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Monitoring of changes in lipid profiles during PLK1 knockdown in cancer cells using DESI MS

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Abstract

The importance of the polo-like kinase 1 (PLK1) gene is increasing substantially both as a biomarker and as a target for highly specific cancer therapy. This is due to its involvement in multiple points of cell progression and carcinogenesis. PLK1 inhibitors’ efficacy in treating human cancers has been limited due to the lack of a specific targeting strategy. Here, we describe a method of targeted downregulation of PLK1 in cancer cells and the concomitant rapid detection of surface lipidomic perturbations using desorption electrospray ionization mass spectrometry (DESI MS). The efficient delivery of siRNA targeting PLK1 gene selectively to the cancer cells is achieved by targeting overexpressed cell surface epithelial cell adhesion molecule (EpCAM) by the EpDT3 aptamer. The chimeric aptamer (EpDT3-siPLK1) showed the knockdown of PLK1 gene expression and PLK1 protein levels by quantitative PCR and western blotting, respectively. The abundant surface lipids, phosphatidylcholines (PCs), such as PC(32:1) (m/z 754.6), PC(34:1) (m/z 782.6), and PC(36:2) (m/z 808.6), were highly expressed in MCF-7 and WERI-RB1 cancer cells compared to normal MIO-M1 cells and they were observed using DESI MS. These overexpressed cell surface lipids in the cancer cells were downregulated upon the treatment of EpDT3-siPLK1 chimera indicating a novel role of PLK1 to regulate surface lipid expression in addition to the efficient selective cancer targeting ability. Our results indicate that DESI MS has a potential ability to rapidly monitor aptamer-mediated cancer therapy and accelerate the drug discovery process.

Binding of aptamer chimera to the cells and changes in lipid profile

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References

  1. Beljebbar A, Dukic S, Amharref N, Bellefqih S, Manfait M. Monitoring of biochemical changes through the C6 gliomas progression and invasion by Fourier transform infrared (FTIR) imaging. Anal Chem. 2009;81:9247–56.

    Article  CAS  Google Scholar 

  2. Shimma S, Sugiura Y, Hayasaka T, Hoshikawa Y, Noda T, Setou M. MALDI-based imaging mass spectrometry revealed abnormal distribution of phospholipids in colon cancer liver metastasis. J Chromatogr B. 2007;855:98–103.

    Article  CAS  Google Scholar 

  3. Ackerstaff E, Glunde K, Bhujwalla ZM. Choline phospholipid metabolism: a target in cancer cells? J Cell Biochem. 2003;90:525–33.

    Article  CAS  Google Scholar 

  4. Glunde K, Jie C, Bhujwalla ZM. Molecular causes of the aberrant choline phospholipid metabolism in breast cancer. Cancer Res. 2004;64:4270–6.

    Article  CAS  Google Scholar 

  5. Gillies RJ, Morse DL. In vivo magnetic resonance spectroscopy in cancer. Annu Rev Biomed Eng. 2005;7:287–326.

    Article  CAS  Google Scholar 

  6. Glunde K, Artemov D, Penet M-F, Jacobs MA, Bhujwalla ZM. Magnetic resonance spectroscopy in metabolic and molecular imaging and diagnosis of cancer. Chem Rev. 2010;110:3043–59.

    Article  CAS  Google Scholar 

  7. Monge ME, Harris GA, Dwivedi P, Fernandez FM. Mass spectrometry: recent advances in direct open air surface sampling/ionization. Chem Rev. 2013;113:2269–308.

    Article  CAS  Google Scholar 

  8. Takats Z, Wiseman JM, Gologan B, Cooks RG. Mass spectrometry sampling under ambient conditions with desorption electrospray ionization. Science. 2004;306:471–3.

    Article  CAS  Google Scholar 

  9. Pasilis SP, Kertesz V, Van Berkel GJ, Schulz M, Schorcht S. HPTLC/DESI-MS imaging of tryptic protein digests separated in two dimensions. J Mass Spectrom. 2008;43:1627–35.

