Introduction. Alors qu’elle constitue le seul traitement curatif du cholangiocarcinome intrahépatique (CCIH), la résection reste associée à un taux de récidive supérieur à 60% et un taux de survie réelle à 5 ans inférieur à 20%. Une estimation fiable du pronostic ainsi qu’une meilleure compréhension de la biologie tumorale est essentielle pour améliorer le pronostic.Méthodes. A l’appui des données clinico-biologiques de deux larges cohortes de patients avec CCIH réséqué (MSKCC, n=189 et AFC, n=522), trois objectifs ont été explorés. Tout d’abord, définir quel modèle pronostique publié est le plus performant. Ensuite, définir la fiabilité de l’évaluation pronostique préopératoire à partir de, respectivement, l’imagerie, des microARN (miR) circulants diagnostiques et du profil génomique tumoral. Enfin, évaluer l’impact pronostique de la survenue d’événements périopératoires tels que transfusion et morbidité.Résultats. Premièrement, les nomogrammes apportaient une meilleure estimation pronostique en comparaison à la classification AJCC 7ème édition. Deuxièmement, la taille et la multifocalité tumorale sur l’imagerie préopératoire permettaient de différencier deux groupes de patients de pronostic clairement distincts (p<0,001). L’existence d’une mutation d’un gène de remodelage de la chromatine (BAP1, ARID1A, PBRM1) tendait à être associé à une survie sans récidive plus favorable qu’en l’absence de mutation (p=0,09). Alors qu’ayant un potentiel comme marqueur diagnostique circulant, miR21 et miR221 n’étaient pas associé à la survie. Troisièmement, la transfusion peropératoire n’impactait pas la survie à long terme alors que la survenue d’une complication sévère (grade Dindo-Clavien > 2) était indépendamment associée à une survie globale plus courte (p=0,002).Conclusion. Alors que les nomogrammes postopératoires apportent une meilleure estimation pronostique, le développement de modèles pronostiques préopératoires est faisable notamment à partir de l’imagerie et de marqueurs biologiques tumoraux complémentaires. / Introduction. Complete resection stands as the only curative option for intrahepatic cholangiocarcinoma (IHCC). Still, prognosis remains poor after resection due to a recurrence rate over 60% leading to actual 5-year survival rates below 20%. Reliable prognostic estimation and better understanding of tumor biology would be of interest for improving IHCC prognosis.Methods. Using clinical and biological data from two large cohort of resected IHCC (MSKCC, n=189 and AFC, n=522), three objectives have been explored. First, assessing the performances of different published prognostic models. Second, defining the reliability of preoperative prognostic estimation using imaging, tumoral genomic profiling and circulating tumoral microRNA (miR). Third, evaluating the prognostic impact of perioperative events such as blood transfusion and morbidity.Results. First, nomograms displayed better prognostic accuracy over the AJCC 7th edition staging system. Second, tumor size and multifocality on preoperative imaging allowed patient stratification in groups statistically different regarding prognosis (p<0.001). Further, the presence of chromatine remodeling gene mutations (BAP1, ARID1A, PBRM1) tended towards longer recurrence-free survical (p=0,09). Some diagnostic circulating miR such as miR21 and miR221 were not associated with survival. Third, in contrast with intraoperative transfusion, the occurrence of severe morbidity (Dindo-Clavien grade > 2) was independently associated with shorter overall survival (p=0.002).Conclusion. Nomograms outperform conventional staging sytem. Preoperative prognostic estimation is feasible and reliable using imaging. Identifying new prognostic biomarkers would help refining preoperative prognostic estimation.

Cholangiocarcinoma (CCA) is a fatal primary malignancy that is formed in the bile ducts. Cancer-associated myofibroblasts play a crucial role in CCA proliferation and invasion. Furthermore, there is a growing interest in the role of the exosome in the interaction between the cancer-associated myofibroblasts and cholangiocarcinoma which lead to CCA growth. However how cholangiocarcinoma-derived exosome affect the cancer-associated myofibroblasts in the tumor microenvironment remain unknown. In this study, we examined whether exosome produced by cholangiocarcinoma could involve in the prompt of CCA cells growth by regulation of myofibroblast. We found that cholangiocarcinoma-derived exosome could prompt elevated α-smooth muscle actin and stromal cell-derived factor one expression that induces myofibroblast proliferation. We then demonstrated that cholangiocarcinoma-derived exosome upregulated periostin expression that plays an important role in cancer metastasis. In 3D organotypic rat CCA coculture model, TCA and S1P considerably increase the growth of CCA cell. Conclusion: cholangiocarcinoma-derived exosome trigger cancer-associated myofibroblasts proliferation in the tumor microenvironment that leads to prompt CCA growth.

