CK-18 Fragments Research Articles

Association of human leukocyte antigen class II alleles with epithelial cell apoptosis and extracellular matrix production in acute COVID-19.

Pathogenic mechanisms and long-term consequences of COVID-19 require attention in studies on SARS-CoV-2. The association of the severity of COVID-19 with genetic factors, such as human leukocyte antigen (HLA) genes, remains underexplored. Our study assessed the relationships between HLA class II alleles and COVID-19 severity and blood-based indicators of systemic inflammation and organ damage, serum markers of epithelial cell apoptosis such as caspase-cleaved CK18 fragment M30 (CK18-M30) and the extracellular matrix product hyaluronic acid (HA). The study included 101 hospitalized COVID-19 patients (mean age 60 ±14 years). Clinical tests were performed at admission to the hospital. The levels of CK18-M30 and HA were detected in serum by enzyme-linked immunosorbent assay (ELISA). HLA typing was performed in HLA-DRB1, -DQA1, and -DQB1 loci by the polymerase chain reaction with low-resolution sequence-specific primers. Sixty-one patients had a non-severe and 40 had a severe or critical disease course (following the WHO definition). The severity was associated with older age, male gender, higher HA, CK18-M30, and some indicators of inflammation. Despite the lack of direct association between HLA alleles and the severity of COVID-19, the presence of HLA-DRB1*04 and 12 alleles in the genotype was associated with lowered or elevated HA, respectively. The HLA-DQB1*03:01 allele was associated with lowered CK18-M30, aspartate aminotransferase, and ferritin. In addition, HLA-DQB1*06:01 was associated with elevated alanine aminotransferase. Associations of HLA class II alleles with markers of epithelial cell apoptosis and extracellular matrix production indirectly support the influence of HLA genes on acute COVID-19 severity.

Surrogate markers in non-alcoholic steatohepatitis.

Hepatic steatosis with inflammation, inflated hepatocytes, and potential fibrosis defines non-alcoholic steatohepatitis (NASH), which can possibly lead to liver cirrhosis. Although liver biopsy is still the gold standard for diagnosing NASH, numerous non-invasive surrogate markers have been investigated to reduce the need for this invasive technique. In this review we present several currently assessed biomarkers, scores, and indexes in assessing NASH. A search in the main medical literature databases was conducted. We searched for observational studies evaluating non-invasive markers, scores, and panels in predicting NASH. Several proinflammatory markers, inflammation and apoptosis biomarkers, as well as complex models have been studied in predicting NASH. Proinflammatory markers include C-reactive protein, ferritin, tumor necrosis factor-α, interleukin-6, pentraxin-3, and neutrophil extracellular traps. Inflammation and apoptosis biomarkers include cytokineratin-18, adipocytokines, lipid oxidation panels, plasminogen activator inhibitor-1, and products of free radical-mediated oxidation of linoleic acid. Moreover, several studied complex models such as NashTest, NashTest-2, pairing CK18 fragments with other biomarkers such as ALT and the presence of MetS, the HAIR model, acNASH, NAFIC score, Visceral Adiposity Index have also been studied. A variety of diagnostic panels have shown good predictive values for diagnosing NASH. Nevertheless, non-invasive surrogate markers are currently unable to replace liver biopsy. However, their clinical significance is mainly in triaging patients for liver biopsy, reducing the financial burden associated with the procedure.

Assessment of Fatty Liver Diseases Based on Serum Ck18 Fragment, Gamma Glutamyl Transferase and Aspartate Aminotransfease /Alanine Aminotransferase Ratio

Assessment of Fatty Liver Diseases Based on Serum Ck18 Fragment, Gamma Glutamyl Transferase and Aspartate Aminotransfease /Alanine Aminotransferase Ratio

Changes in Cytokeratin 18 during Neoadjuvant Chemotherapy of Breast Cancer: A Prospective Study.

