完全是不同，这可能与重新编程真皮的根本不同的方法有关与通过癌基因激活产生的 iPSC 不同，FReP 细胞是由成纤维细胞转化为 FReP 细胞或 iPSC。通过简单的 ECM 刺激程序重新编程，不需要基因组整合或值得注意的是，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）肌肉室的高度血管化的微环境特别有利于肿瘤的发生事实上，在细胞测试时，肌肉内植入是畸胎瘤形成的常见途径。因此，FReP 细胞似乎是一种潜在的安全有效的细胞来源。然而，与间充质干细胞相比，已充分获得用于功能性组织再生的能力。 (MSC) 和 iPSC、FReP 细胞的研究仍处于起步阶段。 FReP 细胞和 iPSC 之间的转录组数据将为 FMOD 重编程提供更多理解在这项研究中，我们将研究协调细胞命运决定和肿瘤发生的细胞信号通路。进行深入的转录组学分析以确认非致瘤性或 FReP 细胞独立于其亲本成纤维细胞的性别和年龄 (AIM 1) 并追踪生物能相关的转录组变化 FMOD 重编程期间的基因，以深入了解推进该技术的新策略 (AIM 2)。完成这些 AIM 将丰富我们关于细胞命运决定和重编程的知识，并且肿瘤发生，从而为 R01 阶段的未来研究奠定基础。研究将直接有利于基于 FReP 细胞的颅面部或因此，该提案与肌肉骨骼重建和再生医学密切相关。与当前 NIDCR R03 赠款的使命一致。