    Article  CAS  Google Scholar 

  10. Chipuk JE, Gelb MH, Brodbelt JS. Rapid and selective screening for sulfhydryl analytes in plasma and urine using surface-enhanced transmission mode desorption electrospray ionization mass spectrometry. Anal Chem. 2010;82:4130–9.

    Article  CAS  Google Scholar 

  11. Wiseman JM, Ifa DR, Venter A, Cooks RG. Ambient molecular imaging by desorption electrospray ionization mass spectrometry. Nat Protoc. 2008;3:517–24.

    Article  CAS  Google Scholar 

  12. Vismeh R, Waldon DJ, Teffera Y, Zhao Z. Localization and quantification of drugs in animal tissues by use of desorption electrospray ionization mass spectrometry imaging. Anal Chem. 2012;84:5439–45.

    Article  CAS  Google Scholar 

  13. Eberlin LS, Norton I, Orringer D, Dunn IF, Liu X, Ide JL, et al. Ambient mass spectrometry for the intraoperative molecular diagnosis of human brain tumors. Proc Natl Acad Sci U S A. 2013;110:1611–6.

    Article  Google Scholar 

  14. Eberlin LS, Tibshirani RJ, Zhang J, Longacre TA, Berry GJ, Bingham DB, et al. Molecular assessment of surgical-resection margins of gastric cancer by mass-spectrometric imaging. Proc Natl Acad Sci U S A. 2014;111:2436–41.

    Article  CAS  Google Scholar 

  15. Srimany A, Ifa DR, Naik HR, Bhat V, Cooks RG, Pradeep T. Direct analysis of camptothecin from Nothapodytes nimmoniana by desorption electrospray ionization mass spectrometry (DESI-MS). Analyst. 2011;136:3066–8.

    Article  CAS  Google Scholar 

  16. Ifa DR, Srimany A, Eberlin LS, Naik HR, Bhat V, Cooks RG, et al. Tissue imprint imaging by desorption electrospray ionization mass spectrometry. Anal Methods. 2011;3:1910–2.

    Article  CAS  Google Scholar 

  17. Mohana Kumara P, Srimany A, Ravikanth G, Uma Shaanker R, Pradeep T. Ambient ionization mass spectrometry imaging of rohitukine, a chromone anti-cancer alkaloid, during seed development in Dysoxylum binectariferum Hook.f (Meliaceae). Phytochemistry. 2015;116:104–10.

    Article  CAS  Google Scholar 

  18. Hemalatha RG, Pradeep T. Understanding the molecular signatures in leaves and flowers by desorption electrospray ionization mass spectrometry (DESI MS) imaging. J Agric Food Chem. 2013;61:7477–87.

    Article  CAS  Google Scholar 

  19. Glover DM, Hagan IM, Tavares ÁAM. Polo-like kinases: a team that plays throughout mitosis. Gene Dev. 1998;12:3777–87.

    Article  CAS  Google Scholar 

  20. Glover DM, Ohkura H, Tavares A. Polo kinase: the choreographer of the mitotic stage? J Cell Biol. 1996;135:1681–4.

    Article  CAS  Google Scholar 

  21. Nigg EA, Blangy A, Lane HA. Dynamic changes in nuclear architecture during mitosis: on the role of protein phosphorylation in spindle assembly and chromosome segregation. Exp Cell Res. 1996;229:174–80.

    Article  CAS  Google Scholar 

  22. Kumagai A, Dunphy WG. Purification and molecular cloning of Plx1, a Cdc25-regulatory kinase from Xenopus egg extracts. Science. 1996;273:1377–80.

    Article  CAS  Google Scholar 

  23. Nigg EA. Polo-like kinases: positive regulators of cell division from start to finish. Curr Opin Cell Biol. 1998;10:776–83.

