Glial cell proteome using targeted quantitative methods for potential multi-diagnostic biomarkers
Tóm tắt
Glioblastoma is one of the most malignant primary brain cancer. Despite surgical resection with modern technology followed by chemo-radiation therapy with temozolomide, resistance to the treatment and recurrence is common due to its aggressive and infiltrating nature of the tumor with high proliferation index. The median survival time of the patients with glioblastomas is less than 15 months. Till now there has been no report of molecular target specific for glioblastomas. Early diagnosis and development of molecular target specific for glioblastomas are essential for longer survival of the patients with glioblastomas. Development of biomarkers specific for glioblastomas is most important for early diagnosis, estimation of the prognosis, and molecular target therapy of glioblastomas. To that end, in this study, we have conducted a comprehensive proteome study using primary cells and tissues from patients with glioblastoma. In the discovery stage, we have identified 7429 glioblastoma-specific proteins, where 476 proteins were quantitated using Tandem Mass Tag (TMT) method; 228 and 248 proteins showed up and down-regulated pattern, respectively. In the validation stage (20 selected target proteins), we developed quantitative targeted method (MRM: Multiple reaction monitoring) using stable isotope standards (SIS) peptide. In this study, five proteins (CCT3, PCMT1, TKT, TOMM34, UBA1) showed the significantly different protein levels (t-test: p value ≤ 0.05, AUC ≥ 0.7) between control and cancer groups and the result of multiplex assay using logistic regression showed the 5-marker panel showed better sensitivity (0.80 and 0.90), specificity (0.92 and 1.00), error rate (10 and 2%), and AUC value (0.94 and 0.98) than the best single marker (TOMM34) in primary cells and tissues, respectively. Although we acknowledge that the model requires further validation in a large sample size, the 5 protein marker panel can be used as baseline data for the discovery of novel biomarkers of the glioblastoma. For the discovery of multi-diagnostic biomarker, we have conducted a comprehensive proteome study using primary cells from patients with glioblastoma. In this study, 7429 glioblastoma-specific proteins were identified and then 20 selected target proteins were verified using MRM method. Finally, five proteins (CCT3, PCMT1, TKT, TOMM34, UBA1) showed the significantly different protein levels (t-test: p value ≤ 0.05, AUC ≥ 0.7) between control and cancer groups and the result of multiplex assay using logistic regression showed the 5-marker panel showed better sensitivity (0.80 and 0.90), specificity (0.92 and 1.00), error rate (10 and 2%), and AUC value (0.94 and 0.98) than the best single marker (TOMM34) in primary cells and tissues, respectively.
Tài liệu tham khảo
Wesseling P, Capper D. WHO 2016 classification of gliomas. Neuropathol Appl Neurobiol. 2018;44:139–50.
Louis DN, Perry A, Reifenberger G, von Deimling A, et al. The 2016 world Health organization classification of tumors of the central nervous system: a summary. Acta Neuropathol. 2016;131:803–20.
Rahmawati D, Marhaendraputro EA, Kurniawan SN, Wirathmawati A. Extracranial metastasis of glioblastoma: a rare case. MNJ. 2019;2019(5):4.
Nizamutdinov D, Stock EM, Dandashi JA, Vasquez EA, et al. Prognostication of survival outcomes in patients diagnosed with glioblastoma. World Neurosurg. 2018;109:e67–74.
Stupp R, Mason WP, van den Bent MJ, Weller M, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352:987–96.
Scott JN, Rewcastle NB, Brasher PMA, Fulton D, et al. Which glioblastoma multiforme patient will become a long-term survivor? A Popul-Based Study Ann Neurol. 1999;46:183–8.
Donato V, Papaleo A, Castrichino A, Banelli E, et al. Prognostic implication of clinical and pathologic features in patients with glioblastoma multiforme treated with concomitant radiation plus temozolomide. Tumori. 2007;93:248–56.
Chamberlain MC. Radiographic patterns of relapse in glioblastoma. J Neurooncol. 2011;101:319–23.
Dobelbower MC, Burnett Iii OL, Nordal RA, Nabors LB, et al. Patterns of failure for glioblastoma multiforme following concurrent radiation and temozolomide. J Med Imaging Radiat Oncol. 2011;55:77–81.