A análise das proteínas transportadoras de compostos biliares, antes restrita à fisiologia e à fisiopatologia de colestases, recentemente passou a incluir neoplasias hepato-biliares. O presente estudo teve como objetivo caracterizar a expressão das proteínas ABC de transporte biliar BSEP, MDR3, MRP2 e MRP3 em amostras retrospectivamente colecionadas de 80 casos de autópsias de carcinoma hepatocelular (CHC) e 56 casos de ressecção cirúrgica de colangiocarcinoma (CC). Áreas representativas das neoplasias foram organizadas em tissue microarrays e submetidas à pesquisa imuno-histoquímica (IHQ) com o anticorpo policlonal anti-BSEP (HPA019035) e os anticorpos monoclonais anti-BSEP (F6), anti-MDR3 (P3 II-26), anti-MRP2 (M2 III-6) e anti-MRP3 (DTX-1) com amplificação de sinal mediante uso de sistema de polímeros curtos conjugados à peroxidase. A comparação entre a positividade das reações imuno-histoquímicas para cada anticorpo e as variáveis anatomopatológicas foi realizada através dos testes de qui-quadrado de Pearson ou Exato de Fisher. A positividade das reações IHQ cujos anticorpos propiciaram melhor distinção do sinal positivo vs coloração inespecífica de fundo e detecção de casos positivos e/ou melhor capacidade de discriminar as duas neoplasias hepáticas foi comparada com a positividade observada para as reações IHQ com os anticorpos anti-CEA policlonal, anti-Hep-par-1 e anti-Arginase-1. A expressão canalicular de BSEP nos CHC foi observada em 77,3% (58/75) com o anticorpo monoclonal e 75,9% (60/79) com o anticorpo policlonal. Não foi detectada associação significativa da expressão de BSEP em relação ao tamanho, número dos nódulos e grau de diferenciação de CHC, tendo apenas sido significativamente reduzida (P < 0,05) tal reação nos casos de padrão arquitetural mais complexo. A reatividade dos CHC para o anticorpo monoclonal anti-BSEP foi aparentemente menor que a obtida com a expressão canalicular de CEA, Hep-par-1 e Arginase-1 no CHC, mas esses valores não atingiram significância estatística. Todos os casos de colangiocarcinoma foram negativos para reações IHQ para pesquisa de BSEP, resultado significativamente diferente (P=0,0001) do obtido com uso do Ac policlonal anti-CEA (padrão circunferencial) e Hep-par-1, não tendo sido demonstrada diferença significativa (P=0,222) da expressão de BSEP e de Arginase-1. A expressão canalicular de MDR3 foi observada em 56,4% (44/78) dos casos de CHC, não tendo sido detectada associação significativa quanto ao tamanho e número de nódulos. Foi observada expressão significativamente menor de MDR3 nos casos de CHC de padrão mais complexo (P=0,009), e nos casos de maior grau histológico (P=0,005). A expressão de MDR3 em CHC foi significativamente menor que a de CEA, Hep-par-1 e Arginase-1 (P < 0,05). Todos os casos de colangiocarcinoma foram negativos para a avaliação da expressão de MDR3, diferindo significativamente em relação a expressão de CEA (P=0,001), mas não em comparação a Hep-par-1 e Arginase-1 (P > 0,05). As reações IHQ para detecção de MRP2 exibiram positividade canalicular em 92,3% dos casos de CHC e em 96,3% nos casos de CC. A detecção da alta expressão de MRP2 no CHC foi constante (P > 0,05) em comparação ao tamanho, número dos nódulos, padrão arquitetural e grau histológico de diferenciação de CHC assim como, também não apresentou associação (P > 0,05) com a localização, padrão de crescimento e grau de diferenciação do CC. A reação IHQ para MRP3 resultou positiva em 15/80 casos de CH (18,8%). A reatividade IHQ para MRP foi detectada em 24/54 (44,5%) de CC. Diferente dos transportadores descritos acima, a expressão de MRP3 foi preferencialmente basolateral. A positividade para MRP3 não variou (P > 0,05) em relação ao número, tamanho dos nódulos, padrão arquitetural (inclusive os sólidos), e grau de diferenciação (inclusive os menos diferenciados). A proteína MRP3 esteve expressa regularmente (P > 0,05) em todos os casos de CC, apresentando-se reduzida apenas no subtipo histológico ductular (P=0,023). Em conclusão, o excelente contraste de reação, a frequência razoavelmente alta de positividade de CHC e a plena negatividade de CC para BSEP levam-nos a recomendar a introdução do anticorpo monoclonal anti-BSEP no painel adotado para o diagnóstico diferencial dessas duas neoplasias. A alta expressão de MRP2 no CHC e no CC é conservada independentemente dos parâmetros