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2 Abstracts / Digestive and

RTERIAL AND PERIBILIARY VASCULOGENESIS DURINGIVER DEVELOPMENT IS MODULATED BY ANGIOGENICROWTH FACTORS EXPRESSED BY DUCTAL PLATE CELLSND HEPATOBLASTS

. Fabris a,b, M. Cadamuro a,b, L. Libbrecht c, C. Spirlı a,d, R. Fiorotto a,b,d,

. Okolicsanyi b, T. Roskams c, M. Strazzabosco a,d

Center for Liver Research (CeLiveR), Ospedali Riuniti di Bergamo, Berg-mo, ItalyDepartment of Surgical and Gastroenterological Sciences, Universita diadova, ItalyDepartment of Pathology, University-Hospital St Rafael, Leuven, BelgiumLiver Center and Department of Internal Medicine, Yale University, Newaven, CT, USA

ackground and aim. Intrahepatic bile ducts (BD) maintain close anatom-cal relationships with hepatic arteries (HA). Hepatic artery developments thought to be driven by signals originating from the bile ducts, but theature of these signals is unknown. We recently demonstrate that VEGF andngiopoietin-1 stimulate both peribiliary angiogenesis and cholangiocyteroliferation. We hypothesized that angiogenic factors generated by ductallate cells may direct the development of the portal arterioles.aterials. We studied the immunohistochemical expression of vascular

ndothelial growth factor (VEGF), angiopoietin-1 and -2 (Ang-1, Ang-2)nd their receptors (VEGFR-1, VEGFR-2, Tie-2) in 16 foetal liver spec-mens (gestational age from 10 to 36 weeks); 320 portal spaces werenalyzed according to their level of BD maturation in: ductal plate, migra-ory and incorporated bile duct stage. The following phenotypic markersere studied: cytokeratin-7 (biliary marker), CD34 (endothelial marker),

mooth muscle actin (mural cells) in preparations double immunostainedor angiogenic factors.esults. Throughout the different developmental stages the developing BD

nd hepatoblasts express VEGF and Ang-1, respectively. Precursors ofndothelial and mural cells expressed VEGFR-2 and Tie-2, respectively.ndothelial cells (EC) of immature HA express VEGFR-1 and mural cellsxpressed both Tie-2 and Ang-2. In mature HA, EC expressed Tie-2 andEGFR-1. During the migratory stage small clumps of CD34-positive

ells expressing VEGFR-1 are recruited close to VEGF-positive BD toorm a capillary network likely representing the peribiliary vascular plexusPBP).onclusions. These data suggest that, in liver vasculogenesis, VEGF andng-1 mediate the cross-talk between liver epithelial cells and developingascular structures. Specifically, VEGF production by BD may influencerterial and PBP vasculogenesis to provide blood flow to the developingpithelium, and Ang-1 signalling from hepatoblasts may contribute to HAemodelling.

oi:10.1016/j.dld.2006.12.023

HE HEPATIC STEM CELL NICHE FORMS IN LIVER INJURY:TS CELLULAR COMPOSITION AND ORIGIN

. Lorenzini a,b, T.G. Bird a, H. Caldwell a, P. Vig b, P. Andreone b, J.P.redale a, M.R. Alison c, S.J. Forbes a

MRC/Centre for Inflammation Research, The Queen’s Medical Researchnstitute, University of Edinburgh, UKHepatology Section, Imperial College, London, UKCentre for Diabetes and Metabolic Medicine, Queen Mary’s School ofedicine and Dentistry, Institute of Cell and Molecular Science, London,K

ntroduction. Stem cell niches in many tissues regulate stem/progenitorell behaviour through local signalling and by promoting differentiationnd proliferation in response to injury. In liver, the stem/progenitor cellompartment is activated in response to injury from the canals of Hering.p to now, these oval/progenitor cells have been intensively studied but there

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isease 39 (2007) A1–A28

s little data regarding the “niche cells” surrounding them. In this study, weave evaluated the cellular composition and the origin of the hepatic stemell niche in rodents and humans.

