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A Novel Imaging Biomarker Extracted from Fluorescence Microscopic Imaging of TRA-8/DR5 Oligomers Predicts TRA-8 Therapeutic Efficacy in Breast and Pancreatic Cancer Mouse Models

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Abstract

Purpose

The aim of the study was to develop a reliable quantitative imaging biomarker from fluorescence microscopic imaging of TRA-8/death receptor 5 (DR5) oligomer to predict TRA-8 therapeutic efficacy in human breast and pancreatic cancer mouse models.

Procedures

Two breast (2LMP, SUM159) and two pancreatic (MIA PaCa-2, PANC1) cancer cell lines were used. 105 cells per cell line were placed in a culture dish and treated with Cy5.5-labeled TRA-8 overnight in vitro. Three fluorescence microphotographs (×20) were acquired from randomly selected areas, and about 300 cells were analyzed per cell line. Two-dimensional (2D) fluorescence signal distribution of Cy5.5-TRA-8 on each cell was measured. Gaussian curve fitting to the distribution was determined by the least square regression method, and the coefficient of determination (R 2) of the fitting was found. In addition, two features of the best fitting Gaussian curve such as peak amplitude and the volume under the curve (VUC) were retrieved. A novel image biomarker was extracted by correlating the combination of R 2 value, peak amplitude, and the VUC with the logarithmic values of the half maximal inhibitory concentrations (IC50) of TRA-8 for the four cell lines or the percentage of tumor growth inhibition (%TGI) at a week of TRA-8 treatment in animal models.

Results

Cy5.5-TRA-8 binding to DR5 receptors resulted in an oligomer on each cell membrane, and its fluorescence signal distribution followed Gaussian curve. Peak amplitude of fluorescence signal in the oligomeric region, R 2 value of the Gaussian fitting, and the VUC in TRA-8-sensitive cells were significantly higher than those in resistant cells (p < 0.05). The novel imaging biomarker was significantly correlated with either log10(IC50) or %TGI (p < 0.001).

Conclusion

The imaging biomarker extracted from the cellular distribution pattern of Cy5.5-TRA-8 may serve as a predictive biomarker of TRA-8 therapy for cancer patients.

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References

  1. Ichikawa K, Liu W, Zhao L et al (2001) Tumoricidal activity of a novel anti-human DR5 monoclonal antibody without hepatocyte cytotoxicity. Nat Med 7:954–960

    Article  CAS  PubMed  Google Scholar 

  2. Forero-Torres A, Shah J, Wood T et al (2010) Phase I trial of weekly tigatuzumab, an agonistic humanized monoclonal antibody targeting death receptor 5 (DR5). Cancer Biother Radiopharm 25:13–19

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Buchsbaum DJ, Zhou T, Grizzle WE et al (2003) Antitumor efficacy of TRA-8 anti-DR5 monoclonal antibody alone or in combination with chemotherapy and/or radiation therapy in a human breast cancer model. Clin Cancer Res 9:3731–3741

    CAS  PubMed  Google Scholar 

  4. Oliver PG, LoBuglio AF, Zhou T et al (2012) Effect of anti-DR5 and chemotherapy on basal-like breast cancer. Breast Cancer Res Treat 133:417–426

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Estes JM, Oliver PG, Straughn JM Jr et al (2007) Efficacy of anti-death receptor 5 (DR5) antibody (TRA-8) against primary human ovarian carcinoma using a novel ex vivo tissue slice model. Gynecol Oncol 105:291–298

    Article  CAS  PubMed  Google Scholar 

  6. Frederick PJ, Kendrick JE, Straughn JM Jr et al (2009) Effect of TRA-8 anti-death receptor 5 antibody in combination with chemotherapy in an ex vivo human ovarian cancer model. Int J Gynecol Cancer 19:814–819

    Article  PubMed  Google Scholar 

  7. Bevis KS, McNally LR, Sellers JC et al (2011) Anti-tumor activity of an anti-DR5 monoclonal antibody, TRA-8, in combination with taxane/platinum-based chemotherapy in an ovarian cancer model. Gynecol Oncol 121:193–199

    Article  CAS  PubMed  Google Scholar 

  8. Oliver PG, LoBuglio AF, Zinn KR et al (2008) Treatment of human colon cancer xenografts with TRA-8 anti-death receptor 5 antibody alone or in combination with CPT-11. Clin Cancer Res 14:2180–2189

