Histological remission has arisen as the optimal treatment outcome in ulcerative colitis (UC). The aim of this retrospective study was to explore the diagnostic performance of the noninvasive fecal biomarkers calprotectin (FC) and lactoferrin (FL) compared to the histological indices Nancy Index (NI) and Riley Index (RI).This study is a retrospective diagnostic accuracy study based on secondary analysis of patient data from 2002 to 2017 extracted from medical registries of our clinics in Essen-Mitte, Germany. Patients with UC underwent a colonoscopy, with biopsies taken from the rectum and the sigmoid scored by 2 experienced pathologists according to NI and RI and provided a stool sample within 7 days pre- or post-colonoscopy. Diagnostic accuracy of recommended cutoffs for FC (>50 μg/g) and FL (≥7.25 μg/g) were tested against our reference standard (NI ≥2) in terms of specificity, sensitivity, positive predictive value, negative predictive value, and accuracy (effectiveness).The number of patients with UC recruited was n = 226, aged 45.2 (SD 13.3). Histological indices were highly correlated (r = 0.980, P < 0.001). Fecal biomarkers correlated moderately with NI (FC: r = 0.383, P < 0.001; FL: r = 0.420, P < 0.001) and RI (FC: r = 0.395, P < 0.001; FL: r = 0.424, P < 0.001). Fecal biomarker concentrations were increased in patients with active histological disease (NI ≥2), median [IQR], FC 69.72 [20.07-254.38], FL 18.59 [6.06-44.42], compared to those with inactive disease (NI ≤1), FC 12.35 [3.89 - 32.16], FL 3.14 [0.75-11.05], z = -6.60, P < 0.001. Fecal biomarker concentrations differed significantly across NI grades 0-4 (FC: H4 = 45.2; FL: H4 = 47.5, both P < 0.001). Patients with grade 0 had significantly lower concentrations of fecal biomarkers than those with grade 3 (median; FC 10.94 vs 72.22; FL 2.30 vs 29.10; both P < 0.001) or grade 4 (FC 10.94 vs 67.00; FL 2.30 vs 27.64; both P < 0.001), as well as grade 2 for FC only (10.94 vs 56.22, P = 0.001). Concentrations were also lower in patients with grade 1 compared to those with grade 3 (FC 17.49 vs 72.22; FL 4.24 vs. 29.10; both P ≤ 0.001) or grade 4 (FC 17.49 vs 67.00; FL 4.24 vs 27.64; both P < 0.001).Receiver operating characteristics area under the curve showed moderate diagnostic accuracy for both FC 0.76 (95% confidence interval [CI] 0.70-0.83) and FL 0.73 (95% CI 0.66-0.80). Optimized cutoffs for both FC (≥34.29) and FL (≥5.85 μg/g) had slightly improved accuracy, compared with the manufacturer's cutoffs (FC: 69.9% vs 65.9%; FL: 71.7% vs 69.0%).Fecal biomarkers calprotectin and lactoferrin correlate with histological disease activity and differentiate between patients in histological remission from those with evidence of moderate to severe disease activity. Their noninvasiveness, in addition to being inexpensive, supports their use in the clinical monitoring of patients with UC.
Introduction: Cold agglutinins (CA) are commonly found cold reacting autoantibodies which reversibly interact with red blood cell antigens at low temperatures. Unsuspected complications such as hemolysis and intravascular coagulation can occur in patients with high titer and high termal amplitude undergoing on-pump cardiac surgery. We determined the incidence of CA in all patients undergoing on-pump cardiac surgery to verify the clinical relevance.
