[Show abstract][Hide abstract] ABSTRACT: Our goal was to optimize the radiosensitizing potential of anti-epidermal growth factor receptor (EGFR) monoclonal antibodies, when given concomitantly with preoperative radiotherapy in KRAS wild-type locally advanced rectal cancer (LARC). Based on pre-clinical studies conducted by our group, we designed a phase II trial in which panitumumab (6 mg/kg/q2 weeks) was combined with preoperative radiotherapy (45 Gy in 25 fractions) to treat cT3-4/N + KRAS wild-type LARC. The primary endpoint was complete pathologic response (pCR) (H0 = 5 %, H1 = 17 %, α = 0.05, β = 0.2). From 19 enrolled patients, 17 (89 %) were evaluable for pathology assessment. Although no pCR was observed, seven patients (41 %) had grade 3 Dworak pathological tumor regression. The regimen was safe and was associated with 95 % of sphincter-preservation rate. No NRAS, BRAF, or PI3KCA mutation was found in this study, but one patient (5 %) showed loss of PTEN expression. The quantification of plasma EGFR ligands during treatment showed significant upregulation of plasma TGF-α and EGF following panitumumab administration (p < 0.05). At surgery, patients with important pathological regression (grade 3 Dworak) had higher plasma TGF-α (p = 0.03) but lower plasma EGF (p = 0.003) compared to those with grade 0-2 Dworak. Our study suggests that concomitant panitumumab and preoperative radiotherapy in KRAS wild-type LARC is feasible and results in some tumor regression. However, pCR rate remained modest. Given that the primary endpoint of our study was not reached, we remain unable to recommend the use of panitumumab as a radiosensitizer in KRAS wild-type LARC outside a research setting.
[Show abstract][Hide abstract] ABSTRACT: Tryptophan catabolism by indoleamine 2,3-dioxygenase 1 (IDO1) plays a key role in tumoral resistance to immune rejection. In humans, constitutive expression of IDO1 has been observed in several tumor types. However, a comprehensive view of its expression in normal and tumor tissues is still required in order to anticipate the risks and potential benefits of IDO1 inhibitors. Using a new validated monoclonal antibody to human IDO1, we performed an extensive immunohistochemical analysis of IDO1 expression in normal and tumor tissues. In normal tissues, IDO1 was expressed by endothelial cells in placenta and lung, and by epithelial cells in the female genital tract. It was also expressed in lymphoid tissues, in cells with a phenotype of mature dendritic cells (CD83+, DC-LAMP+, langerin-, CD123-, CD163-) distinct from plasmacytoid dendritic cells. Importantly, IDO1-expressing dendritic cells were not enriched in tumor-draining lymph nodes, in contrast with previously reported findings. IDO1-expressing cells were observed in a large fraction (505/866, 58%) of human tumors. These cells were tumor cells, endothelial cells and stromal cells, in proportions that varied depending on the tumor type. Tumors showing the highest proportions of IDO1-immunolabelled samples included carcinomas of endometrium and cervix, followed by kidney, lung, and colon. This hierarchy was confirmed by gene expression data mined from the TCGA database. Our observations indicate that the relevant IDO1 for tumoral immune escape is expressed in the tumor itself rather than in its draining lymph node. They also help selecting the most likely tumors to benefit from targeted therapy with IDO1 inhibitors.
[Show abstract][Hide abstract] ABSTRACT: This practical atlas aims to help liver and non liver pathologists to recognize benign hepatocellular nodules on resected specimen. Macroscopic and microscopic views together with immunohistochemical stains illustrate typical and atypical aspects of focal nodular hyperplasia and of hepatocellular adenoma, including hepatocellular adenomas subtypes with references to clinical and imaging data. Each step is important to make a correct diagnosis. The specimen including the nodule and the non-tumoral liver should be sliced, photographed and all different looking areas adequately sampled for paraffin inclusion. Routine histology includes HE, trichrome and cytokeratin 7. Immunohistochemistry includes glutamine synthase and according to the above results additional markers such as liver fatty acid binding protein, C reactive protein and beta catenin may be realized to differentiate focal nodular hyperplasia from hepatocellular adenoma subtypes. Clues for differential diagnosis and pitfalls are explained and illustrated.
