Genetic and Dietary Interactions in MMR Deficient Colon Tumorigenesis

MMR 缺陷性结肠肿瘤发生中的遗传和饮食相互作用

• 批准号: 10405006
• 负责人: LEONARD H AUGENLICHT
• 金额: $ 59.58万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-05 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10405006
• 关键词: ATAC-seq; Address; Back; Benign; CRISPR/Cas technology; Carcinoma; Cells; Characteristics; Chromatin; Chromatin Structure; Clinical; Collaborations; Colon; Colonic Adenoma; Colonic Neoplasms; Colorectal Cancer; Critiques; DNA; DNA Repair; Data; Databases; Defect; Detection; Development; Diet; Dietary Factors; Dissection; Early Diagnosis; Engineering; Epidemiology; Epigenetic Process; Epithelial Cells; Etiology; Gene Expression; General Population; Genes; Genetic; Genetic Models; Hereditary Nonpolyposis Colorectal Neoplasms; Histopathology; Homeostasis; Human; Incidence; Individual; Inherited; Intake; Intestinal Neoplasms; Intestines; Link; Mismatch Repair; Modeling; Modification; Molecular; Mucinous; Mucous Membrane; Mus; Mutation; Nutrient; Oligonucleotides; Oxidative Phosphorylation; Pathogenesis; Pathology; Pathway interactions; Patients; Penetrance; Prevention; Prevention strategy; Public Health; Publishing; Relative Risks; Risk; Risk Marker; Rodent; Sentinel; Site; Small Intestinal Neoplasm; Societies; Somatic Mutation; Specificity; Stomach; Stretching; Time; Tissues; Transforming Growth Factor beta; WNT Signaling Pathway; Wild Type Mouse; Work; adenoma; base; beta catenin; colon tumorigenesis; design; dietary; dietary control; exome sequencing; feeding; genome-wide analysis; histone modification; individualized medicine; mouse genetics; mouse model; novel; response; stem cell function; stem cell population; stem cells; transcriptome sequencing; tumor; tumorigenesis; villin; western diet

Mouse genetic models of Lynch syndrome (LS)/defective MMR (dMMR) driven tumorigenesis produce small intestinal tumors. However, long-standing collaborative work of the Co-PIs has now developed a new mouse model that causes LS/dMMR tumorigenesis in the mouse colon by integrating the genetic and environmental etiology that mimics that of the human. In the Villin-cre, Msh2loxp/loxp, TgfbRIIhu/hu mice (VMshThu), homozygous inactivation of Msh2 in intestinal and colonic epithelial cells causes tissue tissue-specific dMMR. Further, “G” residues in an otherwise “A” stretch in the mouse TgfbRll gene were altered by CRISPR/Cas9 to be oligoA, replicating the oligoA in the human gene that makes TgbRll a key target of dMMR. The mice develop intestinal tumors with the unique histopathologies of flat adenoma progressing to mucinous invasive carcinoma characteristic of LS/dMMR tumors, and every tumor has a mutation in the oligo A of the “humanized” TgfbRII gene. Feeding a purified rodent western-style diet (NWD1) to VMshThu mice shifts tumor penetrance into the colon, producing the same unique pathologies as in human LS/dMMR. The NWD1 recapitulates intake for the mouse of several nutrients at levels of each epidemiologically linked in western-societies to higher incidence of colorectal cancer. This new model has very high relevance to human LS, including historical data linking dietary shift in human LS to colon penetrance, as well as for somatically linked dMMR and sporadic colon tumorigenesis in the general population. Our published and submitted data establish that feeding NWD1 to wild-type mice profoundly alters function of intestinal stem cells in homeostasis and tumorigenesis, with underlying changes in stem cell programming and accumulation, spectra and signature of mutations. Therefore, we hypothesize shift to colon tumor penetrance by feeding NWD1 to VMshThu mice is due to altered colonic stem cell function. Aim 1 dissects alterations in function, programming, epigenetic and genetic changes induced by NWD1 in VMshThu mice in Lgr5hi, Bmi1+ and Aldh1+ colonic stem cell populations. Emphasis is on the Tgfb, Wnt signaling, DNA repair and oxidative phosphorylation pathways, and extended globally. Aim 2 addresses the fundamental public health issue of persistence of specific programming, epigenetic and genetic changes in long-lived colonic stem cells and their progeny upon switching the protumorigenic NWD1 back to purified control diet. Aim 3 dissects molecular changes that characterize the progression of the unique benign flat adenomas to mucinous invasive carcinomas. Further, using unique RNAseq data bases of LS patients compared to average risk individuals for the uninvolved mucosa, and for adenomas and carcinomas of LS and dMMR patients, correspondence between genes and pathways in the mouse model and in human will be determined. This will prioritize targets for development of markers of risk and progression and for further mechanistic dissection.

林奇综合征（LS）/有缺陷的MMR（DMMR）驱动肿瘤发生的小鼠遗传模型产生的小 肠道肿瘤。但是，Co-Pis的长期合作工作现在已经开发了新的鼠标 通过整合遗传和环境，导致小鼠结肠中LS/DMMR肿瘤发生的模型 模仿人类的病因。在Villin-Cre中 MSH2在肠道和结肠上皮细胞中的失活会导致组织组织特异性DMMR。此外，“ G” 小鼠TGFBRLL基因中原本“ A”中的残留物被CRISPR/CAS9改变为Oligoa， 复制人类基因中的寡核酸，这使TGBRLL成为DMMR的关键靶标。小鼠发育肠道 具有扁平腺瘤的独特组织病理学的肿瘤发展为粘液浸润性癌 LS/DMMR肿瘤的特征，每个肿瘤在“人源化” TGFBRII的寡核A中具有突变 基因。将纯化的啮齿动物西方风格饮食（NWD1）喂给VMSHTHU小鼠，将肿瘤的渗透转移到 结肠，产生与人类LS/DMMR相同的独特病理。 NWD1概括了 在西方占地的每个流行病学水平上，几种营养素的小鼠与更高的发生率 结直肠癌。这个新模型与人类LS具有很高的相关性，包括与饮食联系的历史数据 人体LS转移到结肠渗透性，以及与体体连接的DMMR和零星结肠肿瘤发生 在一般人群中。 我们已发表和提交的数据建立，将NWD1喂给野生型小鼠的数据深刻改变了功能 体内平衡和肿瘤发生中的肠道干细胞，干细胞编程的潜在变化和 突变的积累，光谱和标志。因此，我们假设转移到结肠肿瘤的渗透率 通过将NWD1喂入VMSHTHU小鼠是由于结肠干细胞功能的改变。 AIM 1剖析改变 NWD1在LGR5HI中，NWD1引起的功能，编程，表观遗传和遗传变化，BMI1+ 和ALDH1+结肠干细胞群体。重点是TGFB，Wnt信号传导，DNA修复和氧化 磷酸化途径，并在全球范围内扩展。 AIM 2解决了基本的公共卫生问题 长期寿命的结肠干细胞及其其特定编程，表观遗传和遗传变化的持久性 后代将杂种NWD1切换回纯化的对照饮食。 AIM 3解剖分子 特征的变化是独特的良性扁平腺瘤向粘液浸润的进展的变化 癌。此外，使用LS患者的唯一RNASEQ数据库与平均风险个人相比 未参与的粘膜，对于LS和DMMR患者的腺瘤和癌， 将确定小鼠模型和人中的基因和途径。这将优先考虑 发展风险和进展标记以及进一步的机械剖析。