Colorectal cancer is one of the leading malignancies and still accounts for almost 25 000 deaths in Germany each year. Although there is accumulating data on the molecular basis, treatment and clinical outcome of patients within clinical trials evidence from the real-world setting is mostly lacking. We started the molecular registry trial Colopredict Plus in 2013 to collect clinical and molecular data from a real-world cohort of patients with early colon cancer stage II and III in 70 German colon cancer centers focusing on the prognostic impact of high microsatellite instability. In this interim report, we characterize a clinical cohort of 2615 patients, of whom 1787 tissue probes were analyzed. Microsatellite status was assessed using immunhistochemistry and fragment length analysis, with a concordance of 91.4 %. These established histopathological methods are sensitive and cost-effective. The median age was 72 years, significantly higher compared to clinical trial populations, with a median Charlson Comorbidity Index of 3. The stage-dependent incidence of microsatellite instability was 23.7 % and was associated with female gender, BRAF-mutation, UICC stage II and localization in the right colon. Survival calculated in disease free, relapse free and overall survival significantly differed between MSI-H and MSS, in favor of MSI-H patients. Multivariate age-adjusted analyses of relapse-free survival, disease-free survival, and overall survival highlighted microsatellite instability as a robust and positive prognostic marker for early colon cancer independent of age.Das kolorektale Karzinom ist weiterhin eine der häufigsten Tumorerkrankungen mit einer Mortalität von fast 25 000 pro Jahr in Deutschland. Obwohl wir mehr und mehr Daten zur molekularen Basis, zu neuen Behandlungsmethoden und Verläufen von Patienten im Rahmen klinischer Studien generieren, gibt es nur wenige Informationen aus der klinischen Versorgungsrealität. Wir starteten die molekulare Registerstudie Colopredict Plus im Jahre 2013, um klinische sowie molekulare Daten von einer „Real-World“-Kohorte von Patienten mit Kolonkarzinomen im UICC-Stadium II und III aus 70 deutschen Darmkrebszentren zu dokumentieren. Der Fokus liegt hier auf dem prognostischen Wert einer hochgradigen Mikrosatelliteninstabilität. In dieser Interimsanalyse charakterisieren wir das klinische Kollektiv von 2615 Patienten, von denen 1787 Tumorproben analysiert wurden. Der Mikrosatellitenstatus wurde via Immunhistochemie und Fragmentlängenanalyse mit einer Konkordanz von 91,4 % durchgeführt, was eine kosteneffektive histopathologische Nachweismethodik darstellt. Das mediane Alter lag bei 72 Jahren und damit deutlich höher als üblicherweise in klinischen Studien, der mediane Charlson Comorbidity Index lag bei 3. Die stadienabhängige Mikrosatelliteninstabilität-Inzidenz lag bei 23,7 % und war assoziiert mit dem weiblichen Geschlecht, BRAF-Mutationen, UICC-Stadium II und der Primärtumorlokalisation im rechten Kolon. Das Überleben berechnet in krankheitsfreiem, rezidivfreiem sowie Gesamtüberleben unterschied sich signifikant zugunsten der MSI-H-Patienten. Multivariate altersadjustierte Analysen des rezidivfreien und krankheitsfreien Überlebens sowie des Gesamtüberlebens bestätigten eine hochgradige Mikrosatelliteninstabilität als robusten und positiven prognostischer Marker für frühe Kolonkarzinome unabhängig vom Alter.
