Mitochondrial complex I activity and NAD+/NADH balance regulate breast cancer progression.

Despite advances in clinical therapy, metastasis remains the leading cause of death in breast cancer patients. Mutations in mitochondrial DNA, including those affecting complex I and oxidative phosphorylation, are found in breast tumors and could facilitate metastasis. This study identifies mitochondrial complex I as critical for defining an aggressive phenotype in breast cancer cells. Specific enhancement of mitochondrial complex I activity inhibited tumor growth and metastasis through regulation of the tumor cell NAD+/NADH redox balance, mTORC1 activity, and autophagy. Conversely, nonlethal reduction of NAD+ levels by interfering with nicotinamide phosphoribosyltransferase expression rendered tumor cells more aggressive and increased metastasis. The results translate into a new therapeutic strategy: enhancement of the NAD+/NADH balance through treatment with NAD+ precursors inhibited metastasis in xenograft models, increased animal survival, and strongly interfered with oncogene-driven breast cancer progression in the MMTV-PyMT mouse model. Thus, aberration in mitochondrial complex I NADH dehydrogenase activity can profoundly enhance the aggressiveness of human breast cancer cells, while therapeutic normalization of the NAD+/NADH balance can inhibit metastasis and prevent disease progression.

[1]  A. Ashworth,et al.  Synthetic lethality of PARP and NAMPT inhibition in triple-negative breast cancer cells , 2012, EMBO molecular medicine.

[2]  W. Lam,et al.  Mitochondrial DNA mutations in respiratory complex‐I in never‐smoker lung cancer patients contribute to lung cancer progression and associated with EGFR gene mutation , 2012, Journal of cellular physiology.

[3]  J. Kirkland.  Niacin requirements for genomic stability. , 2012, Mutation research.

[4]  H. T. Kang,et al.  Nicotinamide-induced Mitophagy , 2012, The Journal of Biological Chemistry.

[5]  P. Bénit,et al.  Mitochondrial complex I deficiency of nuclear origin I. Structural genes. , 2012, Molecular genetics and metabolism.

[6]  P. Bénit,et al.  Mitochondrial complex I deficiency of nuclear origin II. Non-structural genes. , 2012, Molecular genetics and metabolism.

[7]  Angeliki Chalkiadaki,et al.  Sirtuins mediate mammalian metabolic responses to nutrient availability , 2012, Nature Reviews Endocrinology.

[8]  Yidong Bai,et al.  Mitochondrial respiratory complex I dysfunction promotes tumorigenesis through ROS alteration and AKT activation. , 2011, Human molecular genetics.

[9]  T. Flotte,et al.  No Immune Responses by the Expression of the Yeast Ndi1 Protein in Rats , 2011, PloS one.

[10]  Kornelia Polyak,et al.  Heterogeneity in breast cancer. , 2011, The Journal of clinical investigation.

[11]  Y. Tu,et al.  Mitochondria of highly metastatic breast cancer cell line MDA-MB-231 exhibits increased autophagic properties. , 2011, Biochimica et biophysica acta.

[12]  G. Garcia-Manero,et al.  K-rasG12V transformation leads to mitochondrial dysfunction and a metabolic switch from oxidative phosphorylation to glycolysis , 2011, Cell Research.

[13]  H. Klocker,et al.  Accumulation of mutations over the entire mitochondrial genome of breast cancer cells obtained by tissue microdissection , 2011, Breast Cancer Research and Treatment.

[14]  Stefan Fischer,et al.  Compartmentation of NAD+‐dependent signalling , 2011, FEBS letters.

[15]  G. Bhanot,et al.  Autophagy Suppresses Tumorigenesis through Elimination of p62 , 2011, Cell.

[16]  Jia Wei,et al.  Evaluating mitochondrial DNA in patients with breast cancer and benign breast disease , 2011, Journal of Cancer Research and Clinical Oncology.

[17]  H. Wu,et al.  NAMPT overexpression in prostate cancer and its contribution to tumor cell survival and stress response , 2011, Oncogene.