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Fiveash JB, Gillespie GY, Oliver PG et al (2008) Enhancement of glioma radiotherapy and chemotherapy response with targeted antibody therapy against death receptor 5. Int J Radiat Oncol Biol Phys 71:507–516

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Kim H, Morgan DE, Buchsbaum DJ et al (2008) Early therapy evaluation of combined anti-death receptor 5 antibody and gemcitabine in orthotopic pancreatic tumor xenografts by diffusion-weighted magnetic resonance imaging. Cancer Res 68:8369–8376

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Kim H, Zhai G, Samuel SL et al (2012) Dual combination therapy targeting DR5 and EMMPRIN in pancreatic adenocarcinoma. Mol Cancer Ther 11:405–415

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Derosier LC, Buchsbaum DJ, Oliver PG et al (2007) Combination treatment with TRA-8 anti death receptor 5 antibody and CPT-11 induces tumor regression in an orthotopic model of pancreatic cancer. Clin Cancer Res 13:5535s–5543s

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Derosier LC, Vickers SM, Zinn KR et al (2007) TRA-8 anti-DR5 monoclonal antibody and gemcitabine induce apoptosis and inhibit radiologically validated orthotopic pancreatic tumor growth. Mol Cancer Ther 6:3198–3207

    Article  CAS  PubMed  Google Scholar 

  14. Rahman M, Davis SR, Pumphrey JG et al (2009) TRAIL induces apoptosis in triple-negative breast cancer cells with a mesenchymal phenotype. Breast Cancer Res Treat 113:217–230

    Article  PubMed  PubMed Central  Google Scholar 

  15. Forero-Torres A, Varley KE, Abramson VG et al (2015) TBCRC 019: a phase II trial of nanoparticle albumin-bound paclitaxel with or without the anti-death receptor 5 monoclonal antibody tigatuzumab in patients with triple-negative breast cancer. Clin Cancer Res 21:2722–2729

    Article  CAS  PubMed  Google Scholar 

  16. Forero-Torres A, Infante JR, Waterhouse D et al (2013) Phase 2, multicenter, open-label study of tigatuzumab (CS-1008), a humanized monoclonal antibody targeting death receptor 5, in combination with gemcitabine in chemotherapy-naive patients with unresectable or metastatic pancreatic cancer. Cancer Med 2:925–932

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Gajate C, Mollinedo F (2001) The antitumor ether lipid ET-18-OCH(3) induces apoptosis through translocation and capping of Fas/CD95 into membrane rafts in human leukemic cells. Blood 98:3860–3863

    Article  CAS  PubMed  Google Scholar 

  18. Cremesti A, Paris F, Grassme H et al (2001) Ceramide enables fas to cap and kill. J Biol Chem 276:23954–23961

    Article  CAS  PubMed  Google Scholar 

  19. Delmas D, Rebe C, Lacour S et al (2003) Resveratrol-induced apoptosis is associated with Fas redistribution in the rafts and the formation of a death-inducing signaling complex in colon cancer cells. J Biol Chem 278:41482–41490

    Article  CAS  PubMed  Google Scholar 

  20. Scott FL, Stec B, Pop C et al (2009) The Fas-FADD death domain complex structure unravels signalling by receptor clustering. Nature 457:1019–1022

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Poh TW, Huang S, Hirpara JL, Pervaiz S (2007) LY303511 amplifies TRAIL-induced apoptosis in tumor cells by enhancing DR5 oligomerization, DISC assembly, and mitochondrial permeabilization. Cell Death Differ 14:1813–1825

    Article  CAS  PubMed  Google Scholar 

  22. Umiker WO, Gadebusch HH (1960) Fluorescence microscopy: a review. Med Bull 26:199–211

    CAS  Google Scholar 

  23. Kim H, Samuel SL, Zhai G et al (2014) Combination therapy with anti-DR5 antibody and tamoxifen for triple negative breast cancer. Cancer Biol Ther 15:1053–1060

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Dougherty G (2009) Digital image processing for medical applications. Cambridge University Press, New York

    Google Scholar 

  25. Bates DM, Watts DG (1988) Nonlinear regression analysis and its applications. Wiley, New York

    Book  Google Scholar 

  26. Kim H, Samuel S, Totenhagen JW et al (2015) Dynamic contrast enhanced magnetic resonance imaging of an orthotopic pancreatic cancer mouse model. J Vis Exp 18:98

    Google Scholar 

  27. Neter J, Kutner MH, Nachtsheim JC, Wasserman W (1996) Applied linear statistical models. The McGraw-Hill Companies, Inc., Columbus

