AND-gated Synthetic Biomarkers for Early Detection of Liver Metastasis

用于早期检测肝转移的 AND 门控合成生物标志物

• 批准号: 10330265
• 负责人: Gabriel A Kwong
• 金额: $ 59.96万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-23 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330265
• 关键词: Amplifiers; Bacteria; Bar Codes; Benchmarking; Biodistribution; Biological; Biological Markers; Biology; Biosensor; Blood; Blood Tests; Cancer Biology; Cancer Diagnostics; Cancerous; Carcinoembryonic Antigen; Cells; Classification; Cleaved cell; Colorectal Cancer; Complex; Coupled; Cyclic Peptides; Data; Detection; Development; Diagnostic; Diagnostic radiologic examination; Disease; Drug Kinetics; Early Diagnosis; Enzymes; Excision; FDA approved; Fluorogenic Substrate; Future; Gene Expression; Genes; Genetic; Genetic Engineering; Genetic Transcription; Growth; Human; Hydrolysis; Image; In Situ; Inbred BALB C Mice; Kinetics; LS174T colon cancer cell line; Lead; Liquid substance; Liver; Logic; Longitudinal Studies; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Metastatic Neoplasm to the Liver; Modeling; Molecular; Monitor; Neoplasm Metastasis; Noise; Nude Mice; Patient-Focused Outcomes; Peptide Fragments; Peptide Hydrolases; Peptides; Performance; Play; Preclinical Testing; Production; Proteins; Recombinants; Reporter; Role; Safety; Sampling; Screening for cancer; Serum; Shapes; Signal Transduction; Site; Specificity; T cell response; T cell therapy; Testing; Thermal Ablation Therapy; Tissues; Tumor Antigens; Urine; Validation; Xenograft Model; Xenograft procedure; antigen test; base; clinically relevant; colorectal cancer screening; design; detection limit; differential expression; early detection biomarkers; engineered T cells; fluorophore; immunogenicity; improved; in vivo; iron oxide nanoparticle; mathematical model; microCT; models and simulation; mouse model; neoplastic cell; pre-clinical; preclinical evaluation; predictive modeling; programs; sensor; synthetic biology; systemic toxicity; tumor; tumor specificity; validation studies

PROJECT SUMMARY Advances in synthetic biology will play a fundamental role in shaping the future of cancer diagnostics toward earlier and more specific detection of disease. For example, whole-cell biosensors such as bacteria have been genetically engineered to perform complex functions such as signal amplification to detect clinically relevant biomarkers in human urine and serum. In mammalian cells, sense-and-respond components that employ Boolean logic have been demonstrated for multiplexed control of engineered T cell therapies, thereby increasing the specificity of tumor sensing and reducing systemic toxicity. These advances highlight the promise of synthetic biology when applied to cancer, yet the vast majority of these strategies rely on genetic circuits and make use of non-mammalian protein components. Such circuits are complex and raise safety and immunogenicity concerns for regulatory approval, especially in the context of in vivo early cancer detection where repeated administrations of biosensors are likely needed to monitor for nascent disease. This proposal seeks to develop a new class of diagnostics called AND-gated synthetic biomarkers for early detection of cancer metastasis. Synthetic biomarkers are an emerging class of activatable biological sensors that are designed to be administered systemically, query sites of early disease, and harness tumor-dependent activation mechanisms, such as dysregulated protease activity, to drive production of a reporter. These reporters can then be detected noninvasively from blood, urine, or other bodily fluid samples. Proteases play key biological roles across the major hallmarks of metastasis and are particularly potent molecular amplifiers by catalyzing the irreversible hydrolysis of peptide bonds, allowing a single protease to turnover thousands of substrates. AND-gated synthetic biomarkers will be applied for early detection of colorectal cancer (CRC) liver metastasis. Although the liver is a common site for metastatic spread from primary CRC, regional resection of liver-isolated metastases can lead to potentially curative results. Yet early detection of CRC liver metastases at a size when they are most responsive to therapy (1–2 mm) remains challenging by radiographic imaging such as CT and FDA-approved blood test such as the carcinoembryonic antigen (CEA) test. To design AND-gated synthetic biomarkers for CRC liver metastasis, pairwise combinations of proteases will be selected based on differential RNA expression in CRC liver metastases compared to healthy liver tissue. Multivariate mathematical models will be developed to understand how design parameters enhance specificity and cooperativity compared to experimental results. Syngeneic and xenograft models of CRC liver metastasis will be used for preclinical validation studies to allow benchmarking against CT and CEA. This proposal will lay the groundwork for earlier detection of cancer metastasis by programmable synthetic biomarkers.

项目摘要 合成生物学的进步将在塑造癌症诊断的未来方面发挥基本作用 早期，更具体地检测疾病。例如，诸如细菌之类的全细胞生物传感器已经 一般设计以执行复杂功能，例如信号放大以检测临床相关的 人类尿液和血清中的生物标志物。在哺乳动物细胞中，采用的感官和回答成分 已经证明了布尔逻辑以用于对工程T细胞疗法的多路复用控制，从而增加 肿瘤感应和降低全身毒性的特异性。这些进步突出了合成的希望 生物学应用于癌症时，这些策略中的绝大多数都依赖于遗传回路并利用 非哺乳动物蛋白成分。这样的电路很复杂，并提高了安全性和免疫原性问题 为了获得监管批准，尤其是在体内早期癌症检测的背景下，重复管理 可能需要生物传感器来监测新生疾病。该建议旨在开发一个新的类 称为诊断和门控的合成生物标志物，用于早期检测癌症转移。合成的 生物标志物是一类新兴的可激活的生物传感器，旨在施用 从系统上，早期疾病的查询部位和线束依赖肿瘤的激活机制，例如 失调的蛋白酶活性，以驱动记者的产生。然后可以检测到这些记者 从血液，尿液或其他体液样品中无创的。蛋白酶在遍布关键的生物学角色 转移的主要标志，并且是通过催化不可逆的，特别是潜在的分子放大器 胡椒键的水解，使单个蛋白酶的营业额数千个底物。和门控合成 生物标志物将用于早期检测结直肠癌（CRC）肝转移。虽然肝脏是 从原发性CRC中转移传播的常见部位，区域切除型转移可以引导 潜在的治疗结果。然而，当它们最多的时候，早期发现CRC肝转移的大小 对治疗的反应（1-2毫米）仍然受到射线照相成像的挑战，例如CT和FDA批准 血液测试，例如癌胚抗原（CEA）测试。为CRC设计和门控的合成生物标志物 肝转移，将根据差异RNA表达选择蛋白酶的成对组合 CRC肝转移与健康的肝组织相比。多元数学模型将开发为 与实验结果相比，了解设计参数如何增强特异性和协调。 CRC肝转移的合成性和异种移植模型将用于临床前验证研究，以允许 针对CT和CEA进行基准测试。该提案将为早期检测到癌症奠定基础 可编程合成生物标志物的转移。