[18]  Chengqun Huang,et al.  Xenotransplantation of Mitochondrial Electron Transfer Enzyme, Ndi1, in Myocardial Reperfusion Injury , 2011, PloS one.

[19]  Robin Mathew,et al.  Autophagy in tumorigenesis and energy metabolism: friend by day, foe by night. , 2011, Current opinion in genetics & development.

[20]  C. Deng,et al.  Redox-dependent Brca1 transcriptional regulation by an NADH-sensor CtBP1 , 2010, Oncogene.

[21]  D. Sabatini,et al.  mTOR: from growth signal integration to cancer, diabetes and ageing , 2010, Nature Reviews Molecular Cell Biology.

[22]  H. Yonekawa,et al.  Generation of trans‐mitochondrial mito‐mice by the introduction of a pathogenic G13997A mtDNA from highly metastatic lung carcinoma cells , 2010, FEBS letters.

[23]  K. Gardner,et al.  Transcriptional regulation of BRCA1 expression by a metabolic switch , 2010, Nature Structural &Molecular Biology.

[24]  T. Yagi,et al.  Successful Amelioration of Mitochondrial Optic Neuropathy Using the Yeast NDI1 Gene in a Rat Animal Model , 2010, PloS one.

[25]  H. Jacobs,et al.  Expression of the yeast NADH dehydrogenase Ndi1 in Drosophila confers increased lifespan independently of dietary restriction , 2010, Proceedings of the National Academy of Sciences.

[26]  A. Sauve,et al.  Vitamin B3, the nicotinamide adenine dinucleotides and aging , 2010, Mechanisms of Ageing and Development.

[27]  C. Kenific,et al.  Autophagy and metastasis: another double-edged sword. , 2010, Current opinion in cell biology.

[28]  T. Uzu,et al.  Calorie restriction enhances cell adaptation to hypoxia through Sirt1-dependent mitochondrial autophagy in mouse aged kidney. , 2010, The Journal of clinical investigation.

[29]  L. Guarente,et al.  Aging and disease: connections to sirtuins , 2010, Aging cell.

[30]  P. Robbins,et al.  SIRT1 Negatively Regulates the Mammalian Target of Rapamycin , 2010, PloS one.

[31]  L. Alberghina,et al.  Mitochondrial Complex I decrease is responsible for bioenergetic dysfunction in K-ras transformed cells. , 2010, Biochimica et biophysica acta.

[32]  N. Sundaresan,et al.  Exogenous NAD Blocks Cardiac Hypertrophic Response via Activation of the SIRT3-LKB1-AMP-activated Kinase Pathway* , 2009, The Journal of Biological Chemistry.

[33]  T. Yagi,et al.  Neuroprotective effect of long-term NDI1 gene expression in a chronic mouse model of Parkinson disorder. , 2009, Rejuvenation research.

[34]  Keshav K. Singh,et al.  Cancer cell mitochondria confer apoptosis resistance and promote metastasis , 2009, Cancer biology & therapy.

[35]  A. Chambers,et al.  MDA-MB-435 and M14 cell lines: identical but not M14 melanoma? , 2009, Cancer research.

[36]  L. Cantley,et al.  Understanding the Warburg Effect: The Metabolic Requirements of Cell Proliferation , 2009, Science.

[37]  A. LeBeau,et al.  Targeting the cancer stroma with a fibroblast activation protein-activated promelittin protoxin , 2009, Molecular Cancer Therapeutics.

[38]  N. Ruderman,et al.  SIRT1 Modulation of the Acetylation Status, Cytosolic Localization, and Activity of LKB1 , 2008, Journal of Biological Chemistry.

[39]  J. Hayashi,et al.  ROS-Generating Mitochondrial DNA Mutations Can Regulate Tumor Cell Metastasis , 2008, Science.

[40]  D. Meierhofer,et al.  Loss of Complex I due to Mitochondrial DNA Mutations in Renal Oncocytoma , 2008, Clinical Cancer Research.