anatomopatológicos avaliados. A expressão do transportador MRP3 mostrou variação dentre os subtipos histológicos de CC, aspecto que torna promissoras pesquisas futuras para avaliação mais detalhada da expressão deste marcador nos colangiocarcinomas / The assessment of biliary transporters, previously restricted to the physiology and pathophysiology of cholestasis, has recently included hepato-biliary neoplasms. The present study aimed to characterize the expression of BSEP, MDR3, MRP2 and MRP3 biliary transport proteins in retrospectively collected samples from 80 cases of autopsy of hepatocellular carcinoma (HCC) and 56 cases of surgical resection of cholangiocarcinoma (CC). Representative areas of the neoplasms were organized into tissue microarrays and submitted to immunohistochemical (IHC) reaction with polyclonal antibody anti-BSEP (HPA019035) and monoclonal antibodies anti-BSEP (F6), MDR3 (P3 II-26), MRP2 (M2 III-6) and MRP3 (DTX-1). Signal amplification was achieved with a short polymer system conjugated to peroxidase. The comparison between the positivity of the immunohistochemical reactions for each antibody and the pathological variables was performed using the Pearson chi-square test or the Fisher\'s exact test. The performance of antibodies which provided a better distinction of the positive signal vs nonspecific background staining and yield better discrimination between the two hepatic neoplasms was compared with that achieved with the already accepted HCC markers polyclonal anti-CEA, Hep-par-1 and Arginase-1. The canalicular expression of BSEP in HCC was observed in 77.3% (58/75) with the monoclonal antibody and 75.9% (60/79) with the polyclonal antibody. BSEP expression levels were not significantly different according to tumor size, number of nodules and degree of differentiation. The frequency of positive reaction of HCC cases with the monoclonal anti-BSEP was apparently lower than that achieved with the canalicular expression of CEA, Hep-par-1 and Arginase-1, but these values did not reach statistical significance. All cases of cholangiocarcinoma were negative for IHC reactions to BSEP, which was significantly different (P=0.0001) from the results obtained with polyclonal anti-CEA (circumferential pattern) and Hep-par-1, but not from the resultas achieved with Arginase-1 (P=0.222). The canalicular expression of MDR3 was observed in 56.4% (44/78) of HCC cases. Among histological variables, only the finding of more complex architecture (P=0.009) and higher histological grade (P=0.005) of HCC yielded, significantly lower expression of MDR3. The expression of MDR3 in HCC was significantly lower than that of CEA, Hep-par-1 and Arginase-1 (P < 0.05). All cases of cholangiocarcinoma were negative for the evaluation of MDR3 expression, differing significantly with that achieved with polyclonal anti-CEA (P=0.001) but not with that achieved with Hep-par-1 or with Arginase-1 (P > 0.05). The IHC reactions with the MRP2 antibody exhibited canalicular positivity in 92.3% of HCC cases and 96.3% in CC cases. High expression of MRP2 in HCC was constant (P > 0.05) despite changes in size, number of nodules, architectural pattern and histological degree of HCC differentiation, as well as no association (P > 0.05) with the location, pattern of growth and degree of differentiation of CC. The IHC reaction for MRP3 was positive in 15/80 cases of HCC (18.8%) and in 24/54 (44.5%) of CC. Unlike the carriers described above, the hepatocellular expression of MRP3 was preferentially basolateral. Positivity for MRP3 did not vary (P > 0.05) in relation to number, nodule size, architectural standard (including solids), and degree of differentiation. The MRP3 protein was expressed regularly (P > 0.05) in different presentations of CC, but significant lower frequency of positivity was found in the ductular histological subtype (P=0.023). In conclusion, the excellent signal-to-noise ratio, reasonably high frequency of HCC positivity and full negativity of CC to BSEP lead us to recommend the introduction of the anti-BSEP monoclonal antibody in the panel adopted for the differential diagnosis of these two neoplasms. The high expression of MRP2 in HCC and in CC is conserved independently of the pathological parameters evaluated herein. The frequency of expression of the MRP3 transporter varied among the histological subtypes of CC, which makes promising future research for a more detailed assessment of the expression of this marker in the cholangiocarcinomas