Background & Objective:Prediction of response to neoadjuvant treatment is an important part of treatment of patients with breast cancer. This study aimed to assess changes in serum levels of Cytokeratin 18 during neoadjuvant chemotherapy in patients with locally advanced breast cancer and its association with neoadjuvant treatments. Methods:This research was performed on newly diagnosed breast cancer patients referred to Omid Radiotherapy Center and radiotherapy and oncology departments of Emam Reza and Ghaem hospitals, in Mashhad, Iran. Serum levels of M30 and M65 fragments of Cytokeratin 18 were measured before and 24 hours after the first course of neoadjuvant chemotherapy. Changes in serum levels of Cytokeratin 18 and its fragments and their correlation with pathologic response were analyzed.Results:Pre- and post-chemotherapy levels of M30 were respectively 223.9±18.94 and 250.7±23.92 U/L (P=0.24). For M65, these levels were respectively 301.5±313.9 and 330.2±352.2 U/L (P=0.1). Changes in M30 level during chemotherapy in patients with and without pathologic complete response were -20±92.69 and 43.1±106.5, respectively (P=0.1). For M65, these changes were respectively -247±55 and 76±240 (P=0.1). Baseline levels of M30 and M65 had no relation with menopausal status, tumor grade, hormone receptor status, Ki67 expression, molecular subtype, and stage. Conclusion:Our findings showed statistically insignificant changes in the level of Caspase-cleaved- (M30) and uncleaved- (M65) cytokeratin 18 fragments (apoptotic and necrotic indicators, respectively) during neoadjuvant chemotherapy in patients with breast cancer. There was no notable relationship between tumor-related factors and either baseline levels or serum changes of CK18 fragments. Also, there was no correlation between M30/M65 level and pathologic response to neoadjuvant chemotherapy.

The Role of Noninvasive Tests for Differentiating NASH From NAFL and Diagnosing Advanced Fibrosis Among Patients With NAFLD.

Assessing for the presence of non-alcoholic steatohepatitis (NASH) and the presence of advanced fibrosis is vital among patients with non-alcoholic fatty liver disease (NAFLD) as each is predictive of disease outcomes. A liver biopsy is the gold standard method for doing so but is impossible to perform among all patients with NAFLD. Reliable methods for noninvasively detecting for the presence of NASH and advanced fibrosis are thus a pressing need. The search for noninvasive tests has been more successful for advanced fibrosis than for NASH. Clinical prediction models and elastography have acceptable accuracy for ruling out advanced fibrosis; when used together, as in a fibrosis prediction algorithm presented in this review, it can avoid the need for liver biopsy among many patients with NAFLD. Several biomarkers for identifying the presence of NASH have been studied but none are sufficiently accurate or validated. Of those studied, the most promising include CK-18 fragments, lipodomic and metabolomics candidates, and magnetic resonance elastography with proton density fat fraction. However, none are ready for clinical use and ultimately large multicenter prospective cohort studies are needed to validate select novel biomarkers.

Genetic determinants of hepatic steatosis and serum cytokeratin-18 fragment levels in Taiwanese children.

There are substantial genetic components contributing to the susceptibility of nonalcoholic fatty liver disease (NAFLD). It has recently been reported that the rs641738 C>T variant in the membrane-bound O-acyltransferase domain-containing protein 7 (MBOAT7) gene increased severity of NAFLD in adults of European descent. We aimed to test the hypothesis that MBOAT7 rs641738 variant would increase hepatic steatosis and hepatocellular injury in obese children. A total of 831 obese children aged 7-15years were recruited. Hepatic steatosis was measured by ultrasonography. Because PNPLA3 rs738409, GCKR rs780094 and TM6SF2 rs58542926 variants are known to confer susceptibility to NAFLD, we assessed the influence of MBOAT7 rs641738 on hepatic steatosis, and serum levels of CK-18 fragment (a biomarker of hepatocellular injury and apoptosis for NAFLD) after adjusting the effects of PNPLA3, GCKR and TM6SF2 polymorphisms. Of the recruited obese children, 22.7% had hepatic steatosis. PNPLA3 rs738409, GCKR rs780094 and TM6SF2 rs58542926 variants were independent risk factors of hepatic steatosis and elevated ALT levels. In contrast, MBOAT7 rs641738 variants, neither heterozygous nor homozygous genotypes, were not associated with hepatic steatosis, insulin resistance, lipid levels and liver enzymes. The multiple linear regression model revealed that after adjusting for age, gender, body mass index z score, PNPLA3 rs738409 and GCKR rs780094 variants, but not MBOAT7 rs641738, were associated with serum levels of CK-18 fragment. The variant MBOAT7 rs641738 genotype is not associated with hepatic steatosis and serum levels of CK-18 fragment in obese Taiwanese children.

