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Supplementary Data from A Novel Polymorphism in the Promoter Region of ERBB4 Is Associated with Breast and Colorectal Cancer Risk

Purpose: The receptor tyrosine kinase ERBB4/HER4 plays a role in cell division, migration, differentiation, as well as apoptosis, and is frequently overexpressed in breast and colorectal tumors. To understand the role of genetic variations in the regulation of ERBB4 expression, we identified new polymorphisms and investigated their functional implication and risk association with breast and colorectal cancer.Experimental Design: We screened colorectal tumors from 92 patients for genetic variants at the ERBB4 ATG −1000 bp 5′-regulatory region by denaturing high-performance liquid chromatography and sequencing. Variants were subjected to DNA-protein interaction analyses (electrophoretic mobility shift assay), reporter gene assays in breast cancer cell lines MDA134 and MDA157, and immunohistochemical analyses of breast tumors. We established genotype frequencies within a breast cancer case-control collection (1,021 cases, 1,015 population-based controls) and a colorectal cancer case-control collection (459 cases, 569 blood donors) using matrix-assisted laser desorption ionization/time of flight mass spectrometry. Adjusted odds ratios (OR) and 95% confidence intervals (CI) were assessed by multivariate logistic regression.Results: We identified five new germ line variants −815 A>T, −782 G>T, −638 insTC, −267 C>G, and −219 del10bp. Two variants showed in vitro functional effects. The −782T allele showed lower protein binding affinity and lower promoter activity compared with the −782G allele, however, the −815T allele showed higher protein binding affinity and higher promoter activity. The −782T variant was identified as a risk allele for breast and colorectal cancer (OR, 1.59; 95% CI, 1.06-2.34 and OR, 2.21; 95% CI, 1.22-3.99, respectively).Conclusion: The ERBB4 −782 G>T polymorphism, by virtue of its in vitro functional implication and incidence, is a risk factor for breast and colorectal cancer. more...

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ZusammenfassungAutoimmune Lebererkrankungen umfassen ein Spektrum idiopathischer fortschreitender Leberentzündungen mit im Einzelfall histologisch kennzeichnenden Befunden. Hierzu gehören bei der autoimmunen Hepatitis (AIH) das Bild einer chronischen Hepatitis mit plasmazellreicher dominierender Grenzflächenaktivität, Rosettierung der Hepatozyten und Emperipolese, bei der primären biliären Cholangitis (PBC) chronische, nichteitrige, destruierende Gallengangsläsionen, bei der primären sklerosierenden Cholangitis (PSC) die zwiebelschalenartige periduktale Fibrose bis zur verödenden Gangsklerose. Histopathologische Schnittmengenbefunde dieser Entitäten untereinander kennzeichnen Varianten der AIH bzw. Overlapsyndrome. Die Diagnose autoimmuner Lebererkrankungen beruht auf der Gesamtkonstellation klinischer, enzymserologischer, immunserologischer und bildgebender Befunde, zu denen fakultativ oder obligat die Leberbiopsie hinzugezogen wird. Der Leberbiopsie kommt eine gewichtige Rolle zu bei der Diagnose der AIH und ihrer Varianten, der Small-Duct-PSC, der AMA-negativen PBC, der IgG4-assoziierten Entzündungen und der Abgrenzung gegenüber konkurrierenden Lebererkrankungen insbesondere der medikamentös-toxischen Leberschädigung. Damit leistet die histologisch gestützte Diagnose einen gewichtigen Beitrag zu einer differenzierten Therapie autoimmun-entzündlicher Lebererkrankungen. more...

Das klinische Bild sinusoidaler Erkrankungen der Leber ist eher uncharakteristisch. Es ausert sich in einem breiten klinischen Manifestationsspektrum wie Hepatomegalie, portaler Hypertonus oder als chronisches Leberversagen unbekannter Ursache. Im Gegensatz zu anderen Lebererkrankungen fehlt es an spezifischen serologischen, immunserologischen und rontgenologischen Befunden. Haufig weisen erst bioptische Befunde auf eine sinusoidale Erkrankung hin. Diese Ubersicht mochte einen praktisch orientierten Uberblick uber Leberumbauprozesse, hamodynamische Storungen und vaskulare Tumoren geben. more...

