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.

项目总结/摘要 尽管其总体发生率下降，但结直肠癌（CRC）仍然是第四大常见原因 在美国癌症死亡人数。不幸的是，流行病学研究表明，inci- 在50岁以下的人群中，没有进行常规筛查。此外，CRC检测是困难的 在高危人群中，包括具有遗传倾向的人群（如家族性腺瘤性息肉病）， 特征（例如炎症性肠病）或某些人口统计学特征（例如黑人）。由此可见，有一 迫切需要开发创新的解决方案，以早期发现CRC和预防 从腺瘤到CRC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，以识别早期CRC发展，监测 并报告腺瘤和CRC微环境的变化，防止癌症进展。实现 为了达到上述目的，工程菌必须植入并定殖在不利的管腔环境中， 并区分异常环境信号，计算该信号，并表达报告基因或治疗基因， 剂然而，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。 为癌症研究做贡献。重要的是，CRC与E.大肠杆菌，我们有许多合成的 生物学工具此外，我们初步的概念验证研究表明，原生E。大肠杆菌可以是 被设计为永久地定殖完全常规的（即非微生物组耗尽的）宿主，并执行功能， 感兴趣的内容，例如，管腔胆汁酸的去结合。去共轭胆汁酸和所得的法尼醇X re-binding 受体（FXR）激动可以抑制CRC的发展，表明工程改造的FXR具有潜在的治疗用途。 原生细菌基于我们强有力的支持性初步结果，我们将确定天然E。大肠杆菌， 腺瘤和CRC组织，并对其进行工程改造以检测和治疗CRC， 到癌症微环境。此外，我们将描述不同肿瘤环境的影响， 影响工程天然E.大肠杆菌在结肠类器官模型中， 芯片上器官的数学建模的支持，从而识别特定的CRC信号的程序， 观察工程天然E.大肠杆菌作为CRC报告基因和治疗剂。最后，我们将工程师 通过定量报告CRC的水平来检测和减弱CRC的进展， 相关的半胱氨酸蛋白酶并选择性地抑制它们的活性。本提案中所述的研究将 产生新的、急需的合成生物学载体，可开发为生物传感器和治疗方法 腺瘤和CRC，以及许多其他疾病。此外，该项目将丰富我们的基础知识， 关于CRC-微生物组关系的知识，并阐明半胱氨酸蛋白酶在CRC前体中的作用。 回归与治疗