Background: The association of body mass index (BMI) and long-term prognosis and outcome of patients with perihilar cholangiocarcinoma (pCCA) has not been well defined. The aim of this study was to evaluate clinicopathologic and oncologic outcomes with pCCA undergoing resection, according to their BMI. Methods: Patients undergoing liver resection in curative intention for pCCA at a tertiary German hepatobiliary (HPB) center were identified from a prospective database. Patients were classified as normal weight (BMI 18.5–24.9 kg/m2), overweight (BMI 25.0–29.9 kg/m2) and obese (>30 kg/m2) according to their BMI. Impact of clinical and histo-pathological characteristics on recurrence-free survival (RFS) were assessed using Cox proportional hazard regression analysis among patients of all BMI groups. Results: Among a total of 95 patients undergoing liver resection in curative intention for pCCA in the analytic cohort, 48 patients (50.5%) had normal weight, 33 (34.7%) were overweight and 14 patients (14.7%) were obese. After a median follow-up of 4.3 ± 2.9 years, recurrence was observed in totally 53 patients (56%). The cumulative recurrence probability was higher in obese and overweight patients than normal weight patients (5-year recurrence rate: obese: 82% versus overweight: 81% versus normal weight: 58% at 5 years; p = 0.02). Totally, 1-, 3-, 5- and 10-year recurrence-free survival rates were 68.5%, 44.6%, 28.9% and 13%, respectively. On multivariable analysis, increased BMI (HR 1.08, 95% CI: 1.01–1.16; p = 0.021), poor/moderate tumor differentiation (HR 2.49, 95% CI: 1.2–5.2; p = 0.014), positive lymph node status (HR 2.01, 95% CI: 1.11–3.65; p = 0.021), positive resection margins (HR 1.89, 95% CI:1.02–3.4; p = 0.019) and positive perineural invasion (HR 2.92, 95% CI: 1.02–8.3; p = 0.045) were independent prognostic risk factors for inferior RFS. Conclusion: Our study shows that a high BMI is significantly associated with an increased risk of recurrence after liver resection in curative intention for pCCA. This factor should be considered in future studies to better predict patient’s individual prognosis and outcome based on their BMI.

Objectives: In our study, we evaluated and compared the prognostic value and performance of the 6th, 7th, and 8th editions of the American Joint Committee on Cancer (AJCC) staging system in patients undergoing surgery for perihilar cholangiocarcinoma (PHC). Methods: Patients undergoing liver surgery with curative intention for PHC between 2002 and 2019 were identified from a prospective database. Histopathological parameters and stage of the PHC were assessed according to the 6th, 7th, and 8th editions of the tumor node metastasis (TNM) classification. The prognostic accuracy between staging systems was compared using the area under the receiver operating characteristic curve (AUC) model. Results: Data for a total of 95 patients undergoing liver resection for PHC were analyzed. The median overall survival time was 21 months (95% CI 8.1–33.9), and the three- and five-year survival rates were 46.1% and 36.2%, respectively. Staging according to the 8th edition vs. the 7th edition resulted in the reclassification of 25 patients (26.3%). The log-rank p-values for the 7th and 8th editions were highly statistically significant (p ≤ 0.01) compared to the 6th edition (p = 0.035). The AJCC 8th edition staging system showed a trend to better discrimination, with an AUC of 0.69 (95% CI: 0.52–0.84) compared to 0.61 (95% CI: 0.51–0.73) for the 7th edition. Multivariate survival analysis revealed male gender, age >65 years, positive resection margins, presence of distant metastases, poorly tumor differentiation, and lymph node involvement, such as no caudate lobe resection, as independent predictors of poor survival (p < 0.05). Conclusions: In the current study, the newly released 8th edition of AJCC staging system showed no significant benefit compared to the previous 7th edition in predicting the prognosis of patients undergoing liver resection for perihilar cholangiocarcinoma. Further research may help to improve the prognostic value of the AJCC staging system for PHC—for instance, by identifying new prognostic markers or staging criteria, which may improve that individual patient’s outcome.

