Elevation of serum fortilin levels is specific for apoptosis and signifies cell death in vivo
Patuma Sinthujaroen, Nattaporn Wanachottrakul, Decha Pinkaew, John R Petersen, Amornrat Phongdara, Melinda Sheffield-Moore, Ken Fujise, Patuma Sinthujaroen, Nattaporn Wanachottrakul, Decha Pinkaew, John R Petersen, Amornrat Phongdara, Melinda Sheffield-Moore, Ken Fujise
Abstract
Background: Billions of cells undergo apoptosis each day in the average normal adult. The ability to readily assess the degree of apoptosis in human diseases is hampered by the lack of sensitive and specific serum biomarkers of apoptosis. Fortilin is a novel prosurvival molecule that protects cells against various noxious stimuli. While fortilin is secreted into the extracellular space under certain conditions, the relationship between the serum concentration of fortilin and the presence and extent of apoptosis in vivo remains unknown.
Methods & results: Using a newly developed fortilin ELISA system, we show here that fortilin exists in the normal human and mouse circulation. We further demonstrate that fortilin serum levels are significantly elevated in patients with solid cancer, in response to anti-cancer chemo- or radiation therapy. The elevation of fortilin serum levels is more robust and sensitive than that of such previously-reported serum biomarkers of apoptosis as fragmented cytokeratin-18, cytochrome c, and nucleosomal DNA. In addition, targeted apoptotic liver damage induced by Jo2 anti-Fas (CD95) antibody consistently and significantly increased serum fortilin levels in C57BL/6J mice. Finally, when challenged by anti-human-Fas IgM antibody, Jurkat leukemic T cells apoptosed and released fortilin into the medium before plasma membrane integrity was compromised.
Conclusions: Taken together, these data suggest that serum fortilin levels reflect the degree and extent of apoptosis occurring in vivo.
General significance: Fortilin is a viable serum biomarker of in vivo apoptosis and can be utilized to noninvasively assess the status of in vivo apoptosis in humans.
Keywords: Apoptosis; Biomarker; Fortilin; Programmed Cell Death.
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References
- Reed J.C. Dysregulation of apoptosis in cancer. J. Clin. Oncol. Off. J. Am. Soc. Clin. Oncol. 1999;17:2941–2953.
- Dupont-Versteegden E.E. Apoptosis in muscle atrophy: relevance to sarcopenia. Exp. Gerontol. 2005;40:473–481.
- Schwartz L.M. Atrophy and programmed cell death of skeletal muscle. Cell Death Differ. 2008;15:1163–1169.
- Ulukaya E., Yilmaztepe A., Akgoz S., Linder S., Karadag M. The levels of caspase-cleaved cytokeratin 18 are elevated in serum from patients with lung cancer and helpful to predict the survival. Lung Cancer. 2007;56:399–404.
- Ward T.H., Cummings J., Dean E., Greystoke A., Hou J.M., Backen A., Ranson M., Dive C. Biomarkers of apoptosis. Br. J. Cancer. 2008;99:841–846.
- Caulin C., Salvesen G.S., Oshima R.G. Caspase cleavage of keratin 18 and reorganization of intermediate filaments during epithelial cell apoptosis. J. Cell Biol. 1997;138:1379–1394.
- Tamkovich S.N., Cherepanova A.V., Kolesnikova E.V., Rykova E.Y., Pyshnyi D.V., Vlassov V.V., Laktionov P.P. Circulating DNA and DNase activity in human blood. Ann. N. Y. Acad. Sci. 2006;1075:191–196.
- Renz A., Berdel W.E., Kreuter M., Belka C., Schulze-Osthoff K., Los M. Rapid extracellular release of cytochrome c is specific for apoptosis and marks cell death in vivo. Blood. 2001;98:1542–1548.
- Jemmerson R., LaPlante B., Treeful A. Release of intact, monomeric cytochrome c from apoptotic and necrotic cells. Cell Death Differ. 2002;9:538–548.
- Gross B., Gaestel M., Bohm H., Bielka H. cDNA sequence coding for a translationally controlled human tumor protein. Nucleic Acids Res. 1989;17:8367.