World journal of hepatology. 08/2014; 6(8):580-95.
[Show abstract][Hide abstract] ABSTRACT: We report here the case of a 56-year-old man with well-documented cardiac light chain amyloidosis successfully imaged with F-flutemetamol PET/CT in 2011. A matched pair healthy volunteer was imaged in the same conditions, and no specific heart uptake could be detected. The present report suggests the potential benefit of this new pharmaceutical in this indication, as it has been recently reported for C-Pittsburgh compound B, but with the main advantage to benefit from the more practical half-life of F. Further prospective trials would, however, be required to define the potential impact of F-flutemetamol PET/CT in cardiac amyloidosis.
[Show abstract][Hide abstract] ABSTRACT: Progressive liver allograft fibrosis (LAF) is well known to occur long term, as shown by its high prevalence in late posttransplant liver biopsies (LBs). To evaluate the influence of clinical variables and immunosuppression on LAF progression, LAF dynamic was assessed in 54 pediatric liver transplantation (LT) recipients at 6 months, 3 and 7 years post-LT, reviewing clinical, biochemical data and protocol LBs using METAVIR and the liver allograft fibrosis score, previously designed and validated specifically for LAF assessment. Scoring evaluations were correlated with fibrosis quantification by morphometric analysis. Progressive LAF was found in 74% of long-term patients, 70% of whom had unaltered liver enzymes. Deceased grafts showed more fibrosis than living-related grafts (p = 0.0001). Portal fibrosis was observed in correlation with prolonged ischemia time, deceased grafts and lymphoproliferative disease (p = 0.001, 0.006 and 0.012, respectively). Sinusoidal fibrosis was correlated with biliary complications (p = 0.01). Centrilobular fibrosis was associated with vascular complications (p = 0.044), positive autoantibodies (p = 0.017) and high gamma-globulins levels (p = 0.028). Steroid therapy was not associated with reduced fibrosis (p = 0.83). LAF could be viewed as a dynamic process with mostly progression along the time. Peri- and post-LT-associated factors may condition fibrosis development in a specific area of the liver parenchyma.
American Journal of Transplantation 06/2014; · 6.19 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The molecular alterations that drive tumorigenesis in intrahepatic cholangiocarcinoma (ICC) remain poorly defined. We sought to determine the incidence and prognostic significance of mutations associated with ICC among patients undergoing surgical resection.
Annals of Surgical Oncology 06/2014; · 4.12 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Non-alcoholic fatty liver (steatosis) and steatohepatitis (NASH) are hepatic complications of metabolic syndrome. Endoplasmic reticulum (ER) stress is proposed as a crucial disease mechanism in obese and insulin resistant animals (such as ob/ob mice) with simple steatosis but its role in NASH remains controversial. We therefore evaluated the role of ER stress as a disease mechanism in foz/foz mice, which develop both the metabolic and the histological features that mimic human NASH. We explored ER stress markers in the liver of foz/foz mice in response to high-fat diet (HFD) after several time points. We then evaluated the effect of treatment with ER stress inducer tunicamycin, or conversely with ER protectant tauro-ursodeoxycholic acid (TUDCA) on the metabolic and hepatic features. Foz/foz mice are obese, glucose intolerant and develop NASH characterized by steatosis, inflammation, ballooned hepatocytes and apoptosis from 6 weeks of HFD feeding. This was not associated with activation of the upstream unfolded protein response (phospho-eIF2α, IRE1α activity, spliced Xbp1). Activation of JNK and up-regulation of Atf4 and Chop transcripts were however compatible with a "pathologic" response to ER stress. We tested it by intervention experiments. Induction of chronic ER stress failed to worsen obesity, glucose intolerance and NASH pathology in HFD-fed foz/foz mice. In addition, ER protectant TUDCA, although reducing steatosis, failed to improve glucose intolerance, hepatic inflammation and apoptosis in HFD-fed foz/foz mice. These results show that signals driving hepatic inflammation, apoptosis and insulin resistance are independent of ER stress in obese, diabetic mice with steatohepatitis.