BackgroundhTERT promotor mutation represents a common and early event in hepatocarcinogenesis. Its linkage with the morphologic status of the underlying liver tissue is not really understood. We analysed the connection between the histopathological changes of tumour bearing liver tissue and the occurrence of hTERT promotor mutation in hepatocellular carcinoma, correlated with clinical data. MethodsThe study cohort comprised 160 histologically confirmed hepatocellular carcinomas (HCC) in liver cirrhosis and non-cirrhosis which were investigated for hTERT promotor mutation. Frequency of hTERT promotor mutation in HCC in cirrhosis and non-cirrhosis was evaluated and correlated with potential clinical and histopathological drivers. Especially, tumour bearing non-cirrhotic liver tissue was examined regarding inflammation, modified histological activity index (mHAI), fibrosis and steatosis and its correlation with the frequency of hTERT promotor mutation in HCC. Overall survival analysis via multivariate Cox regression analysis was performed. Furthermore, hTERT antibody immunohistochemistry and molecular hTERT promotor mutation analysis of both HCC and background liver tissue were compared.ResultshTERT promotor mutation was especially related to HCC in cirrhosis compared to non-cirrhotic liver tissue (p < 0.001) and independently from cirrhosis to HCC in patients ≥ 60 (p = 0.005). Furthermore, hTERT promotor mutation was associated with liver cirrhosis caused by alcohol toxicity and hepatitis C virus infection (HCV). In non-cirrhotic liver tissue the frequency of hTERT promotor mutated HCC increased with the degree of inflammation and fibrosis. Nevertheless, 25% of hTERT promotor mutated HCC developed in normal liver tissue without HCC risk factors. Multivariate Cox regression analysis did not show an influence of hTERT promotor mutation in HCC on overall survival of 3, 5 and 16 years. Immunohistochemical analysis with the hTERT antibodies LS-B95 and 2D8 hTERT promotor mutated HCC and wildtype HCC showed a mildly stronger immunoreaction compared to the tumour bearing liver tissue (LS-B95: p < 0.01, 2D8: p < 0.01).Conclusions Our study reveals a connection between pathological changes of tumour bearing liver tissue and hTERT promotor mutation in most HCC, even in non-cirrhotic liver tissue. Immunohistochemical hTERT antibodies do not discriminate between hTERT promotor mutated and wildtype HCC. more...

Objective: The aim of the study was to introduce our new modification of the Indiana pouch with a refluxing ureteral anastomosis in a tubular afferent ileal segment of the ileo-caecal urinary reservoir. Patients and Methods: Between February 2008 and December 2020, we performed a total of 37 modified continent ileo-caecal pouches for urinary diversion when orthotopic bladder substitution was not possible. Hereby, we modified the Indiana pouch procedure with a new refluxing end-to-end ureteral anastomosis into an 8-cm afferent tubular ileal segment. Results: We performed the modified Indiana pouch in 27 women (73%) and 10 men (27%). The median age of the patients at time of operation was 64 years (43–80 years). To date, the average follow-up is 69 months (3–156 months). In 32/37 cases, we performed the new pouch procedure after radical cystectomy for muscle-invasive bladder cancer and in 1/37 cases after radical cystectomy for locally advanced prostate cancer. In 4 cases, the procedure was performed after total exenteration of the pelvis due to locally advanced bladder, colorectal, or gynaecological cancers. Ureteral anastomotic strictures were seen in 2/37 patients (5.4%) or 2/72 (2.8%) of renal units. Conclusions: Our modification of the Indiana pouch cutaneous continent urinary diversion with the ureteral anastomosis to a tubular segment of the pouch is easy to perform and effective in reducing the rate of ureteral anastomotic strictures. By lengthening, the afferent tubular ileal segment, it additionally allows easy ureteral replacement. more...

The early and definitive diagnosis of malignant bile duct stenoses is essential for a timely and adequate therapy. However, tissue sampling with transpapillary brush cytology (BC) or forceps biopsy (FB) remains challenging. With this study, we aimed to compare the effectiveness and safety of different tissue sampling modalities (BC, FB without/after previous balloon dilatation). Standardized database research identified all patients, who underwent endoscopic retrograde cholangiography with BC and/or FB for indeterminate bile duct stenosis between January 2010 and April 2018 and with a definitive diagnosis. 218 patients were enrolled (149 cases with malignant and 69 with benign disease). FB had a significant higher sensitivity than BC (43% vs. 16%, p more...

Autoimmune liver diseases comprise a spectrum of progredient idiopathic inflammatory diseases. Typical histological features of autoimmune hepatitis (AIH) include the pattern of chronic hepatitis with predominant plasma cell-rich interface activity, rosetting of hepatocytes, and emperipolesis. Florid bile duct lesions are the key feature of primary biliary cholangitis (PBC); onion-like periductal fibrosis characterizes the primary sclerosing cholangitis (PSC). Variants of AIH, or overlap syndromes, show intersecting histomorphologic findings with PBC or PSC. The diagnosis of the different autoimmune inflammatory liver diseases is based on clinical presentation, a hepatitic or cholestatic pattern of liver enzymes, immuno-serological findings, image analysis in PSC, and liver biopsy as a facultative or obligatory adjunct. Liver biopsy plays a major role in the diagnosis of AIH, small-duct PSC, AMA-negative PBC, IgG4-related diseases, overlap syndrome, and in the recognition of concurrent liver diseases, especially drug-induced liver diseases. Herewith pathologists can help clinicians find adequate therapy for different autoimmune inflammatory liver diseases. more...

This review covers a spectrum of pathologic changes and diseases involving hepatic sinusoids. In the majority of patients, clinical findings are rather uncharacteristic such as hepatomegaly, portal hypertension, or lingering liver failure of unknown origin. In contrast to more common hepatic disorders, characteristic clinical, serological, immunoserological, and radiographical findings are lacking. In these cases, biopsy findings may be crucial to guide treatment decisions. This review covers a variety of hepatic disorders that practicing pathologists may encounter in their clinical routine. more...

