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.

项目摘要/摘要长期以来，MMP已被认为是癌症治疗的潜在靶标，但是开发出来的药物靶向这些酶没有成功。主要原因是选择性不足，因为大多数MMP 抑制剂不能区分驱动癌症进展的MMP和其他预防癌症的MMP 进展。我们最近开发了一种新的方法，专业知识和方法，用于工程基于人类蛋白质，金属蛋白酶2的组织抑制剂-2 （TIMP2）。在我们最近发表的作品中，我们创建了TIMP2 N终端的工程变体域（N-TIMP2）对MMP-9的选择性大大提高，这种酶严重参与了三重阴性乳腺癌（TNBC）进展和转移。在初步研究中，我们发现该原型抑制剂显示出增强的活性，可阻止TNBC细胞侵袭。我们建议进一步设计n- 提高对MMP-9的选择性的TIMP2，也可以增强对α3β1整合素的亲和力，第二 TIMP2的自然目标TIMP2介导了肿瘤生长的抑制作用。我们将定义结构基于设计性的N-TIMP2变体的MMP结合以实现更大的分子改进，我们将评估这些工程蛋白的治疗潜力 TNBC的互补临床前模型。在AIM 1中，我们将结合结构见解，计算设计和酵母表面显示（YSD）技术，用于工程N-TIMP2，进一步优化对MMP-9的选择性并增强有益整合素的结合活性。在AIM 2中，我们将阐明使用X射线晶体学的靶标和抗目标MMP的工程蛋白质结构，以发现工程选择性的结构性基础，并促进我们的工程学的改进平台和我们的选择性MMP-9抑制剂。在AIM 3中，我们将使用完整的鼠标原位， TNBC的转基因和患者衍生的Xenographotic（PDX）模型，以评估工程N-TIMP2的实用性变体是TNBC中一种治疗策略，并确定候选生物标志物的反应生物标志物，并具有潜力将这种治疗方法指导给最大程度地从中受益的患者。我们的建议在概念上都是在技术上创新的方法是生成新型蛋白质疗法的方法。这拟议的研究非常重要，因为它具有开发全新的巨大潜力通过选择性抑制MMP-9的靶向治疗方法，MMP-9是一种具有关键作用的验证靶标的方法在肿瘤生长，侵袭，转移和血管生成中。