Lung Cancer-Associated Keratin 19 Fragments: Development and Biochemical Characterisation of the New Serum Assay Enzymun-Test<sup>®</sup> Cyfra 21–1

Bodenmüller, Heinz; Ofenloch-Hähnle, Beatus; Lane, E. Birgitte; Dessauer, Andreas; Böttger, Volker; Donié, F

doi:10.1177/172460089400900203

Cited by 60 publications

(33 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Detection of this Asp396 neo-epitope by the M30 ELISA is, therefore, only possible following caspasedependent apoptosis. It has been postulated that necrosis of epithelial cells releases full-length cytokeratins into the extracellular space following breakdown of the cell membrane (Bodenmüller et al, 1994). In vitro studies have shown that induction of necrosis in a human breast epithelial cell line is associated with a significant increase in the concentration of extracellular CK18, as recorded by the M65 ELISA (Kramer et al, 2004).…”

Section: Histopathological Assessment Of Necrosis and Apoptosis In Rementioning

confidence: 99%

Considerations for the use of plasma cytokeratin 18 as a biomarker in pancreatic cancer

et al. 2010

View full text Add to dashboard Cite

BACKGROUND: Enzyme-linked immunoassays of full-length (M65) and/or caspase-cleaved (M30) cytokeratin 18 (CK18) released from epithelial cells undergoing necrosis and/or apoptosis, respectively, may have prognostic or predictive biomarker utility in a range of solid tumour types. Characterisation of baseline levels of circulating full length and cleaved CK18 specifically in patients with pancreatic cancer. METHODS: Plasma samples from 103 patients with pancreatic cancer stored at À80 1C were assayed for M65 and M30 levels. The median (inter-quartile range (IQR)) duration of plasma storage was 34 (23 -57) months. Patients with metastatic disease (n ¼ 19) were found to have greater median (IQR) M65 levels (1145 (739 -1698) U l À1 ) compared with the locally advanced (n ¼ 20; 748 (406 -1150) U l À1 ) and resected (n ¼ 64; 612 (331 -987) U l À1 ) patients (P ¼ 0.002). Elevated M65 levels were associated with poorer overall survival on univariate (Po0.001) but not multivariate (P ¼ 0.202) analysis. M65 concentrations also exhibited significant associations with concurrent serum -bilirubin levels (Po0.001) and the duration of plasma storage (Po0.001). CONCLUSIONS: Baseline plasma CK18 levels in pancreatic cancer are affected by the presence of obstructive jaundice and prolonged plasma storage. Clinical biomarker studies utilising serial CK18 levels are warranted in pancreatic cancer, provided consideration is given to these potentially confounding factors. Pancreatic ductal adenocarcinoma is a leading cause of cancer deaths (http://www-dep.iarc.fr/) wherein, because of its characteristically late presentation, only 10 -15% of patients present with resectable tumours (Alexakis et al, 2004). For the remaining 85 -90% of patients presenting with inoperable disease, administration of systemic chemotherapy represents the most widely utilised palliative treatment modality (Yip et al, 2006). Conventional chemotherapy regimens incorporating either 5-FU or gemcitabine typically result in only modest improvement in overall survival (Sultana et al, 2007). One of the key challenges in accelerating improved cancer therapeutics is the identification of biomarkers to enable early objective assessment of tumour responses to chemotherapy (Kummar et al, 2007).Necrosis of malignant and normal epithelial cells cause release of cytokeratin 18 (CK18) (Kramer et al, 2004), a type I intermediate filament protein and a component of the intracellular cytoskeleton (Fuchs and Weber, 1994). Caspase-mediated cleavage of the CK18 contributes to the degradation of the intracellular cytoskeleton if epithelial cells undergo apoptosis (Leers et al, 1999). Enzymelinked immunosorbent assays (ELISAs) have been developed to measure circulating concentrations of both full-length CK18 (M65) and caspase-cleaved CK18 fragments (M30) (Kramer et al, 2004). Thus, the M65 ELISA reports necrotic and apoptotic epithelial cell death, whereas the M30 ELISA reports levels of epithelial apoptosis specifically (Kramer et al, 2004). Early studies suggest that these assays ...

