Characterization of microbial and host requirements for colonic biofilm assembly and biofilm-mediated colon tumorigenesis

结肠生物膜组装和生物膜介导的结肠肿瘤发生的微生物和宿主需求的表征

基本信息

批准号：
10456150
负责人：
Julia L Drewes
金额：
$ 24.9万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-08-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10456150
关键词：
Acute Address Age Animal Model Antibodies ApcMin/+ mice Apoptosis Bacteria Bacterial Model Bacteroidetes Biological Response Modifiers Biology Cancer Etiology Cells Cessation of life Clinical Research Colon Colonoscopy Colorectal Cancer Communities Data Data Set Development Distal E-Cadherin Epidemiology Epithelial Epithelial Cells Familial colorectal cancer Flow Cytometry Germ-Free Grant Health Histologic Human Immune Immune response Immune signaling In Situ Nick-End Labeling Individual Interleukin-17 Interleukin-6 Intestinal Neoplasms Knock-out Knockout Mice Laboratories Lead Left Malignant Neoplasms Mass Spectrum Analysis Mediating Mediator of activation protein Metabolic Methods Microbe Microbial Biofilms Modification Mucous Membrane Mus Normal tissue morphology Patients Phenotype Play Polyamines Production Prognosis Quantitative Reverse Transcriptase PCR Role Sampling Side Signal Pathway Stains Suggestion Testing Therapeutic Time Tissues Virulence Factors base carcinogenesis colon cancer patients colon hepatic flexure colon tumorigenesis colorectal cancer prevention colorectal cancer risk cytokine experimental study global health gut microbiome inhibitor mass spectrometric imaging metabolomics microbial microbial host microbiome mouse model mucosal biofilms novel prospective response screening single-cell RNA sequencing therapeutic biomarker therapeutic target tool tumor tumor progression tumorigenesis tumorigenic

项目摘要

Sporadic colorectal cancer (CRC) remains a global health burden as one of the leading causes of cancer and cancer-related deaths, and, strikingly, is also emerging as an acute health crisis in younger patients in the US under the age of 50. These data suggest that novel efforts to identify contributors to the changing landscape of CRC are urgently needed. In this R00 proposal, we seek to dissect the assembly and tumorigenic mechanisms behind invasive, polymicrobial colonic biofilms, which were previously established in the Sears laboratory to be a nearly universal feature of CRC tumors proximal to the hepatic flexure. Biofilms were also identified on a subset (10-15%) of healthy patients without CRC, in whom biofilms were associated with early, procarcinogenic changes, positing a role for bacterial biofilms in CRC development. These biofilms were also associated with elevations in polyamine metabolites in mass spectrometry-based analyses and induced a strong Interleukin-17 (IL-17) response in the mice by one week, a cytokine signature associated with a worse prognosis in human CRC. Tissue slurries made from human biofilms re-assembled into biofilms in the distal mouse colon; some - but not all – of these slurries were directly tumorigenic in germ-free ApcMin/+ mice by 10-15 weeks. Extensive culturing of one tumorigenic slurry following inoculation into mice revealed 35 bacterial isolates that, when re-inoculated into additional mice, recapitulated the tumorigenesis of the original patient tumor slurry. Our specific aims seek to build on these preliminary studies. Specific Aim (SA) 1 will identify the critical bacterial species within the 35 isolate mixture that are responsible for biofilm formation and tumorigenesis. SA2 will test the hypothesis that IL-17 and other immune parameters are critical mediators of biofilm formation and biofilm-mediated tumorigenesis utilizing a combination of single-cell RNA sequencing, flow cytometry, and qRT-PCR. Therapeutic modulation with an IL-17 antibody or knockout mice will be used to confirm the requirement of IL-17 in biofilm formation and biofilm-mediated tumorigenesis. SA3 will utilize advanced mass spectrometry methods to determine metabolites associated with biofilms in both patient and mouse tissues and the localization of these metabolites in select samples. The experiments in this proposal will provide valuable information regarding the host and microbial requirements for biofilm formation and biofilm-mediated tumorigenesis that may lead to potential therapeutic targets or biomarkers for biofilms.

散发性结直肠癌（CRC）作为癌症的主要原因之一，仍然是全球健康负担和与癌症相关的死亡，而且令人惊讶的是，它也正在成为年轻患者的严重健康危机 50 岁以下的美国。这些数据表明，旨在确定变革贡献者的新努力在这个 R00 提案中，我们迫切需要 CRC 的景观。侵入性、多种微生物结肠生物膜背后的致瘤机制，这些生物膜以前被认为是 Sears 实验室确定这是近肝结直肠癌肿瘤的几乎普遍特征在一组（10-15%）无 CRC 的健康患者中也发现了生物膜。生物膜与早期致癌变化相关，表明细菌生物膜在结直肠癌中发挥作用这些生物膜还与多胺代谢物质量的升高有关。一种基于光谱分析的方法，在小鼠中诱导了强烈的白细胞介素 17 (IL-17) 反应周，细胞因子特征与人类结直肠癌组织浆液的较差预后相关。人类生物膜在小鼠远端结肠中重新组装成生物膜；其中一些（但不是全部）浆液在无菌 ApcMin/+ 小鼠中广泛培养 10-15 周后可直接致瘤。接种小鼠后的致瘤浆液显示出 35 种细菌分离株，当重新接种时进入其他小鼠体内，重现了原始患者肿瘤浆液的肿瘤发生，我们的具体目标。寻求以这些初步研究为基础，具体目标 (SA) 1 将确定关键细菌种类。 SA2 将测试负责生物膜形成和肿瘤发生的 35 种分离混合物。假设 IL-17 和其他免疫参数是生物膜形成的关键介质生物膜介导的肿瘤发生利用单细胞 RNA 测序、流式细胞术和使用 IL-17 抗体或基因敲除小鼠进行的治疗性调节将用于确认 IL-17 在生物膜形成和生物膜介导的肿瘤发生中的需求将利用先进的 SA3。质谱法测定患者和小鼠生物膜相关代谢物本提案中的实验将研究组织和这些代谢物在选定样本中的定位。提供有关生物膜形成的宿主和微生物需求的有价值的信息生物膜介导的肿瘤发生可能导致生物膜的潜在治疗靶点或生物标志物。