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: 10700076
• 负责人: Amir Zarrinpar
• 金额: $ 61.32万
• 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/10700076
• 关键词: Acceleration; Address; Affect; Age; Animal Genetics; Animal Model; Attenuated; Bacteria; Bile Acids; Biological Response Modifier Therapy; Biosensor; Black American; Cancer Biology; Cancer Detection; Cancer Etiology; Caspase; Cells; Cessation of life; Colon; Colorectal Cancer; Data; Development; Diagnosis; Disease; Disease Progression; Early identification; Engineering; Engraftment; 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; Prevention; Progress Reports; 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; Visualization; Work; adenoma; anticancer research; 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; 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.

项目摘要/摘要 尽管总体下降次数减少，但结直肠癌（CRC）仍然是第四大最常见原因 美国的癌症死亡。不幸的是，流行病学研究表明，令人震惊 在50岁以下的人口中的病情，他们没有经常筛选。此外，CRC检测很困难 在高风险组中，包括具有遗传易感性的疾病（例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（例如黑人）。因此，有一个 为开发创新解决方案以早期检测和预防创新解决方案的重要需求 从腺瘤到CRC的过渡。为了满足这一需求，我们的跨学科研究团队将发展 使用合成生物学方法鉴定早期CRC开发，监测的基因工程细菌 并报告腺瘤和CRC微环境的变化，并防止癌症进展。实现 上述目标，工程细菌必须植入和定居敌对的腔环境， 并区分异常的环境信号，计算此信号并表达记者或治疗方法 代理人。但是，与这些特征的适当向量仍然缺乏，限制合成生物学应用 癌症研究的研究。重要的是，CRC与大肠杆菌高度相关，我们有许多合成 生物学工具。此外，我们的初步概念验证研究表明，本地大肠杆菌可以是 设计为永久地定居完全常规（即非微生物组耗尽）主机，并执行func- 感兴趣的象征，例如，腔胆酸的解偶。脱糖的胆汁酸和由此产生的Farnesoid X重新 CEPTOR（FXR）激动剂可以抑制CRC的发展，表明潜在的治疗用途 天然细菌。在我们强大的支持性初步结果的基础上，我们将确定健康的大肠杆菌 CRC遗传模型的腺瘤和CRC组织，并设计其检测和治疗CRC以响应 到癌症微环境。此外，我们将表征不同肿瘤环境的影响 关于在结肠器官模型中的定植和表现的因素 在数学建模的支持下，芯片的器官，从而确定了程序的特定CRC信号 - 将工程的本地大肠杆菌作为CRC记者和治疗剂的反应。最后，我们将设计 通过定量报告CRC- 相关的半胱氨酸蛋白酶并有选择地抑制其活性。该提案中描述的研究将 产生新的，急需的合成生物学媒介，可以作为生物传感器和治疗剂开发 腺瘤和CRC以及许多其他疾病。此外，该项目将丰富我们的基本 有关CRC-微生物组关系的知识，并阐明了半胱氨酸蛋白酶在CRC Pro- gression和治疗。