    Google Scholar 

  28. Rodgers JL, Nicewander WA (1988) Thirteen ways to look at the correlation coefficient. Am Stat 42:59–66

    Article  Google Scholar 

  29. Kim H, Chaudhuri TR, Buchsbaum DJ et al (2007) High-resolution single-photon emission computed tomography and X-ray computed tomography imaging of Tc-99m-labeled anti-DR5 antibody in breast tumor xenografts. Mol Cancer Ther 6:866–875

    Article  CAS  PubMed  Google Scholar 

  30. Del Monte U (2009) Does the cell number 10(9) still really fit one gram of tumor tissue? Cell Cycle 8:505–506

    Article  PubMed  Google Scholar 

  31. Sugarbaker EV, Thornthwaite JT, Temple WT, Ketcham AS (1979) Flow cytometry: general principles and applications to selected studies in tumor biology. Int Adv Surg Oncol 2:125–153

    CAS  PubMed  Google Scholar 

  32. Kim H, Chaudhuri TR, Buchsbaum DJ et al (2007) Single-photon emission computed tomography imaging with a humanized, apoptosis-inducing antibody targeting human death receptor 5 in pancreas and breast tumor xenografts. Cancer Biol Ther 6:1396–1402

    CAS  Google Scholar 

  33. Versteeg HH, Spek CA, Peppelenbosch MP, Richel DJ (2004) Tissue factor and cancer metastasis: the role of intracellular and extracellular signaling pathways. Mol Med 10:6–11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Chen H, Zhu G, Li Y et al (2009) Extracellular signal-regulated kinase signaling pathway regulates breast cancer cell migration by maintaining slug expression. Cancer Res 69:9228–9235

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Kohno M, Tanimura S, Ozaki K (2011) Targeting the extracellular signal-regulated kinase pathway in cancer therapy. Biol Pharm Bull 34:1781–1784

    Article  CAS  PubMed  Google Scholar 

  36. Williams SA, Anderson WC, Santaguida MT, Dylla SJ (2013) Patient-derived xenografts, the cancer stem cell paradigm, and cancer pathobiology in the 21st century. Lab Invest 93:970–982

    Article  PubMed  Google Scholar 

  37. Fiebig HH, Maier A, Burger AM (2004) Clonogenic assay with established human tumour xenografts: correlation of in vitro to in vivo activity as a basis for anticancer drug discovery. Eur J Cancer 40:802–820

    Article  CAS  PubMed  Google Scholar 

  38. Kim H, Samuel SMW et al. (2015) Dynamic contrast enhanced magnetic resonance imaging evaluates therapeutic mechanism of nab-paclitaxel in pancreatic cancer patient derived xenograft mouse models. Proceeding of the International Society of Magnetic Resonance in Medicine:Abstract nr 3874

  39. Mayeux R (2004) Biomarkers: potential uses and limitations. NeuroRx 1:182–188

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

The authors thank Dr. Tong Zhou for providing TRA-8 and Dr. Janet F. Eary for editing this manuscript. The authors also thank Ms. Sharon Samuel and Ms. Marie Taylor for cell culturing, animal modeling, CT imaging, and fluorescence imaging and Dr. Guihua Zhai for small animal MRI. This study was financially supported by Komen for the Cure Promise Grant KG090969, NIH grants 2P30CA013148 and P50CA101955, and a Research Initiative Pilot Award from the Department of Radiology at UAB.

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Authors and Affiliations

  1. Department of Radiology, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Harrison Kim & Kurt R. Zinn

  2. Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Harrison Kim

  3. Department of Radiation Oncology, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Donald J. Buchsbaum

  4. Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, AL, 35294, USA

    Harrison Kim & Kurt R. Zinn

  5. G082C5 Volker Hall, 1670 University Blvd., Birmingham, AL, 35294-0019, USA

    Harrison Kim

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  1. Harrison Kim
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Correspondence to Harrison Kim.

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Conflict of Interest

Dr. Donald J. Buchsbaum has intellectual property related to TRA-8.

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Kim, H., Buchsbaum, D.J. & Zinn, K.R. A Novel Imaging Biomarker Extracted from Fluorescence Microscopic Imaging of TRA-8/DR5 Oligomers Predicts TRA-8 Therapeutic Efficacy in Breast and Pancreatic Cancer Mouse Models. Mol Imaging Biol 18, 325–333 (2016). https://doi.org/10.1007/s11307-015-0913-x

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  • DOI: https://doi.org/10.1007/s11307-015-0913-x

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