Serum Cell Death Biomarker Mirrors Liver Cancer Regression after Transarterial Chemoembolisation

Background Hepatocellular carcinoma (HCC) represents an increasing health problem with limited therapeutic options. In patients with intermediate disease stage, transarterial chemoembolisation (TACE) is widely applied. Treatment response is routinely assessed by imaging techniques according to the international response evaluation criteria in solid tumours (RECIST), which consider tumour regression or additionally tumour necrosis (modified RECIST). Evaluation of treatment response, however, by these methods is time- and cost-intensive and usually performed at earliest several months following TACE. Aim To investigate the suitability of novel non-invasive cell death biomarkers for an earlier prediction of TACE response. Methods We analysed activation of pro-apoptotic caspases and the proteolytic cleavage of the caspase substrate CK-18 in liver tissues and sera from HCC patients by immunohistochemistry, a luminometric substrate assay and ELISA. Results Both caspase activity and caspase-cleaved CK-18 fragments were elevated in HCC patients compared to healthy controls. CK-18 serum levels significantly increased during the first 3 days and peaked at day two following TACE. Interestingly, we found significant differences in CK-18 levels between patients with and without tumour regression. Detection of CK-18 fragments revealed a promising performance for the early prediction of TACE response with an area under the curve value of 0.76. Conclusions Caspase-cleaved CK-18 levels mirror liver cancer regression and allow an earlier prediction of TACE response. The concordance with mRECIST suggests that the detection of CK-18 levels immediately after TACE might be used as a short-term decision guide to continue or change HCC therapy.

Serum cell death biomarker mirrors liver cancer regression after transarterial chemoembolisation.

Hepatocellular carcinoma (HCC) represents an increasing health problem with limited therapeutic options. In patients with intermediate disease stage, transarterial chemoembolisation (TACE) is widely applied. Treatment response is routinely assessed by imaging techniques according to the international response evaluation criteria in solid tumours (RECIST), which consider tumour regression or additionally tumour necrosis (modified RECIST). Evaluation of treatment response, however, by these methods is time- and cost-intensive and usually performed at earliest several months following TACE. To investigate the suitability of novel non-invasive cell death biomarkers for an earlier prediction of TACE response. We analysed activation of pro-apoptotic caspases and the proteolytic cleavage of the caspase substrate CK-18 in liver tissues and sera from HCC patients by immunohistochemistry, a luminometric substrate assay and ELISA. Both caspase activity and caspase-cleaved CK-18 fragments were elevated in HCC patients compared to healthy controls. CK-18 serum levels significantly increased during the first 3 days and peaked at day two following TACE. Interestingly, we found significant differences in CK-18 levels between patients with and without tumour regression. Detection of CK-18 fragments revealed a promising performance for the early prediction of TACE response with an area under the curve value of 0.76. Caspase-cleaved CK-18 levels mirror liver cancer regression and allow an earlier prediction of TACE response. The concordance with mRECIST suggests that the detection of CK-18 levels immediately after TACE might be used as a short-term decision guide to continue or change HCC therapy.

