He Yanpu, Hong Celestine, Irvine Darrell J, Li Jiahe, Hammond Paula T
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Bio Protoc. 2021 Feb 5;11(3):e3905. doi: 10.21769/BioProtoc.3905.
Activating the STING (stimulator of interferon genes) signaling pathway via administration of STING agonist cyclic GMP-AMP (cGAMP) has shown great promise in cancer immunotherapy. While state-of-the-art approaches have predominantly focused on the encapsulation of cGAMP into liposomes or polymersomes for cellular delivery, we discovered that the recombinant STING protein lacking the transmembrane domain (STINGΔTM) could be used as a functional carrier for cGAMP delivery and elicit type I IFN expression in STING-deficient cell lines. Using this approach, we generated anti-tumoral immunity in mouse melanoma and colon cancer models, providing a potential translatable platform for STING agonist-based immunotherapy. Here, we report the detailed STING activation protocols with cGAMP-STINGΔTM complex to assist researchers in further development of this approach. This protocol can also be easily expanded to other applications related to STING activation, such as control of various types of infections.
通过给予STING（干扰素基因刺激因子）激动剂环鸟苷酸-腺苷酸（cGAMP）激活STING信号通路在癌症免疫治疗中显示出巨大潜力。虽然目前的先进方法主要集中于将cGAMP封装到脂质体或多囊泡体中用于细胞递送，但我们发现缺乏跨膜结构域的重组STING蛋白（STINGΔTM）可作为cGAMP递送的功能性载体，并在STING缺陷细胞系中引发I型干扰素表达。利用这种方法，我们在小鼠黑色素瘤和结肠癌模型中产生了抗肿瘤免疫，为基于STING激动剂的免疫治疗提供了一个潜在的可转化平台。在此，我们报告使用cGAMP-STINGΔTM复合物进行STING激活的详细方案，以协助研究人员进一步开发这种方法。该方案也可轻松扩展到与STING激活相关的其他应用，如控制各种类型的感染。