Le cholangiocarcinome (CC) est un cancer des voies biliaires qui représente environ 15% des cancers primitifs du foie,mais de pronostic redoutable en raison d'un diagnostic tardif faute de marqueurs spécifiques.La présence d'auto anticorps (Ac) est rapportée comme marqueurs diagnostiques précoce de certains cancers.La présence d'auto-Ac dans le CC n'a pas été signalée, et aucun biomarqueur immunologique de cette maladie n'a été identifié. L'objectif de notre étude était d'identifier des auto-Ac potentiellement utilisables comme biomarqueur de CC, par analyse sérologique du protéome. Des immunoblots ont été réalisés à partir de la séparation par électrophorèse 2D de protéines de lignées tumorales de CC, CCSW1 et CCLP1, de 5 pièces d'hépatectomie ac leur partie tumorale et non tumorale,ainsi que de foie normal de neuropathie amyloïde. Les sérums de 13 patients atteints de CC et un pool de 10 sujets sains ont été testés sur ces immunoblot.La comparaison informatique des profils des protéines immunomarquées par les sérums des patients comparés aux profils des contrôles a permis de définir des spots immunoréactifs d'intérêt.Ces spots d'intérêt marqués par plus d'1/3 de sérums ont été ensuite identifiés par spectrométrie de masse de type Orbitrap®.Ainsi, nous avons identifié 10 protéines d'intérêt de CCSW1,11 protéines de CCLP1, 9 de la partie tumorale des foies, 14 des parties non-tumoral et 16 protéines appartenant au foie normal.Une extrême variabilité était observée selon les sérums pour un même Ag.Différents profils de réactivité étaient observés sur le même extrait antigénique en fonction des sérums testés, et pour un même sérum selon l'extrait antigénique utilisé.Il en résulte qu'un AC d'intérêt donné ne peut être considéré comme biomarqueur de CC que pour une faible proportion de patients.Pr cette raison, il faut envisager la combinaison de plusieurs anticorps pour avoir un test avec une sensibilité et une spécificité utilisable en clinique. Les protéines identifiées ont été classées par bio-informatique (logiciel Panther®) selon la description des gènes et de leurs produits selon une ontologie commune à toutes les espèces : fonctions moléculaires effectuées, processus biologiques assurés et localisation subcellulaire.Dans cette classification, 2 profils d'immunoréactivité se distinguent. La grande majorité des protéines cibles d'intérêt avec une fonction catalytique étaient présentes dans le foie normal ou dans les parties non tumorales des exérèses. L'autre profil était celui des protéines-cibles avec une fonction de protéines structurale et étaient présentes dans les lignées cellulaires tumorales ainsi que des parties tumorales des hépatectomies. Les protéines identifiées avec une activité catalytique étaient:l'alpha-énolase, le fructose biphosphate aldolase B et la glyceraldedyde 3-phosphate déshydrogénase, toutes troisréactives avec+de 50% des sérums de CC. Les protéines de structure identifiées par+de 60% des sérums de CC provenaient des lignées cellulaires et des tissus tumoraux.Il s'agissait de la vimentine, des prélamines A / C, de l'annexine A2 et de l'actine.Enfin, la sérotranferrine, protéines de transport, est reconnues par 100% des sérums CC en utilisant comme antigène des tissus tumoraux.Une sensibilité importante et une spécificité élevée sont des caractéristiques princeps d'un Ac pour pouvoir l'utiliser comme biomarqueur.La plupart des auto-Ac détectés dans cette étude avaient déjà été rapportées dans d'autres cancers et maladies auto-immunes. Pour trouver des protéines antigéniques spécifiques du CC, une combinaison de plusieurs semble nécessaire afin de permettre le développement de nouveaux biomarqueurs pour le diagnostic et le pronostic des CC.En conclusion, les biomarqueurs potentiels proposés dans cette étude doivent être testés en différentes combinaisons avec un panel en nb significatif de patients et en utilisant le substrat antigénique le plus approprié comme défini au cours de cette