Necrotic cell death and suppression of T-cell immunity characterized acute liver failure due to drug-induced liver injury

Background & aims: The aim of this study was to investigate the clinical characteristics and pathophysiology of drug-induced liver injury (DILI) – acute liver failure (ALF). Methods: The patients with acute liver injury (ALI) including ALF from 2009 to 2014 were analyzed. The hepatic encephalopathy (HE) development rate was compared with the findings from a national survey in Japan. The serum cytokines levels and the findings of a liver function test were evaluated in the DILI patients. Results: The HE development rate substantially decreased for autoimmune hepatitis (AIH) – and undetermined cause-induced ALI owing to the early prediction system, but not in DILI-ALI. Among the DILI-ALF and AIH-ALF cases, the CK-18 fragment (1480.1U/L, 3945.4U/L), IL-8 (82.9pg/mL, 207.5pg/mL), IP-10 (1379.6pg/mL, 3731.2pg/mL) and MIP-1β (1017.7pg/mL, 2273.3pg/mL) levels were lower in the DILI-ALF cases. Among the DILI-ALI and DILI-ALF cases, IL-4 (19.8pg/mL, 25.4pg/mL) and RANTES (14028.0pg/mL, 17804.7pg/mL) were higher in DILI-ALI, and HMGB-1 (397.1pg/μL, 326.2pg/μL) and HGF (2.41ng/mL, 0.55ng/mL) were higher in DILI-ALF. We observed that HGF independently associated with DLI-ALF development. Conclusions: Despite the low grade apoptosis and inflammation, DILI patients progressed to ALF comparable with that of the AIH patients.

Cytokeratin (CK)-18 and CK18 Fragments As Sensitive and Specific Biomarkers of Acute Hepato-Intestinal Gvhd: Results of a Prospective Observational Study

Introduction:Preliminary data have suggested that CK18, a highly abundant protein in liver and gut epithelial cells, and its apoptotic degradation product CK18 fragments (CK18F) could indicate apoptotic end organ damage induced by acute hepato-intestinal GVHD and, thus, may be used as an acute GVHD biomarker (Luft et al, Blood 2011; 118:1685). Here, we present the first results of a clinical study aiming at prospective validation of the capacity of CK18/ CK18F to predict acute hepato-intestinal GvHD activity and its response to treatment.Patients and methods:Main endpoints of the study were (1) prediction of imminent acute hepato-intestinal GvHD by comparing CK18/CK18F serum levels measured 7 to 14 days prior to symptom onset to with baseline levels (pre-conditioning, at transplant and one week after alloSCT) of the same subject; and (2) prediction of response to GVHD treatment by measuring CK18/CK18F kinetics 3, 7 and 14 days after start of immunosuppressive therapy for hepato-intestinal GvHD. Eligible were all adult patients undergoing allogeneic stem cell transplantation (alloSCT) at our institution from December 2008 onwards. Target sample size was 100. Blood samples were collected in the study population prior to the initiation of the conditioning regimen and from the day of transplantation on a weekly basis for one year. The M30-Apoptosense ELISA kit was used for measuring serum levels of CK18 fragments, CK18 was analyzed with the M65 (EpiDeath) ELISA Kit (both provided by PEVIVA, Nacka, Sweden). Serum levels measured at the pre-specified landmarks were compared by Wilcoxon's signed rank test. The study was registered at ClinicalTrials.gov (Identifier: NCT00935324).Results:Between December 2008 to February 2011, 109 patients were enrolled. The median age was 53 years (range 19 - 69), 72 were male. The underlying disease was acute myeloid leukemia in 39 patients, myeloproliferative / myelodysplastic disorders in 18 patients, acute lymphoblastic leukemia in 10 patients, and lymphoproliferative malignancies in 42 patients. Donors were matched related (36), matched unrelated (52), or mismatched unrelated (21). Stem cell source was peripheral blood in 100 patients and bone marrow in 9 patients. Reduced intensity conditioning protocols (n = 89) prevailed over myeloablative (n = 7) and aplasia conditioning protocols (n = 13). Immunosuppression was based on a calcineurin inhibitor (CNI) backbone with mycophenolate or short-course methotrexate, plus ATG in mismatched or unrelated transplants. Patients received pravastatin (20 mg daily dose) for routine prophylaxis of CNI-induced vascular complications as per institutional policy as of 2010.Acute hepato-intestinal GVHD grade II-IV developed in 38 patients (34,9 %) at a median of 53 days post transplant and was steroid-refractory in 12 patients. In patients with biopsy-proven acute hepato-intestinal GVHD, CK18 significantly raised 7 to 14 days before clinical GVHD onset and can therefore serve as an early predictor of impending acute hepato-intestinal GVHD (median at baseline vs preGVHD, 149 ng/ml vs 527 ng/ml; p=0.012), and peaked at symptom onset (median 1281 ng/ml; p=0.003, compared to baseline). CK18F kinetics paralleled that of CK18 but was less pronounced. In contrast, patients with isolated acute skin GVHD, patients without acute GVHD (cut-off day+53), and patients with GVHD-typical symptoms but a negative biopsy did not show any increase CK18 or CK18F in serum (Figure 1). However, CK18/CK18F levels were not correlated with acute GVHD severity. Neither marker predicted refractory GVHD or non-relapse mortality after acute GVHD at any of the time points measured.Conclusions:This prospective study validates CK18 and CK18F as specific biomarkers suitable for prediction and diagnosis of suspected imminent and clinically manifest acute hepato-intestinal GVHD. In contrast, neither CK18 nor CK18F can be used to predict the prognosis of an ongoing GVHD episode. Interventional trials employing CK18 as trigger for pre-emptive GVHD therapy seem to be warranted. [Display omitted] DisclosuresHegenbart:Janssen: Honoraria, Other: travel support. Ho:Genzyme/Sanofi-Aventis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees. Luft:Immundiagnostik AG: Research Funding.

