Engineering mesenchymal stromal cells to treat muscle fibrosis

项目摘要 间充质基质细胞（MSC）已经在近一千个临床试验中进行了测试，主要是因为 它们能够分泌能够改变宿主环境的因子。例如，MSC可以适应它们的生态位， 重塑细胞外基质虽然MSC的这种特性可能有利于治疗纤维化， 并促进组织再生，利用这种特性一直具有挑战性，因为特定的信号， 使MSC能够重塑基质仍有待于在基于MSC的治疗中定义和利用。这里我们 描述了一种高效的方法，将单个细胞包封在工程化的凝胶涂层中， 明确的生物物理和生物化学线索。我们已经开发了这种方法，以显示软，薄的凝胶涂层， 是一种激活因子，可增加可溶性间质胶原酶的产生， 坏死因子-α（TNFα）。重要的是，我们的初步数据表明，凝胶包被的MSC减少胶原蛋白， 沉积在鼠肌营养不良模型中。在本K99/R 00建议书中，我将基于这些结果， 测试使用专门工程化的微凝胶编程MSC激活潜在的 抑制肌肉纤维化。在目标1中，我们将确定工程凝胶涂层在TNFα- 诱导MSC活化以产生高水平的间质胶原酶并降解胶原，和 了解这个过程背后的机制。在目标2中，我们将研究凝胶包被的MSC在以下方面的作用： 抑制肌肉纤维化和恢复肌肉功能。这项建议的成功将导致一个有效的 这是一种治疗肌肉纤维化的策略，这是一个主要的未满足的临床需求。在K99阶段，我将与 我的顾问委员会，以提高我在生物材料设计，细胞机械生物学， 纤维化的病理生理学，以及先进的微技术，成像和计算方法， 有效地推动这项研究项目。这些经验以及职业发展活动描述了 这份建议书将确保我顺利过渡到一个独立的调查员之间的接口， 机械生物学和生物工程，以开发新的治疗策略，肌肉疾病。

Project Summary Mesenchymal stromal cells (MSCs) have been tested in nearly one thousand clinical trials, mostly because of their ability to secrete factors that can modify host environments. For instance, MSCs can adapt to their niches and remodel the extracellular matrix. While this property of MSCs can potentially be beneficial to treat fibrosis and promote tissue regeneration, leveraging this property has been challenging because specific signals that enable MSCs to remodel the matrix remain to be defined and leveraged in MSC-based therapeutics. Here, we describe a highly efficient approach to encapsulate individual cells in engineered gel coatings with specifically defined biophysical and biochemical cues. We have developed this approach to show that soft, thin gel coating is an enabling cue that increases the production of soluble interstitial collagenases in response to tumor necrosis factor-α (TNFα). Importantly, our preliminary data show that gel-coated MSCs decrease collagen deposition in a murine muscular dystrophy model. In this K99/R00 proposal, I will build upon these results to test the hypothesis that programming of MSCs using specifically engineered microgels activates the potential of MSCs to inhibit muscle fibrosis. In Aim 1, we will determine the role of engineered gel coating in TNFα- induced activation of MSCs to produce high levels of interstitial collagenases and degrade collagen, and understand mechanisms behind this process. In Aim 2, we will investigate the role of gel-coated MSCs in inhibiting muscle fibrosis and restoring muscle functions. Success of this proposal will lead to an effective strategy to treat muscle fibrosis, which is a major unmet clinical need. During the K99 phase, I will work with my advisory committee to enhance my knowledge in biomaterial design, cellular mechanobiology, pathophysiology of fibrosis, as well as advanced microtechnologies, imaging, and computational approaches to effectively drive this research project. These experiences together with career development activities described in this proposal will ensure my smooth transition to an independent investigator at the interface between mechanobiology and bioengineering to develop novel therapeutic strategies for muscle disorders.