The finding of new diagnostic and prognostic markers of local radiation injury, and particularly of the cutaneous radiation syndrome, is crucial for its medical management, in the case of both accidental exposure and radiotherapy side effects. Especially, a fast high-throughput method is still needed for triage of people accidentally exposed to ionizing radiation. In this study, we investigated the impact of localized irradiation of the skin on the early alteration of the serum proteome of mice in an effort to discover markers associated with the exposure and severity of impending damage. Using two different large-scale quantitative proteomic approaches, 2D-DIGE-MS and SELDI-TOF-MS, we performed global analyses of serum proteins collected in the clinical latency phase (days 3 and 7) from non-irradiated and locally irradiated mice exposed to high doses of 20, 40 and 80 Gy which will develop respectively erythema, moist desquamation and necrosis. Unsupervised and supervised multivariate statistical analyses (principal component analysis, partial-least square discriminant analysis and Random Forest analysis) using 2D-DIGE quantitative protein data allowed us to discriminate early between non-irradiated and irradiated animals, and between uninjured/slightly injured animals and animals that will develop severe lesions. On the other hand, despite a high number of animal replicates, PLS-DA and Random Forest analyses of SELDI-TOF-MS data failed to reveal sets of MS peaks able to discriminate between the different groups of animals. Our results show that, unlike SELDI-TOF-MS, the 2D-DIGE approach remains a powerful and promising method for the discovery of sets of proteins that could be used for the development of clinical tests for triage and the prognosis of the severity of radiation-induced skin lesions. We propose a list of 15 proteins which constitutes a set of candidate proteins for triage and prognosis of skin lesion outcomes.
References
[1]
Hopewell, J.W. The skin: Its structure and response to ionizing radiation. Int. J. Radiat. Biol. 1990, 57, 751–773, doi:10.1080/09553009014550911.
[2]
Gottlober, P.; Bezold, G.; Weber, L.; Gourmelon, P.; Cosset, J.M.; Bahren, W.; Hald, H.J.; Fliedner, T.M.; Peter, R.U. The radiation accident in Georgia: Clinical appearance and diagnosis of cutaneous radiation syndrome. J. Am. Acad. Dermatol. 2000, 42, 453–458, doi:10.1016/S0190-9622(00)90218-4.
[3]
Peter, R.U.; Steinert, M.; Gottlober, P. The cutaneous radiation syndrome: Diagnosis and treatment. Radioprotection 2001, 49, 451–457.
[4]
Muller, K.; Meineke, V. Advances in the management of localized radiation injuries. Health Phys. 2010, 98, 843–850, doi:10.1097/HP.0b013e3181adcba7.
[5]
Meineke, V. The role of damage to the cutaneous system in radiation-induced multi-organ failure. Br. J. Radiol. Suppl. 2005, 27, 85–99.
[6]
Peter, R.U. Cutaneous radiation syndrome in multi-organ failure. Br. J. Radiol. 2005, 27, 180–184, doi:10.1259/bjr/56925969.
[7]
Ainsbury, E.A.; Bakhanova, E.; Barquinero, J.F.; Brai, M.; Chumak, V.; Correcher, V.; Darroudi, F.; Fattibene, P.; Gruel, G.; Guclu, I.; et al. Review of retrospective dosimetry techniques for external ionising radiation exposures. Radiat. Prot. Dosimetry 2011, 147, 573–592, doi:10.1093/rpd/ncq499.
[8]
Lataillade, J.J.; Doucet, C.; Bey, E.; Carsin, H.; Huet, C.; Clairand, I.; Bottollier-Depois, J.F.; Chapel, A.; Ernou, I.; Gourven, M.; et al. New approach to radiation burn treatment by dosimetry-guided surgery combined with autologous mesenchymal stem cell therapy. Regen. Med. 2007, 2, 785–794, doi:10.2217/17460751.2.5.785.
[9]
Bey, E.; Prat, M.; Duhamel, P.; Benderitter, M.; Brachet, M.; Trompier, F.; Battaglini, P.; Ernou, I.; Boutin, L.; Gourven, M.; et al. Emerging therapy for improving wound repair of severe radiation burns using local bone marrow-derived stem cell administrations. Wound Repair Regen. 2010, 18, 50–58, doi:10.1111/j.1524-475X.2009.00562.x.