Panel of MiRNAs -122,-192 and -21 correlate with CK-18 fragment level in serum and hold potential as non-invasive NASH biomarkers

Panel of MiRNAs -122,-192 and -21 correlate with CK-18 fragment level in serum and hold potential as non-invasive NASH biomarkers

Serum levels of chemokines CCL4 and CCL5 in cirrhotic patients indicate the presence of hepatocellular carcinoma.

Background:Most hepatocellular carcinomas (HCCs) are diagnosed at an advanced stage. The prognostic value of serum tumour markers alpha-fetoprotein (AFP) and des-gamma-carboxy prothrombin (DCP) is limited. The aim of our study is to evaluate the diagnostic value of serum growth factors, apoptotic and inflammatory mediators of cirrhotic patients with and without HCC.Methods:Serum samples were collected from cirrhotic potential liver transplant patients (LTx) with (n=61) and without HCC (n=78) as well as from healthy controls (HCs; n=39). Serum concentrations of CRP, neopterin and IL-6 as markers of inflammation and thrombopoietin (TPO), GCSF, FGF basic and VEGF, HMGB1, CK-18 (M65) and CK18 fragment (M30) and a panel of proinflammatory chemokines (CCL2, CCL3, CCL4, CCL5, CXCL5 and IL-8) were measured. Chi square, Fisher exact, Mann–Whitney U-tests, ROC curve analysis and forward stepwise logistic regression analyses were applied.Results:Patients with HCC had higher serum TPO and chemokines (P<0.001 for TPO, CCL4, CCL5 and CXCL5) and lower CCL2 (P=0.008) levels than cirrhotic patients without HCC. Multivariate forward stepwise regression analysis for significant parameters showed that among the studied parameters CCL4 and CCL5 (P=0.001) are diagnostic markers of HCC. Serum levels of TPO and chemokines were lower, whereas M30 was significantly higher in cirrhotic patients than in HCs.Conclusions:High serum levels of inflammatory chemokines such as CCL4 and CCL5 in the serum of cirrhotic patients indicate the presence of HCC.

