Encapsulation of mesenchymal stromal cells in engineered microgels for resolution of lung fibrosis

项目摘要 骨髓间充质干细胞（MSCs）适应其组织生态位和重塑细胞外基质的能力 (ECM)在治疗纤维化疾病方面可能是有益的。然而，利用这一特性， 这是具有挑战性的，因为MSC一旦递送就迅速从组织中清除。因此，特定的途径 诱导MSC重塑ECM，这些的效用仍不清楚。我们在这里描述了一个高度 将单细胞包封在具有预定生化和生物活性的工程微凝胶中的有效方法 生物物理线索我们已经对这种方法进行了微调，以显示MSC的清除明显延迟。 当它们以薄的微凝胶形式被递送到肺内时。重要的是，我们的初步数据显示， 这种治疗减少了鼠肺纤维化模型中的胶原积累。我们在这些支持中显示， 研究了当细胞被包裹在工程化的 具有可调刚度的水凝胶。在我的第一个RO 1提案中，我将基于这些结果来测试 假设使用特别工程化的微凝胶编程MSC激活MSC的潜力 以促进纤维化的消退，从而恢复肺功能。在目标1中，我们将确定 微囊化在控制MSC在肺中的保留和促进肺纤维化的解决中的作用。在目标2中， 我们将确定炎性细胞因子肿瘤坏死因子-α（TNFα）在 修饰微囊化MSC的表型以产生大量胶原酶， 解决肺纤维化的潜力。我们预测，选择性激活包封在微囊中的MSC中的TNF受体2， 软微凝胶可以解决肺纤维化。该项目是高度多学科的，因为它将采用一个 结合生物材料、肺纤维化、生物物理学、遗传学和小鼠体内方法方面的专业知识， 解决具体目标。这些结果将有助于确定MSC如何通过以下方式促进纤维化的解决： 延长它们在肺中的体内停留时间，并通过包封修饰它们的表型， 工程化和可调的微凝胶，以优化其胶原酶的生产。因此，我们希望开发新的 基于MSC的方法来重塑异常细胞外基质和治疗肺纤维化。 !

PROJECT SUMMARY The ability of mesenchymal stromal cells (MSCs) to adapt to their tissue niche and remodel extracellular matrix (ECM) could be therapeutically beneficial in treating fibrotic diseases. However, leveraging this property has been challenging because MSCs are rapidly cleared from tissue once delivered. Thus, specific pathways to induce MSCs to remodel the ECM and the utility of these remain unclear. We described herein a highly efficient approach to encapsulate single cells in engineered microgels with predefined biochemical and biophysical cues. We have fine-tuned this approach to show that clearance of MSCs is significantly delayed when they are delivered in thin microgels intratracheally to lungs. Importantly, our preliminary data show that this treatment decreases collagen accumulation in a murine lung fibrosis model. We show in these supporting studies amplified ability of MSCs to degrade collagen-I when the cells are encapsulated in engineered hydrogels with tunable stiffness. In this my first RO1 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 promote resolution of fibrosis and thus restore lung function. In Aim 1, we will determine the role of microencapsulation in controlling retention of MSCs in lungs and facilitating resolution of lung fibrosis. In Aim 2, we will determine the potentially important role of the inflammatory cytokine tumor necrosis factor-α (TNFα) in modifying the phenotype of microencapsulated MSCs to produce high amounts collagenases with enhanced potential to resolve lung fibrosis. We predict that selectively activating TNF receptor 2 in MSCs encapsulated in soft microgels can resolve lung fibrosis. The project is highly multidisciplinary in that it will employ a combination of expertise in biomaterials, lung fibrosis, biophysical, genetic, and mouse in vivo approaches to address the specific aims. The results will help to define how MSCs can facilitate resolution of fibrosis through prolonging their in vivo residence time in lungs, and modifying their phenotype through encapsulation in engineered and tunable microgels to optimize their production of collagenases. We thus hope to develop novel MSC based approaches to remodel aberrant extracellular matrix and to treat lung fibrosis. !