[Show abstract][Hide abstract] ABSTRACT: To investigate the activity and expression of EAAT2 glutamate transporter in both in vitro and in vivo models of cholestasis.
This study was conducted on human hepatoblastoma HepG2 cell cultures, the liver of bile duct ligated rats and human specimens from cholestatic patients. EAAT2 glutamate transporter activity and expression were analyzed using a substrate uptake assay, immunofluorescence, reverse transcription-polymerase chain reaction, and immunohistochemistry, respectively.
In HepG2 cells, cholestasis was mimicked by treating cells with the protein kinase C activator, phorbol 12-myristate 13-acetate. Under such conditions, EAAT2 transporter activity was decreased both at the level of substrate affinity and maximal transport velocity. The decreased uptake was correlated with intracellular translocation of EAAT2 molecules as demonstrated using immunofluorescence. In the liver of bile duct ligated rats, an increase in EAAT2 transporter protein expression in hepatocytes was demonstrated using immunohistochemistry. The same findings were observed in human liver specimens of cholestasis in which high levels of γ-glutamyl transpeptidase were documented in patients with biliary atresia and progressive familial intrahepatic cholestasis type 3.
This study demonstrates the alteration in glutamate handling by hepatocytes in liver cholestasis and suggests a potential cross-talk between glutamatergic and bile systems.
World Journal of Gastroenterology 02/2014; 20(6):1554-64. · 2.55 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A 1-year-old girl born to consanguineous parents presented with unexplained liver failure, leading to transplantation at 19 months. Subsequent partial splenectomy for persistent cytopenia showed the presence of foamy cells, and Gaucher disease was confirmed by homozygosity for the p.Leu483Pro mutation in the GBA gene. She was treated by enzyme replacement therapy (ERT). Clinical follow-up showed mild developmental delay, strabismus, nystagmus and oculomotor apraxia. Biochemical studies revealed multiple respiratory chain deficiencies and a mosaic pattern of deficient complex IV immunostaining in liver and fibroblast. Molecular analysis identified a mtDNA depletion syndrome due to the homozygous p.Pro98Leu mutation in MPV17. A younger sister unaffected by mtDNA depletion, presented with pancytopenia and hepatosplenomegaly. ERT for Gaucher disease resulted in visceral normalization without any neurological symptom. A third sister, affected by both conditions, had marked developmental delay, strabismus and ophthalmoplegia but no liver cirrhosis. In conclusion, intrafamilal variability occurs in MPV17-related disease. The combined pathological effect of Gaucher and mitochondrial diseases can negatively impact neurological and liver functions and influence the outcome in consanguineous families. The immunocytochemical staining of OXPHOS protein in tissues and cultured cells is a powerful tool revealing mosaic pattern of deficiency pointing to mtDNA-related mitochondrial disorders.
Molecular Genetics and Metabolism Reports. 01/2014; 1:223–231.
[Show abstract][Hide abstract] ABSTRACT: To report the co-existence of inflammatory hepatocellular adenoma (HCA) and HNF1 inactivated HCA (H-HCA) coming from a multicentric study.
We report 9 cases with co-existence of inflammatory HCA and HNF1 inactivated HCA (H-HCA); 8 occurred in women and 1 in a man. The number of nodules and the sizes of the largest and smallest HCA were variable. In one case, the nodules of the 2 different subtypes were even discovered at different times. In all women, HCAs were histologically typical regardless of their subtype; while H-HCA in the man differed histologically from classical H-HCA.