Association of Serum Cytokeratin-18 Fragment Concentration in Patients with Different Types of Nonalcoholic Fatty Liver Disease

Nonalcoholic fatty liver disease (NAFLD) is one of the most common forms of chronic liver diseases worldwide. The prevalence of NAFLD was 15% in the Asian population. In a study, NASH was observed in 42.4% of NAFLD cases in Bangladesh. It is a serious clinical problem because of its worldwide distribution and potential adverse sequel. Cytokeratin-18 fragment is a marker of hepatocyte apoptosis. Measurement of serum cytokeratin-18 fragment concentration is a noninvasive, quick and simple procedure that can be carried out easily in peripheral hospital. It can be very much helpful for early detection and frequent monitoring of disease progression and response to therapy. This cross sectional study was conducted in the Department of Clinical Pathology in collaboration with Department of Hepatology and Department of Pathology, BSMMU, Dhaka from March 2014 to February 2015. Forty patients who fulfilled the criteria of NAFLD were determined in our study. The concentration of serum CK-18 fragment was measured to evaluate its association in different types of NAFLD by enzyme linked immune sorbent assay (ELISA). In this study, we observed that the serum CK-18 fragment concentration was increased in relation to Nonalcoholic fatty liver disease Activity Score (NAS) (r=+0.7134, P<0.05). To assess diagnostic performance of CK-18 fragment were used receiver-operator characteristic (ROC) curve. The area under ROC was 0.918 for identification of NASH. Using a cutoff value of 150 U/L the sensitivity and specificity of CK-18 fragment were 85.7% and 80.8% respectively. The positive predictive value, negative predictive value and accuracy of CK-18 fragment were 66.67%, 90.91% and 80.00% respectively. Our data revealed that serum CK-18 fragment was strongly associated with Nonalcoholic fatty liver disease Activity Score (NAS). Serum CK-18 fragment may be a useful noninvasive tool for assessing NASH in patients with NAFLD.

Su1045 The Combined Ck-18 Fragments and Noninvasive Scoring Systems Do Not Perform Better Than Those of Scoring System Alone in Identifying NASH

Su1045 The Combined Ck-18 Fragments and Noninvasive Scoring Systems Do Not Perform Better Than Those of Scoring System Alone in Identifying NASH

Zytokeratin-18 als Marker zur nichtinvasiven Diagnostik und Prognose akuter und chronischer Lebererkrankungen

Currently liver biopsy represents the gold standard to assess severity and fibrosis grade in liver diseases. Since this laborious, costly, and invasive procedure is associated with possible complications, non-invasive methods and biomarkers, which allow for an easy, reliable, and repeatable assessment of liver disease are warranted. Cytokeratin (CK) 18 is an intermediary filament protein, expressed in hepatocytes, which is proteolytically cleaved during liver damage. The resultant CK-18 fragments are released by hepatocytes and can be detected in serum. A selective literature search in PubMed for original publications about the detection of CK-18 cell death markers in liver diseases was undertaken. Assessment of CK-18 cell death biomarkers allows for the early detection of liver damage in acute and chronic liver diseases. This is even feasible when transaminases are in the normal ranges. Detection of CK-18 biomarkers can also hint at disease activity and severity. For example, patients with non-alcoholic steatohepatitis exhibit elevated serum cell-death markers compared to those with simple steatosis. Furthermore, in patients with relevant fibrosis higher CK-18 values are found as compared to those with low fibrosis. In acute liver failure, cell death biomarkers may assist decision finding for the necessity of liver transplantation. Due to promising results of various studies, CK-18 cell death markers could be applied in clinical routine soon.

A first-in-human phase I study of oral pan-CDK inhibitor BAY 1000394 in patients with advanced solid tumors: Dose escalation with an intermittent 3 days on/4 days off schedule.

