ALOX15 regulation of colon cancer invasiveness via PI3P-linoleic acid metabolism

ALOX15 通过 PI3P-亚油酸代谢调节结肠癌侵袭力

基本信息

批准号：
10339182
负责人：
Imad Shureiqi
金额：
$ 66.6万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10339182
关键词：
ALOX15 gene APC mutation Adenocarcinoma Adverse event Affect American diet Animals Arachidonate 15-Lipoxygenase Automobile Driving Azoxymethane Benign Biological Assay Biological Markers Breeding CD44 gene CDX2 gene Cancer Etiology Cell membrane Cellular Membrane Cessation of life Chemicals Chemopreventive Agent Colon Carcinoma Colonic Polyps Colorectal Colorectal Cancer Complex Consumption Corn Oil Cyclin D1 DNA Sequence Alteration Data Development Diet Disease Doxycycline Endosomes Enzymes Event Genes Goals Human Immunodeficient Mouse In Vitro Incidence Induced Mutation Intake Integration Host Factors Intervention KRAS2 gene KRASG12D Knowledge LDL-Receptor Related Protein 1 LGR5 gene Lead Lentivirus Linoleic Acids Lipoxygenase 1 Liquid Chromatography Mass Spectrum Analysis Metabolism Mucous Membrane Mus Mutation Omega-6 Fatty Acids Organoids Phospholipids Polyunsaturated Fatty Acids Recycling Regulation Reporting Research Resected Resolution Risk Rodent Role Sampling Signal Transduction System Testing Tetanus Helper Peptide Tissues Transgenic Organisms United States adenoma aldehyde dehydrogenase 1 beta catenin c-myc Genes cancer invasiveness colon cancer risk colon carcinogenesis dietary dietary excess drinking water gain of function in vivo insight lipoprotein receptor related protein 5 loss of function mouse model novel novel chemoprevention organoid transplantation peroxidation phosphatidylinositol 3-phosphate prevent receptor stem cell self renewal stem cells stemness therapy development tumor tumorigenesis villin

项目摘要

Colorectal cancer is the third leading cause of cancer deaths in the United States. The long-term goal of our research is to develop novel interventions to prevent colorectal carcinogenesis (CRC). CRC invasiveness, a critical adverse step during CRC progression, requires a combination of certain genetic mutations (e.g. APC, KRAS and Trp53 mutations), which are the key events to drive CRC. However, CRC progression also requires additional factors which increase aberrant Beta-catenin (B-catenin) activation above levels induced by APC/B- catenin mutations. Linoleic acid (LA), the most commonly consumed omega-6 polyunsaturated fatty acids in humans, increases both chemically (AOM)– and APC mutation– induced CRC tumorigenesis in mice. Nonetheless, human studies have been inconclusive regarding the impact of dietary LA on CRC. Determination of LA's role in CRC is important because American diets are enriched with LA while expression of the main metabolizing enzyme for LA,15-lipoxygenase-1 (ALOX15), is lost in human CRC. Recently, we found that 1) high dietary levels of LA promoted CRC by increasing phosphatidylinositol 3-phosphate (PI3P) containing LA (PI3P_LA), which increases LRP5 membranous recycling and subsequently B-catenin activation; 2) ALOX15- induced conversion of PI3P_LA to PI3P_13-HODE suppresses; LRP5 membranous recycling, B-catenin activation, CRC stemness and LA promotion of CRC, especially formation of large tumors, associated with CRC invasiveness; 3) ALOX15 loss of function (LOF) promotes large CRC tumor formation by azoxymethane in 12/15LOX-KO-12LOX (ALOX15-LOF) mice. Whether loss of ALOX15 expression promotes CRC invasiveness remains unknown. Our preliminary data show that ALOX15-LOF mice increased CRC invasiveness and targeted APC mutation into Lgr5+ colorectal stem cells induced CRC in the mice, which was blocked by transgenic ALOX15 expression. We therefore hypothesize that ALOX15 loss of function promotes CRC invasiveness by increasing PI3P_LA levels, which enhances LRP5 membranous recycling, thus potentiating Wnt/B-catenin signaling and subsequently stemness. Aim 1 will determine the effects of ALOX15 gain of function and ALOX15 LOF on LRP5, B-catenin activation, CRC stemness and invasiveness using CRC mouse models in which CRC invasiveness is promoted by either a combination of APC, KRASG12D and Trp53R172H mutations or Trp53R172H mutation with AOM induced B-catenin and KRAS mutations. Aim 2 will determine the effects of ALOX15 LOF on PI3P-LA, LRP5, B-catenin activation, stemness and invasiveness in human CRCs and examine the effects of ALOX15 re-expression via lentivirus Tet-on inducible system in human CRC-derived organoids on invasiveness in-vitro and in-vivo studies. The proposed studies are expected to provide important mechanistic insights into whether colonic ALOX15 expression as a host factor affects CRC invasiveness risk especially with high dietary LA intake. This gained knowledge could inform subjects with colorectal ALOX15 LOF to avoid high LA intake and spur development of interventions to target ALOX15 for re-expression to prevent invasive CRC.

