Understanding the AGL metastasis suppressor for therapeutic gain

了解 AGL 转移抑制因子的治疗效果

基本信息

批准号：
9223676
负责人：
DAN THEODORESCU
金额：
$ 43.54万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-12-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9223676
关键词：
American Autophagocytosis Biological Assay Biological Markers Bladder Bladder Neoplasm Bladder Urothelium CD44 gene Cancer Biology Cancer Death Rates Cells Cessation of life Clinical Clinical Trials Data Death Rate Dependence Development Diagnosis Disease Enzymes Exhibits Federal Government Foundations Future Gene Components Gene Targeting Genes Genomics Glucose Glycogen Glycolysis Goals Growth Guanosine Triphosphate Phosphohydrolases HMMR gene Human Hyaluronic Acid In Vitro Individual Knockout Mice Knowledge Lead Libraries Link Location Malignant Neoplasms Malignant neoplasm of urinary bladder Medicare Membrane Metabolic Modeling Molecular Target Mus Neoplasm Metastasis Pathway interactions Patients Pharmacology Phenotype Predictive Value Primary Neoplasm Production RNA Interference Receptor Inhibition Recurrence Resources Role SLC2A1 gene Signal Transduction Source Testing Therapeutic Translating Tumor Cell Line Tumor Suppressor Genes Urothelium Visceral Work Xenograft procedure biomarker panel biomarker-driven cancer cell cancer recurrence cancer type carcinogenesis cell growth chemical carcinogenesis cost expectation experimental study extracellular glucosidase glycogenolysis high risk in vivo inhibitor/antagonist innovation killings neoplastic cell new therapeutic target novel novel therapeutics outcome forecast pre-clinical prognostic assays prognostic value public health relevance receptor research clinical testing small hairpin RNA therapeutic development therapeutic target tumor tumor growth tumor initiation tumor metabolism tumor progression tumorigenesis whole genome

项目摘要

DESCRIPTION (provided by applicant): Bladder cancer kills 15,000 annually and because of few therapeutic advances there is a need for innovation. This project has discovered suppressors of bladder tumor growth and metastasis and shown that low expression of AGL, an enzyme involved in glycogenolysis, and RhoGDI2, an inhibitor of multiple GTPases, is associated with cancer recurrence and death. The Goal of this continuation project is to understand how reduced AGL levels result in aggressive bladder cancer and to translate this knowledge into novel therapies. We found bladder cancer cells with low AGL express elevated levels of Hyaluronic Acid (HA) synthase 2 (HAS2), and its extracellular product HA, and have activated RhoC and Rac1, known drivers of tumor progression. We demonstrated RhoC/Rac1 activity is suppressed by RhoGDI2, linking the actions of the two metastasis suppressors. Cells with low AGL also exhibit increased glycolysis, glucose import and autophagy. When either glucose import or autophagy are blocked, low AGL tumor cells show greater reduction of in vitro growth than cells with higher AGL levels, exposing a vulnerability of these aggressive cells. These data support an innovative Hypothesis: Tumor cells with low AGL generate growth and metastatic signals via production of HA, activation of its receptors CD44 and RHAMM, and downstream effectors such as RhoC and Rac1. For these signals to be actualized, sufficient energy needs to be provided via enhanced glucose import and autophagy. Three Specific Aims will test this hypothesis. 1) Evaluate impact of HA signaling on aggressiveness of bladder cancers with low AGL. We will determine if HA drives tumor progression via its receptors CD44 and RHAMM and effectors RhoC and Rac1 that are inhibited by RhoGDI2. Experiments will use RNAi and receptor inhibition in human xenografts. 2) Determine role of the AGL signaling network in bladder cancer development and progression. We will evaluate the independent predictive value of tumor suppressor genes and components of the AGL signaling network in a large panel of annotated human tumors, to define a predictive multigene biomarker signature and possible new therapeutic targets. We will use full and conditional (bladder urothelium) AGL knockout mice in a chemical carcinogenesis model that generates spontaneous visceral metastases to test whether lack of AGL makes urothelium more susceptible to carcinogenesis and resulting tumors more aggressive. 3) Target molecular vulnerabilities of bladder cancers with low AGL expression. Here we test whether blocking glucose import and/or autophagy reduces in vivo bladder tumor growth. We will use shRNA and pharmacologic agents already in preclinical/clinical testing. We will screen shRNA libraries to metabolic and autophagy genes in cells with low AGL to discover novel genes essential to the aggressive tumor phenotype, with the expectation that some of these will become therapeutic targets. We will evaluate combined inhibition of HA signaling and energy production pathways with the expectation this will have synergistic detrimental effects on tumor progression. Impact: This project will lay the foundation for future biomarker-driven personalized clinical trials that target critical vulnerabilities in bldder cancer while informing us on how AGL executes its unanticipated role in cancer.