étude. / Cholangiocarcinoma (CC) is a rare but fatal primary liver cancer and accounts for an estimated 15% of primary liver cancer worldwide. It is associated with high mortality due to the lack of established diagnostic approaches. Autoantibodies can be used clinically as diagnostic markers for early cancer detection of cholangiocarcinoma. Studies, indicating the presence of auto-antibodies (AAbs) in CC have not been reported yet. No immunological biomarker, correlated to the disease, has been identified. The objective of our study was to identify cellular proteins from liver tissues (tumoral and non tumoral) and cholangiocarcinoma cell lines which could be recognized by antibody of CC patients. We used serological proteome analysis (SERPA) technique which leads us to suggest some molecules as potential biomarkers for the early diagnosis of CC. Proteins from different origins were 2DE separated: CCSW1 and CCLP1 tumor cell lines, five different samples of hepatectomies for CC with respect to their tumoral and non-tumoral counterparts and a normal liver from amyloid neuropathy. Sera from 13 CC patients and a pool of 10 healthy subjects were probed on immunoblot performed with these different separations. Comparison of immunoblotting patterns given by patient’s sera compared to patterns given by controls allowed to define immunoreactive spots of interest and those reacting with more than one-third of sera were identified by orbitrap type mass spectrometry. In this way we identified 10, 11, 9, 14 and 16 proteins from CCSW1, CCLP1, tumor part, non-tumor counterpart and normal liver antigenic extracts respectively. Different patterns of reactivity were observed according to sera on the same antigenic extract, and for a same serum, according to the antigenic extract, even though few common patterns were also observed. This widespread of reactivity is not unusual and reported earlier in several studies of this sort. It is indicated that a single AAb have an ability to identify only a small proportion of patient. For this reason, several antibodies in combination must be used to ensure sensitivity and specificity of assays used in the daily clinic.Identified proteins were then categorized by gene ontology analysis by which they fall into three main groups; biological process and molecular functions, protein class and molecular pathway and cellular component, according to the Panther classification. By Gene Ontology classification, two different patterns of targeted antigens were observed. The vast majority of targeted-proteins with catalytic activity were found in normal liver or non-tumor specimens. The second pattern was mainly represented by targeted proteins categorized as structural proteins extracted from CC cell lines and tumor tissues. Proteins identified with catalytic activity were: alpha-enolase, fructose biphosphate aldolase B and glyceraldedyde 3-phosphate dehydrogenase; which were reactive with more than 50% of CC sera. Proteins identified with structural activity, and detected with high rates by using cell lines and tumor tissues, were: vimentin, prelamine A/C, annexin A2 and actin; reactivity of each protein was higher than 62% with CC sera. Serotranferrin, identified under the category of transfer/carrier proteins, recognized by 100% of CC sera by using tumor tissues.High sensitivity and specificity is a prime requisite of AAbs that might be used as CC biomarkers for CC diagnosis. Most of the AAbs detected in this study had previously been reported in other cancers and auto-immune disorders. Hence it is essential to prove the specificity of antigenic proteins, a combination of various antigens therefore needs to be tested to enable the development of new biomarkers for the diagnosis and prognosis of CC.In conclusion, the proposed potential biomarkers need to be tested in a variety of different combinations with a panel of significant number of patients and using the most appropriate substrate defined during this study.