Die hochauflosende radiologische Schnittbildtechnik fuhrt dazu, dass auch kleine, hochdifferenzierte hepatozellulare Tumoren zur leberbioptischen Diagnostik gelangen. In Leberzirrhosen sind regeneratorische und dysplastische Knoten gegenuber einem hepatozellularen Karzinom abzugrenzen. In nichtzirrhotischem Lebergewebe besteht die Differenzialdiagnose zum hepatozellularen Adenom, regeneratorischen Knoten, leberzellahnlichen lebereigenen Tumoren sowie zu Lebermetastasen. Die Diagnostik umfasst die Matrixanalyse des tumortragenden Lebergewebes, die histologische Strukturanalyse von Zytomorphologie, Trabekel- und Gefasbettarchitektur, die immunhistochemische Prufung einer hepatozellularen Differenzierung und die Typisierung leberzellahnlicher Fremdzellen. Molekularpathologische Untersuchungen konnen die Dignitatseinschatzung untermauern. Die vorliegende Ubersicht liefert einen Leitfaden zur Diagnose hepatozellularer Karzinome und leberzellahnlicher Tumoren auch an kleinen Biopsien. more...

Fibrous dysplasia is a benign bone disease, presenting as monostotic or polyostotic lesions, or as part of a syndrome (McCune-Albright/Mazabraud). Its clinical course shows a variegated picture and the progression of its growth is unpredictable. In the workup of 39 fibrous dysplasia cases in the cranio-facial area, four cases presented fast growth tendencies, of which two patients with McCune-Albright syndrome showed malignant-like rapid growth. This local aggressive form is extremely rare, and the concept of this issue has not been clearly defined. With regard to the speed of growth a volumetric-time analysis in one of our cases demonstrated a 74 days tumor doubling rate with an exponential growth curve. According to the literature the aggressive form presented extra-cranially mainly at an adult age, whereas its appearance in our cranio-facial patient collective was much younger. Distinguishing nonmalignant and malignant aggressive forms is difficult and highly inconsistent in the literature. We therefore implemented a quantitative growth measure analysis to define aggressive forms based on progression and speed of growth and impartial of type of FD, localization or functional incapacity. Due to our study findings and literature review we state a prevalence of an aggressive form might be possibly about 5 %. more...

Response rates in HER2-overexpressing EBC treated with neoadjuvant chemotherapy and trastuzumab (T) have been improved by addition of pertuzumab (P). The prospective, phase II, neoadjuvant WSG-ADAPT HER2+/HR- trial assessed whether patients with strong early response to dual blockade alone might achieve pathological complete response (pCR) comparable to that of patients receiving dual blockade and chemotherapy. Female patients with HER2+/HR- EBC (M0) were randomized (5:2) to 12 weeks of T + P +/- weekly paclitaxel (pac) at 80 mg/m(2). Early response was defined as proliferation decrease a parts per thousand 30% of Ki-67 (versus baseline) or low cellularity (< 500 invasive tumor cells) in the 3-week biopsy. The trial was designed to test non-inferiority for pCR in early responding patients of the T + P arm versus all chemotherapy-treated patients. From February 2014 to December 2015, 160 patients were screened, 92 were randomized to T + P and 42 to T + P+pac. Baseline characteristics were well balanced (median age 54 versus 51.5 years, cT2 51.1 versus 52.4%, cN0 54.3 versus 61.9%);91.3% of patients completed T + P per protocol and 92.9% T + P+pac. The pCR rate in the T + P+pac arm was 90.5%, compared with 36.3% in the T + P arm as a whole. In the T + P arm, 24/92 were classified as non-responders, and their pCR rate was only 8.3% compared with 44.7% in responders (38/92) and 42.9% in patients with unclassified early response (30/92). No new safety signals were observed in the study population. Addition of taxane monotherapy to dual HER2 blockade in a 12-week neoadjuvant setting substantially increases pCR rates in HER2+/HR- EBC compared with dual blockade alone, even within early responders to dual blockade. Early non-response under dual blockade strongly predicts failure to achieve pCR. more...

Anti-PD-1 treatment increases anti-tumour immune responses in animal models of hepatocellular carcinoma (HCC). Sorafenib, the mainstay of treatment of HCC patients, however, leads to tumour hypoxia and thereby abrogates the efficacy of anti-PD-1 treatment. This served as a rationale to implement CXCR4 inhibition as adjunct to sorafenib and anti-PD-1 treatment in murine HCC models. We studied the relationship between tumour hypoxia, PD-L1 and CXCL12 expression in human HCC, aiming to test the rationale for triple therapy combining sorafenib, PD-1 immune checkpoint inhibitors and CXCR4 inhibitors. Expression of CXCL12, PD-L1 and of surrogate markers for tumour hypoxia was evaluated at messenger RNA (mRNA) level in a cohort of HCC patients from The Cancer Genome Atlas and immunohistochemically in an independent cohort from the University Hospital of Bonn. Retrospective survival analyses were conducted. CXCL12 mRNA level significantly correlated with markers indicating tumour hypoxia in HCC (HIF1-α ρ = 0.104, p = 0.047). PD-L1 expression was significantly increased in tumours with a high number of tumour-infiltrating lymphocytes (ρ = 0.533, p < 0.001). In Cox proportional hazard analyses, high PD-L1 expression and loss of nuclear CXCL12 expression showed significant prognostic value in terms of overall survival (hazard ratio (HR) = 3.35 [95%CI 1.33–8.46], p = 0.011 for PD-L1; HR = 2.64 [95%CI 1.18–5.88], p = 0.018 for CXCL12, respectively). This study supports the rationale to combine CXCR4 inhibitors and PD-1 immune checkpoint inhibitors in patients with HCC, as sorafenib-induced tumour hypoxia leads to upregulation of PD-L1 and CXCL12. more...