描述（由适用提供）：膀胱癌每年杀死15,000人，由于几乎没有治疗进展，需要创新。该项目发现了叶叶肿瘤生长和转移的补充剂，表明AGL的低表达是一种参与糖原分解的酶，而多个GTPase的抑制剂RhoGDI2与癌症复发和死亡有关。该延续项目的目的是了解降低的AGL水平如何导致侵袭性的膀胱癌，并将这些知识转化为新的疗法。我们发现，具有低AGL的Blader癌细胞表达透明质酸（HA）合酶2（HAS2）及其细胞外产物HA的水平升高，并激活了RHOC和RAC1，这是已知的肿瘤进展驱动因素。我们证明了RHOC/RAC1活性被RHOGDI2抑制，将两个转移细胞的作用与低AGL相关联，也暴露了糖酵解，葡萄糖进口和自噬。当葡萄糖进口或自噬被阻断时，低的AGL肿瘤细胞比具有更高水平的细胞显示出更大的体外生长降低，从而暴露了这些侵袭性细胞的脆弱性。这些数据支持创新的假设：通过产生HA，其受体CD44和RHAMM的激活以及诸如RHOC和RAC1之类的下游效应，AGL较低的肿瘤细胞会产生生长和转移信号。为了实现这些信号，需要通过增强的葡萄糖进口和自噬提供足够的能量。三个具体目标将检验这一假设。 1）评估HA信号传导对AGL低的膀胱癌的攻击性的影响。我们将确定HA是否通过其受体CD44和RHAMM和效应驱动肿瘤进展。 RHOGDI2抑制的RHOC和RAC1。实验将在人异种移植物中使用RNAi和受体抑制。 2）确定AGL信号网络在膀胱癌发展和进展中的作用。我们将评估大型注释的人类肿瘤中AGL信号网络的肿瘤抑制基因和成分的独立预测值，以定义预测性的多基因生物标志物和可能的新治疗靶标。我们将在化学癌变模型中使用完全有条件的（膀胱尿液）AGL基因敲除小鼠，该模型会产生自发的内脏转移，以测试缺乏AGL是否会使尿液尿液更容易受到致癌作用，并导致肿瘤更具侵略性。 3）具有低AGL表达的膀胱癌的靶标分子脆弱性。在这里，我们测试阻断葡萄糖进口和/或自噬是否会减少体内膀胱肿瘤生长。我们将在临床前/临床测试中使用SHRNA和药理学剂。我们将筛查shRNA文库中的SHRNA文库中的AGL低细胞中的代谢和自噬基因，以发现对侵袭性肿瘤表型必不可少的新型基因，并期望其中一些将成为治疗靶标。我们将评估对HA信号传导和能源生产途径的综合抑制作用，并预期这将对肿瘤进展产生协同的有害影响。影响：该项目将为未来的生物标志物驱动的个性化临床试验奠定基础，这些临床试验针对大厦癌症中的关键脆弱性，同时向我们告知AGL如何在癌症中扮演意外角色。