McDonald MW, Shu H-KG, Curran WJ, Crocker IR. Pattern of failure after limited margin radiotherapy and temozolomide for glioblastoma. Int J Radiation. 2011;79:130–6.
Minniti G, Amelio D, Amichetti M, Salvati M, et al. Patterns of failure and comparison of different target volume delineations in patients with glioblastoma treated with conformal radiotherapy plus concomitant and adjuvant temozolomide. Radiother Oncol. 2010;97:377–81.
Shankar GM, Balaj L, Stott SL, Nahed B, Carter BS. Liquid biopsy for brain tumors. Expert Rev Mol Diagn. 2017;17:943–7.
Saenz-Antoñanzas A, Auzmendi-Iriarte J, Carrasco-Garcia E, Moreno-Cugnon L, et al. Liquid biopsy in glioblastoma: opportunities applications and challenges. Cancers. 2019. https://doi.org/10.3390/cancers11070950.
Jovčevska I, Kočevar N, Komel R. Glioma and glioblastoma—how much do we (not) know? (Review). Mol Clin Oncol. 2013;1:935–41.
Baehring JM, Bi WL, Bannykh S, Piepmeier JM, Fulbright RK. Diffusion MRI in the early diagnosis of malignant glioma. J Neurooncol. 2007;82:221–5.
Huang Z, Ma L, Huang C, Li Q, Nice EC. Proteomic profiling of human plasma for cancer biomarker discovery. Proteomics. 2017. https://doi.org/10.1002/pmic.201600240.
Gollapalli K, Ray S, Srivastava R, Renu D, et al. Investigation of serum proteome alterations in human glioblastoma multiforme. Proteomics. 2012;12:2378–90.
Min H, Han D, Kim Y, Cho JY, et al. Label-free quantitative proteomics and N-terminal analysis of human metastatic lung cancer cells. Mol Cells. 2014;37:457–66.
Jin J, Son M, Kim H, Kim H, et al. Comparative proteomic analysis of human malignant ascitic fluids for the development of gastric cancer biomarkers. Clin Biochem. 2018;56:55–61.
Silantyev AS, Falzone L, Libra M, Gurina OI, et al. Current and future trends on diagnosis and prognosis of glioblastoma: from molecular biology to proteomics. Cells. 2019. https://doi.org/10.3390/cells8080863.
Hegi ME, Liu L, Herman JG, Stupp R, et al. Correlation of O6-methylguanine methyltransferase (MGMT) promoter methylation with clinical outcomes in glioblastoma and clinical strategies to modulate MGMT activity. J Clin Oncol. 2008;26:4189–99.
Xue J, Sang W, Su LP, Gao HX, et al. Proteomics reveals protein phosphatase 1γ as a biomarker associated with Hippo signal pathway in glioma. Pathol Res Pract. 2020;216: 153187.
Kalinina J, Peng J, Ritchie JC, Van Meir EG. Proteomics of gliomas: initial biomarker discovery and evolution of technology. Neuro Oncol. 2011;13:926–42.
Miyauchi E, Furuta T, Ohtsuki S, Tachikawa M, et al. Identification of blood biomarkers in glioblastoma by SWATH mass spectrometry and quantitative targeted absolute proteomics. PLoS ONE. 2018;13: e0193799.
Arora A, Patil V, Kundu P, Kondaiah P, et al. Serum biomarkers identification by iTRAQ and verification by MRM: S100A8/S100A9 levels predict tumor-stroma involvement and prognosis in glioblastoma. Sci Rep. 2019;9:2749.
Brown KJ, Seol H, Pillai DK, Sankoorikal BJ, et al. The human secretome atlas initiative: implications in health and disease conditions. Biochim Biophys Acta. 2013;1834:2454–61.
Miserocchi G, Mercatali L, Liverani C, De Vita A, et al. Management and potentialities of primary cancer cultures in preclinical and translational studies. J Transl Med. 2017;15:229.
Nesvizhskii AI, Keller A, Kolker E, Aebersold R. A statistical model for identifying proteins by tandem mass spectrometry. Anal Chem. 2003;75:4646–58.
Jin J, Min H, Kim SJ, Oh S, et al. Development of diagnostic biomarkers for detecting diabetic retinopathy at early stages using quantitative proteomics. J Diabetes Res. 2016;2016:6571976.