[41]  Nicholas E. Bruns,et al.  A role for the NAD-dependent deacetylase Sirt1 in the regulation of autophagy , 2008, Proceedings of the National Academy of Sciences.

[42]  John L Cleveland,et al.  Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes , 2008, Autophagy.

[43]  J. Yates,et al.  Adaptation of energy metabolism in breast cancer brain metastases. , 2007, Cancer research.

[44]  C. Maley,et al.  Cancer as an evolutionary and ecological process , 2006, Nature Reviews Cancer.

[45]  P. Schumacker,et al.  Reactive oxygen species in cancer cells: live by the sword, die by the sword. , 2006, Cancer cell.

[46]  Marty C. Brandon,et al.  Mitochondrial mutations in cancer , 2006, Oncogene.

[47]  Jonathan V. Rocheleau,et al.  Redox sensor CtBP mediates hypoxia-induced tumor cell migration. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[48]  D. Wallace.  A Mitochondrial Paradigm of Metabolic and Degenerative Diseases, Aging, and Cancer: A Dawn for Evolutionary Medicine , 2005, Annual review of genetics.

[49]  Max Hasmann,et al.  FK866, a highly specific noncompetitive inhibitor of nicotinamide phosphoribosyltransferase, represents a novel mechanism for induction of tumor cell apoptosis. , 2003, Cancer research.

[50]  Peng Huang,et al.  Mitochondrial defects in cancer , 2002, Molecular Cancer.

[51]  E Gabrielson,et al.  Detection of mitochondrial DNA mutations in primary breast cancer and fine-needle aspirates. , 2001, Cancer research.

[52]  Andrew V. Nguyen,et al.  Colony-Stimulating Factor 1 Promotes Progression of Mammary Tumors to Malignancy , 2001, The Journal of experimental medicine.

[53]  T. Flotte,et al.  Use of the NADH-Quinone Oxidoreductase (NDI1) Gene ofSaccharomyces cerevisiae as a Possible Cure for Complex I Defects in Human Cells* , 2000, The Journal of Biological Chemistry.

[54]  S. Emr,et al.  Autophagy as a regulated pathway of cellular degradation. , 2000, Science.

[55]  A. Matsuno-Yagi,et al.  Molecular remedy of complex I defects: rotenone-insensitive internal NADH-quinone oxidoreductase of Saccharomyces cerevisiae mitochondria restores the NADH oxidase activity of complex I-deficient mammalian cells. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[56]  R. Cardiff,et al.  Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease , 1992, Molecular and cellular biology.

[57]  L. Grivell,et al.  Primary structure and import pathway of the rotenone-insensitive NADH-ubiquinone oxidoreductase of mitochondria from Saccharomyces cerevisiae. , 1992, European journal of biochemistry.

[58]  B. Brinkley,et al.  Variations in cell form and cytoskeleton in human breast carcinoma cells in vitro. , 1980, Cancer research.

[59]  G. S. Johnson,et al.  The effect of growth conditions on NAD+ and NADH concentrations and the NAD+:NADH ratio in normal and transformed fibroblasts. , 1974, The Journal of biological chemistry.

[60]  K. Sargsyan,et al.  Respiratory chain complex I is a mitochondrial tumor suppressor of oncocytic tumors. , 2011, Frontiers in bioscience.

[61]  L. Wong,et al.  Mitochondrial dysfunction in human breast cancer cells and their transmitochondrial cybrids. , 2010, Biochimica et biophysica acta.

[62]  G. Hortobagyi,et al.  Prognosis of women with metastatic breast cancer by HER2 status and trastuzumab treatment: an institutional-based review. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[63]  D. Wallace,et al.  Assessment of mitochondrial oxidative phosphorylation in patient muscle biopsies, lymphoblasts, and transmitochondrial cell lines. , 1996, Methods in enzymology.

[64]  S. Weinhouse.  On respiratory impairment in cancer cells. , 1956, Science.