结直肠癌是美国癌症死亡的第三主要原因。我们的长期目标研究是开发新的干预措施，以防止结直肠癌发生（CRC）。 CRC侵入性，a CRC进展过程中的关键不利步骤需要某些遗传突变的组合（例如APC，， KRAS和TRP53突变），这是驱动CRC的关键事件。但是，CRC的进展也需要增加异常β-catenin（B-catenin）激活的其他因素以上是APC/B-引起的水平 Catenin突变。亚油酸（LA），是最常见的omega-6多不饱和脂肪酸人类在化学上增加（AOM）和APC突变 - 诱导小鼠的CRC肿瘤发生。尽管如此，人类研究在饮食LA对CRC的影响方面尚无定论。决心洛杉矶在CRC中的作用很重要，因为美国饮食充满了洛杉矶，而表达主要 LA，15-脂氧合酶-1（Alox15）的代谢酶在人CRC中丢失。最近，我们发现1）通过增加含有LA的磷脂酰肌醇（PI3P），LA的高饮食水平促进了CRC （PI3P_LA），增加了LRP5膜回收，随后B-catenin激活； 2）Alox15- 将pi3p_la转换为pi3p_13-hode抑制； LRP5膜回收，B-catenin CRC的激活，CRC干和LA促进，尤其是与CRC相关的大肿瘤的形成侵入性； 3）ALOX15功能丧失（LOF）促进了甲氧基烷中的大型CRC肿瘤形成 12/15lox-ko-12lox（Alox15-lof）小鼠。 ALOX15表达的丧失是否促进CRC侵入性仍然未知。我们的初步数据表明，Alox15-洛克小鼠提高了CRC的侵入性并针对目标 APC突变进入LGR5+结直肠干细胞在小鼠中诱导CRC，该突变被转基因阻塞 ALOX15表达。因此，我们假设ALOX15功能的损失促进了CRC的侵入性提高PI3P_LA水平，从而增强LRP5膜回收，从而增强Wnt/B-catenin 信号传导和随后的茎。 AIM 1将确定ALOX15功能和ALOX15的影响 LRP5上的LOF，B-catenin激活，CRC茎和使用CRC模型的侵入性 APC，KRASG12D和TRP53R172H突变或TRP53R172H的组合促进了侵入性用AOM诱导的B-catenin和Kras突变突变。 AIM 2将确定ALOX15 LOF的影响在PI3P-LA，LRP5，B-catenin的激活，STEM和侵入性中，并检查了效果通过慢跑病毒Tet-On诱导系统在人CRC衍生的类器官中通过慢病毒Tet-On诱导系统重新表达的ALOX15 侵入性体内和体内研究。拟议的研究有望提供重要的机械了解结肠ALOX15作为宿主因素是否会影响CRC侵入性风险，尤其是在高饮食LA摄入量。获得的知识可以告知有结直肠ALOX15 LOF的受试者，以避免高 LA摄入量和刺激开发干预措施以靶向ALOX15进行重新表达，以防止侵入性CRC。