Cholangiocarcinoma (CC) is a rare but fatal primary liver cancer and accounts for an estimated 15% of primary liver cancer worldwide. It is associated with high mortality due to the lack of established diagnostic approaches. Autoantibodies can be used clinically as diagnostic markers for early cancer detection of cholangiocarcinoma. Studies, indicating the presence of auto-antibodies (AAbs) in CC have not been reported yet. No immunological biomarker, correlated to the disease, has been identified. The objective of our study was to identify cellular proteins from liver tissues (tumoral and non tumoral) and cholangiocarcinoma cell lines which could be recognized by antibody of CC patients. We used serological proteome analysis (SERPA) technique which leads us to suggest some molecules as potential biomarkers for the early diagnosis of CC. Proteins from different origins were 2DE separated: CCSW1 and CCLP1 tumor cell lines, five different samples of hepatectomies for CC with respect to their tumoral and non-tumoral counterparts and a normal liver from amyloid neuropathy. Sera from 13 CC patients and a pool of 10 healthy subjects were probed on immunoblot performed with these different separations. Comparison of immunoblotting patterns given by patient's sera compared to patterns given by controls allowed to define immunoreactive spots of interest and those reacting with more than one-third of sera were identified by orbitrap type mass spectrometry. In this way we identified 10, 11, 9, 14 and 16 proteins from CCSW1, CCLP1, tumor part, non-tumor counterpart and normal liver antigenic extracts respectively. Different patterns of reactivity were observed according to sera on the same antigenic extract, and for a same serum, according to the antigenic extract, even though few common patterns were also observed. This widespread of reactivity is not unusual and reported earlier in several studies of this sort. It is indicated that a single AAb have an ability to identify only a small proportion of patient. For this reason, several antibodies in combination must be used to ensure sensitivity and specificity of assays used in the daily clinic.Identified proteins were then categorized by gene ontology analysis by which they fall into three main groups; biological process and molecular functions, protein class and molecular pathway and cellular component, according to the Panther classification. By Gene Ontology classification, two different patterns of targeted antigens were observed. The vast majority of targeted-proteins with catalytic activity were found in normal liver or non-tumor specimens. The second pattern was mainly represented by targeted proteins categorized as structural proteins extracted from CC cell lines and tumor tissues. Proteins identified with catalytic activity were: alpha-enolase, fructose biphosphate aldolase B and glyceraldedyde 3-phosphate dehydrogenase; which were reactive with more than 50% of CC sera. Proteins identified with structural activity, and detected with high rates by using cell lines and tumor tissues, were: vimentin, prelamine A/C, annexin A2 and actin; reactivity of each protein was higher than 62% with CC sera. Serotranferrin, identified under the category of transfer/carrier proteins, recognized by 100% of CC sera by using tumor tissues.High sensitivity and specificity is a prime requisite of AAbs that might be used as CC biomarkers for CC diagnosis. Most of the AAbs detected in this study had previously been reported in other cancers and auto-immune disorders. Hence it is essential to prove the specificity of antigenic proteins, a combination of various antigens therefore needs to be tested to enable the development of new biomarkers for the diagnosis and prognosis of CC.In conclusion, the proposed potential biomarkers need to be tested in a variety of different combinations with a panel of significant number of patients and using the most appropriate substrate defined during this study.

Le cholangiocarcinome (CC) est une tumeur maligne au pronostic péjoratif dont le traitement repose sur la résection chirurgicale. Il représente 3 % de l’ensemble des cancers digestifs et il est la deuxième tumeur primitive du foie, en fréquence, derrière le carcinome hépatocellulaire. L’âge moyen est de 70 ans avec une prédominance masculine. On distingue actuellement les cholangiocarcinomes intra et extra-hépatiques. La survie est inférieure à 5% à 5 ans tous stades confondues. 60 à 70 % sont des tumeurs de la convergence des canaux biliaires appelées également tumeurs de Klatskin.Le but de ce travail était de répondre aux interrogations persistantes concernant le cholangiocarcinome péri-hilaire (CCPH) en appliquant différentes méthodes statistiques sur différentes bases de données et revue de la littérature.Les trois axes principaux de ce travail s’articulent selon la temporalité de prise en charge, du diagnostic aux suites post-opératoires en passant par la mise en condition préopératoire.Le premier axe repose sur une base de données locale (registre de cancer digestif du Calvados). Les résultats montrent que le CCPH constitue seulement un tiers des cholangiocarcinomes dans la population générale, que son taux d’incidence est stable avec néanmoins une diminution d’incidence, bien que