// Yuri Tolkach 1 , Diane Goltz 1 , Anika Kremer 1 , Hojjat Ahmadzadehfar 2 , Dominik Bergheim 1 , Markus Essler 2 , Marnix Lam 3 , Bart de Keizer 3 , Hans-Peter Fischer 1 , § and Glen Kristiansen 1 , § 1 Institute of Pathology, University Hospital Bonn, Bonn 53127, Germany 2 Clinic of Nuclear Medicine, University Hospital Bonn, Bonn 53127, Germany 3 Department of Radiology and Nuclear Medicine, University Medical Center Utrecht, Utrecht 3584 CX, the Netherlands § Shared senior co-authorship Correspondence to: Glen Kristiansen, email: glen.kristiansen@ukbonn.de Keywords: FOLH1 ; hepatocellular carcinoma; PET/CT; prostate-specific membrane antigen; theranostic Received: January 01, 2019 Accepted: May 26, 2019 Published: June 25, 2019 ABSTRACT Aim The prostate-specific membrane antigen (PSMA) is currently being established as a potent diagnostic marker in many tumor types. So far, its evidence in hepatocellular carcinoma (HCC) is sparse. The aim of our study was a comprehensive evaluation of PSMA expression and its prognostic role in patients with hepatocellular carcinoma as well as feasibility test of PSMA as an agent for diagnostic imaging. Methods The cohort for immunohistochemistry consisted of 153 patients with HCC. For validation purposes the HCC cohort (n = 359) of The Cancer Genome Atlas was analyzed on transcript level as well. Results On immunohistochemistry, non-tumorous liver tissue showed PSMA expression on canalicular membranes in all cases. In tumor tissue two patterns of expression, with a canalicular (41.1% of tumors) and a neovascular (89.9% of tumors) staining were seen. Completely negative for both two patterns were only 4.1% of tumors; conversely, 79.2% of the tumors showed high levels of PSMA protein expression at any location. At mRNA level higher FOLH1 (PSMA) expression rates were statistically significant and independently associated with longer overall survival times. Additionally, a case report of successful diagnostic 68 Ga-PSMA-11 PET/CT in a patient with HCC progression on multiple therapy lines is provided. Conclusions Majority of hepatocellular carcinomas show high levels of PSMA expression on tumor vessels and on canalicular membrane of the tumor cells. Putative diagnostic, prognostic and therapeutic value of PSMA in HCC warrants further clinically oriented investigations. more...

Angestosen durch neue molekularpathologische Erkenntnisse hat sich die morphologische Diagnostik hepatozellularer Tumoren in den letzten Jahren bedeutend weiterentwickelt. Bildgebende Verfahren filtern insbesondere kleine Tumoren und radiologisch unklare Herde zur bioptischen Abklarung heraus. Das Spektrum dieser Tumoren umfasst im nichtzirrhotischen Lebergewebe hepatozellulare Karzinome, hepatozellulare Adenome, fokale nodulare Hyperplasien und makroregeneratorische Knoten. Primar sollte bei der histopathologischen Analyse dieser Lasionen die Dignitat festgelegt werden. Im 2. Schritt sind nach Ausschluss von Malignitat, einer fokalen nodularen Hyperplasie und makroregeneratorischer Knoten hepatozellulare Adenome auf der Basis immunhistochemischer/molekularpathologischer Kriterien entsprechend der WHO-Klassifikation der Lebertumoren aus dem Jahr 2010 zu subklassifizieren. Die Subtypisierung liefert wesentliche Zusatzinformationen hinsichtlich der Prognose und der Therapieindikation dieser Lasionen. more...

Ein 82-jahriger Patient stellte sich mit einem lachsfarbenen Bindehauttumor am linken Auge vor. Klinisch fiel in der Lasion ein umschriebener, derber, weislicher Anteil auf. Die histopathologische und immunhistochemische Aufarbeitung des Biopsats ergab ein CD20+-Marginalzonenlymphom der Bindehaut mit Amyloidablagerungen. Extranodale Marginalzonenlymphome sind die haufigsten Lymphome im Bereich der okularen Adnexe und machen dort 5–10% der malignen Erkrankungen aus. Eine assoziierte Amyloidproduktion ist sehr selten und hat nach aktuellem Kenntnisstand keine prognostische Relevanz. more...

