Trossmann Vanessa T, Heltmann-Meyer Stefanie, Amouei Hanna, Wajant Harald, Horch Raymund E, Steiner Dominik, Scheibel Thomas
Lehrstuhl Biomaterialien, Fakultät für Ingenieurswissenschaften, Universität Bayreuth, Prof.-Rüdiger-Bormann-Str. 1, Bayreuth 95447, Germany.
Department of Plastic and Hand Surgery and Laboratory for Tissue Engineering and Regenerative Medicine, University Hospital of Erlangen, Friedrich-Alexander-University Erlangen-Nuremberg (FAU), Krankenhaus-Str. 12, Erlangen 91054, Germany.
Biomacromolecules. 2022 Oct 10;23(10):4427-4437. doi: 10.1021/acs.biomac.2c00971. Epub 2022 Sep 6.
Targeted therapies using biopharmaceuticals are of growing clinical importance in disease treatment. Currently, there are several limitations of protein-based therapeutics (biologicals), including suboptimal biodistribution, lack of stability, and systemic side effects. A promising approach to overcoming these limitations could be a therapeutic cell-loaded 3D construct consisting of a suitable matrix component that harbors producer cells continuously secreting the biological of interest. Here, the recombinant spider silk proteins eADF4(C16), eADF4(C16)-RGD, and eADF4(C16)-RGE have been processed together with HEK293 producer cells stably secreting the highly traceable reporter biological TNFR2-Fc-GpL, a fusion protein consisting of the extracellular domain of TNFR2, the Fc domain of human IgG1, and the luciferase of as a reporter domain. eADF4(C16) and eADF4(C16)-RGD hydrogels provide structural and mechanical support, promote HEK293 cell growth, and allow fusion protein production by the latter. Bioink-captured HEK293 producer cells continuously release functional TNFR2-Fc-GpL over 14 days. Thus, the combination of biocompatible, printable spider silk bioinks with drug-producing cells is promising for generating implantable 3D constructs for continuous targeted therapy.
使用生物制药的靶向疗法在疾病治疗中的临床重要性日益增加。目前，基于蛋白质的治疗药物（生物制剂）存在若干局限性，包括生物分布不理想、缺乏稳定性和全身副作用。一种有望克服这些局限性的方法可能是一种负载治疗细胞的3D构建体，它由合适的基质成分组成，其中含有持续分泌感兴趣生物制剂的生产细胞。在此，重组蜘蛛丝蛋白eADF4(C16)、eADF4(C16)-RGD和eADF4(C16)-RGE已与稳定分泌高度可追踪报告生物制剂TNFR2-Fc-GpL的HEK293生产细胞一起进行处理，TNFR2-Fc-GpL是一种融合蛋白，由TNFR2的细胞外结构域、人IgG1的Fc结构域和作为报告结构域的荧光素酶组成。eADF4(C16)和eADF4(C16)-RGD水凝胶提供结构和机械支持，促进HEK293细胞生长，并允许后者生产融合蛋白。生物墨水捕获的HEK293生产细胞在14天内持续释放功能性TNFR2-Fc-GpL。因此，生物相容性、可打印的蜘蛛丝生物墨水与产药细胞的组合有望生成用于连续靶向治疗的可植入3D构建体。
