Engineering mesenchymal stromal cells to treat muscle fibrosis

项目摘要 间充质基质细胞(MSCs)已经在近1000项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，MSCs可以适应自己的利基市场 并重塑细胞外基质。虽然骨髓间充质干细胞的这种特性可能对治疗纤维化有潜在的好处 并促进组织再生，利用这一特性一直具有挑战性，因为特定的信号 使MSCs重塑基质仍有待定义，并在基于MSC的治疗中加以利用。在这里，我们 描述一种将单个细胞包裹在工程凝胶涂层中的高效方法，具体为 明确的生物物理和生化线索。我们开发了这种方法，以显示柔软、薄的凝胶涂层 是一个使能的线索，增加了可溶的间质胶原酶的产生，以响应肿瘤 肿瘤坏死因子α(肿瘤坏死因子α)。重要的是，我们的初步数据显示，凝胶包被的MSCs减少了胶原蛋白。 小鼠肌营养不良症模型中的沉积。在这份K99/R00提案中，我将在这些结果的基础上 验证使用专门设计的微凝胶编程MSCs可以激活潜能的假设 抑制肌肉纤维化的MSCs。在目标1中，我们将确定工程凝胶涂层在肿瘤坏死因子α中的作用。 诱导骨髓间充质干细胞激活，产生高水平的间质胶原酶并降解胶原，以及 了解这一过程背后的机制。在目标2中，我们将研究凝胶包被的间充质干细胞在 抑制肌肉纤维化，恢复肌肉功能。这项提议的成功将带来一个有效的 治疗肌肉纤维化的战略，这是一个主要的未得到满足的临床需求。在K99阶段，我将与 我的顾问委员会，以增强我在生物材料设计，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 有效带动这一研究项目。这些经历与职业发展活动一起描述 在这项提案中将确保我顺利过渡到独立调查员之间的接口 机械生物学和生物工程，以开发肌肉疾病的新治疗策略。

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.