Dietary Risk for Colon Cancer in the Mouse

小鼠结肠癌的饮食风险

基本信息

批准号：
8103612
负责人：
LEONARD H AUGENLICHT
金额：
$ 28.05万
依托单位：
MONTEFIORE MEDICAL CENTER (BRONX, NY)
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-02-11 至 2016-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8103612
关键词：
Aberrant crypt foci Address Affect Alleles Animals Biochemical Markers Biochemistry Calcium Cancer Etiology Cell Count Cell Maturation Cell Separation Cell division Cell physiology Cells Cholecalciferol Clinical Management Colon Colon Carcinoma Colonic Neoplasms Colorectal Cancer Complement Country Data Deoxyglucose Developed Countries Development Diet Dietary Factors Dissection Epithelial Cells Equilibrium Evaluation Fatty acid glycerol esters Frequencies Gene Expression Gene Expression Profiling Gene Mutation General Population Genes Genetic Models Genetic Risk Genetic Transcription Glycolysis Health Histologic Homeostasis Human Image In Situ Incidence Individual Inflammation Inflammatory Response Inflammatory Response Pathway Inflammatory disease of the intestine Intake Interleukin-12 Internal Ribosome Entry Site Intervention Intestinal Cancer Intestinal Mucosa Intestinal Neoplasms Intestines Large Intestine Lesion Link Longevity Mediating Metabolic Pathway Methodology Methods Modeling Mucous Membrane Mus Mutation Normal Cell Nutrient Nutritional Pathway interactions Pattern Physiology Population Positioning Attribute Prevention Probability Production Proliferating Published Comment Publishing Reagent Relative Risks Reporting Risk Risk Factors Rodent Role Screening procedure Serum Signal Transduction Site Small Intestines Somatic Mutation Spatial Distribution Stem cells Stochastic Processes Testing Tissues Transcriptional Activation Vitamin D Weaning base clinical decision-making clinical efficacy cytokine density feeding genetic strain high risk imaging modality macrophage metabolomics molecular marker mouse model mutant neoplastic neutralizing monoclonal antibodies notch protein novel prevent progenitor research study response stem cell population tumor tumorigenesis villin

项目摘要

DESCRIPTION (provided by applicant): "Sporadic" colon cancer refers to >90% of colon cancer in the US and other developed countries. We have shown that this can be modeled by feeding C57Bl/6 mice a "new western- style diet" (NWD1) that combines a number of nutrient risk factors for colon cancer in the US and other western countries: 1) the NWD is formulated on the basis of nutrient density to reflect the higher fat and lower calcium and vitamin D3 levels consumed by large segments of the population; 2) tumors develop in control C57Bl/6 mice fed the diet for 2/3 of their life-span, similar to the 2/3 of the lifespan (ie 50-60 years) before most human sporadic colon cancer develops; 3) tumors develop at the same incidence and frequency as in the general population; 4) elevating calcium and vitamin D3 in the diet to levels associated with lower risk for colon cancer in the human prevents tumor development. We reported that in C57Bl/6 mice fed the NWD1 for 6 months, well before tumors form, there are subtle but significant alterations in gene expression in the flat, histologically normal mucosa of the large and small intestines that significantly overlap with alterations caused by inheritance of a mutant Apc allele, and that can be prevented by elevating calcium and vitamin D3 (NWD2). Thus, the perturbations track with probability of tumor formation. Finally, we have used a novel method of transcriptional imaging to demonstrate that Apc is haploinsufficient in the Apc1638N/+ mouse for regulating the dynamics of intestinal cell maturation, a perturbation of intestinal homeostasis that may also be caused by dietary induced higher risk. Our hypothesis is that development of sporadic colon cancer is a stochastic process the probability of which is determined by adaptation of the intestinal mucosa to patterns of nutrient intake. Aim 1, determines how the higher risk diet alters the number and positional distribution of Lgr5+ intestinal stem cells, allocation of these cells to different lineages, the role of Wnt and Notch signaling and their interaction in perturbations of mucosal homeostasis, and if these are prevented by elevation of calcium and vitamin D3. These experiments make use of novel methodology, including: a mouse genetic strain in which intestinal stem cells are marked and that also allows lineage specific tracing of their progeny; a method of cell isolation as a function of cell position along the crypt-luminal axis; and unique imaging of transcriptional activation of specific genes at the single cell level in situ. Aim 2 employs a model that more closely recapitulates human sporadic colon cancer in that conditional/inducible mutation of the Apc gene will be used to determine how the diets fed for 6 months prime the mucosa for development of preneoplastic and neoplastic lesions upon introduction of heterozygous or homozygous Apc mutation later in the animal's life-span following adaptation to nutritional factors. Aim 3 extends our published gene expression data and recently generated metabolomic data suggesting that both dietary and genetic risk for intestinal cancer cause an early shift towards glycolysis in cells of the normal appearing mucosa, specifically in the progenitor/proliferating cell compartment, long before such shifts that are known to characterize colon tumors. In response to comments made in the prior review, we will inhibit glycolysis with 2-deoxy-D glucose and determine how this modulates the dietary effects on stem cells, cell maturation pathways, and lineage specific allocation, and on inflammatory responses. Aim 4 has been added, again in response to specific comments in the review of the original application. In this aim we characterize the effects of the diets on inflammation and cytokine production of the colon and small intestine. Moreover, based on our recent reports dissecting the role of IL12 in mediating an important regulatory cross-talk between macrophages and colon tumor epithelial cells, and our data that this cytokine is elevated by the NWD1, but that this is prevented by elevating calcium and vitamin D in NWD2, we will use a novel neutralizing monoclonal Ab to IL12 to determine whether this eliminates or modulates the effects of diet on tumor associated alterations in the mucosa.

