Mechanism-Specific Pharmacodynamics of a Novel Complex-I Inhibitor Quantified by Imaging Reversal of Consumptive Hypoxia with [18F]FAZA PET In Vivo

Tumors lack a well-regulated vascular supply of O2 and often fail to balance O2 supply and demand. Net O2 tension within many tumors may not only depend on O2 delivery but also depend strongly on O2 demand. Thus, tumor O2 consumption rates may influence tumor hypoxia up to true anoxia. Recent reports have shown that many human tumors in vivo depend primarily on oxidative phosphorylation (OxPhos), not glycolysis, for energy generation, providing a driver for consumptive hypoxia and an exploitable vulnerability. In this regard, IACS-010759 is a novel high affinity inhibitor of OxPhos targeting mitochondrial complex-I that has recently completed a Phase-I clinical trial in leukemia. However, in solid tumors, the effective translation of OxPhos inhibitors requires methods to monitor pharmacodynamics in vivo. Herein, 18F-fluoroazomycin arabinoside ([18F]FAZA), a 2-nitroimidazole-based hypoxia PET imaging agent, was combined with a rigorous test-retest imaging method for non-invasive quantification of the reversal of consumptive hypoxia in vivo as a mechanism-specific pharmacodynamic (PD) biomarker of target engagement for IACS-010759. Neither cell death nor loss of perfusion could account for the IACS-010759-induced decrease in [18F]FAZA retention. Notably, in an OxPhos-reliant melanoma tumor, a titration curve using [18F]FAZA PET retention in vivo yielded an IC50 for IACS-010759 (1.4 mg/kg) equivalent to analysis ex vivo. Pilot [18F]FAZA PET scans of a patient with grade IV glioblastoma yielded highly reproducible, high-contrast images of hypoxia in vivo as validated by CA-IX and GLUT-1 IHC ex vivo. Thus, [18F]FAZA PET imaging provided direct evidence for the presence of consumptive hypoxia in vivo, the capacity for targeted reversal of consumptive hypoxia through the inhibition of OxPhos, and a highly-coupled mechanism-specific PD biomarker ready for translation.

[1]  M. Protopopova,et al.  An inhibitor of oxidative phosphorylation exploits cancer vulnerability , 2018, Nature Medicine.

[2]  V. Bettinardi,et al.  18F-FAZA PET/CT in the Preoperative Evaluation of NSCLC: Comparison with 18F-FDG and Immunohistochemistry. , 2018, Current radiopharmaceuticals.

[3]  Jamey D. Young,et al.  Lactate Metabolism in Human Lung Tumors , 2017, Cell.

[4]  S. Indraccolo,et al.  Linking metabolic reprogramming to therapy resistance in cancer. , 2017, Biochimica et biophysica acta. Reviews on cancer.

[5]  S. Kreis,et al.  ROS production induced by BRAF inhibitor treatment rewires metabolic processes affecting cell growth of melanoma cells , 2017, Molecular Cancer.

[6]  S. Valable,et al.  In Vivo Relationship Between Hypoxia and Angiogenesis in Human Glioblastoma: A Multimodal Imaging Study , 2017, The Journal of Nuclear Medicine.

[7]  Wei Zhang,et al.  Repositioning chlorpromazine for treating chemoresistant glioma through the inhibition of cytochrome c oxidase bearing the COX4-1 regulatory subunit , 2017, Oncotarget.

[8]  M. Bernaudin,et al.  [18F]-FMISO PET study of hypoxia in gliomas before surgery: correlation with molecular markers of hypoxia and angiogenesis , 2017, European Journal of Nuclear Medicine and Molecular Imaging.

[9]  J. Locasale,et al.  Glutamine Metabolism in Cancer: Understanding the Heterogeneity. , 2017, Trends in cancer.

[10]  Anne Bol,et al.  Evolution of [18F]fluorodeoxyglucose and [18F]fluoroazomycin arabinoside PET uptake distributions in lung tumours during radiation therapy , 2017, Acta oncologica.

[11]  G. Semenza Hypoxia‐inducible factors: coupling glucose metabolism and redox regulation with induction of the breast cancer stem cell phenotype , 2017, The EMBO journal.

[12]  Q. Fu,et al.  Preclinical pharmacokinetics and toxic kinetics study of 2, 4-dinitrophenol (DNP) , 2016 .

