Stichting PAMM, Laboratory for Pathology and Medical Microbiology, De Run 6250, Veldhoven, 5504 DL, The Netherlands.
Institute for Molecules and Materials, Radboud University, Heyendaalseweg 135, Nijmegen, 6525 AJ, The Netherlands.
ACS Synth Biol. 2020 Mar 20;9(3):475-485. doi: 10.1021/acssynbio.9b00404. Epub 2020 Mar 3.
Engineered living materials have the potential for wide-ranging applications such as biosensing and treatment of diseases. Programmable cells provide the functional basis for living materials; however, their release into the environment raises numerous biosafety concerns. Current designs that limit the release of genetically engineered cells typically involve the fabrication of multilayer hybrid materials with submicrometer porous matrices. Nevertheless the stringent physical barriers limit the diffusion of macromolecules and therefore the repertoire of molecules available for actuation in response to communication signals between cells and their environment. Here, we engineer a novel living material entitled "Platform for Adhesin-mediated Trapping of Cells in Hydrogels" (PATCH). This technology is based on engineered that displays an adhesion protein derived from an Antarctic bacterium with a high affinity for glucose. The adhesin stably anchors in dextran-based hydrogels with large pore diameters (10-100 μm) and reduces the leakage of bacteria into the environment by up to 100-fold. As an application of PATCH, we engineered to secrete the bacteriocin lysostaphin which specifically kills with low probability of raising antibiotic resistance. We demonstrated that living materials containing this lysostaphin-secreting inhibit the growth of , including the strain resistant to methicillin (MRSA). Our tunable platform allows stable integration of programmable cells in dextran-based hydrogels without compromising free diffusion of macromolecules and could have potential applications in biotechnology and biomedicine.
工程化活体材料具有广泛的应用潜力,例如生物传感和疾病治疗。可编程细胞为活体材料提供了功能基础;然而,将其释放到环境中会引发许多生物安全问题。目前限制基因工程细胞释放的设计通常涉及制造具有亚微米多孔基质的多层混合材料。然而,严格的物理屏障限制了大分子的扩散,因此可用作响应细胞与其环境之间通信信号的分子的种类有限。在这里,我们设计了一种名为“用于水凝胶中细胞黏附介导捕获的平台(PATCH)”的新型活体材料。这项技术基于一种具有高葡萄糖亲和力的源自南极细菌的黏附蛋白的工程化 。该黏附蛋白将 稳定地锚定在具有大孔径(10-100μm)的葡聚糖基水凝胶中,并将细菌泄漏到环境中的量减少多达 100 倍。作为 PATCH 的应用,我们设计了 来分泌溶葡萄球菌素,它可以特异性杀死 ,而不太可能引起抗生素耐药性。我们证明,含有这种分泌溶葡萄球菌素的活体材料可以抑制 ,包括对甲氧西林(MRSA)有抗性的菌株的生长。我们的可调谐平台允许在不影响大分子自由扩散的情况下将可编程细胞稳定地整合到葡聚糖基水凝胶中,并且在生物技术和生物医学领域可能具有潜在应用。