aterial and methods. In the first group of experiments, single and dou-le immunostainings for oval/hepatic progenitor cells, hepatic stellate cells,ndothelial cells, myofibroblasts, macrophages and basement membraneroteins, have been performed on normal and injured liver tissue, bothn rodents (HBsAg transgenic mice and two acetyl-aminofluorene/partialepatectomy rats) and in humans (viral hepatitis). In the second set of exper-ments, to identify which cells within the niche were of bone marrow (BM)rigin, fluorescent in situ hybridization (FISH) for the Y chromosome haseen performed on liver sections from mice and humans who had received aex-mismatched bone marrow (male into female) or liver transplant (femalento male).esults. In normal liver, stellate cells are in close contact with the small bileucts and canals of Hering but there is no clustering of non-parenchymal cellsround this area. In the injured livers analyzed, single and double immunos-aining showed that the oval/hepatic progenitor cell reaction is intimatelyurrounded by stellate cells, myofibroblasts and macrophages, and to a lesserxtent by endothelial cells, thus constituting an “activated hepatic stem celliche”. Moreover, the basement membrane component, laminin, is highlyxpressed around the niche. FISH analysis for the Y chromosome shows thatproportion of these non-parenchymal niche cells are of BM origin.onclusion. During liver damage a cellular niche forms to surround theval/progenitor cells which links the oval/progenitor cell reaction with cir-ulating BM-derived cells. These niche cells are likely to be critical for thetem/progenitor cell reaction and important in linking tissue damage to stemell mediated tissue repair.

oi:10.1016/j.dld.2006.12.024

NGIOGENESIS DURING TRANS-CATHETHER ARTERIALHEMOEMBOLISATION (TACE) IN HEPATOCELLULARARCINOMA (HCC)

. Cristofori, G. Pivetta, L. Girardi, A. Baldan, R. Cardin, F. Farinati

Department Gastroenterology and Radiology, Padua University, Italy

ackground and aims. TACE is not considered as a curative procedure forCC, despite being an aggressive and effective treatment, particularly foreripheral, well vascularized, large size lesions, but still awaits validation inarge prospective studies. Among the factors potentially impairing its effec-iveness is a hypothetical neo-angiogenetic reaction following the hischemiaue to embolization. With this in mind we evaluated the changes in the circu-ating levels of two angiogenetic factors (vascular endothelial growth factorVEGF and basic fibroblast growth factor-b – FGF) and of one parameter of

umor invasiveness (urokinase plasminogen activator – uPA) in the settingf TACE.ethods. Seventy-one consecutive HCC patients undergoing TACE were

ecruited, after informed consent, and underwent three blood drawings: therst before TACE (t0), the second after 3 days (t1) and the third at 4 weeks,t the time of sCT scanning (t2). The referring radiologist (GP), blindly,valuated tumor size, disease activity and vascularization at t0 and % residualctivity at t2. Clinical parameters (etiology, tumor grading, �fetoproteinevels) were recorded. Statistical analysis was based on linear and Spearmanorrelation analysis, Student’s t-test for paired and unpaired data and Kaplaneier survival curves evaluation. The choice of TACE as treatment was based

n the AASLD-EASL guidelines.esults. Overall, complete response, as assessed by sCT, was recorded in7% of the patients, mean survival was 35 months (31–40 C.L.) and 4 yearsurvival 57%. VEGF levels significantly correlated with the number of nodes,ere increased at t2 in non-responders (p = 0.01), while levels below the

edian significantly predict long term survival (70% survival at 48 months

ersus 26%, p = 0.008). b-FGF correlated with VEGF, tumor size, vascular-zation and residual activity but did not show any correlation with survival.PA correlated with tumor size and grading.

Liver Disease 39 (2007) A1–A28 A3

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Abstracts / Digestive and

onclusions. TACE, when not totally effective, induces a significant neo-ngiogenesis, this potentially affecting patients’ survival. VEGF is singledut as the most reliable prognostic parameter, thus suggesting its determi-ation as a mean of judging TACE effectiveness and prognosis. Finally,nti-angiogenetic drugs could find an indication in TACE-treated HCCatients.

oi:10.1016/j.dld.2006.12.025

URVEILLANCE FOR EARLY DIAGNOSIS OF HEPATOCELLU-AR CARCINOMA (HCC): IS IT EFFECTIVE IN INTERMEDI-TE/ADVANCED CIRRHOSIS?

. Santi, F. Trevisani, A. Gramenzi, R. Casadio, M.A. Di Nolfo, P. Deloggio, L. Benvegnu, G. Rapaccini, F. Farinati, M. Zoli, F. Borzio, E.G.iannini, E. Caturelli, M. Bernardi,

or the Italian Liver Cancer (ITA.LI.CA) group

ackground and aims. Surveillance of cirrhotic patients for early diagnosisf HCC, based on ultrasonography and alpha-fetoprotein (AFP) measure-ent, is a widespread practice. Liver function is crucial for its effectiveness,

ffecting both the feasibility of HCC treatments and the cirrhosis-relatedortality. Child-Pugh class A patients are considered optimal candidates,hile the use of surveillance in more advanced classes is debated. This study