[13]  K. Morten,et al.  The Warburg effect: 80 years on , 2016, Biochemical Society transactions.

[14]  Wei Zhang,et al.  Identification of Small Molecule Inhibitors of Human Cytochrome c Oxidase That Target Chemoresistant Glioma Cells* , 2016, The Journal of Biological Chemistry.

[15]  A. V. van Kuilenburg,et al.  Genotypes Affecting the Pharmacokinetics of Anticancer Drugs , 2016, Clinical Pharmacokinetics.

[16]  R. DePinho,et al.  SF2312 is a natural phosphonate inhibitor of Enolase , 2016, Nature chemical biology.

[17]  C. Heeschen,et al.  Hallmarks of cancer stem cell metabolism , 2016, British Journal of Cancer.

[18]  Navdeep S. Chandel,et al.  Fundamentals of cancer metabolism , 2016, Science Advances.

[19]  D. Jaffray,et al.  Measurement of Tumor Hypoxia in Patients with Advanced Pancreatic Cancer Based on 18F-Fluoroazomyin Arabinoside Uptake , 2016, The Journal of Nuclear Medicine.

[20]  B. Faubert,et al.  Metabolic Heterogeneity in Human Lung Tumors , 2016, Cell.

[21]  Andrea Glasauer,et al.  Targeting mitochondrial complex I using BAY 87-2243 reduces melanoma tumor growth , 2015, Cancer & metabolism.

[22]  C. Heeschen,et al.  MYC/PGC-1α Balance Determines the Metabolic Phenotype and Plasticity of Pancreatic Cancer Stem Cells. , 2015, Cell metabolism.

[23]  R. Steenbakkers,et al.  Assessment of hypoxic subvolumes in laryngeal cancer with (18)F-fluoroazomycinarabinoside ((18)F-FAZA)-PET/CT scanning and immunohistochemistry. , 2015, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[24]  D. Sabatini,et al.  An Essential Role of the Mitochondrial Electron Transport Chain in Cell Proliferation Is to Enable Aspartate Synthesis , 2015, Cell.

[25]  A. Maity,et al.  Molecular Pathways: A Novel Approach to Targeting Hypoxia and Improving Radiotherapy Efficacy via Reduction in Oxygen Demand , 2015, Clinical Cancer Research.

[26]  J. Dick,et al.  AML cells have low spare reserve capacity in their respiratory chain that renders them susceptible to oxidative metabolic stress. , 2015, Blood.

[27]  P. Lambin,et al.  Current preclinical and clinical applications of hypoxia PET imaging using 2-nitroimidazoles. , 2015, The quarterly journal of nuclear medicine and molecular imaging : official publication of the Italian Association of Nuclear Medicine (AIMN) [and] the International Association of Radiopharmacology (IAR), [and] Section of the Society of....

[28]  A. Lane,et al.  Pyruvate carboxylase is critical for non-small-cell lung cancer proliferation. , 2015, The Journal of clinical investigation.

[29]  Kakajan Komurov,et al.  Inhibition of mTORC1/2 overcomes resistance to MAPK pathway inhibitors mediated by PGC1α and oxidative phosphorylation in melanoma. , 2014, Cancer research.

[30]  S. Gambhir,et al.  18F-FAZA PET Imaging Response Tracks the Reoxygenation of Tumors in Mice upon Treatment with the Mitochondrial Complex I Inhibitor BAY 87-2243 , 2014, Clinical Cancer Research.

[31]  Robert J. Gillies,et al.  Pyruvate Induces Transient Tumor Hypoxia by Enhancing Mitochondrial Oxygen Consumption and Potentiates the Anti-Tumor Effect of a Hypoxia-Activated Prodrug TH-302 , 2014, PloS one.

[32]  M. Wuest,et al.  Detecting functional changes with [18F]FAZA in a renal cell carcinoma mouse model following sunitinib therapy , 2014, EJNMMI Research.

[33]  Alexander G. Fletcher,et al.  Oxygen consumption dynamics in steady-state tumour models , 2014, Royal Society Open Science.

[34]  John M. Asara,et al.  Oncogene ablation-resistant pancreatic cancer cells depend on mitochondrial function , 2014, Nature.

[35]  I. Hanamura,et al.  Importance of Glutamine Metabolism in Leukemia Cells by Energy Production Through TCA Cycle and by Redox Homeostasis , 2014, Cancer investigation.