These cases suggest that predisposition to develop multiple adenomas, hypothetically caused by a "benign tumorigenic field effect" while common to all HCA, may result in different genotypes/phenotypes. Even rare, it is anticipated that more cases with co-existence of different genotypes will emerge due to progress in the use of specific immunohistochemistry. This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: Objectives
Assessment of proliferation by the Ki-67 labelling index (Ki67-LI) is an important parameter of pancreatic neuroendocrine tumour (pNET) prognosis on resection specimens. Ki67-LI values for grading are not fully established on endoscopic ultrasound-guided fine needle aspiration (EUS-FNA). The aim of the study was to determine the accuracy of Ki67-LI on EUS-FNA to predict a final grade of pNET and to analyse the relationship between cytological grading and progression-free survival (PFS). Methods
Between 1996 and 2010, 46 pNETs (33 were resected) from 45 patients were diagnosed by EUS-FNA. Ki67-LI was evaluated on cytological and histological material for each tumour and classified according to the 2010 WHO grading system. ResultsA very good inter-observer agreement for Ki67-LI on EUS-FNA and surgical specimens, respectively, were obtained. Discrepancies were observed between histology and cytology, especially in grade 2 (G2) tumours, where cytology underestimated grading owing to tumour heterogeneity. Still, EUS-FNA was able to distinguish a poor prognostic group, as the actuarial PFS of cytological (c) G3 tumours was 10 ± 4 months versus 29 ± 7 and 68 ± 10 for cG2 and cG1 tumours, respectively (P < 0.0001). Conclusion
This study attests the reproducibility of Ki67-LI of pNETs whether counted on cytology or histology with a very good inter-observer correlation. Determination of Ki67-LI on EUS-FNA of pNETs should be included systematically in their prognostic work-up.
[Show abstract][Hide abstract] ABSTRACT: Cholangiocarcinomas are uncommon tumours with a poor prognosis, that frequently present epidermal growth factor receptor overexpression.
In a multi-centre phase II trial, patients with unresectable cholangiocarcinoma, naïve to chemotherapy, received Cetuximab (400 mg/m(2) at week 1, then 250 mg/m(2)/week) and Gemcitabine (1 g/m(2) on day 1, 8 and 15 every 4 weeks). Primary end point was progression-free survival (PFS) rate at 6 months, using a Simon 2-stage design. Moreover, we assessed the impact of KRAS status and skin toxic effect on efficacy.
Forty-four patients (41% locally advanced/59% metastatic) were enrolled. Median age was 61.5 years; ECOG PS was 0 (68%) or 1. Six months PFS reached 47%. Median OS was 13.5 months [95% confidence interval (CI) 9.8-31.8 months]. Nine patients (20.4%) had PR and disease-control rate was 79.5%. Grade 3/4-related toxic effects were haematological (52.2%), skin rash (13.6%) and fatigue (11.4%). KRAS mutations were found in 7 of 27 patients and had no influence on PFS. Skin toxic effect ≥grade 2 was associated with increased PFS (P = 0.05).
Our study met its primary end point, suggesting that Gemcitabine-Cetuximab has activity in cholangiocarcinoma. KRAS status was not associated with PFS, unlike skin toxic effect, which could be used as a surrogate marker for efficacy. ClinicalTrials.gov Identifier: NCT00747097.
[Show abstract][Hide abstract] ABSTRACT: Data on quality control of the pathologic evaluation of total mesorectal excision specimens are scarce. We aimed to assess differences between evaluation by local pathologists participating in PROCARE, a Belgian improvement project on rectal cancer, and by a review panel of experts.
Based on photographic material and histopathology slides, a review committee of gastrointestinal expert pathologists re-evaluated the mesorectal plane, the tumour differentiation grade, the (y)pT stage and the tumour regression grade in 444 cases previously routinely assessed by local pathologists.
The surgical plane was reported in 89% and the circumferential resection margin in 88% of cases by the local pathologist. The median number of lymph nodes harvested in patients undergoing neoadjuvant radiochemotherapy was 11, and 14 in the other patients. The review committee downgraded the surgical plane in 17% from (intra)mesorectal to intramuscular, and upgraded it in 27% from intramuscular to (intra)mesorectal. Tumour differentiation grade, T stage and tumour regression grade differed between local pathologists and review committee in 15%, 10% and 38%, respectively. T stage was upgraded in 8% of cases, mainly from T2 to T3. Tumour regression was judged by the review panel to be less advanced in 15% of cases.