ssesses if patients belonging to class B or C at the time of HCC diagnosisenefit of surveillance.ethods. From the ITA.LI.CA database, including 1834 consecutive

atients with HCC diagnosed in 10 Centres from January 1987 to Decem-er 2004, we selected 468 class B and 140 class C cases. Exclusion criteria:lass A (946 pts), unspecified class (47 pts) or surveillance interval (233 pts).n 252 (41.4%) patients HCC was detected during surveillance (semiannual72, annual 80; Group 1), while in 356 (58.6%) cases outside surveillanceGroup 2). The survival of surveilled patients was corrected for the estimatedead time.esults. Class B: CLIP and UNOS-TNM cancer stage (P < 0.001) and

menability to treatments (P < 0.001) were significantly better in Group 1203 pts) than in Group 2 (265 pts). The survival was higher in Group 1 thann Group 2: median (95% C.I.) 17.1 (13.5–20.6) versus 12.0 (9.4–14.6)

onths (P = 0.022). The survival rates at 1, 3 and 5 years were: 60.4%ersus 49.2%, 26.1% versus 16.1% and 10.7%, versus 4.3%, respectively.FP, gross pathology and treatment of HCC were independent prognostic

actors. Class C: cancer stage (P ≤ 0.002) and amenability to treatmentsP < 0.021) were significantly better in Group 1 (49 pts) than in Group 291 pts). Nonetheless, the survival did not differ: 7.1 (2.1–12.1) versus 6.04.1–7.9) months (P = 0.740).onclusions. These results suggest to enter into surveillance programs classpatients, and maintain on periodic screening class A patients who migrate

o the subsequent class. In class C patients, surveillance improves cancertage and amenability to treatment but not survival because of the hugempact of liver dysfunction on overall mortality and treatment results.

oi:10.1016/j.dld.2006.12.026

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ORTALITY IN PATIENTS WITH HEPATOCELLULAR CARCI-OMA PREDICTED BY SIX SCORING SYSTEMS: NONE IS THEINNER

alogero Camma a, Vito Di Marco a, Giuseppe Cabibbo a, Federica

atteri a, Luigi Sandonato b, Piero Parisi a, Nicola Alessi a, Anna Licata a,assimo Galia c, Marco Enea d, Massimo Attanasio d, Mario A. Latteri b,ntonio Craxı a

Unita di Gastroenterologia ed Epatologia, ItalyUnita di Chirurgia Oncologica, ItalyDipartimento di Radiologia, ItalyDipartimento di metodi quantitativi per le scienze umane, University ofalermo, Italy

ackground. Predicting mortality of patients with hepatocellular carcinomaHCC) is essential in planning active or supportive patient’s care. We com-ared, in a prospective cohort of patients with HCC from one referral Livernit, the ability of six scoring systems (CHILD-PUGH, MELD, OKUDA,LIP, GRECH, BCLC) to predict death.ethods. Four hundred six consecutively patients with cirrhosis andCC (mean age 66.9 ± 7.8; 285/121 male/female; CHILD-PUGH class: A= 214, B n = 136, and C n = 56; 318/406 HCV RNA positive, 41/406 HBsAgositive) were studied. A single lesion was observed in 233/406. Among the06 cases, 178 (44%) were treated, 89/178 (50%) with radiofrequency abla-ion, 64/178 (36%) with transcatheter arterial chemoembolization, 14/1787.8%) with surgical resection and 11/178 (6.1%) underwent liver transplan-ation. Two hundred twenty-eight (56%) subjects received supportive carelone due to advanced disease stage. To compare the performance of the sixrognostic systems by Cox model, the linear trend (LT) χ2 test (discrimina-ory ability) and the likelihood ratio (LR) χ2 test (homogeneity) were used.he area under the receiver-operating characteristic curve (AUROC) wasetermined by logistic regression model.esults. Among treated patients, serum albumin (p < 0.037), ascites

p < 0.006), and ECOG performance status (p < 0.019) were the only inde-endent variables significantly associated with survival by Cox model. TheLIP system had the highest homogeneity LR test (χ2 141.9) and the highestiscriminatory ability LT test (χ2 165.1) when compared with other systemssee Table 1). The BCLC classification had the highest AUROC (0.67; 95%I 0.57–0.77) when predicting the 3-year survival of treated patients.onclusions. The overall predictive ability of the six staging systems weave considered was relatively unsatisfactory and moreover, it was not uni-

orm over the several subgroups of HCC patients considered. None of theodels provided sufficient confidence for the prediction of mortality in

ndividual patients.

oi:10.1016/j.dld.2006.12.027