Molecular dissection of hepatocellular adenomas has brought forward a diversity of well-defined entities. Their distinction is important for routine practice, since prognosis is tightly related to the individual subgroup. Very recent activity has generated new details on the molecular background of hepatocellular adenoma, which this article aims to integrate into the current concepts of taxonomy. more...

Objective Intact atrial septum or highly restrictive inter-atrial communication (I/HRAS) combined with either severe aortic stenosis (SAS) or hypoplastic left heart syndrome (HLHS), respectively, is associated with adverse outcome. This study focusses on changes in alveolo-septal lung parenchyma due to increased left atrial pressure. Method In a retrospective cross-sectional autoptic study, we investigated fetal/neonatal lung specimens of 18 patients with SAS/HLHS with I/HRAS, 11 patients with SAS/HLHS and unrestrictive inter-atrial communications and 18 controls. Pulmonary maturation was investigated by means of morphometric and immunohistochemical analyses. Results In a comparison of all three groups, alveolo-capillary membrane maturation was significantly disturbed in I/HRAS fetuses from week 23 of pregnancy on. I/HRAS lungs showed angiomatoid hyper-capillarisation and significantly wider inter-airspace mesenchyme. Differences in width ranged between 34.58 µm (95% CI: 11.41–57.75 µm) and 46.74 µm (95% CI: 13.97–79.50 µm) in the second and third trimesters. In I/HRAS infants with HLHS, inter-airspace mesenchymal diameters steadily normalised with age; however, significant fibroelastosis of alveolar septae developed. Conclusion Fetal lung maturation with respect to alveolo-capillary membrane formation is severely disordered in patients with SAS/HLHS with I/HRAS. Our findings indicate that, from a morphological point of view, timing of fetal invention in fetuses with I/HRAS should be fixed within the second trimester of pregnancy. © 2015 John Wiley & Sons, Ltd. more...

To compare systematically quantitative MRI, MR spectroscopy (MRS), and different histological methods for liver fat quantification in order to identify possible incongruities. Fifty-nine consecutive patients with liver disorders were examined on a 3 T MRI system. Quantitative MRI was performed using a dual- and a six-echo variant of the modified Dixon (mDixon) sequence, calculating proton density fat fraction (PDFF) maps, in addition to single-voxel MRS. Histological fat quantification included estimation of the percentage of hepatocytes containing fat vesicles as well as semi-automatic quantification (qHisto) using tissue quantification software. In 33 of 59 patients, the hepatic fat fraction was >5 % as determined by MRS (maximum 45 %, mean 17 %). Dual-echo mDixon yielded systematically lower PDFF values than six-echo mDixon (mean difference 1.0 %; P more...

Heterozygous α1-antitrypsin deficiency type PiZ (PiMZ) results in chronic liver injury and predisposes to hepatocellular carcinoma. Gene frequency of the PiZ allele ranges from 0.005 to 0.027 in Western and Central Europe; therefore, there is a substantial risk of coincidence with chronic alcohol abuse. This retrospective case–control study evaluates the impact of PiMZ genotype on the development of chronic liver disease in alcohol consuming patients. Six thousand eight hundred eighty-six consecutive liver specimens were immunohistochemically tested for PiZ-deposits. From 254 PiZ-positive patients, the liver biopsies of 30 PiMZ adults without concomitant liver disease other than alcoholic liver disease (ALD) were selected and matched to PiMM (wild type) patients with respect to age, gender and lifetime daily alcohol ingestion (LDAI). Histomorphological changes were assessed using the SAF score and by digital image analysis. Liver cirrhosis was significantly more frequent in PIMZ patients than in matched PiMM patients (PiMM 9/30 vs. PiMZ 14/30, p = 0.04). Comparison of the extent of fibrosis in PiMZ and PiMM livers by two-way ANOVA indicated that the amount of LDAI has a major effect in PiMZ and PiMM patients (30.04 % of total variation, p more...

BACKGROUND & AIMS: Patients with cirrhosis display hypocontractility of splanchnic vessels because of dysregulation of vasoactive proteins, such as decreased effect of RhoA/ROCK and increased activity of β-Arrestin-2 and eNOS. However, it is unknown whether the dysregulation of vasoactive proteins is displayed in other vessels. We investigated whether expression of vasoactive proteins can be evaluated in gastric mucosa vessels.METHODS: Biopsies from the gastric mucosa of 111 patients with cirrhosis were collected at three different centres and from 13 controls. Forty-nine patients had received TIPS. Portal pressure gradient was measured in 49 patients with TIPS and in 16 patients without TIPS. Biopsies from the antrum were conserved in formaldehyde for immunohistochemistry or shock-frozen for PCR and Western blot.RESULTS: The mucosal transcription of vascular markers (αSMA, CD31) was higher in cirrhotic patients than controls, which was confirmed by immunohistochemistry. On average, relative mucosal levels of RhoA and ROCK were lower, while β-Arrestin-2 levels were higher in cirrhotic patients compared to controls. Transcriptional levels of eNOS increased with presence of ascites and grade of oesophageal varices. Patients with TIPS showed less pronounced markers of vascular dysfunction in gastric mucosa.CONCLUSION: This is the first evidence that the expression of vasoactive proteins in mucosa from the gastric antrum of patients with cirrhosis reflects their vascular dysfunction and possibly changes after therapeutic interventions. more...

