Specific induction of lethal autophagy in triple-negative breast cancer cells

三阴性乳腺癌细胞致死性自噬的特异性诱导

基本信息

批准号：
9324102
负责人：
Terry D. Connell
金额：
$ 17.35万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-03-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9324102
关键词：
4T1 ADP ribosylation Adenylate Cyclase Adverse effects Annexins Apoptosis Apoptotic Area Autophagocytosis Binding Biological Assay Brain Breast Breast Epithelial Cells Cancer Etiology Cancer Intervention Cancer Model Cancer cell line Cardiotoxicity Caspase Cell Membrane Permeability Cells Centers for Disease Control and Prevention (U.S.)Chimera organism Cyclic AMP Diagnosis Disease Disseminated Malignant Neoplasm ERBB2 gene Engineering Ensure Enterotoxins Escherichia coli Estrogens Fatigue Forskolin Frequencies Gangliosides Genetic Goals Growth Factor Hormones Human Immunoblotting In Vitro Individual Induction of Apoptosis Infertility Inhibition of Apoptosis Joints Liver Lung Malignant Neoplasms Mammary gland Mass Spectrum Analysis Measures Methods Micrometastasis Modeling Molecular Molecular Probes Mus Nausea Neoplasm Metastasis Pathway interactions Pharmacology Progesterone Proteins Recombinants Reporter Research Research Personnel Resistance Role Signal Transduction Small Interfering RNA Specialist Staining method Stains Technology Therapeutic Intervention Thin Layer Chromatography Thioguanine Time Tissues Toxin Woman analog cancer cell cancer subtypes cellular transduction chemotherapeutic agent cytotoxic cytotoxicity effective therapy experimental study ganglioside receptor holotoxins in vivo in vivo imaging in vivo imaging system inhibitor/antagonist killings malignant breast neoplasm member men mortality new therapeutic target novel outcome forecast receptor response targeted agent treatment strategy triple-negative invasive breast carcinoma tumor tumor growth tumor progression

项目摘要

Abstract Breast cancer remains a serious global disease associated with high rates of mortality. According to The Centers for Disease Control, 224,147 women and 2,125 men in the U.S. were diagnosed with breast cancer in 2012. Over the next ten years, the CDC estimates that 19.7 million individuals will suffer from breast cancer. Current treatments for the majority of breast cancers often depend upon interfering with hormone (ER, PR) or growth factor (HER2) receptors on those cells. Individuals who receive receptor-targeted treatments have a good prognosis for long term survival. Triple-negative breast cancer (TNBC) is a subtype of cancer caused by proliferation of breast cells that are deficient in expression of those three receptors, thus abrogating the option of targeting of those receptors as a treatment strategy. In contrast to other types of breast cancers, individuals who are diagnosed with TNBC have exceptionally poor treatment options, a higher frequency of metastatic disease, and shorter survival times. Thus, new molecules and pathways in TNBC cells that can be efficiently targeted to treat individuals diagnosed with the disease are urgently needed. In addition to their lower efficacies, current chemotherapeutic agents for TNBC elicit a number of serious side effects that are engendered by off-target activities of the treatment agents. These side effects include overwhelming fatigue, infertility, nausea, and cardiac toxicity. Thus, it is critical that more specific targets in TNBC cells are identified for therapeutic intervention. Recently, we demonstrated that LT-IIc, a bacterial type II ADP-ribosylating toxin (AR-Tx), is lethal for TNBC cells, but has no lasting effects on non- transformed breast epithelial cells. Preliminary observations suggest that the lethal effects of LT-IIc for TNBC cells is due to induction of an irreversible autophagy that is absent in normal breast epithelial cells. We will employ LT-IIc as a PROBE to identify cytotoxic mechanisms that are specifically induced in TNBC cells. A combination of pharmacological, genetic, and fluorescent approaches will be employed to evaluate the contributions of ganglioside-engagement, ADP-ribosylation, and intracellular cAMP on TNBC-specific cytotoxicity. These experiments will be facilitated by the availability of a set of recombinant AR-Tx chimeras that “mix-and-match” the ADP-ribosylating and ganglioside-binding subunits of TNBC-cytotoxic LT-IIc and non-cytotoxic LT-IIb. The roles of preferential ganglioside expression and engagement in TNBC-specific cytotoxic responses, a potentially productive and unexplored area of research, will be evaluated using novel TLC immunoblotting and mass-spectroscopy. The effects of LT-IIc on tumor growth and metastasis will be interrogated using a mouse TNBC tumor model, in vivo imaging, and in vitro culture methods. Our immediate goal is to employ LT-IIc as a molecular and cellular PROBE to uncover new and exploitable TNBC-specific cytotoxic pathways. Our LONG TERM goal is to identify pharmacological agents that target those pathways and to develop new and safer approaches for treatment of TNBC.

抽象的据报道，乳腺癌仍然是一种严重的全球疾病，死亡率很高。美国疾病控制中心的数据显示，2017 年美国有 224,147 名女性和 2,125 名男性被诊断出患有乳腺癌。 2012 年。疾病预防控制中心估计，未来十年将有 1,970 万人罹患乳腺癌。目前大多数乳腺癌的治疗通常依赖于干扰激素（ER、PR）或这些细胞上的生长因子 (HER2) 受体接受受体靶向治疗。三阴性乳腺癌（TNBC）是一种癌症亚型，具有长期生存的良好预后。缺乏这三种受体表达的乳腺细胞的增殖，从而消除了这种选择与其他类型的乳腺癌相比，靶向这些受体作为治疗策略。被诊断患有 TNBC 的人的治疗选择非常差，转移的频率更高因此，TNBC 细胞中的新分子和途径可以有效地发挥作用。迫切需要有针对性地治疗被诊断患有该疾病的个体。除了疗效较低之外，目前的 TNBC 化疗药物还引发了许多问题由于治疗药物的脱靶活性而产生的严重副作用。包括极度疲劳、不孕、恶心和心脏毒性，因此，更具体的研究至关重要。最近，我们证明了 TNBC 细胞中的治疗干预靶点。 II 型细菌 ADP 核糖基化毒素 (AR-Tx) 对 TNBC 细胞具有致命性，但对非初步观察表明 LT-IIc 对 TNBC 具有致死作用。细胞的自噬是由于正常乳腺上皮细胞中不存在的不可逆自噬的诱导而产生的。使用 LT-IIc 作为探针来识别 TNBC 细胞中特异性诱导的细胞毒性机制。将采用药理学、遗传和荧光方法的结合来评估神经节苷脂结合、ADP-核糖基化和细胞内 cAMP 对 TNBC 特异性的贡献一组重组 AR-Tx 嵌合体的可用性将促进这些实验。 “混合匹配”TNBC 细胞毒性 LT-IIc 的 ADP 核糖基化和神经节苷脂结合亚基，非细胞毒性 LT-IIb 神经节苷脂优先表达和参与 TNBC 特异性的作用。细胞毒性反应是一个潜在的富有成效且尚未探索的研究领域，将使用新颖的方法进行评估 TLC 免疫印迹和质谱分析 LT-IIc 对肿瘤生长和转移的影响。使用小鼠 TNBC 肿瘤模型、体内成像和体外培养方法进行研究。我们的近期目标是利用 LT-IIc 作为分子和细胞探针来发现新的和我们的长期目标是确定可利用的 TNBC 特异性细胞毒性途径。针对这些途径并开发新的、更安全的 TNBC 治疗方法。