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(1) Intestinal Organoid Models for APC LOH

(1) APC LOH肠类器官模型

基本信息

批准号：
9172584
负责人：
CHARLES A. GIARDINA
金额：
$ 23.22万
依托单位：
UNIVERSITY OF CONNECTICUT STORRS
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9172584
关键词：
Affect Alleles ApcMin/+ mice Apoptosis Cell Separation Cells Clustered Regularly Interspaced Short Palindromic Repeats Colon Carcinoma Cytokinesis Data Detection Development Diphtheria Toxin Diploid Cells Engineering Environmental Risk Factor Event Failure Frequencies Gene Expression Genes Genetic Genetic Polymorphism Genetic study Genome Stability Genomic Instability Genomics Growth Heterozygote Individual Intestinal Cancer Intestines Lesion Link Loss of Heterozygosity Mitosis Modeling Molecular Profiling Mucous Membrane Mutation Nature Organoids Ploidies Population Premalignant Process Regulatory Pathway Reporter Resistance Risk Role Secretory Cell Signal Transduction Stem cells System TP53 gene Time Tissues Tumor Suppressor Genes abstracting cancer cell cancer prevention cancer risk cancer stem cell diphtheria toxin receptor high risk insight mouse model mutant novel single cell analysis targeted agent transcriptome tumor progression

项目摘要

Project Summary/Abstract A critical step in colon cancer development occurs when a cell within a field of Apc- heterozygote mutant cells undergoes Apc loss of heterozygosity (LOH). Understanding the mechanism and cellular consequences of an Apc LOH event could provide important information on how this event might be suppressed to reduce cancer risk. Critical questions that remain unanswered include: Do the events leading to Apc LOH include a failure of cytokinesis? Are all cells that undergo an LOH event at equivalent risk of progression? What genetic and environmental factors can impact Apc LOH? We propose to develop an intestinal organoid system as a malleable experimental platform to study Apc LOH. We will utilize intestinal organoids generated from ESCs since these “mini-intestines” can be genetically manipulated and grown in the quantities necessary to detect the relatively rare Apc LOH event. We propose to develop this organoid system to study Apc LOH in real-time by linking the ApcMin allele to GFP and the Apc-wild type allele to an RFP reporter and the diphtheria toxin receptor (DTR). We will use these organoids to enumerate and isolate cells that have undergone Apc LOH. We will initially determine how p16 and p53 affect Apc LOH frequency, but envision these organoids being used to assess the impact of a broad range of mutations and polymorphisms. We will study the nature of cells emerging from an Apc LOH event, focusing on their proliferative capacity and their relationship to normal ISCs and cancer stem cells. Finally, we will assess the role of tetraploid cells as precursors for Apc LOH and determine if a novel mitosis-targeting agent that selectively induces apoptosis of tetraploid cells can reduce the frequency of Apc LOH. These exploratory studies are anticipated to establish an experimental system for studying a critical event in colon cancer development; the emergence of Apc-mutant cells in a pre-malignant tissue field. In the long term we envision establishing an integrated experimental pipeline to understand how environmental and genetic factors impact Apc LOH, how cells that have undergone Apc LOH progress to cancer stem cells, and ultimately, how an individual's risk of colon cancer risk can be reduced.

项目总结/文摘