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

基本信息

批准号：
10493339
负责人：
Gabriel A Kwong
金额：
$ 59.16万
依托单位：
GEORGIA INSTITUTE OF TECHNOLOGY
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：
2021-09-23 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10493339
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 Colorectal Cancer Complex Coupled Cyclic Peptides Data Detection Development Diagnostic 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 radiological imaging sensor synthetic biology systemic toxicity tumor tumor specificity validation studies

项目摘要

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.

项目概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础性作用例如，细菌等全细胞生物传感器已被广泛应用。经过基因工程改造以执行复杂的功能，例如信号放大以检测临床相关人类尿液和血清中的生物标志物在哺乳动物细胞中，利用这些成分进行感知和响应。布尔逻辑已被证明可用于工程 T 细胞疗法的多重控制，从而增加肿瘤传感的特异性和降低全身毒性这些都凸显了合成的前景。生物学应用于癌症时，但绝大多数这些策略依赖于遗传电路并利用这种电路很复杂，引起了安全性和免疫原性问题。获得监管批准，特别是在体内早期癌症检测的情况下，重复给药可能需要大量生物传感器来监测新生疾病。该提案旨在开发一类新的传感器。称为 AND 门控合成生物标志物的诊断方法，用于早期检测癌症合成转移。生物标记物是一类新兴的可激活生物传感器，旨在进行管理系统地查询早期疾病的位点，并利用肿瘤依赖性激活机制，例如失调的蛋白酶活性，驱动报告基因的产生，然后可以检测到这些报告基因。无创地从血液、尿液或其他体液样本中提取蛋白酶在整个过程中发挥着关键的生物学作用。转移的主要标志，并且是通过催化不可逆转的特别有效的分子放大器肽键的水解，允许单个蛋白酶转换数千个“与”门控的合成底物。生物标志物将用于结直肠癌（CRC）肝转移的早期检测。原发性 CRC 转移扩散的常见部位，肝隔离转移灶的区域切除可导致早期发现结直肠癌肝转移，以达到潜在的治愈效果。通过放射成像（例如 CT 和 FDA 批准的）对治疗的反应（1-2 毫米）仍然具有挑战性血液测试，例如癌胚抗原 (CEA) 测试设计用于 CRC 的 AND 门控合成生物标志物。肝转移中，将根据差异RNA表达来选择蛋白酶的成对组合将开发 CRC 肝转移与健康肝组织的多变量数学模型。与实验结果相比，了解设计参数如何增强特异性和协同性。 CRC肝转移的同基因和异种移植模型将用于临床前验证研究，以允许该提案将为癌症的早期检测奠定基础。通过可编程合成生物标志物进行转移。