Biofunctional Materials Containing Protein Mimetics

描述（由申请方提供）：大分子生物素在指导宿主-生物材料相互作用和许多疾病病因学方面起着核心作用。我们假设仿生寡肽可以从巨噬细胞活性蛋白（纤连蛋白和白介素-1）的功能结构设计，并用于探测配体-受体相互作用和连接后事件，作为寻求更好地理解参与巨噬细胞行为的分子控制机制的基础。此外，这些肽被用作开发新型生物材料的平台技术的组分，以部分调节体外和体内巨噬细胞功能。在重新提交的竞争性更新申请中，我们假设所观察到的肽介导的巨噬细胞行为是由表面固定后的优先配体构象和调节巨噬细胞功能的多个细胞内事件的存在所贡献的。为实现这些目标，制定了三个相互关联的目标：（1）确定固定到化学上不同的底物上后的肽构象（TCPS，单甲氧基-PEG-单丙烯酸酯-共聚丙烯酸-共聚-四羟甲基丙基丙烯酸酯的共聚物，和含有化学改性明胶和聚乙二醇-二丙烯酸酯的互穿网络），并将该结构与配体-整联蛋白识别和结合相关联，（2）使用单一培养物和共培养物系统鉴定肽介导的粘附巨噬细胞中的细胞内酪氨酸磷酸化蛋白质、备选的连接后细胞内途径、连接后基因表达和细胞功能，（3）对巨噬细胞活性寡肽与整联蛋白的结合动力学进行定量和建模，并在肽-整联蛋白复合时与鉴定的磷酸化整联蛋白相关蛋白相关联。我们在探索材料与巨噬细胞相互作用的分子机制方面的持续努力为吞噬细胞功能的配体-受体结合和调节途径提供了见解，这对于理解生物医学设备的宿主-材料相互关系至关重要。

DESCRIPTION (provided by applicant): Macrophages are central in directing host-biomaterial interaction and many disease etiologies. We hypothesized that biomimetic oligopeptides can be designed from the functional structure of macrophage-active proteins (fibronectin and intedeukin-1) and employed to probe the ligand-receptor interaction and post-ligation events as a basis for seeking a greater understanding in the molecular control mechanism involved in macrophage behavior. Furthermore, these peptides were utilized as a component of a platform technology in the development of novel biomaterials to partly modulate macrophage function in vitro and in vivo. In this resubmission of the competing renewal application, we hypothesize that the observed peptide-mediated macrophage behavior is contributed by the preferential ligand conformation upon surface immobilization and the presence of multiple intracellular events in regulating macrophage function. To address these goals, three interrelated aims are formulated: (1) to determine the peptide conformation upon immobilization onto chemically distinct substrates (TCPS, copolymer of monomethoxy-PEG-monoacrylate-colacrylic acid-co-tetramethylolpropylacrylate, and interpenetrating networks containing chemically modified gelatin and polyethyleneglycol-diacrylate) and to correlate this structure with ligand-integrin recognition and binding, (2) to identify intracellular tyrosine phosphorylated proteins, alternative post-ligation intracellular pathways, post-ligation gene expression and cellular function in peptide-mediated adherent macrophages using mono- and co-culture systems, (3) to quantify and model the binding kinetics of macrophage-active oligopeptides with integrins, and to correlate with the identified phosphorylated integrin-associated proteins upon peptide-integrin complexation. Our continuing effort in probing the molecular mechanism of material-macrophage interaction has shed insights on ligand-receptor association and regulation pathways for phagocytic cell function, that are fundamental in understanding the host-material interrelationship for biomedical devices.