[10]
Akita, S.; Akino, K.; Hirano, A.; Ohtsuru, A.; Yamashita, S. Mesenchymal stem cell therapy for cutaneous radiation syndrome. Health Phys. 2010, 98, 858–862, doi:10.1097/HP.0b013e3181d3d52c.
[11]
Benderitter, M.; Gourmelon, P.; Bey, E.; Chapel, A.; Clairand, I.; Prat, M.; Lataillade, J.J. New emerging concepts in the medical management of local radiation injury. Health Phys. 2010, 98, 851–857, doi:10.1097/HP.0b013e3181c9f79a.
[12]
Forcheron, F.; Agay, D.; Scherthan, H.; Riccobono, D.; Herodin, F.; Meineke, V.; Drouet, M. Autologous adipocyte derived stem cells favour healing in a minipig model of cutaneous radiation syndrome. PLoS One 2012, 7, e31694.
[13]
Friesecke, I.; Beyrer, K.; Fliedner, T.M. How to cope with radiation accidents: The medical management. Br. J. Radiol. 2001, 74, 121–122.
[14]
Fliedner, T.M.; Friesecke, I.; Beyrer, K. Medical Management of Radiation Accident: Manual on the Acute Radiation Syndrome; British Institute of Radiology: Oxford, UK, 2001; pp. 1–66.
[15]
Guipaud, O.; Benderitter, M. Protein biomarkers for radiation exposure: Towards a proteomic approach as a new investigation tool. Ann. Ist. Super Sanita 2009, 45, 278–286.
[16]
Sharma, M.; Moulder, J.E. The urine proteome as a radiation biodosimeter. Adv. Exp. Med. Biol. 2013, 990, 87–100, doi:10.1007/978-94-007-5896-4_5.
[17]
Guipaud, O. Serum and plasma proteomics and its possible use as detector and predictor of radiation diseases. Adv. Exp. Med. Biol. 2013, 990, 61–86, doi:10.1007/978-94-007-5896-4_4.
[18]
Guipaud, O.; Holler, V.; Buard, V.; Tarlet, G.; Royer, N.; Vinh, J.; Benderitter, M. Time-course analysis of mouse serum proteome changes following exposure of the skin to ionizing radiation. Proteomics 2007, 7, 3992–4002, doi:10.1002/pmic.200601032.
[19]
Chaze, T.; Slomianny, M.C.; Milliat, F.; Tarlet, G.; Lefebvre-Darroman, T.; Gourmelon, P.; Bey, E.; Benderitter, M.; Michalski, J.C.; Guipaud, O. Alteration of the serum N-glycome of mice locally exposed to high doses of ionizing radiation. Mol. Cell Proteomics 2013, 12, 283–301, doi:10.1074/mcp.M111.014639.
[20]
Petricoin, E.F.; Ardekani, A.M.; Hitt, B.A.; Levine, P.J.; Fusaro, V.A.; Steinberg, S.M.; Mills, G.B.; Simone, C.; Fishman, D.A.; Kohn, E.C.; et al. Use of proteomic patterns in serum to identify ovarian cancer. Lancet 2002, 359, 572–577, doi:10.1016/S0140-6736(02)07746-2.
Fung, E.T.; Yip, T.T.; Lomas, L.; Wang, Z.; Yip, C.; Meng, X.Y.; Lin, S.; Zhang, F.; Zhang, Z.; Chan, D.W.; et al. Classification of cancer types by measuring variants of host response proteins using SELDI serum assays. Int. J. Cancer 2005, 115, 783–789, doi:10.1002/ijc.20928.
[23]
Zhang, Z.; Bast, R.C., Jr.; Yu, Y.; Li, J.; Sokoll, L.J.; Rai, A.J.; Rosenzweig, J.M.; Cameron, B.; Wang, Y.Y.; Meng, X.Y.; et al. Three biomarkers identified from serum proteomic analysis for the detection of early stage ovarian cancer. Cancer Res. 2004, 64, 5882–5890, doi:10.1158/0008-5472.CAN-04-0746.