    Article  CAS  Google Scholar 

  24. Cunningham JT, Ruggero D. New connections between old pathways: PDK1 signaling promotes cellular transformation through PLK1-dependent MYC stabilization. Cancer Discov. 2013;3:1099–102.

    Article  CAS  Google Scholar 

  25. Wang G, Chen Q, Zhang X, Zhang B, Zhuo X, Liu J, et al. PCM1 recruits Plk1 to the pericentriolar matrix to promote primary cilia disassembly before mitotic entry. J Cell Sci. 2013;126:1355–65.

    Article  CAS  Google Scholar 

  26. Cholewa BD, Liu X, Ahmad N. The role of polo-like kinase 1 in carcinogenesis: cause or consequence? Cancer Res. 2013;73:6848–55.

    Article  CAS  Google Scholar 

  27. Bhola NE, Jansen VM, Bafna S, Giltnane JM, Balko JM, Estrada MV, et al. Kinome-wide functional screen identifies role of PLK1 in hormone-independent, ER-positive breast cancer. Cancer Res. 2015;75:405–14.

    Article  CAS  Google Scholar 

  28. Zhang Z, Hou X, Shao C, Li J, Cheng J-X, Kuang S, et al. Plk1 inhibition enhances the efficacy of androgen signaling blockade in castration-resistant prostate cancer. Cancer Res. 2014;74:6635–47.

    Article  CAS  Google Scholar 

  29. Denholt CL, Hansen PR, Pedersen N, Poulsen HS, Gillings N, Kjær A. Identification of novel peptide ligands for the cancer-specific receptor mutation EFGRvIII using a mixture-based synthetic combinatorial library. Biopolymers. 2009;91:201–6.

    Article  CAS  Google Scholar 

  30. Kim M, Shin D-S, Kim J, Lee Y-S. Substrate screening of protein kinases: detection methods and combinatorial peptide libraries. Biopolymers. 2010;94:753–62.

    Article  CAS  Google Scholar 

  31. Li J, Tan S, Chen X, Zhang CY, Zhang Y. Peptide aptamers with biological and therapeutic applications. Curr Med Chem. 2011;18:4215–22.

    Article  CAS  Google Scholar 

  32. Pirogova E, Istivan T, Gan E, Cosic I. Advances in methods for therapeutic peptide discovery, design and development. Curr Pharm Biotechnol. 2011;12:1117–27.

    Article  CAS  Google Scholar 

  33. Aina OH, Liu R, Sutcliffe JL, Marik J, Pan C-X, Lam KS. From combinatorial chemistry to cancer-targeting peptides. Mol Pharmaceutics. 2007;4:631–51.

    Article  CAS  Google Scholar 

  34. Zhu Z, Song Y, Li C, Zou Y, Zhu L, An Y, et al. Monoclonal surface display SELEX for simple, rapid, efficient, and cost-effective aptamer enrichment and identification. Anal Chem. 2014;86:5881–8.

    Article  CAS  Google Scholar 

  35. Subramanian N, Raghunathan V, Kanwar JR, Kanwar RK, Elchuri SV, Khetan V, et al. Target-specific delivery of doxorubicin to retinoblastoma using epithelial cell adhesion molecule aptamer. Mol Vis. 2012;18:2783–95.

    CAS  Google Scholar 

  36. Gilboa-Geffen A, Hamar P, Le MTN, Wheeler LA, Trifonova R, Petrocca F, et al. Gene knockdown by EpCAM aptamer-siRNA chimeras suppresses epithelial breast cancers and their tumor-initiating cells. Mol Cancer Ther. 2015;14:2279–91.

    Article  CAS  Google Scholar 

  37. Srimany A, Jayashree B, Krishnakumar S, Elchuri S, Pradeep T. Identification of effective substrates for the direct analysis of lipids from cell lines using desorption electrospray ionization mass spectrometry. Rapid Commun Mass Spectrom. 2015;29:349–56.

