Design of de novo interleukin mimics for targeted immunotherapy

用于靶向免疫治疗的从头白细胞介素模拟物的设计

基本信息

批准号：
10475003
负责人：
DAVID BAKER
金额：
$ 33.9万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10475003
关键词：
Affinity Binding Biological Assay Biological Markers Biophysics Categories Cell surface Cells Clinic Cytokine Signaling DNA Dose-Limiting ERBB2 gene Effector Cell Elements Engineering Epidermal Growth Factor Receptor Escherichia coli Extracellular Domain Flow Cytometry Goals Human Immune Immune system Immunologics Immunotherapeutic agent Immunotherapy In Vitro Interferometry Interleukin Receptor Interleukin-12 Interleukin-2 Interleukin-4 Interleukins Label Malignant Neoplasms Measures Mission Modeling Molecular Sieve Chromatography Monitor Mus Oncogenes Peptide Synthesis Phosphorylation Population Property Protein Engineering Proteins Public Health Regenerative Medicine Renal Cell Carcinoma Research Proposals Signal Transduction Structure Surface T-Lymphocyte Therapeutic Toxic effect Transactivation Tumor Markers United States National Institutes of Health X-Ray Crystallography Yeasts anti-cancer base cancer cell cancer immunotherapy cancer therapy clinical application combat cytokine deep sequencing design disability extracellular frontier immunogenicity improved in vivo innovation interleukin-21 manufacturability melanoma mimetics nanobodies novel programmed cell death ligand 1 receptor reconstitution tumor tumor growth tumor xenograft

项目摘要

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.

项目摘要尽管可以使用免疫刺激性细胞因子来对抗癌症，但它们的稳定性差和远离目标较高毒性限制了其在诊所中的应用。该项目的长期目标是产生目标抗癌细胞因子模仿毒性降低。总体目标是在DE中应用最新的突破 NOVO蛋白设计可产生新的靶向，无毒的免疫刺激性蛋白质。中央假设是，可以将天然细胞因子的有益刺激作用进行设计为从头设计设计的蛋白质不参与脱靶结合，从而规避限制剂量的毒性在天然/重新设计细胞因子的临床应用中可见。具体目的是：（1）使用从头蛋白设计生成白介素-2，-4，-12，-15和-21的高级生物活性模仿者在体内与（即结合）白细胞介素受体接合；（2）将这些模仿分成不活跃的部分可以通过体内重新组装来恢复活动；（3）将这些不活动部分融合到特定的目标域，从而产生有条件活跃的细胞因子模拟物，仅通过重新组装来刺激T细胞靶细胞的表面（即显示两个特定表面生物标志物的癌细胞）。作为证明原理，第一个从头设计的细胞因子模拟物已产生，分裂并证明是为了减少小鼠的肿瘤无参与毒性或免疫原性。该研究建议是创新的，因为它试图解决癌症免疫疗法的长期障碍（即基于细胞因子的疗法）通过从头开始设计一类新的无毒细胞因子模拟物。提案很重要，因为它将是计算蛋白设计的第一个例子，该蛋白质设计产生了目标，生物癌治疗。最终，这种分子有可能治疗广泛的癌症，包括恶性黑色素瘤，肾细胞癌等。