Mechanism of Activity of Lonidamine

氯尼达明的活性机制

• 批准号: 9071364
• 负责人: JERRY D GLICKSON
• 金额: $ 54.23万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9071364
• 关键词: Address; Alkylating Agents; Anthracyclines; Antineoplastic Agents; Biochemical Pathway; Biochemistry; Bioenergetics; Bioreactors; Cardiac; Cell membrane; Cells; Characteristics; Clinical; Collaborations; Data; Detection; Development; Disease; Doxorubicin; Electron Transport; Energy Metabolism; Funding; Glycolysis; Glycolysis Inhibition; Goals; Grant; Health; Heart; Hexokinase 2; Human; Isolated limb perfusion; Isotopes; Kinetics; Label; Laboratories; Lactic acid; Liver; Liver Mitochondria; Lonidamine; Magnetic Resonance Spectroscopy; Malignant Neoplasms; Measures; Mechlorethamine; Mediating; Melanoma Cell; Melphalan; Metabolic; Metabolism; Methods; Mitochondria; Monitor; Monocarboxylic Acid Transporters; Mustard; Nude Mice; Oxidation-Reduction; Oxidative Phosphorylation; Pharmaceutical Preparations; Pharmacologic Substance; Phospholipid Metabolism; Production; Protein Isoforms; Pyruvate; Radiation Tolerance; Rana; Respiration; Skin Cancer; Staging; Systemic Therapy; Techniques; Testing; Therapeutic Agents; Time; Tissues; Translations; Tumor Oxygenation; Universities; Virulent; Warburg Effect; Xenograft procedure; Xenopus laevis; base; cancer cell; cancer therapy; chemotherapeutic agent; chemotherapy; clinical practice; design; egg; glucose metabolism; improved; in vivo; inhibitor/antagonist; liquid chromatography mass spectrometry; melanoma; metabolomics; mitochondrial metabolism; monolayer; neoplastic cell; network models; novel; pyruvate carrier; research clinical testing; response; targeted treatment; tumor; tumor metabolism; tumor microenvironment; tumor xenograft

DESCRIPTION (provided by applicant): Melanoma remains one of the deadliest of human cancers with no effective method for treating the disseminated disease. While targeted therapies have shown some efficacy, they have thus far proven to be noncurative, which points to the need for other forms of systemic therapy. This proposal is part of a long-term effort to develop methods for systemic therapy of melanoma. We have recently shown that administration of lonidamine (LND, 100 mg/kg) substantially enhances the activity of melphalan and have now extended this finding to doxorubicin treatment. As a key step towards eventual clinical translation, we will now examine the detailed mechanism of LND activity. We and others have shown that LND produces tumor specific intracellular acidification and a substantial decrease in tumor energy status (NTP/Pi). A number of mechanisms have been proposed for LND: 1) inhibition of hexokinase II, 2) interference with mitochondrial electron transport, 3) inhibition of cellular lactate export through the monocarboxylate transporters (MCT). We are proposing a fourth mechanism to explain the bioenergetic decline of the tumor following treatment with LND: 4) inhibition of the putative mitochondrial pyruvate carrier (MPC), which would deplete the tumor cells of a key substrate for oxidative phosphorylation and induce an increase in glycolytic metabolism to compensate for decreased oxidative ATP production. We have recently validated a novel extension of isotopomer analysis called cumomer analysis and applied it to perfused DB1 melanoma cells. We believe that this is the first validated metabolic network model of tumor energy metabolism. We propose to test the hypotheses that 1) selective tumor acidification results from inhibition of MCT1, and 2) tumor de-energization is caused by inhibition of the MPC. In Aim 1 of this proposal, we propose to use 13C magnetic resonance spectroscopy (MRS) and liquid chromatography-mass spectrometry (LC-MS) in conjunction with bonded cumomer analysis to test these hypotheses in perfused DB1 and DB8 melanoma cells and in in vivo xenografts of these tumor lines. In Aim 2 we will directly measure the inhibitory effect of LND on MCT1 and MCT4 expressed in Xenopus laevis and will also evaluate the effect of LND on isolated liver and cardiac mitochondria and on permeabilized DB1 and DB8 cells. All of the criticisms of the previous review have been addressed including the critical issue of clinical translation for which we assembled a team of leading experts on melanoma who recommended: 1) define the mechanism of LND (the aim of this proposal), 2) first incorporate LND into hyperthermic isolated limb perfusion of melanoma with melphalan (a method already in clinical practice), and 3) then proceed to systemic therapy of cancers that are already treated with N-mustards or doxorubicin. This project will have a major impact on elucidating the mechanism of activity of a new class of drugs that like LND inhibit MCTs and other key transporters in tumor cells and thereby modify the tumor microenvironment to augment the activity of conventional anticancer agents. It will also develop novel metabolomic methods for the study of tumor metabolism and mechanisms of cancer drug activity.

描述(申请人提供)：黑色素瘤仍然是人类最致命的癌症之一，目前还没有有效的方法来治疗这种播散性疾病。虽然靶向治疗显示出了一些效果，但到目前为止，它们被证明是不能治愈的，这表明需要其他形式的系统治疗。这项建议是开发系统治疗黑色素瘤方法的长期努力的一部分。我们最近已经证明，服用LND(100 mg/kg)大大增强了马法兰的活性，现在已经将这一发现扩展到阿霉素治疗。作为最终临床翻译的关键一步，我们现在将研究LND活动的详细机制。我们和其他人已经证明，LND产生肿瘤特异性的细胞内酸化和肿瘤能量状态(NTP/PI)的大幅降低。LND的机制有许多：1)抑制己糖激酶II，2)干扰线粒体电子传递，3)通过单羧酸转运体(MCT)抑制细胞乳酸输出。我们提出了第四种机制来解释LND治疗后肿瘤的生物能量下降：4)抑制假定的线粒体丙酮酸载体(MPC)，这将耗尽肿瘤细胞氧化磷酸化的关键底物，并诱导糖酵解代谢增加，以补偿氧化ATP产生的减少。我们最近验证了一种称为累积聚体分析的新的同位素分析扩展，并将其应用于灌流的DB1黑色素瘤细胞。我们认为，这是第一个经过验证的肿瘤能量代谢的代谢网络模型。我们建议检验这样的假设：1)选择性肿瘤酸化是由于抑制MCT1，2)肿瘤失能是由抑制MPC引起的。在这项建议的目标1中，我们建议使用13C磁共振波谱(MRS)和液质联用(LC-MS)结合键合累积分析来在灌流的DB1和Db8黑色素瘤细胞以及这些肿瘤系的体内异种移植瘤中测试这些假设。在目标2中，我们将直接检测LND对非洲爪哇MCT1和MCT4表达的抑制作用，并评价LND对分离的肝脏和心脏线粒体以及通透性DB1和DB8细胞的影响。对先前综述的所有批评都得到了解决，包括临床翻译的关键问题，为此我们组建了一个领先的黑色素瘤专家团队，他们建议：1)确定LND的机制(本建议的目的)，2)首先将LND纳入含有马法兰的黑色素瘤的热隔离肢体灌注中(一种已在临床实践中的方法)，3)然后对已用N-MUSTARDS或阿霉素治疗的癌症进行系统治疗。该项目将对阐明一类新型药物的活性机制产生重大影响，这些药物如LND可以抑制肿瘤细胞中的MCTs和其他关键转运蛋白，从而改变肿瘤微环境以增强传统抗癌药物的活性。它还将开发新的代谢方法来研究肿瘤代谢和抗癌药物活性的机制。