[24]
Wadsworth, J.T.; Somers, K.D.; Cazares, L.H.; Malik, G.; Adam, B.L.; Stack, B.C., Jr.; Wright, G.L., Jr.; Semmes, O.J. Serum protein profiles to identify head and neck cancer. Clin. Cancer Res. 2004, 10, 1625–1632, doi:10.1158/1078-0432.CCR-0297-3.
[25]
Koopmann, J.; Zhang, Z.; White, N.; Rosenzweig, J.; Fedarko, N.; Jagannath, S.; Canto, M.I.; Yeo, C.J.; Chan, D.W.; Goggins, M. Serum diagnosis of pancreatic adenocarcinoma using surface-enhanced laser desorption and ionization mass spectrometry. Clin. Cancer. Res. 2004, 10, 860–868, doi:10.1158/1078-0432.CCR-1167-3.
[26]
Vlahou, A.; Laronga, C.; Wilson, L.; Gregory, B.; Fournier, K.; McGaughey, D.; Perry, R.R.; Wright, G.L., Jr.; Semmes, O.J. A novel approach toward development of a rapid blood test for breast cancer. Clin. Breast Cancer 2003, 4, 203–209.
[27]
Petricoin, E.F., 3rd; Ornstein, D.K.; Paweletz, C.P.; Ardekani, A.; Hackett, P.S.; Hitt, B.A.; Velassco, A.; Trucco, C.; Wiegand, L.; Wood, K.; et al. Serum proteomic patterns for detection of prostate cancer. J. Natl. Cancer Inst. 2002, 94, 1576–1578.
[28]
Lin, Q.; Peng, Q.; Yao, F.; Pan, X.F.; Xiong, L.W.; Wang, Y.; Geng, J.F.; Feng, J.X.; Han, B.H.; Bao, G.L.; et al. A classification method based on principal components of SELDI spectra to diagnose of lung adenocarcinoma. PLoS One 2012, 7, e34457.
[29]
Bonneterre, J.; Revillion, F.; Desauw, C.; Blot, E.; Kramar, A.; Fournier, C.; Hornez, L.; Peyrat, J.P. Plasma and tissue proteomic prognostic factors of response in primary breast cancer patients receiving neoadjuvant chemotherapy. Oncol. Rep. 2013, 29, 355–361.
[30]
Menard, C.; Johann, D.; Lowenthal, M.; Muanza, T.; Sproull, M.; Ross, S.; Gulley, J.; Petricoin, E.; Coleman, C.N.; Whiteley, G.; et al. Discovering clinical biomarkers of ionizing radiation exposure with serum proteomic analysis. Cancer Res. 2006, 66, 1844–1850, doi:10.1158/0008-5472.CAN-05-3466.
[31]
Benjamini, Y.; Yekutieli, D. The control of false discovery rate under dependency. Ann. Stat. 2001, 61, 1165–1188.
[32]
Guipaud, O.; Guillonneau, F.; Labas, V.; Praseuth, D.; Rossier, J.; Lopez, B.; Bertrand, P. An in vitro enzymatic assay coupled to proteomics analysis reveals a new DNA processing activity for Ewing sarcoma and TAF(II)68 proteins. Proteomics 2006, 6, 5962–5972, doi:10.1002/pmic.200600259.
[33]
Quintana, M.; Palicki, O.; Lucchi, G.; Ducoroy, P.; Chambon, C.; Salles, C.; Morzel, M. Inter-individual variability of protein patterns in saliva of healthy adults. J. Proteomics 2009, 72, 822–830, doi:10.1016/j.jprot.2009.05.004.
[34]
Poullet, P.; Carpentier, S.; Barillot, E. myProMS, a web server for management and validation of mass spectrometry-based proteomic data. Proteomics 2007, 7, 2553–2556, doi:10.1002/pmic.200600784.
[35]
Nikitin, A.; Egorov, S.; Daraselia, N.; Mazo, I. Pathway studio—The analysis and navigation of molecular networks. Bioinformatics 2003, 19, 2155–2157, doi:10.1093/bioinformatics/btg290.