e15086 Background: High microsatellite instability (MSI-H) is a prognostic marker in resected colon cancer (CC) identified in post-hoc analysis of multiple trials. However, validation in real-life cohorts and its association with clinical and molecular markers is lacking. Methods: In Sep 2013 we started a platform trial in 49 German cancer centers recruiting patients with UICC stage II and III CC diagnosed between 2008 to 2016. MSI was tested by immunohistochemistry (IHC) of mismatch repair proteins MLH1, MSH2, MSH6 and PMS2. In case of any loss of protein expression (IHC neg), fragment length analysis (FLA) was performed, defining MSI high tumors (MSI-H) and MS stable tumors (MSS). Besides, mutations in known prognostic factors for CC such as RAS, BRAF, PI3K and others were determined by next generation sequencing (NGS). Results: By end of 2016, 1249 patients have been recruited into the trial: median age 73yrs., stage II/III: 686/563 pts. So far, tissue was analysed in 512 pts. Of these, 182 pts. were IHC neg with 116 pts. tested MSI-H upon FLA (22.7%). Median age was 73yrs., female/male: 260/252 pts., stageII/III: 292/220 pts. Association of MS status with clinical factors is shown in Table 1. Upon NGS analysis we found 16.3% BRAF mutations, 39% KRAS mutations, 2.8% NRAS mutations and 22.6% PI3K mutations. MSI-H status was significantly associated with BRAF mutation and wildtype status of RAS. Conclusions: We found a higher percentage of MSI-H cancers in our registry compared to reported data from randomised trials, possibly related to a higher median age in this real-life cohort. MSI-H was associated with female sex, primary tumor site, and distinct molecular markers and should therefore be considered a heterogeneous subgroup of CC. Updated analysis including NGS data and survival will be presented at the meeting. Clinical trial information: AIO- KRK-0413, DKRS:00004305. [Table: see text]
e15024 Background: We recently demonstrated improved objective response rate in untreated all-RAS wildtype mCRC with the addition of the anti-EGFR-antibody pmab to FOLFOXIRI. In this subgroup analysis we focused on histopathological response as a predictive marker for PFS. Additionally we analyzed chemotherapy induced steatosis hepatitis (CASH) in both treatment arms. Methods: Tissue samples from pts. achieving secondary resection of liver metastases in VOLFI were analyzed. We defined a cut-off for very good histopathological response at 20% of residual tumor cells in proportion to the total tumor area. For CASH sinusoidal obstructive syndrome, ballooning, steatosis, cholestasis, fibrosis and inflammation were determined. PFS was estimated using LIFETEST procedure. Results: Tissue of 14/19 resected pts. was evaluable (pmab-FOLFOXIRI/FOLFOXIRI: 11/3). All showed partial remission by RECIST. Median age was 56 yrs. (32–67), male/female: 7/7. All primary tumors were located in the left colon. Molecular analysis detected 1 BRAF (V600E) mutation and 1 MSI-H tumor. Median treatment duration until resection in this cohort: pmab-FOLFOXIRI 7 cycles (3-12)/FOLFOXIRI 9.5 cycles (7-11).7 pts. achieved very good histopathological response with vital tumor cells ≤20% (pmab-FOLFOXIRI/FOLFOXIRI 5/2) and 7 pts. showed vital tumor cells >20% (pmab-FOLFOXIRI/FOLFOXIRI 6/1). The cut-off correlated with an improved PFS in the group ≤20 vs >20% (median PFS 12.40; confidence interval (CI) 6.43-51.22 vs PFS 9.88; CI 6.17-15.26 months). The severity of CASH was not increased by the addition of pmab. Conclusions: In this preliminary analysis of VOLFI histopathological response seems to correlate with a better PFS after secondary resection of liver metastases. There was no relevant difference in CASH between pmab-FOLFOXIRI vs. FOLFOXIRI alone. The trial is registered with ClinicalTrials.gov, NCT01328171. Clinical trial information: AIO-KRK-0109.
