Li Jie, Chen Mingkuan, Liu Zilei, Zhang Linda, Felding Brunie H, Moremen Kelley W, Lauvau Gregoire, Abadier Michael, Ley Klaus, Wu Peng
Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California 92037, United States.
Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States.
ACS Cent Sci. 2018 Dec 26;4(12):1633-1641. doi: 10.1021/acscentsci.8b00552. Epub 2018 Dec 7.
Employing live cells as therapeutics is a direction of future drug discovery. An easy and robust method to modify the surfaces of cells directly to incorporate novel functionalities is highly desirable. However, genetic methods for cell-surface engineering are laborious and limited by low efficiency for primary cell modification. Here we report a chemoenzymatic approach that exploits a fucosyltransferase to transfer bio-macromolecules, such as an IgG antibody (MW∼ 150 KD), to the glycocalyx on the surfaces of live cells when the antibody is conjugated to the enzyme's natural donor substrate GDP-Fucose. Requiring no genetic modification, this method is fast and biocompatible with little interference to cells' endogenous functions. We applied this method to construct two antibody-cell conjugates (ACCs) using both cell lines and primary cells, and the modified cells exhibited specific tumor targeting and resistance to inhibitory signals produced by tumor cells, respectively. Remarkably, Herceptin-NK-92MI conjugates, a natural killer cell line modified with Herceptin, exhibit enhanced activities to induce the lysis of HER2+ cancer cells both and in a human tumor xenograft model. Given the unprecedented substrate tolerance of the fucosyltransferase, this chemoenzymatic method offers a general approach to engineer cells as research tools and for therapeutic applications.
利用活细胞作为治疗手段是未来药物研发的一个方向。人们非常渴望有一种简单且可靠的方法来直接修饰细胞表面，以赋予其新的功能。然而，用于细胞表面工程的基因方法既费力，又受限于原代细胞修饰效率低下。在此，我们报告一种化学酶法，该方法利用岩藻糖基转移酶将生物大分子，如IgG抗体（分子量约150 KD），转移到活细胞表面的糖萼上，前提是该抗体与该酶的天然供体底物GDP-岩藻糖偶联。该方法无需基因修饰，快速且具有生物相容性，对细胞内源性功能几乎没有干扰。我们应用此方法使用细胞系和原代细胞构建了两种抗体-细胞偶联物（ACC），修饰后的细胞分别表现出特异性肿瘤靶向性和对肿瘤细胞产生的抑制信号的抗性。值得注意的是，赫赛汀-NK-92MI偶联物，即一种用赫赛汀修饰的自然杀伤细胞系，在体外和人肿瘤异种移植模型中均表现出增强的诱导HER2+癌细胞裂解的活性。鉴于岩藻糖基转移酶前所未有的底物耐受性，这种化学酶法为将细胞工程化为研究工具和用于治疗应用提供了一种通用方法。