non-significative, chez les femmes ayant un CCPH et que le sexe féminin est un facteur pronostic négatif pour la survie à 5 ans. Le deuxième axe concernait la prise en charge préopératoire des patients, notamment l’optimisation préopératoire du foie restant par le drainage biliaire. Ce travail repose sur deux méta-analyses. Il a permis de mettre en évidence la supériorité de la voie radiologique sur la voie endoscopique concernant les complications liées à la procédure mais en revanche, l’absence de différence significative sur la morbi-mortalité post-résection hépatique, la survie à 5 ans, la survie sans récidive et le taux de dissémination liée à la procédure quand les procédures sont étudiées en intention de traiter. Nos résultats suggèrent qu’un mauvais choix de voie d’abord pour réaliser le drainage biliaire conduit à des échecs répétés qui influencent la récidive tumorale et donc la survie. Le troisième axe s’intéressait aux facteurs pronostiques de morbi-mortalité immédiates post-résection hépatique à partir d’une base de données Européenne (base de l’association Française de chirurgie). Les résultats montrent que la surface corporelle ≥ 1.82 m², l’hyperbilirubinémie > 50 µmol/l et la résection hépatique droite sont des facteurs prédictifs indépendants influençant la mortalité post-opératoire à 30 jours. / Cholangiocarcinoma (CC) is a malignant tumor with a poor prognosis. Its treatment is based on surgical resection. It accounts for 3% of all digestive cancers and is the second primary tumor of the liver, in frequency, after hepatocellular carcinoma. The average age is 70 years old with male predominance. At present intra and extrahepatic cholangiocarcinomas are distinguished. Survival rate is less than 5% at 5 years in all stages. 60 to 70% are tumors of the biliary convergence also called Klatskin tumors.The aim of this work was to answer persistent questions about peri-hilar cholangiocarcinoma (PHCC) by applying different statistical methods on different databases and review of the literature.The three main axes of this work are articulated according to the temporality of management, from the diagnosis to the postoperative follow-up, going through the preoperative setting.The first axis is based on a local database (registry of digestive cancer of Calvados). The results show that PHCC accounts for only one third of cholangiocarcinomas in the general population, that its incidence rate is stable with a decrease in incidence, although not significant, in women having PHCC and that female gender is a negative prognostic factor for 5-year survival. The second axis concerned the preoperative management of patients, including preoperative optimization of the remaining liver by biliary drainage. This work is based on two Meta-analyzes. It made it possible to highlight the superiority of the radiological way in the endoscopic way concerning the complications related to the procedure but on the other hand, the absence of significant difference on the morbi-mortality post hepatic resection, the survival at 5 years, the recurrence free survival and the rate of dissemination related to the procedure when the procedures are studied in intent to treat. Our results suggest that a poor choice of pathway for achieving biliary drainage leads to repeated failures that influence tumor recurrence and thus survival. The third axis was concerned with the prognostic factors of immediate morbidity and mortality after hepatic resection from a European database (base of the French association of surgery). The results show that body surface area ≥ 1.82 m², hyperbilirubinemia > 50 μmol / l and right hepatic resection are independent predictors influencing post-operative mortality at 30 days.

Background: Tumor-associated macrophages (TAMs) promote tumor progression and have an effect on survival in human cancer. However, little is known regarding their influence on tumor progression and prognosis in human hilar cholangiocarcinoma. Methods: We analyzed surgically resected tumor specimens of hilar cholangiocarcinoma (n = 47) for distribution and localization of TAMs, as defined by expression of CD68. Abundance of TAMs was correlated with clinicopathologic characteristics, tumor recurrence and patients’ survival. Statistical analysis was performed using SPSS software. Results: Patients with high density of TAMs in tumor invasive front (TIF) showed significantly higher local and overall tumor recurrence (both ρ < 0.05). Furthermore, high density of TAMs was associated with decreased overall (one-year 83.6 % vs. 75.1 %; three-year 61.3 % vs. 42.4 %; both ρ < 0.05) and recurrence-free survival (one-year 93.9 % vs. 57.4 %; three-year 59.8 % vs. 26.2 %; both ρ < 0.05). TAMs in TIF and tumor recurrence, were confirmed as the only independent prognostic variables in the multivariate survival analysis (all ρ < 0.05). Conclusions: Overall survival and recurrence free survival of patients with hilar cholangiocarcinoma significantly improved in patients with low levels of TAMs in the area of TIF, when compared to those with a high density of TAMs. These observations suggest their utilization as valuable prognostic markers in routine histopathologic evaluation, and might indicate future therapeutic approaches by targeting TAMs.

No description available.