Identifying hMSC interaction pathways in soluble interferon-gamma and stratified collagen/heparin coatings for the manufacturing of therapeutic cells

人类间充质间质细胞（hMSC）有望治疗致命的不治之症。hMSC生产的主要限制之一是在生物反应器中生长功能齐全的细胞。干扰素γ (IFN)治疗增强了hMSCs的治疗效力，但也抑制了其生长。用胶原蛋白（COL）和肝素（HEP）层包裹hMSCs，在保持治疗增强的同时减少了抑制，但尚不清楚这些层是如何导致这种情况发生的。该项目将确定COL/HEP涂层增强hMSC对IFN反应的具体途径。这些知识将支持设计用于制造高治疗性细胞的最佳生物反应器平台。来自代表性不足群体的学生将从事工程学和生物学交叉领域的研究。这将有助于发展一支强大而多样化的生物制造劳动力队伍。该项目的中心假设是，COL/HEP涂层与可溶性IFN协同作用，调节调节增殖和免疫抑制特性的信号通路。这一假设在很大程度上是基于初步数据提出的，这些数据表明，COL/HEP涂层：诱导碱性成纤维细胞生长因子（FGF2）的表达，降低IFN的抗增殖作用，同时增强免疫调节细胞因子的分泌。具体目标是：(1)建立膜层与hMSCs中FGF2通路激活的关系及其与增殖的关系；(2)建立IFN活化COL/HEP膜培养的hMSCs的机制；(3)确定COL/HEP涂层转化为悬浮生物反应器微载体时保持一致性的程度。该项目将直接影响细胞制造领域，通过将聚合材料与生化信号相结合，产生一种增强hMSCs激活和治疗效力的技术。该项目由细胞与生物化学工程项目和促进竞争性研究的既定项目（EPSCoR）共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Human mesenchymal stromal cells (hMSC) show promise for treating deadly incurable diseases. One of the main limitations of hMSC production is the growth of fully functional cells in bioreactors. Treatment with interferon gamma (IFN) enhances the therapeutic potency of hMSCs but also inhibits growth. Coating hMSCs with layers of collagen (COL) and heparin (HEP) reduces the inhibition while maintaining the therapeutic enhancement, but it is unknown how the layers cause this to occur. This project will identify the specific pathways by which COL/HEP coatings enhance hMSC response to IFN. This knowledge will support the design of optimal bioreactor platforms for the manufacturing of highly therapeutic cells. Students from underrepresented populations will engage in research at the interface between engineering and biology. This will help develop a strong and diverse biomanufacturing workforce.The central hypothesis of this project is that COL/HEP coatings work synergistically with soluble IFN to modulate signaling pathways regulating proliferation and immunosuppressive properties. This hypothesis was formulated, in large part, based on preliminary data indicating that COL/HEP coatings: induce expression of basic fibroblast growth factor (FGF2) and reduce the anti-proliferative effect of IFN while enhancing immunomodulatory cytokine secretion. The specific objectives are: (1) To establish the relationship between the coatings and the activation of the FGF2 pathway in hMSCs, and its relation to proliferation; (2) To establish the mechanism by which IFN activates hMSCs cultured on COL/HEP coatings; and (3) To determine the extent to which COL/HEP coatings remain consistent when translated to microcarriers for suspension-based bioreactors. This project will directly impact the cell manufacturing field by merging polymeric materials with biochemical signals to produce a technology that enhances hMSCs activation and therapeutic potency.This project is jointly funded by the Cellular and Biochemical Engineering Program and the Established Program to Stimulate Competitive Research (EPSCoR).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.