Modifications in cellular viability, DNA damage and stress responses inflicted in cancer cells by copper-64 ions

Due to combined therapeutical emissions, a high linear energy transfer Auger-electrons with the longer ranged β− particles, 64Cu-based radiopharmaceuticals raise particular theragnostic interest in cancer, by joined therapeutic and real-time PET imaging properties. The in vitro study aimed to investigate the biological and molecular background of 64CuCl2 therapy by analyzing the damages and stress responses inflicted in various human normal and tumor cell lines. Colon (HT29 and HCT116) and prostate carcinoma (DU145) cell lines, as well as human normal BJ fibroblasts, were treated up to 72 h with 2–40 MBq/mL 64CuCl2. Radioisotope uptake and retention were assessed, and cell viability/death, DNA damage, oxidative stress, and the expression of 84 stress genes were investigated at various time points after [64Cu]CuCl2 addition. All the investigated cells incorporated 64Cu ions similarly, independent of their tumoral or normal status, but their fate after exposure to [64Cu]CuCl2 was cell-dependent. The most striking cytotoxic effects of the radioisotope were registered in colon carcinoma HCT116 cells, for which a substantial decrease in the number of metabolically active cells, and an increased DNA damage and oxidative stress were registered. The stress gene expression study highlighted the activation of both death and repair mechanisms in these cells, related to extrinsic apoptosis, necrosis/necroptosis or autophagy, and cell cycle arrest, nucleotide excision repair, antioxidant, and hypoxic responses, respectively. The in vitro study indicated that 40 MBq/mL [64Cu]CuCl2 delivers a therapeutic effect in human colon carcinoma, but its use is limited by harmful, yet lower effects on normal fibroblasts. The exposure of tumor cells to 20 MBq/mL [64Cu]CuCl2, might be used for a softer approach aiming for a lower radiotoxicity in normal fibroblasts as compared to tumor cells. This radioactive concentration was able to induce a persistent decrease in the number of metabolically active cells, accompanied by DNA damage and oxidative stress, associated with significant changes in stress gene expression in HCT116 colon cancer cells.

[1]  C. Ur,et al.  Cyclotron production of 64Cu by proton irradiation of enriched 64Ni target: Validation of Geant4 simulation parameters through experimental data , 2022, Frontiers in Physics.

[2]  A. Lu,et al.  DNA binding by the Rad9A subunit of the Rad9-Rad1-Hus1 complex , 2022, PloS one.

[3]  Xuejun Jiang,et al.  Grb2 interacts with necrosome components and is involved in rasfonin-induced necroptosis , 2022, Cell death discovery.

[4]  S. D. do Carmo,et al.  Production of GMP-Compliant Clinical Amounts of Copper-61 Radiopharmaceuticals from Liquid Targets , 2022, Pharmaceuticals.

[5]  V. Oliveri.  Selective Targeting of Cancer Cells by Copper Ionophores: An Overview , 2022, Frontiers in Molecular Biosciences.

[6]  S. Kalkhof,et al.  Osmotic Stress Interferes with DNA Damage Response and H2AX Phosphorylation in Human Keratinocytes , 2022, Cells.

[7]  Junying Yuan,et al.  Genetic Regulation of RIPK1 and Necroptosis. , 2021, Annual review of genetics.

[8]  E. Moon,et al.  The importance of hypoxia in radiotherapy for the immune response, metastatic potential and FLASH-RT , 2021, International journal of radiation biology.

[9]  A. Elorza,et al.  Role of Copper on Mitochondrial Function and Metabolism , 2021, Frontiers in Molecular Biosciences.

[10]  A. Rezaeian,et al.  ATM: Main Features, Signaling Pathways, and Its Diverse Roles in DNA Damage Response, Tumor Suppression, and Cancer Development , 2021, Genes.

[11]  D. Ross,et al.  The diverse functionality of NQO1 and its roles in redox control , 2021, Redox biology.

[12]  D. Niculae,et al.  Biological Pathways as Substantiation of the Use of Copper Radioisotopes in Cancer Theranostics , 2021, Frontiers in Physics.

[13]  C. L. da Silva,et al.  Copper-64 Chloride Exhibits Therapeutic Potential in Three-Dimensional Cellular Models of Prostate Cancer , 2020, Frontiers in Molecular Biosciences.

[14]  J. Slabbert,et al.  Estimating the Relative Biological Effectiveness of Auger Electron Emitter 123I in Human Lymphocytes , 2020, Frontiers in Physics.

[15]  L. Milković,et al.  The NRF2, Thioredoxin, and Glutathione System in Tumorigenesis and Anticancer Therapies , 2020, Antioxidants.

