Diagnostic peptide-nanoparticle probes for profiling tumor protease activity

用于分析肿瘤蛋白酶活性的诊断肽纳米颗粒探针

• 批准号: 8319688
• 负责人: Gabriel A Kwong
• 金额: $ 5.01万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8319688
• 关键词: Aftercare; Amino Acids; Anatomy; Animals; Autoimmunity; Bacterial Infections; Basement membrane; Biochemical; Biological Assay; Biopsy; Biotechnology; Blood Circulation; Blood Clot; Blood Coagulation Disorders; Blood coagulation; Cardiovascular Diseases; Comparative Study; Complex; Detection; Diagnostic; Disease; Drug Formulations; Enzyme Precursors; Event; Extracellular Matrix; Fluorescent Probes; Fluorogenic Substrate; Goals; Growth; Growth Factor Receptors; Half-Life; Image; Immunoglobulin Joining Region; Kidney; Ligands; Literature; Liver Fibrosis; Location; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Methods; Metric; Modeling; Molecular; Molecular Chaperones; Monitor; Mus; Neoplasm Metastasis; Optical reporter; Parasitic Diseases; Pathology; Patient Care; Peptide Fragments; Peptide Hydrolases; Peptide Library; Peptides; Positioning Attribute; Preclinical Drug Evaluation; Property; Protease Inhibitor; Proteolysis; Proteolytic Processing; Reaction; Regulation; Renal clearance function; Reporter; Reporting; Signal Transduction; Site; Specificity; Staging; Surveys; System; Techniques; Technology; Testing; Therapeutic; Tissues; Tumor-Associated Process; Urine; Virus Diseases; Work; Xenograft procedure; angiogenesis; base; clinical Diagnosis; cofactor; design; improved; in vivo; inhibitor/antagonist; insight; iron oxide; mouse model; nanoparticle; prognostic; response; stable isotope; stem; transcriptomics; tumor; tumor progression

DESCRIPTION (provided by applicant): Aberrant protease activity is essential in many complex tumor processes including growth, invasion and metastasis. Proteolytic events are responsible for unveiling cryptic ECM signaling domains, degrading basement membranes, and activating a suite of molecules including receptors and growth factors. Despite their importance, few technologies exist for detecting and monitoring activities in vivo. Traditional ex vivo techniques like tumor biopsy followed by zymography assays are invasive and provide limited insight since protease activity is highly contextual and tightly regulated in vivo. Recent work with activity-based fluorescent probes are limited by the small number of tissue-penetrating reporters available, precluding studies of proteolytic networks and detection of tumor activity at sites deep within tissue. Here, we propose to circumvent these challenges by constructing long circulating peptide-nanoparticle probes that can survey, sense and remotely report on tumor activity through the urine. In this strategy, iron oxide nanoparticles are utilized as chaperones to deliver protease-specific peptide libraries to tumors whereupon selective cleavage by active proteases releases peptide fragments that are cleared by the renal system into urine. These peptide fragments are predesigned with photo-labile triggers to uncage isobaric peptide "reporters" optimized for multiplexed LC MS/MS quantification. We hypothesize that mass spectrometric analysis of urine in comparative studies involving tumor-bearing and tumor-free mice will uncover unique reporter signatures that can be correlated with protease activity in vivo. Quantified protease signatures will be utilized as a metric for detecting, monitoring and evaluating tumor responses to anti-tumor therapies. This strategy of quantifying proteolytic activity through mass-encoded remote reporters will enable a high degree of multiplexing and will permit the detection of tumors at sites independent of anatomical position. We expect this platform to have broad utility and amenable to a number of protease-dependent diseases such as cardiovascular diseases, coagulopathies, and liver fibrosis.

描述（由申请人提供）：在许多复杂的肿瘤过程中，异常蛋白酶活性至关重要，包括生长，侵袭和转移。蛋白水解事件负责揭示隐秘的ECM信号传导域，降解基底膜，并激活一套分子，包括受体和生长因子。尽管它们的重要性，但在体内检测和监测活动的技术很少。传统的离体技术，例如肿瘤活检，然后进行同志分析，因此具有侵入性，并且提供有限的见解，因为蛋白酶活性是高度的上下文，并且在体内受到严格调节。最新的基于活性的荧光探针的工作受到了少量的组织渗透记者的限制，排除了蛋白水解网络的研究以及在组织深处的部位检测肿瘤活性。在这里，我们建议通过构建长循环肽 - 纳米粒子探针来避免这些挑战，这些探针可以通过尿液进行调查，感知和远程报道肿瘤活性。在这种策略中，将氧化铁纳米颗粒用作伴侣蛋白将蛋白酶特异性的肽库传递到肿瘤中，从而通过活性蛋白酶的选择性切割释放了肾脏系统清除为尿液的肽片段。这些肽片段是用光比勒触发器预先设计的，以优化用于多路复用LC MS/MS定量的Uncabile肽“ Reporter”。我们假设在涉及肿瘤和无肿瘤小鼠的比较研究中对尿液的质谱分析将发现可以与体内蛋白酶活性相关的独特记者特征。定量的蛋白酶特征将被用作检测，监测和评估对抗肿瘤疗法的肿瘤反应的度量。通过质量编码的远程记者来量化蛋白水解活性的策略将使高度的多重速度进行，并允许在与解剖位置无关的部位检测肿瘤。我们期望这个平台具有广泛的效用，并且可以适应许多蛋白酶依赖性疾病，例如心血管疾病，凝血病和肝纤维化。