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.

Public Health Relevance

Melanoma, the most virulent form of skin cancer, is not responsive to chemotherapy. We have recently shown that a cancer drug, lonidamine (LND), substantially enhances the activity of a class of cancer drugs called nitrogen mustards against melanoma and are now extending this observation to another class of drugs, the anthracylines. The purpose of this proposal is to elucidate the mechanism by which LND is enhancing the activities of conventional cancer drugs like nitrogen mustards and anthracyclines. This information will provide a basis for clinical testing of this concept and for development of an entire new class of anticancer agents now being produced by pharmaceutical companies that selectively modify the metabolic characteristics of cancer cells to make them more susceptible to chemotherapy. We are also introducing a new method called bonded cumomer analysis to quantitatively study the effects of LND and other drugs on tumor metabolism.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA172820-05
Application #
9269066
Study Section
Medical Imaging Study Section (MEDI)
Program Officer
Chen, Weiwei
Project Start
2013-08-01
Project End
2018-05-31
Budget Start
2017-06-01
Budget End
2018-05-31
Support Year
5
Fiscal Year
2017
Total Cost
$542,325
Indirect Cost
$203,372
Name
University of Pennsylvania
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Xu, He N; Feng, Min; Nath, Kavindra et al. (2018) Optical Redox Imaging of Lonidamine Treatment Response of Melanoma Cells and Xenografts. Mol Imaging Biol :
Abbruzzese, Claudia; Matteoni, Silvia; Signore, Michele et al. (2017) Drug repurposing for the treatment of glioblastoma multiforme. J Exp Clin Cancer Res 36:169
Nath, Kavindra; Nelson, David S; Roman, Jeffrey et al. (2017) Effect of Lonidamine on Systemic Therapy of DB-1 Human Melanoma Xenografts with Temozolomide. Anticancer Res 37:3413-3421
Nancolas, Bethany; Guo, Lili; Zhou, Rong et al. (2016) The anti-tumour agent lonidamine is a potent inhibitor of the mitochondrial pyruvate carrier and plasma membrane monocarboxylate transporters. Biochem J 473:929-36
Shestov, Alexander A; Mancuso, Anthony; Lee, Seung-Cheol et al. (2016) Bonded Cumomer Analysis of Human Melanoma Metabolism Monitored by 13C NMR Spectroscopy of Perfused Tumor Cells. J Biol Chem 291:5157-71
Nath, Kavindra; Nelson, David S; Putt, Mary E et al. (2016) Comparison of the Lonidamine Potentiated Effect of Nitrogen Mustard Alkylating Agents on the Systemic Treatment of DB-1 Human Melanoma Xenografts in Mice. PLoS One 11:e0157125
Shestov, Alexander A; Lee, Seung-Cheol; Nath, Kavindra et al. (2016) (13)C MRS and LC-MS Flux Analysis of Tumor Intermediary Metabolism. Front Oncol 6:135
Nath, Kavindra; Guo, Lili; Nancolas, Bethany et al. (2016) Mechanism of antineoplastic activity of lonidamine. Biochim Biophys Acta 1866:151-162
Guo, Lili; Worth, Andrew J; Mesaros, Clementina et al. (2016) Diisopropylethylamine/hexafluoroisopropanol-mediated ion-pairing ultra-high-performance liquid chromatography/mass spectrometry for phosphate and carboxylate metabolite analysis: utility for studying cellular metabolism. Rapid Commun Mass Spectrom 30:1835-45
Guo, Lili; Shestov, Alexander A; Worth, Andrew J et al. (2016) Inhibition of Mitochondrial Complex II by the Anticancer Agent Lonidamine. J Biol Chem 291:42-57

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