[16]  G. Manda,et al.  Preclinical Evaluation of NHS-Activated Gold Nanoparticles Functionalized with Bombesin or Neurotensin-Like Peptides for Targeting Colon and Prostate Tumours , 2020, Molecules.

[17]  S. Mondal,et al.  Matrix metalloproteinase-9 (MMP-9) and its inhibitors in cancer: A minireview. , 2020, European journal of medicinal chemistry.

[18]  M. Xie,et al.  Role of hypoxia in cancer therapy by regulating the tumor microenvironment , 2019, Molecular Cancer.

[19]  U. Haberkorn,et al.  Development of Novel PSMA Ligands for Imaging and Therapy with Copper Isotopes , 2019, The Journal of Nuclear Medicine.

[20]  K. Seong,et al.  Cellular Stress Responses in Radiotherapy , 2019, Cells.

[21]  Lijiang Hu,et al.  Unraveling the Potential Role of Glutathione in Multiple Forms of Cell Death in Cancer Therapy , 2019, Oxidative medicine and cellular longevity.

[22]  Gul-e-Saba Chaudhry,et al.  Understanding Apoptosis and Apoptotic Pathways Targeted Cancer Therapeutics , 2019, Advanced pharmaceutical bulletin.

[23]  G. E. Atilla‐Gokcumen,et al.  Membrane disruption by very long chain fatty acids during necroptosis. , 2019, ACS chemical biology.

[24]  T. Riss,et al.  Cytotoxicity Assays: In Vitro Methods to Measure Dead Cells , 2019 .

[25]  A. Dzutsev,et al.  Cell‐Type‐Specific Responses to Interleukin‐1 Control Microbial Invasion and Tumor‐Elicited Inflammation in Colorectal Cancer , 2019, Immunity.

[26]  M. de Ridder,et al.  Hypoxic Radioresistance: Can ROS Be the Key to Overcome It? , 2019, Cancers.

[27]  L. Galluzzi,et al.  Linking cellular stress responses to systemic homeostasis , 2018, Nature Reviews Molecular Cell Biology.

[28]  Y. Inagaki,et al.  Glucose starvation induces LKB1-AMPK-mediated MMP-9 expression in cancer cells , 2018, Scientific Reports.

[29]  P. Cobine,et al.  The mitochondrion: a central architect of copper homeostasis. , 2017, Metallomics : integrated biometal science.

[30]  E. Azzam,et al.  Genomic instability induced in distant progeny of bystander cells depends on the connexins expressed in the irradiated cells , 2017, International journal of radiation biology.

[31]  Hao Chen,et al.  Pilot Study of 64Cu(I) for PET Imaging of Melanoma , 2017, Scientific Reports.

[32]  Chuanqin Xia,et al.  Establishing Reliable Cu-64 Production Process: From Target Plating to Molecular Specific Tumor Micro-PET Imaging , 2017, Molecules.

[33]  I. Navarro-Teulon,et al.  Localized Irradiation of Cell Membrane by Auger Electrons Is Cytotoxic Through Oxidative Stress-Mediated Nontargeted Effects , 2016, Antioxidants & redox signaling.

[34]  Ming-Rong Zhang,et al.  (64)Cu-ATSM therapy targets regions with activated DNA repair and enrichment of CD133(+) cells in an HT-29 tumor model: Sensitization with a nucleic acid antimetabolite. , 2016, Cancer letters.

[35]  Mary G. Lin,et al.  Structure and function of the ULK1 complex in autophagy. , 2016, Current opinion in cell biology.

[36]  Sergey V. Gudkov,et al.  Targeted Radionuclide Therapy of Human Tumors , 2015, International journal of molecular sciences.

[37]  Tilman Grune,et al.  Clinical Relevance of Biomarkers of Oxidative Stress , 2015, Antioxidants & redox signaling.

[38]  Y. Fujibayashi,et al.  Validation of 64Cu-ATSM damaging DNA via high-LET Auger electron emission , 2015, Journal of radiation research.

[39]  A. Shelat,et al.  Nuclear ULK1 promotes cell death in response to oxidative stress through PARP1 , 2015, Cell Death and Differentiation.

[40]  C. E. Erdahl,et al.  Practical experience implementing the Comecer ALCEO Metal solid targetry system , 2015 .

[41]  M. Sauer,et al.  Hypotonic Activation of the Myo-Inositol Transporter SLC5A3 in HEK293 Cells Probed by Cell Volumetry, Confocal and Super-Resolution Microscopy , 2015, PloS one.