show abstract

Section: Histopathological Assessment Of Necrosis and Apoptosis In Rementioning

confidence: 99%

Considerations for the use of plasma cytokeratin 18 as a biomarker in pancreatic cancer

et al. 2010

View full text Add to dashboard Cite

show abstract

“…Furthermore, a kit (CYFRA 21-1, Roche Diagnostics) is currently being marketed in some parts of Europe for specific detection of a lung cancer-associated K19 fragment (48,49). The assays used for these measurements have employed antibodies that recognize both the intact and the fragmented forms of K8, K18, and K19.…”

Section: Discussionmentioning

confidence: 99%

Cancer-associated Cleavage of Cytokeratin 8/18 Heterotypic Complexes Exposes a Neoepitope in Human Adenocarcinomas

Ditzel¹,

Strik²,

Kobæk-Larsen³

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

The intermediate filament network in simple glandular epithelial cells predominantly consists of heterotypic complexes of cytokeratin 8 (K8) and cytokeratin 18 (K18). In contrast to other cytokeratins, K8 and K18 are persistently expressed during malignant transformation, but changes in cell morphology are accompanied by alterations in the intermediate filament network. To study molecular changes, K8 and K18 were purified from surgically removed colon cancer and normal epithelia tissues. Western blotting and amino acid sequencing revealed the presence of abundant K8 and K18 fragments, truncated at the N terminus, from cancerous, but not normal, epithelial cells. The fragmentation pattern indicates proteolysis mediated by several enzymes, including trypsin-like enzymes. The cancer-associated forms of K8 and K18 are specifically recognized by the human antibody, COU-1, cloned from the B cells of a cancer patient. We demonstrate that COU-1 recognizes a unique conformational epitope presented only by a complex between K8 and K18. The epitope is revealed after proteolytic removal of the head domain of either K8 or K18. A large panel of recombinant K8 and K18 fragments, deleted N-or C-terminally, allowed for the localization of the COU-1 epitope to the N-terminal part of the rod domains. Using surface plasmon resonance, the affinity of COU-1 for this epitope was determined to be 10 9 M ؊1 , i.e. more than 2 orders of magnitude higher than for intact heterotypic K8/K18 complexes. The cellular distribution of truncated K8/K18 heterotypic complexes in viable adenocarcinomas cells was probed using COU-1 showing small fibrillar structures distinct from those of intact K8/K18 complexes. Previously we demonstrated the binding and subsequent internalization of recombinant Fab COU-1 to live cancer cells. We have thus characterized a cancer neoepitope recognized by the humoral immune system. The results have biological as well as clinical implications.The 10 nm intermediate filaments (IF) 1 provide resilience in response to mechanical stress by forming a stable network attached to specific desmosome cell-cell contacts (1). IF can be classified into groups expressed in higher eukaryotes in a tissue-specific and cell type-restricted pattern (2, 3). In epithelial cells, IFs consist of stoichiometrically equal amounts of type I (smaller and acidic) and type II (larger, neutral, or basic) cytokeratin polypeptides, which form strongly interacting heterodimers (4 -6). Cytokeratin 8 (K8) and cytokeratin 18 (K18), type I and II, respectively, are the major components of IFs of simple or single layer epithelia, such as those of the intestine, the liver, and the breast ducts, although cytokeratin 19 (K19) is also present and forms heterodimers with K8 (2, 5). The assembly of IF involves several association steps where the ␣-helical rod domain of the cytokeratin molecules plays the central role (7-9). The head and the tail domains are not thought to be part of the filamentous backbone but to protrude laterally and contribute to profilame...