The tumor proliferation marker, Ki-67 index, is a well-established prognostic marker in gastroenteropancreatic neuroendocrine neoplasms (NENs). Noninvasive molecular imaging allows whole-body metabolic characterization of metastatic disease. We investigated the prognostic impact of 18F-FDG PET in inoperable multifocal disease. Methods: Retrospective, dual-center analysis was performed on 89 patients with histologically confirmed, inoperable metastatic gastroenteropancreatic NENs undergoing 18F-FDG PET/CT within the staging routine. Metabolic (PET-based) grading was in accordance with the most prominent 18F-FDG uptake (reference tumor lesion): mG1, tumor-to-liver ratio of maximum standardized uptake value ≤ 1.0; mG2, 1.0–2.3; mG3, >2.3. Other potential variables influencing overall survival, including age, tumor origin, performance status, tumor burden, plasma chromogranin A (≥600 μg/L), neuron-specific enolase (≥25 μg/L), and classic grading (Ki-67–based) underwent univariate (log-rank test) and multivariate analysis (Cox proportional hazards model), with a P value of less than 0.05 considered significant. Results: The median follow-up period was 38 mo (95% confidence interval [CI], 27–49 mo); median overall survival of the 89 patients left for multivariate analysis was 29 mo (95% CI, 21–37 mo). According to metabolic grading, 9 patients (10.2%) had mG1 tumors, 22 (25.0%) mG2, and 57 (64.8%) mG3. On multivariate analysis, markedly elevated plasma neuron-specific enolase (P = 0.016; hazard ratio, 2.9; 95% CI, 1.2–7.0) and high metabolic grade (P = 0.015; hazard ratio, 4.7; 95% CI, 1.2–7.0) were independent predictors of survival. Conclusion: This study demonstrated the feasibility of prognostic 3-grade stratification of metastatic gastroenteropancreatic NENs by whole-body molecular imaging using 18F-FDG PET. more...

Background & Aims The genetic background of alcoholic liver diseases and their complications are increasingly recognized. A common polymorphism in the neurocan ( NCAN ) gene, which is known to be expressed in neuronal tissue, has been identified as a risk factor for non-alcoholic fatty liver disease (NAFLD). We investigated if this polymorphism may also be related to alcoholic liver disease (ALD) and hepatocellular carcinoma (HCC). Methods We analysed the distribution of the NCAN rs2228603 genotypes in 356 patients with alcoholic liver cirrhosis, 126 patients with alcoholic HCC, 382 persons with alcohol abuse without liver damage, 362 healthy controls and in 171 patients with hepatitis C virus (HCV) associated HCC. Furthermore, a validation cohort of 229 patients with alcoholic cirrhosis (83 with HCC) was analysed. The genotypes were determined by LightSNiP assays. The expression of NCAN was studied by RT-PCR and immunofluorescence microscopy. Results The frequency of the NCAN rs2228603 T allele was significantly increased in patients with HCC due to ALD (15.1%) compared to alcoholic cirrhosis without HCC (9.3%), alcoholic controls (7.2%), healthy controls (7.9%), and HCV associated HCC (9.1%). This finding was confirmed in the validation cohort (15.7% vs. 6.8%, OR=2.53; 95%CI: 1.36–4.68; p =0.0025) and by multivariate analysis (OR=1.840; 95%CI: 1.22–2.78; p =0.004 for carriage of the rs2228603 T allele). In addition, we identified and localised NCAN expression in human liver. Conclusions NCAN is not only expressed in neuronal tissue, but also in the liver. Its rs2228603 polymorphism is a risk factor for HCC in ALD, but not in HCV infection. more...

Purpose: Perivascular epitheloid cell tumour [PEComa] is a rare neoplasm entity, characterized by perivascular epitheloid cells with a coexpression of smooth muscle and melanocytic markers. PEComas are found in a variety of localizations, though lesions within the liver are still scarcely found. Although the majority of these tumours are recognized as benign, there are some reports about advanced and aggressive tumours even with fatal outcome. By means of this case report and literary review including other 21 published cases, potential treatment modalities concerning clinical diagnostics, therapy and the follow-up care should be discussed. Methods: The following report presents the case of a 53-year old woman with a known liver lesion, since four years under regularly sonographic controls. Finally, after a haemorrhage episode, the lesion was resected and the diagnosis found. For the literary review a systematic search for case reports published between January 1, 1999 and May 1, 2012 was performed on Pubmed. Results: The only way, till now, of confirming the diagnosis is through immunohistochemical examinations. The already published Malignancy criteria by Folpe et al. must be taken carefully in question, as there are cases of malignant behaviour, that do not exactly coincide with these. Conclusion: Primary PEComa of the liver must be treated as potential malignant and therefore a close follow-up is demanded. more...