[36]  V. Grégoire,et al.  A prospective clinical study of 18 F-FAZA PET-CT hypoxia imaging in head and neck squamous cell carcinoma before and during radiation therapy , 2014, European Journal of Nuclear Medicine and Molecular Imaging.

[37]  Timothy W. Secomb,et al.  Quantitative Mapping of Hemodynamics in the Lung, Brain, and Dorsal Window Chamber‐Grown Tumors Using a Novel, Automated Algorithm , 2013, Microcirculation.

[38]  M. Dewhirst,et al.  Catabolism of Exogenous Lactate Reveals It as a Legitimate Metabolic Substrate in Breast Cancer , 2013, PloS one.

[39]  Michael R. Green,et al.  Metabolic signatures uncover distinct targets in molecular subsets of diffuse large B cell lymphoma. , 2012, Cancer cell.

[40]  Marit Holden,et al.  Hypoxia-induced gene expression in chemoradioresistant cervical cancer revealed by dynamic contrast-enhanced MRI. , 2012, Cancer research.

[41]  Anne Bol,et al.  Hypoxia imaging with the nitroimidazole 18F-FAZA PET tracer: a comparison with OxyLite, EPR oximetry and 19F-MRI relaxometry. , 2012, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[42]  John L. Humm,et al.  Image-Guided Po2 Probe Measurements Correlated with Parametric Images Derived from 18F-Fluoromisonidazole Small-Animal PET Data in Rats , 2012, The Journal of Nuclear Medicine.

[43]  Steen Jakobsen,et al.  FAZA PET/CT hypoxia imaging in patients with squamous cell carcinoma of the head and neck treated with radiotherapy: results from the DAHANCA 24 trial. , 2012, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[44]  L. Chin,et al.  Passenger Deletions Generate Therapeutic Vulnerabilities in Cancer , 2012, Nature.

[45]  K. Houkin,et al.  18F-Fluoromisonidazole positron emission tomography may differentiate glioblastoma multiforme from less malignant gliomas , 2012, European Journal of Nuclear Medicine and Molecular Imaging.

[46]  Jan J Wilkens,et al.  Theoretical analysis of the dose dependence of the oxygen enhancement ratio and its relevance for clinical applications , 2011, Radiation oncology.

[47]  Xiao-Hong Zhu,et al.  In vivo oxygen-17 NMR for imaging brain oxygen metabolism at high field. , 2011, Progress in nuclear magnetic resonance spectroscopy.

[48]  C. Dang,et al.  Otto Warburg's contributions to current concepts of cancer metabolism , 2011, Nature Reviews Cancer.

[49]  Bradford D Henderson,et al.  Highlighting the Versatility of the Tracerlab Synthesis Modules. Part 1: Fully Automated Production of [F]Labelled Radiopharmaceuticals using a Tracerlab FX(FN). , 2011, Journal of labelled compounds & radiopharmaceuticals.

[50]  Chi V. Dang,et al.  Otto Warburg's contributions to current concepts of cancer metabolism , 2011, Nature Reviews Cancer.

[51]  Richard Pötter,et al.  Evaluating repetitive 18F-fluoroazomycin-arabinoside (18FAZA) PET in the setting of MRI guided adaptive radiotherapy in cervical cancer , 2010, Acta oncologica.

[52]  M. Roth,et al.  Inhibition of Iron Uptake Is Responsible for Differential Sensitivity to V-ATPase Inhibitors in Several Cancer Cell Lines , 2010, PloS one.

[53]  Jun Xu,et al.  Oligomycin-induced Bioenergetic Adaptation in Cancer Cells with Heterogeneous Bioenergetic Organization , 2010, The Journal of Biological Chemistry.

[54]  E. Graves,et al.  Pharmacologically Increased Tumor Hypoxia Can Be Measured by 18F-Fluoroazomycin Arabinoside Positron Emission Tomography and Enhances Tumor Response to Hypoxic Cytotoxin PR-104 , 2009, Clinical Cancer Research.

[55]  Terence A. Riauka,et al.  Initial results of hypoxia imaging using 1-α-d-(5-deoxy-5-[18F]-fluoroarabinofuranosyl)-2-nitroimidazole (18F-FAZA) , 2009, European Journal of Nuclear Medicine and Molecular Imaging.

[56]  Jayne E. Telford,et al.  Complex I Is Rate-limiting for Oxygen Consumption in the Nerve Terminal* , 2009, Journal of Biological Chemistry.