- Kim M., Jung Y., Lee K., Kim C. Identification of the calcium binding sites in translationally controlled tumor protein. Arch. Pharm. Res. 2000;23:633–636.
- Graidist P., Yazawa M., Tonganunt M., Nakatomi A., Lin C., Phongdara A., Fujise K. Fortilin binds Ca2 + and blocks Ca2 +-dependent apoptosis in vivo. Biochem. J. 2007;408:181–191.
- Yarm F.R. Plk phosphorylation regulates the microtubule-stabilizing protein TCTP. Mol. Cell. Biol. 2002;22:6209–6221.
- Kashiwakura J.I., Ando T., Matsumoto K., Kimura M., Kitaura J., Matho M.H., Zajonc D.M., Ozeki T., Ra C., Macdonald S.M., Siraganian R.P., Broide D.H., Kawakami Y., Kawakami T. Histamine-releasing factor has a proinflammatory role in mouse models of asthma and allergy. J. Clin. Invest. 2012;122:218–228.
- Amzallag N., Passer B.J., Allanic D., Segura E., Thery C., Goud B., Amson R., Telerman A. TSAP6 facilitates the secretion of translationally controlled tumor protein/histamine-releasing factor via a nonclassical pathway. J. Biol. Chem. 2004;279:46104–46112.
- MacDonald S.M., Rafnar T., Langdon J., Lichtenstein L.M. Molecular identification of an IgE-dependent histamine-releasing factor. Science. 1995;269:688–690.
- Fujita T., Felix K., Pinkaew D., Hutadilok-Towatana N., Liu Z., Fujise K. Human fortilin is a molecular target of dihydroartemisinin. FEBS Lett. 2008;582:1055–1060.
- Graidist P., Phongdara A., Fujise K. Antiapoptotic protein partners fortilin and MCL1 independently protect cells from 5-FU-induced cytotoxicity. J. Biol. Chem. 2004;279:40868–40875.
- Koide Y., Kiyota T., Tonganunt M., Pinkaew D., Liu Z., Kato Y., Hutadilok-Towantana N., Phongdara A., Fujise K. Embryonic lethality of fortilin-null mutant mice by BMP-pathway overactivation. Biochim. Biophys. Acta. 2009;1790:326–338.
- Li F., Zhang D., Fujise K. Characterization of fortilin, a novel anti-apoptotic protein. J. Biol. Chem. 2001;276:47542–47549.
- Tulis D.A., Mnjoyan Z.H., Schiesser R.L., Shelat H.S., Evans A.J., Zoldhelyi P., Fujise K. Adenoviral gene transfer of fortilin attenuates neointima formation through suppression of vascular smooth muscle cell proliferation and migration. Circulation. 2003;107:98–105.
- Zhang D., Li F., Weidner D., Mnjoyan Z.H., Fujise K. Physical and functional interaction between MCL1 and fortilin. The potential role of MCL1 as a fortilin chaperone. J. Biol. Chem. 2002;277:37430–37438.
- Chen Y., Fujita T., Zhang D., Doan H., Pinkaew D., Liu Z., Wu J., Koide Y., Chiu A., Jun-Lin C., Chang J.Y., Ruan K.H., Fujise K. The physical and functional antagonism between p53 and fortilin, an anti-apoptotic molecule. J. Biol. Chem. 2011;286:32575–32585.
- Marriott H.M., Bingle C.D., Read R.C., Braley K.E., Kroemer G., Hellewell P.G., Craig R.W., Whyte M.K., Dockrell D.H. Dynamic changes in Mcl-1 expression regulate macrophage viability or commitment to apoptosis during bacterial clearance. J. Clin. Invest. 2005;115:359–368.
- Steimer D.A., Boyd K., Takeuchi O., Fisher J.K., Zambetti G.P., Opferman J.T. Selective roles for antiapoptotic MCL-1 during granulocyte development and macrophage effector function. Blood. 2009;113:2805–2815.
- Cerne K., Erman A., Veranic P. Analysis of cytotoxicity of melittin on adherent culture of human endothelial cells reveals advantage of fluorescence microscopy over flow cytometry and haemocytometer assay. Protoplasma. 2013;250:1131–1137.