Background & Aims Regulatory CD4 + T cells (Tregs) are considered to affect outcomes of HCV infection, because they increase in number during chronic hepatitis C and can suppress T-cell functions. Methods Using microarray analysis, in situ immunofluorescence, ELISA, and flowcytometry, we characterised functional differentiation and localisation of adaptive Tregs in patients with chronic hepatitis C. Results We found substantial upregulation of IL-8 in Foxp3 + CD4 + Tregs from chronic hepatitis C. Activated GARP-positive IL-8 + Tregs were particularly enriched in livers of patients with chronic hepatitis C in close proximity to areas of fibrosis and their numbers were correlated with the stage of fibrosis. Moreover, Tregs induced upregulation of profibrogenic markers TIMP1, MMP2, TGF-beta1, alpha-SMA, collagen, and CCL2 in primary human hepatic stellate cells (HSC). HSC activation, but not Treg suppressor function, was blocked by adding a neutralizing IL-8 antibody. Conclusions Our studies identified Foxp3 + CD4 + Tregs as an additional intrahepatic source of IL-8 in chronic hepatitis C acting on HSC. Thus, Foxp3 + CD4 + Tregs in chronic hepatitis C have acquired differentiation as regulators of fibrogenesis in addition to suppressing local immune responses. more...

Small ubiquitin-like modifier (SUMO) proteins are covalently attached to target proteins to modify their function. SUMO conjugation participates in processes tightly linked to tumorigenesis. Recently USPL1 (ubiquitin-specific peptidase-like (1) was identified as a SUMO isopeptidase. We report here on the first exploratory study investigating the relationship between genetic variability in USPL1 and breast cancer. Three potentially functional nonsynonymous coding SNPs (rs3742303, rs17609459, rs7984952) were genotyped in 1,021 breast cancer cases and 1,015 controls from the population-based GENICA study. We took advantage of multiple genotype imputation based on HapMap and the 1000 Genomes Project data to refine the association screening in the investigated region. Public genetic databases were also used to investigate the relationship with USPL1 expression in lymphoblastoid cell lines and breast tissue. Women homozygous for the minor C allele of rs7984952 showed a lower risk of Grade 3 breast tumors compared to TT homozygotes (OR 0.50, 95% CI 0.30-0.81). Case-only analyses confirmed the association between rs7984952 and tumor grade (OR 0.60, 95% CI 0.39-0.93). Imputation results in a 238 kb region around rs7984952 based on HapMap and the 1000 Genomes Project data were similar. No imputed variant showed an association signal stronger than rs7984952. USPL1 expression in tumor breast tissue increased with the number of C alleles. The present study illustrates the contribution of multiple imputation of genotypes using public data repositories to standard genotyping laboratory. The provided information may facilitate the design of independent studies to validate the association between USPL1 rs7984952 and risk of Grade 3 breast tumors. more...

Eine hypothalamisch-hypophysare Dysfunktion kann mit einer ungewohnlich rasch progredienten nichtalkoholischen Steatohepatitis (NASH) assoziiert sein. Zwei adoleszente Patientinnen zeigten 19 bzw. 5 Jahre nach Exstirpation eines Kraniopharyngeoms mit konsekutiver hypothalamisch-hypophysarer Dysfunktion eine weit fortgeschrittene Leberfibrose/Zirrhose auf dem Boden einer floriden NASH. Bei jungen Patienten und Kindern mit einer floriden Steatohepatitis sollte die Moglichkeit einer Grunderkrankung des Hypothalamus-Hypophysen-Systems bedacht werden. In diesen Fallen kann eine Steatohepatitis wesentlich aggressiver als gewohnlich verlaufen und rasch zur Leberzirrhose fortschreiten. more...

Ein 66-jahriger Patient mit vorausgegangener ausgedehnter Exzision und Bestrahlung eines maxillaren Ameloblastoms stellte sich mit einem Rezidiv des Tumors vor. Klinisch zeigte sich eine Infiltration der knochernen Orbita und des intraorbitalen Weichteilgewebes sowie der Nasennebenhohlen und Schadelbasis. Die histologische Untersuchung des exzidierten Gewebes bestatigte die Diagnose eines Ameloblastoms. Es zeigte sich eine starke Expression von CD56 in den Tumorzellen. Obwohl das Ameloblastom gemeinhin ein niedrigmaligner Tumor ist, werden zuweilen lokal aggressive Formen beobachtet. more...

We investigated circumscribed cell proliferations in healthy livers in comparison to non-cirrhotic livers bearing hepatocellular carcinoma. Using histochemical staining for cytochrome c oxidase, the fourth complex of the respiratory chain, we visualized patch-forming descendents of regeneratively active liver cells. The clonal nature of these patches was verified by laser-capture microdissection and Sanger sequencing of the enzyme's core subunits in patches carrying marker mutations on the mtDNA. We demonstrate a highly significant increase of the patch size and also a highly significant increase in the number of patches carrying marker mutations between hepatocellular carcinoma-free and -bearing livers. Thus, the carcinoma-bearing livers accumulated more genetic damage on mtDNA than the control group. Furthermore, for the first time, we present evidence in hepatocellular carcinoma-bearing non-cirrhotic livers of a significantly reduced pool of regeneratively active liver cells that are genetically and functionally altered. The analogy to ageing-related changes is suggestive of premature ageing of stem cells in non-cirrhotic hepatocellular carcinoma-bearing liver as an early step to hepatocarcinogenesis. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. more...