562 Background: Patients with muscle-invasive bladder cancer (MIBC) that underwent neoadjuvant chemotherapy (NAC) prior to radical cystectomy (RC) show improved overall survival. Those with a pathological complete response (pCR) usually have the best prognosis. In the literature, improved response to NAC has been associated with basal tumor characteristics in MIBC so far. The aim of the present study was to examine the association of luminal (KRT20) and basal (KRT5) mRNA expression patterns at transurethral resection (TUR) with pCR at RC after NAC in a contemporary cohort of consecutive MIBC patients. Methods: Clinical Data and formalin fixed paraffin embedded tumor tissue samples from TUR and RC of 49 patients with MIBC were retrospectively analyzed. Using RT-PCR KRT20 and KRT5 mRNA expression were measured in 40-∆Ct values and normalized against the control gene CALM2. Statistical analyses comprised nonparametric and chi 2 testing, partition models and spearman correlation analyses. Results: The study cohort had a median age of 63 years and consisted of 38/49 (78%) males. After NAC, 17/49 (35%) patients had cPR. Using partition models, we found that patients with high-KRT20 (≥39.5 ∆C t ) had a higher chance of pCR (57% vs. 26%, p=0.04). Using a cutoff for KRT5 at <38.1 ∆C t within the subgroup of patients with low-KRT20 (<39.5 ∆C t , n=35), we found poorest response among low-KRT20/low-KRT5 compared to low-KRT20/high-KRT5 and high-KRT20 (13% vs. 37% vs. 57%, p=0.29), respectively. For low-KRT20/low-KRT5, low-KRT20/high-KRT5 and high-KRT20 median KRT5 was 34.8 vs. 39.5 vs. 34.1 ∆C t ( p=0.001) and median KRT20 was 37.9 vs. 32.9 vs. 40.1 ∆C t ,( p=0.001), respectively. Conclusions: Patients with MIBC showing high expression of KRT20 were more likely to show pCR at RC after NAC. Moreover, we were able to identify a high risk group of patients with lowKRT20/lowKRT5 that was less likely to achieve pCR at RC after NAC. Our findings are contradicting previous studies and need further verification in larger cohorts. However, our results might be useful for treatment stratification in MIBC patients.
Histological healing in ulcerative colitis (UC) may be a better predictor than macroscopic appearance or clinical criteria for time to relapse. Indicators of acute mucosal inflammation which are used in the Riley Score or the Nancy index are associated with a two- to threefold increase in the risk of UC relapse during 12 months’ follow-up. However histologic assessment requires invasive endoscopy and gaining of biopsy. Non-invasive surrogates of mucosal healing would help to lower risks, costs and might increase patient acceptance. Fecal biomarkers are frequently used in UC and might be of help in this endeavour. The study aimed to investigate the performance of non-invasive faecal biomarkers compared with the Riley Score and Nancy index in patients with UC. Colonoscopy was performed in every patient and a faecal sample was harvested within 72 h. Three biopsies were taken from the macroscopically most inflamed location or random from each colonic segment and Riley Score and Nancy index were calculated. For each patient the highest calculated score was compared with the faecal biomarkers Lactoferrin (LF), Calprotectin (CALPREST - CalP), PMN elastase (PMN-E), S100 calcium-binding protein A12 (S100A12) and Eosinophil-derived Neurotoxin (EDN). It was evaluated if the median levels of the fecal markers differ significantly between the grades 0–4 of the Nancy index. 50 patients (32 female), mean age 42.9 ± 12.3 years (range 23–67) with diagnosed UC were included. The Riley score and the Nancy index correlated with EDN (r (49) = 0.56; p = .001/ r (49) = 0.45; p = .001), S100A12 (r (49) = 0.33; p = .022/ r (49) = 0.35; p = .015), PMN-e (r (49) = 0.31; p = .028/ r (49) = 0.29; p = .044) and LF (r (49) = 0.45; p = .001/ r (49) = 0.35; p = .015), but not with CalP (p > .05). The median levels of the faecal markers of the Nancy index and the results of the Kruskal–Wallis test are presented in Table 1. LF, EDN and CalP differed significantly between the grades. Post-hoc tests showed that LF differed significantly between the grades 2 and 3 (z = −3.13, p = .011), EDN between the grades 2 and 4 (z = −3.01, p = .016) and S100A12 between the grades 2 and 3 (z = −3.97, p = .000) and 2 and 4 (z = −3.07, p = .013). The fecal biomarkers LF, EDN, S100A12 and PMN-e were correlated significantly with the Riley and Nancy index, LF, EDN and CalP differed significantly between the grades of the Nancy index. The results support the utility of fecal biomarkers for detecting active histologic inflammation in patients with ulcerative colitis.