[57]  Julien Verrax,et al.  Targeting lactate-fueled respiration selectively kills hypoxic tumor cells in mice. , 2008, The Journal of clinical investigation.

[58]  Kenneth A Krohn,et al.  Molecular Imaging of Hypoxia , 2008, Journal of Nuclear Medicine.

[59]  P. Workman,et al.  Use of pharmacokinetic/pharmacodynamic biomarkers to support rational cancer drug development. , 2007, Biomarkers in medicine.

[60]  D. Hedley,et al.  Effect of distributional heterogeneity on the analysis of tumor hypoxia based on carbonic anhydrase IX , 2007, Laboratory Investigation.

[61]  N. Denko,et al.  HIF-1 mediates adaptation to hypoxia by actively downregulating mitochondrial oxygen consumption. , 2006, Cell metabolism.

[62]  C. Solbach,et al.  Preparation of the hypoxia imaging PET tracer [18F]FAZA: reaction parameters and automation. , 2005, Applied radiation and isotopes : including data, instrumentation and methods for use in agriculture, industry and medicine.

[63]  N. Denko,et al.  Cellular reaction to hypoxia: sensing and responding to an adverse environment. , 2005, Mutation research.

[64]  M. Holness,et al.  Regulation of pyruvate dehydrogenase complex activity by reversible phosphorylation. , 2003, Biochemical Society transactions.

[65]  H. Leong,et al.  Glycolysis and pyruvate oxidation in cardiac hypertrophy--why so unbalanced? , 2003, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[66]  P. Price,et al.  PET for in vivo pharmacokinetic and pharmacodynamic measurements. , 2002, European journal of cancer.

[67]  G. Semenza,et al.  HIF-1, O2, and the 3 PHDs How Animal Cells Signal Hypoxia to the Nucleus , 2001, Cell.

[68]  Z L Gokaslan,et al.  A multivariate analysis of 416 patients with glioblastoma multiforme: prognosis, extent of resection, and survival. , 2001, Journal of neurosurgery.

[69]  P. Vaupel,et al.  Tumor hypoxia: definitions and current clinical, biologic, and molecular aspects. , 2001, Journal of the National Cancer Institute.

[70]  D. Voehringer,et al.  Understanding and exploiting the mechanistic basis for selectivity of polyketide inhibitors of F(0)F(1)-ATPase. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[71]  P. Vaupel,et al.  Blood supply, oxygenation status and metabolic micromilieu of breast cancers: characterization and therapeutic relevance. , 2000, International journal of oncology.

[72]  J P Logue,et al.  Tumour oxygenation levels correlate with dynamic contrast-enhanced magnetic resonance imaging parameters in carcinoma of the cervix. , 2000, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[73]  A Brahme,et al.  Development of Radiation Therapy Optimization , 2000, Acta oncologica.

[74]  M. Dewhirst,et al.  Microvascular studies on the origins of perfusion-limited hypoxia. , 1996, The British journal of cancer. Supplement.

[75]  G L Rosner,et al.  Arteriolar oxygenation in tumour and subcutaneous arterioles: effects of inspired air oxygen content. , 1996, The British journal of cancer. Supplement.

[76]  M. Dewhirst,et al.  Tumor oxygenation: a matter of supply and demand. , 1996, Anticancer research.

[77]  O. J. Dunn Multiple Comparisons among Means , 1961 .

[78]  W. Scholz The Contribution of Patho‐Anatomical Research to the Problem of Epilepsy , 1959 .

[79]  D. Binns,et al.  Imaging of tumor hypoxia with [124I]IAZA in comparison with [18F]FMISO and [18F]FAZA--first small animal PET results. , 2007, Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques.

[80]  Morand Piert,et al.  Hypoxia-specific tumor imaging with 18F-fluoroazomycin arabinoside. , 2005, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[81]  C. Koch,et al.  Measurement of absolute oxygen levels in cells and tissues using oxygen sensors and 2-nitroimidazole EF5. , 2002, Methods in enzymology.

[82]  J F Gross,et al.  Analysis of the effects of oxygen supply and demand on hypoxic fraction in tumors. , 1995, Acta oncologica.

[83]  John F. Nunn,et al.  Respiratory Physiology—the essentials , 1975 .

[84]  H. Krebs The Pasteur effect and the relations between respiration and fermentation. , 1972, Essays in biochemistry.