3012 Background: BAY 1000394 (BAY) is an oral pan-CDK inhibitor targeting CDKs 1,2,4, 7 and 9 in the low nanomolar range. A phase I dose escalation study was initiated to determine the maximum tolerated dose (MTD), pharmacokinetics (PK), and pharmacodynamics (PD) in patients (pts) with advanced solid tumors. Methods: BAY was administered twice daily in a 3 days on / 4 days off schedule (cycle length 21 days, 3+3 design). PK was evaluated on cycle 1 day 1 and day 10. Response rate was assessed according to RECIST 1.1. PD markers included CK18 fragments in plasma. Results: As of Jan 08 2011, 34 pts were treated at doses of 0.6 (3 pts), 1.2 (4), 2.4 (3), 4.8 (3), 9.6 (3), 19.2 (6) mg per day as oral solution and at doses of 10 (4), 15 (6) and 20 (2) mg per day as tablet. Tumor types included 10 colorectal, 4 mesothelioma and 20 others. Cohort 9 (20 mg tablet) is ongoing. Frequent CTCAEv4 grade 1/2 drug related AEs occurring in more than 25% of patients up to cohort 8 were asthenia, diarrhea, nausea, vomiting and anorexia. DLTs (grade 3, 1 pt each) were hyponatremia, aphtous stomatitis at 19.2 mg solution and arterial thrombosis at 15 mg tablet. Aphthous stomatitis (20%) has not been observed with the tablet formulation. Other grade 3 related AEs were asthenia in 2 and nausea and vomiting in one pt each. Nausea and vomiting on treatment days were observed despite antiemetic treatment (aprepitant +/- setron). PK was dose proportional up to 9.6 mg, T1/2 was 10 hours, and relative bioavailability of tablet formulation was excellent; major metabolite levels were low (&lt;10%). Levels of CK18 fragments did not correlate with dose or tumor response. Stable disease (SD) lasting for 2-4 months was observed in 9 patients, among others in 4 of 4 mesothelioma and 2 of 2 ovarian pts. One additional pt with cholangiocarcinoma has ongoing SD lasting for 5 months. One of the ovarian pts had a significant decline of CA125 lasting for 3 months. Conclusions: The tablet formulation of BAY 1000394 was better tolerated than oral solution. So far, doses up to a 15 mg per day with concomitant antiemetic treatment showed an acceptable tolerability. SD was observed in 10 of 25 heavily pretreated pts across cohorts 3 – 8.

A Phase 2, Randomized, Double-Blind, Placebo-Controlled Study of GS-9450 in Subjects With Nonalcoholic Steatohepatitis

In nonalcoholic steatohepatitis (NASH), the extent of hepatocyte apoptosis correlates with disease severity. Reducing hepatocyte apoptosis with the selective caspase inhibitor GS-9450 has a potential for altering the course of the liver disease. In this phase 2, double-blind study, 124 subjects with biopsy-proven NASH were randomized to once-daily placebo or 1, 5, 10, or 40 mg GS-9450 for 4 weeks. Absolute and percent changes from baseline in ALT levels, AST levels, and caspase-3-cleaved cytokeratin (CK)-18 fragments at week 4 were assessed by an analysis of covariance model with adjustment for baseline values. In the 40-mg group, mean (SD) ALT decreased by 47 (43) U/L from baseline to week 4 (P < 0.0001 versus placebo), and the proportion of subjects with normal ALT increased from 0% to 35% at week 4. In the 40-mg group, mean AST decreased by 13 U/L from baseline (not significant), and the proportion with normal AST increased from 20% at baseline to 48% at week 4. By week 4, mean CK-18 fragment levels had decreased to 393 (723) U/L in the GS-9450 10-mg group and 125 (212) U/L in the 40-mg group, but these reductions were not statistically significant. No serious adverse events were reported during treatment, and the percentage of subjects with at least one treatment-emergent grade 3 or 4 laboratory abnormality ranged from 11.5% to 17% across the GS-9450 treatment groups versus 35% in the placebo group. GS-9450 treatment induced significant reductions in ALT levels in NASH patients. Reductions in CK-18 fragment levels also occurred, although they were not statistically significant. At appropriate therapeutic indices, selective caspase inhibitors may be a promising treatment option in patients with NASH.