[42]  L. Stivala,et al.  Biology of the cell cycle inhibitor p21CDKN1A: molecular mechanisms and relevance in chemical toxicology , 2015, Archives of Toxicology.

[43]  J. Kocot,et al.  The many “faces” of copper in medicine and treatment , 2014, BioMetals.

[44]  D. Ehst,et al.  The production, separation, and use of 67Cu for radioimmunotherapy: a review. , 2012, Applied radiation and isotopes : including data, instrumentation and methods for use in agriculture, industry and medicine.

[45]  Michelle R. Campbell,et al.  Identification of novel NRF2-regulated genes by ChIP-Seq: influence on retinoid X receptor alpha , 2012, Nucleic acids research.

[46]  Baohong Zhang,et al.  miRDeepFinder: a miRNA analysis tool for deep sequencing of plant small RNAs , 2012, Plant Molecular Biology.

[47]  Baohong Zhang,et al.  miRDeepFinder: a miRNA analysis tool for deep sequencing of plant small RNAs , 2012, Plant Molecular Biology.

[48]  A. Kassis.  Molecular and cellular radiobiological effects of Auger emitting radionuclides. , 2011, Radiation protection dosimetry.

[49]  P. Raychaudhuri,et al.  DDB2 (Damaged-DNA binding protein 2) in nucleotide excision repair and DNA damage response , 2009, Cell cycle.

[50]  Manuel Bardiès,et al.  Cell Membrane is a More Sensitive Target than Cytoplasm to Dense Ionization Produced by Auger Electrons , 2008, Radiation research.

[51]  John F. Reichard,et al.  Heme oxygenase-1 induction by NRF2 requires inactivation of the transcriptional repressor BACH1 , 2007, Nucleic acids research.

[52]  J. Cook,et al.  How many transcription factors does it take to turn on the heme oxygenase-1 gene? , 2007, American journal of respiratory cell and molecular biology.

[53]  M. Sabet,et al.  An overview of copper radionuclides and production of 61Cu by proton irradiation of (nat)Zn at a medical cyclotron. , 2006, Applied radiation and isotopes : including data, instrumentation and methods for use in agriculture, industry and medicine.

[54]  Peter Vandenabeele,et al.  Necrosis, a well-orchestrated form of cell demise: signalling cascades, important mediators and concomitant immune response. , 2006, Biochimica et biophysica acta.

[55]  Karl Andersson,et al.  Automated, high-resolution cellular retention and uptake studies in vitro. , 2006, Applied radiation and isotopes : including data, instrumentation and methods for use in agriculture, industry and medicine.

[56]  Shelly C. Lu,et al.  Nrf1 and Nrf2 Regulate Rat Glutamate-Cysteine Ligase Catalytic Subunit Transcription Indirectly via NF-κB and AP-1 , 2005, Molecular and Cellular Biology.

[57]  D. Norris,et al.  BMC Biotechnology BioMed Central Methodology article A simple technique for quantifying apoptosis in 96-well plates , 2005 .

[58]  Henry Jay Forman,et al.  Human glutamate cysteine ligase gene regulation through the electrophile response element. , 2004, Free radical biology & medicine.

[59]  E. Hall,et al.  The bystander effect. , 2003, Health physics.

[60]  K. O'Leary,et al.  Preservation of comet assay slides: comparison with fresh slides. , 1999, Mutation research.

[61]  M. D. Berry,et al.  Culture of Adherent Cancer Cell Lines. , 2022, Methods in molecular biology.

[62]  A. Chiriacescu,et al.  Process validation for production of copper radioisotopes in a TR-19 variable energy cyclotron , 2020 .

[63]  V. Narayanankutty,et al.  Glutathione, an antioxidant tripeptide: Dual roles in Carcinogenesis and Chemoprevention. , 2019, Current protein & peptide science.

[64]  R. Uauy,et al.  Biological Aspects of Copper , 2019, Clinical and Translational Perspectives on WILSON DISEASE.

[65]  M. Maes,et al.  Measurement of Apoptotic and Necrotic Cell Death in Primary Hepatocyte Cultures. , 2015, Methods in molecular biology.

[66]  F. Chan,et al.  Detection of necrosis by release of lactate dehydrogenase activity. , 2013, Methods in molecular biology.

[67]  R. Linden,et al.  DNA damage-induced cell death: lessons from the central nervous system , 2008, Cell Research.

[68]  P. Mahadevan,et al.  An overview , 2007, Journal of Biosciences.

[69]  C. Anderson,et al.  Production and applications of copper-64 radiopharmaceuticals. , 2004, Methods in enzymology.

[70]  Robert C. Wolpert,et al.  A Review of the , 1985 .