Acknowledging some shortcomings, this study gives a realistic view of clinical practice. There are differences in interpretation with regard to both macroscopic and microscopic analysis of TME specimens. These findings indicate a need for more objective and reproducible criteria in histopathology. Being aware of this is a first step for improvement. This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: Focal nodular hyperplasia (FNH) and hepatocellular adenoma (HCA) are benign hepatocellular tumors that develop most frequently in females and in non-cirrhotic livers. HCA are prone to bleed and to transform into hepatocellular carcinoma (HCC). Four major subgroups of HCA have been thus far identified: HNF1α mutated HCA, inflammatory HCA (IHCA), β-catenin mutated HCA (b-HCA and b-IHCA), based on mutations in specific oncogenes and tumor suppressors. B-HCA and b-IHCA are strongly associated with HCC transformation. Benign hepatocellular tumors can be classified using immunohistochemistry (LFABP, CRP, GS, b-catenin). Analysis of HCA phenotypes has led to the identification of patients at risk of HCC transformation and therefore improved the indications provided by invasive and non-invasive diagnostic techniques, such as biopsies and MRI. These recent advances have broadened the clinical scope of HCA in various conditions, such as their presence in males, in obese patients, in patients suffering from liver vascular disorders, genetic diseases. However, specific immunohistochemistry has shown limitations particularly for the identification of b-HCA, thereby, outlining the importance of molecular studies to improve the diagnosis/prognosis of HCA. If evaluation of prognosis and treatment has benefited from these advances, much more needs to be done to obtain guidelines for good clinical practice.
Gastroentérologie Clinique et Biologique 07/2013; · 0.80 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background: Alcohol consumption is a major cause of liver disease. It also associates with increased cardiovascular risk and type 2 diabetes. Alcoholic liver diseases (ALD) and non-alcoholic fatty liver diseases (NAFLD) share pathologic features, pathogenic mechanisms and pattern of disease progression. In NAFLD, steatosis, lipotoxicity and liver inflammation participate to hepatic insulin resistance. Our aim here is to verify the effect of alcohol on hepatic insulin sensitivity and to evaluate the role of alcohol-induced steatosis and inflammation on glucose homeostasis. Results : C57BL/6J mice were fed for 20 days a modified Lieber-DeCarli diet in which alcohol concentration was gradually increased up to 35% of daily caloric intake. Alcohol-fed mice show liver steatosis and inflammatory infiltration. OH-fed mice developed insulin resistance in the liver but not in muscles, as demonstrated by euglycemic-hyperinsulinemic clamp and analysis of the insulin signaling cascade. Treatment with the PPAR-α agonist Wy14,643 protected against OH-induced steatosis and Kupffer cell (KC) activation and almost abolished OH-induced insulin resistance. As KC activation may modulate insulin sensitivity, we repeated the clamp studies in mice depleted in KC to decipher the role of macrophages. Depletion of KC using liposomes-encapsuled clodronate in OH-fed mice failed both to improve hepatic steatosis and to restore insulin sensitivity as assessed by clamp. Conclusions : Our study shows that chronic alcohol consumption induces steatosis, Kuffer cell activation and hepatic insulin resistance in mice. PPAR-α agonist treatment that prevents steatosis and dampens hepatic inflammation also prevents alcohol-induced hepatic insulin resistance. However, Kupffer cell depletion little impacts on OH-induced metabolic disturbances.