Background Alcohol is a common cause of hepatic liver injury with steatosis and fibrosis. Cannabinoid receptors (CB) modulate steatosis, inflammation and fibrogenesis. To investigate the differences between CB(1) and CB(2) in the hepatic response to chronic alcohol intake, we examined CB knockout mice (CB(1)(-/-), CB(2)(-/-)). Methods Eight- to 10-week-old CB(1)(-/-), CB(2)(-/-) and wild-type mice received 16% ethanol for 35 weeks. Animals receiving water served as controls. We analysed triglyceride and hydroxyproline contents in liver homogenates. mRNA levels of CBs, pro-inflammatory cytokines [tumour necrosis factor (TNF)-α, monocyte chemotactic protein (MCP)-1, interleukin (IL)-1β] and profibrotic factors [α-smooth muscle actin (α-SMA), procollagen-Ia, platelet-derived growth factor β receptor (PDGFβ-R)] were analysed by reverse transcription-polymerase chain reaction (RT-PCR). Histology (hemalaun and eosin, oil-red O, CD3, CD45R, CD45, F4/80, Sirius red) characterized hepatic steatosis, inflammation and fibrosis. Activation of lipogenic pathways, activation and proliferation of hepatic stellate cell (HSC) were assessed by western blot [fatty acid synthase (FAS), sterol regulatory element binding protein 1c (SREBP-1c), α-SMA, proliferating cell nuclear antigen (PCNA), cathepsin D]. Results Hepatic mRNA levels of the respective CBs were increased in wild-type animals and in CB(1)(-/-) mice after ethanol intake. Ethanol intake in CB(2)(-/-) mice induced much higher steatosis (SREBP-1c mediated) and inflammation (B-cell predominant infiltrates) compared with wild-type animals and CB(1)(-/-) mice. HSC activation and collagen production were increased in all groups after forced ethanol intake, being most pronounced in CB(2)(-/-) mice and least pronounced in CB(1)(-/-) mice. Discussion The fact that CB(2) receptor knockout mice exhibited the most pronounced liver damage after ethanol challenge indicates a protective role of CB(2) receptor expression in chronic ethanol intake. By contrast, in CB(1) knockouts, the effect of ethanol was attenuated, suggesting aggravation of fibrogenesis and SREBP-1c-mediated steatosis via CB(1) receptor expression after ethanol intake. more...

BACKGROUND: CYP3A enzymes, due to their role in the metabolism of steroid hormones, are suggested to affect carcinogenesis of hormone-related cancers. The purpose of the present study was to evaluate the association between polymorphisms located in CYP3A43, breast cancer risk, and tumor characteristics. METHODS: A 3-plex matrix-assisted laser desorption ionization time of flight mass spectrometry assay has been established for CYP3A43_74_delA (CYP3A43*2A), CYP3A43_1018_C>G (CYP3A43*3), and CYP3A43_1047_C>T (CYP3A43*1B) polymorphisms, and 1021 breast cancer cases and 1015 age-matched, population-based controls from the German GENICA collection have been genotyped. RESULTS: No differences in genotype frequencies between cases and controls were observed, indicating that CYP3A43_74_delA is not associated with breast cancer risk. Subgroup analyses showed an association between the CYP3A43_74_delA allele and high-grade tumors (odds ratio, 1.74; 95% confidence interval, 1.14-2.65 [P = .010 and Ptrend = .012]). CONCLUSIONS: The data support the notion that the CYP3A43_74_delA variant may result in decreased protein and/or activity levels, and this may further lead to increased hormone levels to promote tumor cell growth and hinder differentiation. Cancer 2010. © 2010 American Cancer Society. more...

Here we report on a patient with a primary hepatocellular carcinoma in a non-cirrhotic liver, in whom heterozygosity for an AAT-deficiency allele was found (PiMZ). Based on this observation and the current literature, the possible mechanisms for an eventual contribution of a heterozygosity of a heterozygous AAT-deficiency for a hepatocellular carcinoma are discussed. Alpha-1-antitrypsin (AAT)-deficiency (Laurell-Eriksson syndrome) is a genetic disorder, in which individuals who are homozygous for a deficiency allele are at an increased lifetime risk for pulmonary emphysema, liver cirrhosis, and primary hepatocellular carcinoma. It has been controversially discussed whether the heterozygous form (PiMZ) is also associated with an increased risk for liver diseases. Hepatocarcinogenesis for AAT-deficiency is probably based on a series of toxic events. Precipitation of AAT aggregates in hepatocytes is the initial step. These accumulate in the endoplasmic reticulum and cannot be eliminated from all hepatocytes by proteasomal and non-proteasomal mechanisms. AAT aggregates induce proinflammatory pathways and may be a stimulus for hepatocarcinogenesis. This hypothesis is based mostly on studies of individuals homozygous for a deficiency allele (PiZZ). The mechanism may also play a role in heterozygous patients. Since not all patients with precipitates of AAT-aggregates are develop a hepatocellular carcinoma related comorbidities such as chronic hepatitis B, C, chronic alcohol abuse, or so far unknown genetic and environmental factors may be crucial. more...