- Ogasawara J., Watanabe-Fukunaga R., Adachi M., Matsuzawa A., Kasugai T., Kitamura Y., Itoh N., Suda T., Nagata S. Lethal effect of the anti-Fas antibody in mice. Nature. 1993;364:806–809.
- Koide Y., Kiyota T., Tonganunt M., Pinkaew D., Liu Z., Kato Y., Hutadilok-Towatana N., Phongdara A., Fujise K. Embryonic lethality of fortilin-null mutant mice by BMP-pathway overactivation. Biochim. Biophys. Acta. 2009;1790:326–338.
- Leers M.P., Kolgen W., Bjorklund V., Bergman T., Tribbick G., Persson B., Bjorklund P., Ramaekers F.C., Bjorklund B., Nap M., Jornvall H., Schutte B. Immunocytochemical detection and mapping of a cytokeratin 18 neo-epitope exposed during early apoptosis. J. Pathol. 1999;187:567–572.
- John K., Wielgosz S., Schulze-Osthoff K., Bantel H., Hass R. Increased plasma levels of CK-18 as potential cell death biomarker in patients with HELLP syndrome. Cell Death Dis. 2013;4:e886.
- Kakinuma C., Takagaki K., Yatomi T., Nakamura N., Nagata S., Uemura A., Shibutani Y. Acute toxicity of an anti-Fas antibody in mice. Toxicol. Pathol. 1999;27:412–420.
- Osaka A., Hasegawa H., Yamada Y., Yanagihara K., Hayashi T., Mine M., Aoyama M., Sawada T., Kamihira S. A novel role of serum cytochrome c as a tumor marker in patients with operable cancer. J. Cancer Res. Clin. Oncol. 2009;135:371–377.
- Osaka A., Hasegawa H., Tsuruda K., Inokuchi N., Yanagihara K., Yamada Y., Aoyama M., Sawada T., Kamihira S. Serum cytochrome c to indicate the extent of ongoing tumor cell death. Int. J. Lab. Hematol. 2009;31:307–314.
- Barczyk K., Kreuter M., Pryjma J., Booy E.P., Maddika S., Ghavami S., Berdel W.E., Roth J., Los M. Serum cytochrome c indicates in vivo apoptosis and can serve as a prognostic marker during cancer therapy. Int. J. Cancer. 2005;116:167–173.
- Turiak L., Misjak P., Szabo T.G., Aradi B., Paloczi K., Ozohanics O., Drahos L., Kittel A., Falus A., Buzas E.I., Vekey K. Proteomic characterization of thymocyte-derived microvesicles and apoptotic bodies in BALB/c mice. J. Proteome. 2011;74:2025–2033.
- Holdenrieder S., Stieber P., Bodenmuller H., Busch M., Fertig G., Furst H., Schalhorn A., Schmeller N., Untch M., Seidel D. Nucleosomes in serum of patients with benign and malignant diseases. Int. J. Cancer. 2001;95:114–120.
- Oyama K., Fushida S., Kinoshita J., Okamoto K., Makino I., Nakamura K., Hayashi H., Inokuchi M., Nakagawara H., Tajima H., Fujita H., Takamura H., Ninomiya I., Kitagawa H., Fujimura T., Ohta T. Serum cytokeratin 18 as a biomarker for gastric cancer. Clin. Exp. Med. 2013;13:289–295.
- Yaman E., Coskun U., Sancak B., Buyukberber S., Ozturk B., Benekli M. Serum M30 levels are associated with survival in advanced gastric carcinoma patients. Int. Immunopharmacol. 2010;10:719–722.
- Bilici A., Ustaalioglu B.B., Ercan S., Seker M., Yilmaz B.E., Orcun A., Gumus M. The prognostic significance of the increase in the serum M30 and M65 values after chemotherapy and relationship between these values and clinicopathological factors in patients with advanced gastric cancer. Tumour Biol. 2012;33:2201–2208.
- Demiray M., Ulukaya E.E., Arslan M., Gokgoz S., Saraydaroglu O., Ercan I., Evrensel T., Manavoglu O. Response to neoadjuvant chemotherapy in breast cancer could be predictable by measuring a novel serum apoptosis product, caspase-cleaved cytokeratin 18: a prospective pilot study. Cancer Investig. 2006;24:669–676.
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