Shaping the Microenvironment by DPEP1 Facilitates Adenoma Progression

通过 DPEP1 塑造微环境促进腺瘤进展

• 批准号: 10697369
• 负责人: Robert J. Coffey
• 金额: $ 37.95万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10697369
• 关键词: Attention; Behavior; Binding; Biological Assay; Biological Markers; Biological Process; Cell Line; Cells; Cellular biology; Characteristics; Coculture Techniques; Colon; Colonic Adenoma; Colorectal Adenoma; Colorectal Cancer; Communication; Confocal Microscopy; Data; Dipeptidases; Dipeptides; Electrical Resistance; Enzyme-Linked Immunosorbent Assay; Epithelium; Exposure to; Extracellular Matrix; Flow Cytometry; Future; Human; Immune; Immunofluorescence Immunologic; In Vitro; Invaded; Knock-out; Knowledge; Lesion; Leukocyte Elastase; Malignant Neoplasms; Malignant neoplasm of pancreas; Manuscripts; Mediating; Monitor; Mus; Nature; Neoplasm Metastasis; Neutrophil Infiltration; Organoids; Patients; Polyps; Population; Proliferating; Property; Proteins; Reporter; Reporting; Role; Sampling; Selection Criteria; Shapes; Signal Transduction; Source; Specimen; Stains; System; Techniques; Testing; Tissue Microarray; Tissues; Tumor Suppressor Proteins; Western Blotting; adenoma; cancer biomarkers; cancer cell; colon cancer cell line; elastase inhibitor; exosome; extracellular vesicles; healthy volunteer; immunoreactivity; in vivo; interest; microbiota; mouse model; neutrophil; premalignant; protein biomarkers; repository; single-cell RNA sequencing; spatial relationship; stem cells; timeline; transcriptome sequencing; tumor; tumor microenvironment; tumor progression

PROJECT SUMMARY/ABSTRACT Project 1: Shaping the Microenvironment by DPEP1 Facilitates Adenoma Progression Project 1 aims to examine how shaping the microenvironment by DPEP1 facilitates adenoma progression. This will be a basic project. We propose that dipeptidase-1 (DPEP1) marks those adenomas with the potential to progress to colorectal cancer (CRC) through an active bi-directional communication with neutrophils. DPEP1 has two functions: dipeptidase activity and recently identified neutrophil-binding activity. Our hypothesis is that DPEP1, largely through its neutrophil-binding activity, marks adenomas with a predilection for progression. We have found that DPEP1 immunoreactivity is detected in 27% of colorectal adenomas but this increases to 72% of CRCs, consistent with DPEP1 marking the small subset of adenomas that progress to CRC. Utilizing human specimens, a unique Transwell co-culture system of adenoma organoids and freshly isolated neutrophils isolated from healthy volunteers, and an informative mouse model, we will test if DPEP1-expressing adenomas more effectively communicate with neutrophils and create a neutrophil-enriched microenvironment, increasing the likelihood that these adenomas will progress. Exosomes have attracted a great deal of recent attention as a rich source of cargo that may serve as cancer biomarkers. We have found that DPEP1 is released in exosomes from CRC cell lines and that it highly enriched in a subset of exosomes that contain known CRC biomarkers, CEA and EPCAM. Of interest, neutrophils also release small extracellular vesicles (sEVs) that contain neutrophil elastase in a form that cannot be inhibited by elastase inhibitors, and thus it is especially potent in degrading the extracellular matrix, a key step in cancer invasion. We will also use a unique neutrophil reporter mouse to monitor onset and perdurance of neutrophil infiltration, along with the properties of these neutrophils, in an inducible, stem cell-driven mouse model of colonic adenomas. .

项目摘要/摘要 项目1：通过DPEP1塑造微环境，促进腺瘤的进展 项目1旨在研究DPEP1通过DPEP1塑造微环境的促进腺瘤进展。这 将是一个基本项目。我们提出，二肽酶-1（DPEP1）标记了这些腺瘤的潜力 通过与中性粒细胞进行主动双向通信，结直肠癌（CRC）的进展。 DPEP1 具有两个功能：二肽酶活性，最近鉴定出中性粒细胞结合活性。我们的假设是 DPEP1在很大程度上通过其中性粒细胞结合活性，标志着腺瘤具有进展的偏爱。我们 已经发现，在结直肠腺瘤的27％中检测到DPEP1免疫反应性，但这增加到72％ CRC，与DPEP1一致，该dpep1标志着腺瘤的少量子集，这些腺瘤的发展为CRC。利用人类 标本，一种独特的Transwell共培养系统的腺瘤类器官和新鲜分离的中性粒细胞的标本 从健康的志愿者和信息丰富的鼠标模型中，我们将测试表达DPEP1的腺瘤是否更多 有效地与中性粒细胞交流，并创建富含嗜中性粒细胞的微环境，从而增加 这些腺瘤有可能进展的可能性。作为富人，外泌体吸引了最近的关注 可以用作癌症生物标志物的货物来源。我们发现DPEP1在外泌体中发布 CRC细胞系，它高度富集在包含已知CRC生物标志物CEA的外泌体的一部分中 和epcam。有趣的是，中性粒细胞还释放了含有嗜中性粒细胞的小细胞外囊泡（SEV） 弹性蛋白酶以无法抑制弹性酶抑制剂抑制的形式，因此尤其有效地降解 细胞外基质，癌症入侵的关键步骤。我们还将使用独特的中性粒细胞记者鼠标进行监测 中性粒细胞浸润的发作和呈现，以及这些中性粒细胞的特性，在诱导的， 结肠腺瘤的干细胞驱动小鼠模型。 。