A strategy for the selective imaging and inhibition of MT1-MMP to assess its functional roles in breast cancer invasion and metastasis

MT1-MMP 的选择性成像和抑制策略，以评估其在乳腺癌侵袭和转移中的功能作用

• 批准号: 314695690
• 负责人: Dr. Martina Tholen
• 金额: --
• 依托单位: Department of Pathology
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/314695690?language=en
• 关键词: strategy; selective; imaging; inhibition; MT1

Matrix metallo proteases (MMPs) facilitate tumor cell invasion and angiogenesis by mediating degradation of the extracellular matrix, making them promising anti-cancer drug targets. The family of MMPs consists of 23 members with high structural homology but diverse functions in tumor progression. This high degree of structural homology has made the development of specific inhibitors and other chemical tools to study MMPs challenging. Furthermore, in contrast to serine or cysteine proteases, MMPs do not use a protein bound nucleophile to catalyze cleavage of the amide bond. Thus, they lack a nucleophile that can be targeted by inhibitors to covalently modify the enzyme. The group of Prof. Matthew Bogyo has recently developed a new approach that overcomes these limitations and allows specific targeting of individual MMP proteases. The strategy involves the introduction of a functionally silent mutation into a MMP protease to generate a nucleophile for covalent modification by small molecule inhibitors and optical probes that have been designed to target the newly introduced residue. Thus, probes can be generated to be absolutely selective for the engineered MMP allowing specific imaging and selective inhibition of the desired protease with a high degree of temporal control. In addition this protease engineering/probe approach avoids limitations of previous knock out studies targeting MMPs like issues of compensation and lethality. In this proposal I outline plans to apply this newly developed technology to a mouse model of breast cancer to study the function of membrane type 1-matrix metalloproteinase 1 (MT1-MMP) in cancer progression. MT1-MMP is the most potent collagenase among the MMPs and is critical for cancer cell invasion through stromal collagen matrices. This outstanding role among the collagenases is most likely based on its localization at the cell membrane enabling focused ECM remodeling. However, a more complete understanding of MT1-MMP function and especially the consequences of its specific inhibition will be required to both dissect its specific role in tumorigenesis and to validate it as a relevant drug target. To accomplish this goal I will use mice that underwent CRISPR/Cas based genome editing to express the engineered MT1-MMP crossed with a transgenic model for metastasizing breast cancer. This model combined with an engineered inhibitor will allow me to evaluate the effect of specific MT1-MMP inhibition on tumor progression and metastatic seeding. Furthermore, direct visualization of active MT1-MMP localization by a selective probe during primary tumor development and at metastatic sites will provide insight into its primary function in disease pathology. I believe that with the help of these highly novel and powerful chemical tools, it will be possible to revisit MT1-MMP as a drug target in cancer treatment and provide a better understanding of MT1-MMP function so that more effective treatment strategies can be developed.

基质金属蛋白酶（Matrix metallo proteases，MMPs）通过降解细胞外基质促进肿瘤细胞的侵袭和血管生成，是一种很有前途的抗肿瘤药物靶点。MMPs家族由23个成员组成，具有高度的结构同源性，但在肿瘤进展中具有不同的功能。这种高度的结构同源性使得开发特异性抑制剂和其他化学工具来研究MMP具有挑战性。此外，与丝氨酸或半胱氨酸蛋白酶相反，MMP不使用蛋白质结合的亲核试剂来催化酰胺键的裂解。因此，它们缺乏可以被抑制剂靶向以共价修饰酶的亲核试剂。Matthew Bogyo教授的研究小组最近开发了一种新方法，克服了这些限制，并允许特异性靶向个别MMP蛋白酶。该策略涉及将功能沉默突变引入MMP蛋白酶以产生亲核试剂，用于通过小分子抑制剂和光学探针进行共价修饰，所述光学探针被设计为靶向新引入的残基。因此，可以产生对工程化MMP具有绝对选择性的探针，从而允许特异性成像和选择性抑制具有高度时间控制的所需蛋白酶。此外，这种蛋白酶工程/探针方法避免了先前靶向MMP的敲除研究的局限性，如补偿和致死性问题。在这份提案中，我概述了计划将这种新开发的技术应用于小鼠乳腺癌模型，以研究膜型1-基质金属蛋白酶1（MT 1-MMP）在癌症进展中的功能。MT 1-MMP是MMPs中最有效的胶原酶，并且对于癌细胞通过基质胶原基质的侵袭至关重要。胶原酶中的这种突出作用最有可能是基于其在细胞膜上的定位，从而实现集中的ECM重塑。然而，MT 1-MMP功能，特别是其特异性抑制的后果将需要一个更完整的理解，既剖析其在肿瘤发生中的具体作用，并验证它作为一个相关的药物靶点。为了实现这一目标，我将使用经过基于CRISPR/Cas的基因组编辑的小鼠来表达工程改造的MT 1-MMP，并与转移性乳腺癌的转基因模型杂交。该模型结合工程抑制剂将使我能够评估特异性MT 1-MMP抑制对肿瘤进展和转移性种植的影响。此外，在原发性肿瘤发展期间和转移部位通过选择性探针直接可视化活性MT 1-MMP定位将提供对其在疾病病理学中的主要功能的了解。我相信，在这些高度新颖和强大的化学工具的帮助下，将有可能重新审视MT 1-MMP作为癌症治疗的药物靶点，并更好地了解MT 1-MMP的功能，以便开发更有效的治疗策略。

项目成果

Multivariate AND-gate substrate probes as enhanced contrast agents for fluorescence-guided surgery

多元与门底物探针作为荧光引导手术的增强造影剂

• DOI: 10.1101/695403
• 期刊: bioRxiv
• 作者: Widen J.C;Tholen M;Yim J.J;Kerriann M. C;Rogalla S;Bogyo M.
• 通讯作者: Bogyo M.

The clinical drug candidate ebselen attenuates inflammation and promotes microbiome recovery after antibiotic treatment for Clostridium difficile infection

临床候选药物依布硒啉可减轻艰难梭菌感染抗生素治疗后的炎症并促进微生物组恢复

• DOI: 10.1101/827329
• 期刊: bioRxiv
• 作者: Garland M;Hryckowian A;Tholen M;Loscher S;Van Treuren W;Oresic Bender K;Sonnenburg J.L;Bogyo M.
• 通讯作者: Bogyo M.

Chemical Tools for Selective Activity Profiling of Endogenously Expressed MMP-14 in Multicellular Models.

• DOI: 10.1021/acschembio.8b00562
• 发表时间: 2018-09-21
• 期刊: ACS chemical biology
• 影响因子: 4
• 作者: Amara N;Tholen M;Bogyo M
• 通讯作者: Bogyo M