    Article  CAS  Google Scholar 

  38. Dassie JP, Liu XY, Thomas GS, Whitaker RM, Thiel KW, Stockdale KR, et al. Systemic administration of optimized aptamer-siRNA chimeras promotes regression of PSMA-expressing tumors. Nat Biotechnol. 2009;27:839–49.

    Article  CAS  Google Scholar 

  39. Subramanian N, Kanwar JR, Athalya PK, Janakiraman N, Khetan V, Kanwar RK, et al. EpCAM aptamer mediated cancer cell specific delivery of EpCAM siRNA using polymeric nanocomplex. J Biomed Sci. 2015;22:4.

    Article  CAS  Google Scholar 

  40. Liu HY, Gao X. A universal protein tag for delivery of SiRNA-aptamer chimeras. Sci Rep. 2013;3:3129.

    Google Scholar 

  41. Zhou J, Tiemann K, Chomchan P, Alluin J, Swiderski P, Burnett J, et al. Dual functional BAFF receptor aptamers inhibit ligand-induced proliferation and deliver siRNAs to NHL cells. Nucleic Acids Res. 2013;41:4266–83.

    Article  CAS  Google Scholar 

  42. Eberlin LS, Gabay M, Fan AC, Gouw AM, Tibshirani RJ, Felsher DW, et al. Alteration of the lipid profile in lymphomas induced by MYC overexpression. Proc Natl Acad Sci U S A. 2014;111:10450–5.

    Article  CAS  Google Scholar 

  43. Yang L, Cui X, Zhang N, Li M, Bai Y, Han X, et al. Comprehensive lipid profiling of plasma in patients with benign breast tumor and breast cancer reveals novel biomarkers. Anal Bioanal Chem. 2015;407:5065–77.

    Article  CAS  Google Scholar 

  44. Liu R, Peng Y, Li X, Wang Y, Pan E, Guo W, et al. Identification of plasma metabolomic profiling for diagnosis of esophageal squamous-cell carcinoma using an UPLC/TOF/MS platform. Int J Mol Sci. 2013;14:8899–911.

    Article  Google Scholar 

  45. Huang C, Freter C. Lipid metabolism, apoptosis and cancer therapy. Int J Mol Sci. 2015;16:924–49.

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by the Department of Biotechnology, Government of India (Grant no. NO.BT/PR3580/PID/6/625/2011) and the Department of Science and Technology, Government of India. A.S. thanks the Council of Scientific and Industrial Research, Government of India and Indian Institute of Technology Madras for fellowships. The authors also thank Dr. Nithya Subramanian, Ms. Srilatha Jasty, and Ms. Lakshmi for their help and suggestions.

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    Authors and Affiliations

    1. Department of Nanobiotechnology, Vision Research Foundation, Sankara Nethralaya, Chennai, Tamil Nadu, 600006, India

      Balasubramanyam Jayashree, Srinidhi Jayaraman, Anjali Bhutra, Narayanan Janakiraman, Srujana Chitipothu, Subramanian Krishnakumar & Sailaja Elchuri

    2. Centre for Biotechnology, Anna University, Chennai, Tamil Nadu, 600025, India

      Balasubramanyam Jayashree & Lakshmi Subhadra Baddireddi

    3. DST Unit of Nanoscience (DST UNS) and Thematic Unit of Excellence (TUE), Department of Chemistry, Indian Institute of Technology Madras, Chennai, Tamil Nadu, 600036, India

      Amitava Srimany & Thalappil Pradeep

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    Balasubramanyam Jayashree and Amitava Srimany contributed equally to this work.

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    Jayashree, B., Srimany, A., Jayaraman, S. et al. Monitoring of changes in lipid profiles during PLK1 knockdown in cancer cells using DESI MS. Anal Bioanal Chem 408, 5623–5632 (2016). https://doi.org/10.1007/s00216-016-9665-y

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