描述（由申请人提供）：“零星”结肠癌是指美国和其他发达国家的结肠癌> 90％。我们已经表明，这可以通过喂食C57BL/6小鼠来建模，一种“新西方风格饮食”（NWD1），结合了美国和其他西方国家的结肠癌的许多营养危险因素：1）NWD是根据养分较高的脂肪和较低的脂肪和较低的脂肪和维生素d3水平的大型人群来形成的NWD。 2）肿瘤在对照C57BL/6小鼠中喂养2/3的生命周期，类似于大多数人类零星结肠癌的生命周期（即50-60年）的2/3； 3）肿瘤的发病率和频率与普通人群相同； 4）将饮食中的钙和维生素D3升高到与人类中降低结肠癌风险相关的水平，可预防肿瘤的发展。我们报道说，在C57BL/6小鼠中，在肿瘤形成之前喂了NWD1 6个月，在较大和小肠的平坦，组织学正常的粘膜中，基因表达发生了细微但显着的变化，这些粘膜与Mutant APC等位基因的遗传所引起的变化显着重叠，并且可以通过升高钙化和vatimin dyaf carcium and calcium and calcium and calcium and calcium and calcium and calcium and calcium and calcium and calcium and calcium and calcium and calcium and calcium and calcium and calmin d3（nWDDDDDDD）。因此，具有肿瘤形成概率的扰动轨迹。最后，我们使用了一种新型的转录成像方法来证明APC在APC1638/+小鼠中具有一单倍弹性，以调节肠细胞成熟的动力学，这是肠内稳态的扰动，这也可能由饮食诱发的较高的风险引起。我们的假设是，散发性结肠癌的发展是一个随机过程，其概率是由肠粘膜适应营养摄入量的模式来确定的。 AIM 1，确定较高的风险饮食如何改变LGR5+肠干细胞的数量和位置分布，这些细胞分配给不同谱系，Wnt和Notch信号传导的作用以及它们在粘膜稳态扰动中的相互作用，以及是否通过钙和维生素D3的升高来预防这些稳定性。这些实验利用了新方法，包括：标记肠道干细胞的小鼠遗传菌株，还允许对后代的特定谱系追踪；细胞分离的一种方法，沿隐窝 - 延伸轴的细胞位置的函数；以及在原位单细胞水平的特定基因转录激活的独特成像。 AIM 2采用了一个模型，该模型更紧密地概括了人类零星的结肠癌，在该条件/诱导的APC基因突变中将使用该模型来确定在引入杂质和肿瘤病变的粘膜中，在引入杂质和杂质的APC突变因子后来，饮食如何促进粘膜和肿瘤性病变的粘膜。 AIM 3扩展了我们已发表的基因表达数据，并最近产生了代谢组学数据，这表明肠道癌的饮食和遗传风险在正常出现粘膜的细胞中朝着糖酵解的早期转移，特别是在祖细胞/增殖细胞室中，早在已知的转变之前就表征了Clon tumors tumors tumors。为了回应先前综述中的评论，我们将用2-脱氧葡萄糖抑制糖酵解，并确定这如何调节对干细胞，细胞成熟途径以及特定谱系分配以及炎症反应的饮食影响。 AIM 4已被添加，再次响应原始应用程序的审查中的特定评论。在此目标中，我们表征了饮食对结肠和小肠的炎症和细胞因子产生的影响。 Moreover, based on our recent reports dissecting the role of IL12 in mediating an important regulatory cross-talk between macrophages and colon tumor epithelial cells, and our data that this cytokine is elevated by the NWD1, but that this is prevented by elevating calcium and vitamin D in NWD2, we will use a novel neutralizing monoclonal Ab to IL12 to determine whether this eliminates or modulates the饮食对粘膜肿瘤相关改变的影响。