[36]
Holler, V.; Buard, V.; Gaugler, M.H.; Guipaud, O.; Baudelin, C.; Sache, A.; Perez Mdel, R.; Squiban, C.; Tamarat, R.; Milliat, F.; et al. Pravastatin limits radiation-induced vascular dysfunction in the skin. J. Invest. Dermatol. 2009, 129, 1280–1291, doi:10.1038/jid.2008.360.
[37]
Anderson, N.L.; Anderson, N.G. The human plasma proteome: History, character, and diagnostic prospects. Mol. Cell Proteomics 2002, 1, 845–867, doi:10.1074/mcp.R200007-MCP200.
[38]
Robbins, M.E.; Zhao, W. Chronic oxidative stress and radiation-induced late normal tissue injury: A review. Int. J. Radiat. Biol. 2004, 80, 251–259, doi:10.1080/09553000410001692726.
[39]
Zhao, W.; Robbins, M.E. Inflammation and chronic oxidative stress in radiation-induced late normal tissue injury: Therapeutic implications. Curr. Med. Chem. 2009, 16, 130–143, doi:10.2174/092986709787002790.
[40]
Francois, A.; Milliat, F.; Guipaud, O.; Benderitter, M. Inflammation and immunity in radiation damage to the gut mucosa. Biomed. Res. Int. 2013, 2013, e123241.
[41]
Bentzen, S.M. Preventing or reducing late side effects of radiation therapy: Radiobiology meets molecular pathology. Nat. Rev. Cancer 2006, 6, 702–713, doi:10.1038/nrc1950.
[42]
Milliat, F.; Francois, A.; Tamarat, R.; Benderitter, M. Role of endothelium in radiation-induced normal tissue damages. Ann. Cardiol. Angeiol. (Paris) 2008, 57, 139–148.
[43]
Fajardo, L.F.; Berthrong, M. Vascular lesions following radiation. Pathol. Annu. 1988, 23, 297–330.
[44]
Gabay, C.; Kushner, I. Acute-phase proteins and other systemic responses to inflammation. N. Engl. J. Med. 1999, 340, 448–454, doi:10.1056/NEJM199902113400607.
[45]
Magic, Z.; Matic-Ivanovic, S.; Savic, J.; Poznanovic, G. Ionizing radiation-induced expression of the genes associated with the acute response to injury in the rat. Radiat. Res. 1995, 143, 187–193, doi:10.2307/3579156.
[46]
Trutic, N.; Magic, Z.; Urosevic, N.; Krtolica, K. Acute-phase protein gene expression in rat liver following whole body X-irradiation or partial hepatectomy. Comp. Biochem. Physiol. C Toxicol. Pharmacol. 2002, 133, 461–470, doi:10.1016/S1532-0456(02)00174-6.
Goltry, K.L.; Epperly, M.W.; Greenberger, J.S. Induction of serum amyloid A inflammatory response genes in irradiated bone marrow cells. Radiat. Res. 1998, 149, 570–578, doi:10.2307/3579903.
[49]
Chen, C.; Lorimore, S.A.; Evans, C.A.; Whetton, A.D.; Wright, E.G. A proteomic analysis of murine bone marrow and its response to ionizing radiation. Proteomics 2005, 5, 4254–4263, doi:10.1002/pmic.200401295.
[50]
Cengiz, M.; Akbulut, S.; Atahan, I.L.; Grigsby, P.W. Acute phase response during radiotherapy. Int. J. Radiat. Oncol. Biol. Phys. 2001, 49, 1093–1096, doi:10.1016/S0360-3016(00)01426-7.
[51]
Barcellos-Hoff, M.H. How do tissues respond to damage at the cellular level? The role of cytokines in irradiated tissues. Radiat. Res. 1998, 150, S109–S120, doi:10.2307/3579813.
[52]
Muller, K.; Meineke, V. Radiation-induced alterations in cytokine production by skin cells. Exp. Hematol. 2007, 35, 96–104, doi:10.1016/j.exphem.2007.01.017.
[53]
Benderitter, M.; Isoir, M.; Buard, V.; Durand, V.; Linard, C.; Vozenin-Brotons, M.C.; Steffanazi, J.; Carsin, H.; Gourmelon, P. Collapse of skin antioxidant status during the subacute period of cutaneous radiation syndrome: A case report. Radiat. Res. 2007, 167, 43–50, doi:10.1667/RR0577.1.