Enteric-coated cysteamine for the treatment of paediatric non-alcoholic fatty liver disease

Non-alcoholic fatty liver disease (NAFLD) is a common cause of liver disease in children. Hepatic fat accumulation and oxidative stress contribute to its pathogenesis. Cysteamine bitartrate readily traverses cellular membranes and is a potent antioxidant. To evaluate the safety and efficacy of enteric-coated (EC) cysteamine in children with NAFLD. Children, aged ≥10 y, meeting screening criteria with biopsy-proven NAFLD and serum ALT ≥60 IU/L, received twice-daily EC-cysteamine for 24 weeks. Monthly ALT, AST, body mass index (BMI) and gastrointestinal symptom scores were measured. Subjects with >50% reduction or normalisation of ALT achieved the primary endpoint. Of the 13 children enrolled (mean age 14.0 years), 11 completed EC-cysteamine therapy (mean dose 15.2 mg/kg/day) and were included in the final analysis. For these 11 subjects, the mean ALT levels at baseline and 24 weeks were 120.2 and 55 IU/L respectively (P = 0.002), and the AST levels were 60 and 36 IU/L respectively (P = 0.007). The primary endpoint was reached in 7 and normalisation (≤40 IU/L) of ALT in 5. After 24 week therapy, mean adiponectin levels increased (P = 0.009) and CK-18 fragment levels decreased (P = 0.013), insulin levels remained unchanged (P = 0.99). Mean leptin levels were decreased in responders (P = 0.044). Mean BMI was 34.5 at baseline and 34.2 kg/m(2) after treatment (P = 0.35). Mean symptom scores at baseline (1.1) and at 24 weeks (0.7) were similar. No major adverse events were reported. Enteric-coated cysteamine reduces ALT and AST levels in children with NAFLD without reduction in body mass index. Further studies will evaluate optimal cysteamine therapeutic dose and effect on liver histology in NAFLD (Clinicaltrials.gov protocol ID: 07-1699).

Significance of Cytokeratin Fragment M65 and Cytokines IL6, IL8 and IL17A in Bone Marrow Aspirates of Colorectal Cancer Patients

Aims: Soluble cytokeratin (CK) fragments and inflammatory interleukins (ILs) in bone marrow (BM) aspirates of colorectal cancer (CRC) patients are expected to indicate presence of disseminated tumor cells (DTCs) and anticancer response of the host, respectively. The present study investigated the relations of CK18 fragment M65, IL6, IL8, and IL17A in BM samples to the presence of DTCs and prognosis.

An apoptosis panel for nonalcoholic steatohepatitis diagnosis

The extrinsic death receptor-mediated pathway of apoptosis is involved in nonalcoholic steatohepatitis (NASH) development. Our aims were to create and validate a noninvasive prediction model for NASH diagnosis based on specific circulating markers of apoptosis. Our initial cohort consisted of 95 consecutive patients undergoing a liver biopsy for clinically suspected NASH. Blood was obtained from each patient at the time of liver biopsy. Plasma caspase 3 generated cytokeratin-18 fragments (CK-18), soluble Fas (sFas), and soluble Fas ligand (sFasL) were measured. Histology was assessed by an experienced hepatopathologist. The validation cohort consisted of 82 consecutive patients that underwent liver biopsy at the time of bariatric surgery. Patients with NASH had significantly higher levels of CK-18 and sFas than patients in the "not NASH" group [median (25th, 75th percentile): 508 (280, 846) U/L versus 176 (131, 224) U/L (p<0.001), and 11.8 (7.8, 12.5) ng/ml versus 5.9 (4.8, 8.3) ng/ml (p<0.001), respectively]. A significant positive correlation was revealed between the apoptosis markers and liver histopathology independent of other metabolic factors. A prediction model was generated including CK-18 fragments and sFas levels that showed an AUC of 0.93 and 0.79 in the initial and validation cohorts, respectively. A cutoff value using this model predicted NASH with a sensitivity and specificity of 88% and 89%, respectively. Quantification of circulating levels of two apoptotic markers accurately predicts the presence of NASH, supporting the potential usefulness of these markers in clinical practice for noninvasive diagnosis of NASH.