show abstract

“…Selective antibodies raised against simple epithelium type cytokeratin, particularly the acidic (type I) subunit cytokeratin 19, have been shown to react with all histologies of lung cancers (Debus et al, 1984). A fragment of cytokeratin subunit 19 corresponding to epitope sequences lying within the sequences 311 -335 and 346 -367 (Bodenmüller et al, 1994) can be measured in serum by a sandwich assay, CYFRA 21-1, using two mouse monoclonal antibodies, KS 19-1 and BM 19 -21.…”

mentioning

confidence: 99%

CYFRA 21-1 is a prognostic determinant in non-small-cell lung cancer: results of a meta-analysis in 2063 patients

Molinier

Ébert³

et al. 2004

Br J Cancer

167

123

View full text Add to dashboard Cite

The purpose of this study was to determine the prognostic significance of a high pretreatment serum CYFRA 21-1 level (a cytokeratin 19 fragment) adjusted for the effects of well-known co-variables in non-small-cell lung cancer (NSCLC). This meta-analysis based on individual updated data gathered comprehensive databases from published or unpublished controlled studies dealing with the prognostic effect of serum CYFRA 21-1 level at presentation in NSCLC of any stage (nine institutions, 2063 patients). Multivariate regression was carried out with the Cox model. The proportional hazard assumption for each of the selected variables retained in the final model was originally checked by log minus log plots baseline hazard ratio. The follow-up ranged from 25 to 78 months. A total of 1616 events were recorded. In the multivariate analysis performed at the 1-year end point, a high pretreatment CYFRA 21-1 level was an unfavourable prognostic determinant in all centres except one (Hazard ratio (95% confidence interval): 1.88 (1.64 -2.15), Po10 À4 ). Other significant variables were stage of the disease, age and performance status. Within the first 18 months, the procedure disclosed a nearly similar hazard ratio for patients having a high pretreatment serum CYFRA 21-1 level (1.62 (1.42 -1.86), Po10 À4 ). For patients who did not undergo surgery, the hazard ratio during the first year of follow-up was 1.78 (1.54 -2.07), Po10À4 . Finally, in the surgically treated population, at the 2-year end point, a high pretreatment CYFRA 21-1 and a locally advanced stage remained unfavourable prognostic determinants. In conclusion CYFRA 21-1 might be regarded as a putative co-variable in analysing NSCLC outcome inasmuch as a high serum level is a significant determinant of poor prognosis whatever the planned treatment.

show abstract

Lung Cancer-Associated Keratin 19 Fragments: Development and Biochemical Characterisation of the New Serum Assay Enzymun-Test^® Cyfra 21–1

Cited by 60 publications

References 17 publications

Considerations for the use of plasma cytokeratin 18 as a biomarker in pancreatic cancer

Considerations for the use of plasma cytokeratin 18 as a biomarker in pancreatic cancer

Cancer-associated Cleavage of Cytokeratin 8/18 Heterotypic Complexes Exposes a Neoepitope in Human Adenocarcinomas

CYFRA 21-1 is a prognostic determinant in non-small-cell lung cancer: results of a meta-analysis in 2063 patients

Contact Info

Product

Resources

About

Lung Cancer-Associated Keratin 19 Fragments: Development and Biochemical Characterisation of the New Serum Assay Enzymun-Test® Cyfra 21–1

Cited by 60 publications

References 17 publications

Considerations for the use of plasma cytokeratin 18 as a biomarker in pancreatic cancer

Considerations for the use of plasma cytokeratin 18 as a biomarker in pancreatic cancer

Cancer-associated Cleavage of Cytokeratin 8/18 Heterotypic Complexes Exposes a Neoepitope in Human Adenocarcinomas

CYFRA 21-1 is a prognostic determinant in non-small-cell lung cancer: results of a meta-analysis in 2063 patients

Contact Info

Product

Resources

About

Lung Cancer-Associated Keratin 19 Fragments: Development and Biochemical Characterisation of the New Serum Assay Enzymun-Test^® Cyfra 21–1