[Show abstract][Hide abstract] ABSTRACT: β cell plasticity governs the adjustment of β cell mass and function to ensure normoglycemia. The study of how β cell mass is controlled and the identification of alternative sources of β cells are active fields of research. β cell plasticity has been implicated in numerous physiological and pathological conditions. We developed a mice model in which we induced major β cell mass atrophy by implanting insulin pellets (IPI) for 7 or 10 days. The implants were then removed (IPR) to observe the timing and characteristics of β cell regeneration in parallel to changes in glycemia. Following IPR, the endocrine mass was reduced by 60% at day 7 and by 75% at day 10, and transient hyperglycemia was observed, which resolved within 1 week. Five days after IPR, enhanced β cell proliferation and an increased frequency of small islets were observed in 7-day IPI mice. β cell mass was fully restored after an additional 2 days. For the 10-day IPI group, β cell and endocrine mass were no longer significantly different from those of the control group at 2 weeks post-IPR. Furthermore, RT qPCR analysis of endocrine structures isolated by laser capture microdissection (LCM) indicated sequentially enhanced expression of the pancreatic transcription factors Beta2/NeuroD and Pdx-1 post-IPR. Thus, our data suggest this mouse model of β cell plasticity not only relies on replication but also involves enhanced cell differentiation plasticity.
AJP Endocrinology and Metabolism 02/2013; · 4.51 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: BACKGROUND AND AIMS: The objective of this report was to determine the prevalence of underlying nonalcoholic steatohepatitis in resectable intrahepatic cholangiocarcinoma. METHODS: Demographics, comorbidities, clinicopathologic characteristics, surgical treatments, and outcomes from patients who underwent resection of intrahepatic cholangiocarcinoma at one of eight hepatobiliary centers between 1991 and 2011 were reviewed. RESULTS: Of 181 patients who underwent resection for intrahepatic cholangiocarcinoma, 31 (17.1 %) had underlying nonalcoholic steatohepatitis. Patients with nonalcoholic steatohepatitis were more likely obese (median body mass index, 30.0 vs. 26.0 kg/m(2), p < 0.001) and had higher rates of diabetes mellitus (38.7 vs. 22.0 %, p = 0.05) and the metabolic syndrome (22.6 vs. 10.0 %, p = 0.05) compared with those without nonalcoholic steatohepatitis. Presence and severity of hepatic steatosis, lobular inflammation, and hepatocyte ballooning were more common among nonalcoholic steatohepatitis patients (all p < 0.001). Macrovascular (35.5 vs. 11.3 %, p = 0.01) and any vascular (48.4 vs. 26.7 %, p = 0.02) tumor invasion were more common among patients with nonalcoholic steatohepatitis. There were no differences in recurrence-free (median, 17.0 versus 19.4 months, p = 0.42) or overall (median, 31.5 versus 36.3 months, p = 0.97) survival after surgical resection between patients with and without nonalcoholic steatohepatitis. CONCLUSIONS: Nonalcoholic steatohepatitis affects up to 20 % of patients with resectable intrahepatic cholangiocarcinoma.
Journal of Gastrointestinal Surgery 01/2013; · 2.36 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: OBJECTIVES: The most frequent presentation of chemotherapy-related toxicity in colorectal liver metastases (CRLM) is sinusoidal obstruction syndrome (SOS). The purpose of the present study was to identify preoperative factors predictive of SOS and to establish associations between type of chemotherapy and severity of SOS. METHODS: A retrospective study was carried out in a tertiary academic referral hospital. Patients suffering from CRLM who had undergone resection of at least one liver segment were included. Grading of SOS on the non-tumoral liver parenchyma was accomplished according to the Rubbia-Brandt criteria. A total of 151 patients were enrolled and divided into four groups according to the severity of SOS (grades 0-3). RESULTS: Multivariate analysis identified oxaliplatin and 5-fluorouracil as chemotherapeutic agents responsible for severe SOS lesions (P < 0.001 and P = 0.005, respectively). Bevacizumab was identified as having a protective effect against the occurrence of SOS lesions (P = 0.005). Univariate analysis identified the score on the aspartate aminotransferase : platelets ratio index (APRI) as the most significant biological factor predictive of severe SOS lesions. Splenomegaly is also significantly associated with the occurrence of severe SOS lesions. CONCLUSIONS: The APRI score and splenomegaly are effective as factors predictive of SOS. Bevacizumab has a protective effect against SOS.