Engineering Native E. coli to Detect, Report, and Treat Colorectal Cancer

改造天然大肠杆菌来检测、报告和治疗结直肠癌

• 批准号: 10330342
• 负责人: Amir Zarrinpar
• 金额: $ 68.24万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-21 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330342
• 关键词: Acceleration; Address; Affect; African American; Age; Agonist; Animal Genetics; Animal Model; Attenuated; Bacteria; Bile Acids; Biological Response Modifier Therapy; Biosensor; Cancer Biology; Cancer Detection; Cancer Etiology; Caspase; Cells; Cessation of life; Colon; Colorectal Cancer; Data; Development; Diagnosis; Disease; Disease Progression; Engineering; Environment; Escherichia coli; Familial Adenomatous Polyposis Syndrome; Gene Expression; Genetic Engineering; Genetic Models; Genetic Predisposition to Disease; Goals; Growth; Health; Incidence; Inflammatory Bowel Diseases; Interdisciplinary Study; Intervention; Knowledge; Malignant Neoplasms; Methods; Microfluidic Microchips; Mission; Modeling; Monitor; Mus; Organoids; Outcome; Peptide Hydrolases; Performance; Population; Population Dynamics; Protease Inhibitor; Proteins; Proteomics; Public Health; Reporter; Reporting; Research; Research Personnel; Risk; Role; Scientist; Signal Transduction; System; Testing; Therapeutic; Therapeutic Agents; Therapeutic Uses; Time; Tissues; Work; adenoma; anticancer research; base; cancer therapy; colorectal cancer prevention; colorectal cancer progression; colorectal cancer screening; colorectal cancer treatment; demographics; early onset colorectal cancer; epidemiology study; gut microbiota; high risk population; host colonization; host-microbe interactions; improved; innovation; interest; mathematical model; microbial composition; microbiome; microorganism; novel; novel strategies; organ on a chip; predictive test; prevent; receptor; response; sensor; synthetic biology; tool; trait; translational impact; tumor; tumor microenvironment; tumor progression; vector

PROJECT SUMMARY/ABSTRACT Despite its overall decreasing occurrence, colorectal cancer (CRC) remains the fourth most common cause of cancer deaths in the US. Unfortunately, epidemiological studies demonstrate an alarming increase in inci- dence in populations below the age of 50, who are not routinely screened. Furthermore, CRC detection is difficult in high-risk groups, including those with a genetic predisposition (e.g. familial adenomatous polyposis), disease traits (e.g. inflammatory bowel disease), or from certain demographics (e.g. Black-Americans). Thus, there is a significant need for the development of innovative solutions for the early detection of CRC and the prevention of the transition from adenoma to CRC. To address this need, our interdisciplinary research team will develop genetically engineered bacteria using synthetic biology approaches to identify early CRC development, monitor and report changes in the adenoma and CRC microenvironment, and prevent cancer progression. To achieve the above objectives, engineered bacteria have to engraft and colonize the hostile luminal environment, sense and distinguish an abnormal environmental signal, compute this signal, and express a reporter or a therapeutic agent. However, appropriate vectors with these features remain lacking, constraining synthetic biology applica- tions for cancer research. Importantly, CRC is highly associated with E. coli, for which we have many synthetic biology tools. Furthermore, our preliminary proof-of-concept studies have revealed that native E. coli can be engineered to perpetually colonize fully conventional (i.e. non-microbiome depleted) hosts and to execute func- tions of interest, e.g., deconjugation of luminal bile acids. Deconjugated bile acid and resultant farnesoid X re- ceptor (FXR) agonism can suppress CRC development, indicating a potential therapeutic use of engineered native bacteria. Building on our strong supportive preliminary results, we will identify native E. coli from healthy, adenoma, and CRC tissues of a genetic model of CRC and engineer them to detect and treat CRC in response to the cancer microenvironment. Furthermore, we will characterize the effects of different tumor environment factors on the colonization and performances of engineered native E. coli in the colon organoid model in an organ-on-chip with the support of mathematical modeling, thereby identifying specific CRC signals for program- ming the responses of engineered native E. coli as CRC reporters and therapeutics. Finally, we will engineer native bacteria to detect and attenuate the progression of CRC by quantitatively reporting the level of CRC- related cysteine proteases and selectively inhibiting their activity. The research described in this proposal will generate new, much-needed synthetic biology vectors that can be developed as biosensors and therapeutics of adenoma and CRC, as well as many other diseases. Furthermore, this project will enrich our fundamental knowledge about the CRC-microbiome relationship and elucidate the roles of cysteine proteases in CRC pro- gression and treatment.

项目概要/摘要 尽管结直肠癌 (CRC) 的发病率总体下降，但仍然是第四大常见原因 美国癌症死亡人数。不幸的是，流行病学研究表明，突发事件的增加令人震惊。 50 岁以下人群中的流行病，这些人没有进行常规筛查。此外，CRC检测也很困难 在高危人群中，包括具有遗传倾向的人群（例如家族性腺瘤性息肉病）、疾病 特征（例如炎症性肠病），或来自某些人口统计数据（例如美国黑人）。因此，有一个 迫切需要开发创新解决方案来早期发现结直肠癌并预防结直肠癌 从腺瘤到结直肠癌的转变。为了满足这一需求，我们的跨学科研究团队将开发 使用合成生物学方法的基因工程细菌来识别早期结直肠癌的发展、监测 报告腺瘤和结直肠癌微环境的变化，并预防癌症进展。达到 为了实现上述目标，工程细菌必须植入并定植于恶劣的管腔环境中，感知 并区分异常环境信号，计算该信号，并表达报告者或治疗剂 代理人。然而，具有这些特征的合适载体仍然缺乏，限制了合成生物学的应用。 癌症研究的系统蒸发散。重要的是，CRC 与大肠杆菌高度相关，我们有许多合成的大肠杆菌 生物学工具。此外，我们的初步概念验证研究表明，本地大肠杆菌可以 设计用于永久定殖完全传统（即非微生物组耗尽）宿主并执行功能 感兴趣的部分，例如管腔胆汁酸的解离。解离的胆汁酸和所得的法尼醇 X 重新 受体 (FXR) 激动剂可以抑制 CRC 的发展，表明工程化药物的潜在治疗用途 原生细菌。基于我们强有力的支持性初步结果，我们将从健康、 CRC 基因模型的腺瘤和 CRC 组织，并对其进行改造以检测和治疗 CRC 到癌症微环境。此外，我们将表征不同肿瘤环境的影响 影响工程化天然大肠杆菌在结肠类器官模型中定植和性能的因素 芯片上的器官在数学建模的支持下，从而识别特定的CRC信号以进行编程 确定工程化的天然大肠杆菌作为 CRC 报告基因和治疗剂的反应。最后，我们将设计 本地细菌通过定量报告 CRC 水平来检测和减弱 CRC 的进展 相关的半胱氨酸蛋白酶并选择性抑制其活性。本提案中描述的研究将 产生新的、急需的合成生物学载体，可以开发为生物传感器和治疗方法 腺瘤和结直肠癌，以及许多其他疾病。此外，该项目将丰富我们的基础知识 有关 CRC 与微生物组关系的知识，并阐明半胱氨酸蛋白酶在 CRC 中的作用 进展和治疗。