Engineering tissue inhibitor of metalloproteinases-2 (TIMP-2) for triple negative breast cancer therapy

用于三阴性乳腺癌治疗的工程组织金属蛋白酶-2 (TIMP-2) 抑制剂

基本信息

批准号：
10177669
负责人：
Evette S Radisky
金额：
$ 33.66万
依托单位：
MAYO CLINIC JACKSONVILLE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-01 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10177669
关键词：
ART protein Active Sites Affinity Binding Breast Cancer therapy Breast cancer metastasis Cell Surface Receptors Cells Cessation of life Clinical Collection Complex Computer Models Crystallization Dependence Development Directed Molecular Evolution Dose-Limiting Engineering Enzyme Precursors Enzymes Extracellular Matrix Family Gelatinase B Growth Human Individual Integrin Binding Integrin alpha3beta1 Ions Knowledge Libraries Malignant Neoplasms Matrix Metalloproteinase Inhibitor Matrix Metalloproteinases Mediating Metalloproteases Methodology Molecular Mus Musculoskeletal Mutation N-terminal Neoplasm Metastasis Neutrophil Collagenase Patients Peptide Hydrolases Pharmaceutical Preparations Pre-Clinical Model Protease Inhibitor Protein Engineering Proteins Publishing Research Role Site Specificity Structure Surface Technology Tertiary Protein Structure Therapeutic Tissue Engineering Tissue Inhibitor of Metalloproteinases Tissues Transgenic Model Transgenic Organisms Treatment Efficacy Tumor Angiogenesis Variant Work X-Ray Crystallography Yeasts Zinc angiogenesis anti-cancer anti-cancer therapeutic anticancer activity base breast cancer progression cancer therapy candidate marker clinical development design experimental study hormone receptor-negative human model implantation improved improved outcome in vivo in vivo Model inhibitor/antagonist innovation insight malignant breast neoplasm molecular drug target molecular targeted therapies mouse model novel novel anticancer drug novel strategies novel therapeutics patient derived xenograft model preclinical development prevent protein structure prototype receptor response biomarker screening small molecule targeted cancer therapy targeted treatment therapeutic protein therapeutic target triple-negative invasive breast carcinoma tumor tumor growth tumor microenvironment tumor progression

项目摘要

PROJECT SUMMARY/ABSTRACT The MMPs have long been recognized as potential targets for cancer therapy, but drugs developed to target these enzymes have been unsuccessful. A primary reason has been inadequate selectivity, since most MMP inhibitors cannot discriminate among MMPs that drive cancer progression and other MMPs that prevent cancer progression. We have recently developed a new approach, expertise, and methodology for engineering much more highly selective MMP inhibitors based on a human protein, tissue inhibitor of metalloproteinases-2 (TIMP2). In our recently published work, we have created an engineered variant of the TIMP2 N-terminal domain (N-TIMP2) with greatly improved selectivity toward MMP-9, an enzyme critically involved in triple- negative breast cancer (TNBC) progression and metastasis. In preliminary studies, we find that this prototype inhibitor shows enhanced activity for blocking TNBC cellular invasion. We propose to further engineer N- TIMP2 for increased selectivity toward MMP-9 and also for enhanced affinity toward α3β1 integrin, a second natural target of TIMP2 through which TIMP2 mediates inhibition of tumor growth. We will define the structural basis for selective MMP binding of engineered N-TIMP2 variants to enable yet greater molecular improvements, and we will evaluate the therapeutic potential of these engineered proteins in multiple complementary preclinical models of TNBC. In Aim 1, we will use a combination of structural insights, computational design and yeast surface display (YSD) technology to engineer N-TIMP2, further optimizing selectivity toward MMP-9 and enhancing beneficial integrin binding activity. In Aim 2, we will elucidate structures of the engineered proteins with target and anti-target MMPs using X-ray crystallography, to uncover the structural basis for engineered selectivity and to facilitate yet greater refinements of our engineering platform and our selective MMP-9 inhibitors. In Aim 3, we will use complementary mouse orthotopic, transgenic, and patient-derived xenograft (PDX) models of TNBC to evaluate the utility of engineered N-TIMP2 variants as a therapeutic strategy in TNBC, and identify candidate biomarkers of response with potential for directing this therapeutic approach to patients who will most benefit from it. Our proposal is both conceptually and technically innovative in the combination of approaches toward generating novel protein therapeutics. The proposed research is highly significant because it has substantial potential to develop an entirely new approach for targeted treatment of TNBC by selectively inhibiting MMP-9, a well-validated target with key roles in tumor growth, invasion, metastasis, and angiogenesis.

项目概要/摘要 MMP 长期以来一直被认为是癌症治疗的潜在靶点，但针对该靶点开发的药物这些酶不成功的一个主要原因是选择性不足，因为大多数 MMP 都缺乏选择性。抑制剂无法区分促进癌症进展的 MMP 和其他预防癌症的 MMP 我们最近开发了一种新的方法、专业知识和方法论，用于工程设计基于人类蛋白质、金属蛋白酶组织抑制剂-2 的更高选择性 MMP 抑制剂 (TIMP2) 在我们最近发表的工作中，我们创建了 TIMP2 N 端的工程变体。结构域 (N-TIMP2) 对 MMP-9 的选择性大大提高，MMP-9 是一种关键参与三重作用的酶。在初步研究中，我们发现这种原型与阴性乳腺癌（TNBC）的进展和转移有关。抑制剂显示出增强的阻断 TNBC 细胞侵袭的活性。我们建议进一步改造 N-。 TIMP2 可提高对 MMP-9 的选择性，并增强对 α3β1 整合素（第二个）的亲和力 TIMP2 的天然靶标，TIMP2 通过其介导肿瘤生长的抑制。工程 N-TIMP2 变体选择性 MMP 结合的基础，以实现更大的分子改进，我们将评估这些工程蛋白在多种方面的治疗潜力 TNBC 的互补临床前模型在目标 1 中，我们将结合使用结构见解，计算设计和酵母表面显示 (YSD) 技术来设计 N-TIMP2，进一步优化对 MMP-9 的选择性并增强有益的整合素结合活性在目标 2 中，我们将阐明。使用 X 射线晶体学分析具有靶标和抗靶标 MMP 的工程蛋白质的结构，以揭示工程选择性的结构基础，并促进我们工程的进一步完善平台和我们的选择性 MMP-9 抑制剂在目标 3 中，我们将使用互补的小鼠原位， TNBC 转基因和患者来源的异种移植 (PDX) 模型，用于评估工程 N-TIMP2 的实用性变体作为 TNBC 的治疗策略，并确定具有潜在反应的候选生物标志物我们的建议是从概念上指导这种治疗方法给最能从中受益的患者。并在技术上创新结合多种方法来产生新型蛋白质疗法。拟议的研究非常重要，因为它具有开发全新的方法的巨大潜力通过选择性抑制 MMP-9 来靶向治疗 TNBC 的方法，MMP-9 是一个经过充分验证的具有关键作用的靶点肿瘤生长、侵袭、转移和血管生成。