Design of de novo interleukin mimics for targeted immunotherapy

项目总结 虽然免疫刺激细胞因子可以用于抗癌，但它们的稳定性差，靶向性高。 毒性限制了它们在临床上的应用。这个项目的​长期目标是生产有针对性的 毒性较低的抗癌细胞因子模拟物。​的总体目标是将最近的突破应用于​de Novo蛋白质设计以产生一种新的靶向、无毒、免疫刺激蛋白质。​中心 假设天然细胞因子的有益刺激作用可以被改造成​de nevo 设计了不参与脱靶结合的蛋白质，从而绕过了剂量限制的毒性 可见于天然/重组细胞因子的临床应用。​的特定目的是：(1)使用​从头蛋白 设计以产生白细胞介素2、-4、-12、-15和-21的超稳定的生物活性模拟物，其功能是通过 在体内与白细胞介素受体​结合(即结合​)；(2)将这些模拟物分解成非活性部分， 可以通过在体内重组​​来恢复活性；以及(3)将这些非活性部分中的每一个融合到特定的靶向 结构域，从而产生有条件地激活的细胞因子模拟物，通过仅在 靶细胞的表面(即癌细胞显示两个特定的表面生物标志物)。​作为证据 原理​，第一个这样的​从头设计的细胞因子模拟物已经被产生，分裂，并显示出减少 无伴随毒性或免疫原性的小鼠肿瘤。这项研究建议是​的创新，因为 它寻求解决癌症免疫治疗的长期障碍(即， 基于细胞因子的疗法)，从零开始设计一类新的无毒细胞因子模拟物。这个 该提议具有​意义，因为它将是第一个计算蛋白质设计的例子，产生靶向， 生物优势癌症疗法。最终，这种分子有可能治疗多种癌症， 包括恶性黑色素瘤、肾细胞癌等。

PROJECT SUMMARY Although immunostimulatory cytokines can be used to combat cancer, their poor stability and high off-target toxicity has limited their application in the clinic. The ​long-term goal of this project is to produce targeted anti-cancer cytokine mimetics with reduced toxicity. The ​overall objective is to apply recent breakthroughs in ​de novo protein design to yield a new category of targeted, non-toxic, immunostimulatory proteins. The ​central hypothesis is that the beneficial stimulatory effects of natural cytokines can be engineered into ​de novo designed proteins which do not engage in off-target binding, thereby circumventing the dose-limiting toxicities seen in clinical applications of natural/reengineered cytokines. The ​specific aims are to: (1) use ​de novo protein design to generate hyperstable, bioactive mimetics of interleukin-2, -4, -12, -15, and -21 which function by engaging with (i.e. binding to) interleukin receptors ​in vivo​; (2) to split these mimics into inactive parts which can regain activity by reassembling ​in vivo​; and (3) to fuse each of these inactive parts to specific targeting domains, thereby yielding conditionally-active cytokine mimics that stimulate T-cells by reassembling only on the surface of a targeted cells (i.e. cancer cells displaying two specific surface biomarkers). ​As proof of principle​, the first such ​de novo designed cytokine mimetic has been produced, split, and shown to reduce tumors in mice without accompanying toxicity or immunogenicity. This research proposal is ​innovative because it seeks to resolve a long-standing barrier to cancer immunotherapy (namely, the off-target toxicity of cytokine-based therapeutics) by designing from scratch a new class of non-toxic cytokine mimics. The proposal is ​significant because it would be the first example of computational protein design yielding targeted, biosuperior cancer therapeutics. Ultimately, such molecules have the potential to treat a wide range of cancers, including malignant melanoma, renal cell carcinoma, and more.