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：
9796930
负责人：
DAVID BAKER
金额：
$ 35.55万
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/9796930
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 PDCD1LG1 gene 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 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.

项目概要虽然免疫刺激细胞因子可用于对抗癌症，但其稳定性差且脱靶率高毒性限制了它们在临床的应用该项目的长期目标是产生靶向药物。具有降低毒性的抗癌细胞因子模拟物，总体目标是应用最近的突破。 novo 蛋白质设计产生一类新的靶向、无毒、免疫刺激蛋白质。假设天然细胞因子的有益刺激作用可以从头改造设计了不参与脱靶结合的蛋白质，从而避免了剂量限制性毒性在天然/重组细胞因子的临床应用中看到的具体目标是：（1）使用从头蛋白质。设计产生超稳定的、生物活性的白细胞介素-2、-4、-12、-15和-21模拟物，其功能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；可以通过在体内重新组装来恢复活性；以及（3）将这些非活性部分融合到特定的目标上结构域，从而产生有条件活性的细胞因子模拟物，仅通过重新组装来刺激 T 细胞目标细胞的表面（即显示两种特定表面生物标志物的癌细胞）作为证明。，第一个这样的从新原理设计的细胞因子模拟物已经生产出来，分裂，并被证明可以减少这项研究提案具有创新性，因为它没有伴随毒性或免疫原性。它旨在解决癌症免疫治疗的长期障碍（即脱靶毒性）基于细胞因子的疗法）通过从头开始设计一类新型无毒细胞因子模拟物。该提案意义重大，因为它将是计算蛋白质设计产生目标的第一个例子，最终，此类分子具有治疗多种癌症的潜力。包括恶性黑色素瘤、肾细胞癌等。