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Biblio - Bibliographie IAL
Bibliographie IAL

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Recherche bibliographique scientifique

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Accueil > Références bibliographiques

biblio

2016


  • Strassel, C, Bull, A, Moog, S, Receveur, N, Mallo, L, Mangin, P, Eckly, A, Freund, M, Dubart-Kupperschmitt, A, Gachet, C & Lanza, F 2016, « Lentiviral gene rescue of a Bernard-Soulier mouse model to study platelet glycoprotein Ibβ function », Journal of thrombosis and haemostasis: JTH, vol. 14, no. 7, p. 1470-1479.
    Résumé : Essentials A signaling role of glycoprotein (GP)Ibβ is postulated but not formally demonstrated in platelets. Lentiviral-mediated rescue in knock-out mice can be used to evaluate GPIbβ function in vivo. Transduction of the native subunit corrected the main defects associated with GPIb-IX deficiency Deletion of intracellular 159-170 segment increased thrombosis, 150-160 removal increased bleeding. SUMMARY: Background The platelet glycoprotein (GP)Ib-V-IX complex is required for normal hemostasis and megakaryopoiesis. A role in GPIb-dependent responses has been ascribed to the less well characterized GPIbβ subunit using a specific antibody and GPIb-IX transfected cells. Objectives Our aim was to evaluate, in vivo, the role of the GPIbβ in hemostasis and thrombosis. Methods GPIbβ(null) Sca-1(+) progenitors transduced with viral particles harboring hGPIbβ were transplanted into lethally irradiated GPIbβ(-/-) recipient mice. Results hGPIbβ transplanted into the bone marrow of GPIbβ(null) mice rescued GPIb-IX expression in 97% of circulating platelets. These platelets efficiently bound von Willebrand factor (VWF) and extended filopodia on a VWF matrix, demonstrating the restoration of GPIb-dependent adhesive and signaling properties. These mice exhibited less severe macrothrombocytopenia and had normal tail bleeding times as compared with GPIbβ(null) mice. This strategy was employed to manipulate and evaluate the role of the GPIbβ intracellular domain. Removal of the membrane proximal segment (Δ(150-160) ) decreased GPIb-IX expression by 43%, confirming its involvement in receptor assembly and biosynthesis, and resulted in increased bleeding times and decreased thrombosis in a mechanical injury model in the aorta. On the other hand, deletion of the C-flanking 159-170 segment allowed normal GPIb-IX expression, VWF-dependent responses and bleeding times, but resulted in enhanced arterial thrombosis. Conclusion This pointed to a repressor role of GPIbβ in thrombus formation in vivo that was not predicted in studies of heterologous cells. These results highlight the utility of this lentiviral strategy for the structure-function evaluation of GPIb-IX in platelets.
    Mots-clés : Bernard-Soulier syndrome, blood platelets, platelet function tests, platelet glycoprotein GPIb-IX complex, thrombosis, virion.

  • Tranchart, H, Koffi, GM, Gaillard, M, Lainas, P, Poüs, C, Gonin, P, Nguyen, TH, Dubart-Kupperschmitt, A & Dagher, I 2016, « Liver regeneration following repeated reversible portal vein embolization in an experimental model », The British Journal of Surgery, vol. 103, no. 9, p. 1209-1219.
    Résumé : BACKGROUND: Portal vein embolization (PVE) is used routinely to prevent postoperative liver failure as a result of anticipated insufficient future liver remnant volume following resection. The authors have recently developed a technique for temporary PVE. The aim of this study was to assess the effect of repeated reversible PVE on hepatocyte proliferation and subsequent liver hypertrophy in rodents. METHODS: Four treatments were compared (n = 21 rats per group): single reversible PVE, two PVEs separated by 14 days, partial portal vein ligation or sham procedure. The feasibility and tolerance of the procedure were assessed. Volumetric imaging by CT was used to estimate the evolution of liver volumes. After death, the weight of liver lobes was measured and hepatocyte proliferation evaluated by immunostaining. RESULTS: Embolization of portal branches corresponding to 70 per cent of total portal flow was performed successfully in all animals. Repeated PVE induced additional hepatocyte proliferation. Repeated embolization resulted in superior hepatocyte proliferation in the non-occluded segments compared with portal vein ligation (31·1 versus 22·2 per cent; P = 0·003). The non-occluded to total liver volume ratio was higher in the repeated PVE group than in the single PVE and sham groups (P = 0·050 and P = 0·001 respectively). CONCLUSION: Repeated reversible PVE successfully induced additional hepatocyte proliferation and subsequent liver hypertrophy. Surgical relevance Portal vein embolization (PVE) is used routinely to prevent postoperative liver failure as a result of anticipated insufficient future liver remnant volume following resection. In the present study, a technique of repeated temporary PVE was developed in a rat model; this induced additional hepatocyte proliferation and an increase in liver volume compared with single embolization. This novel approach might help induce major hypertrophy of the future remnant liver, which could increase the rate of patients amenable to major liver resections.

  • Zeng, Y, Gao, T, Zhao, G, Jiang, Y, Yang, Y, Yu, H, Kou, Z, Lone, Y, Sun, S & Zhou, Y 2016, « Generation of human MHC (HLA-A11/DR1) transgenic mice for vaccine evaluation », Human Vaccines & Immunotherapeutics, vol. 12, no. 3, p. 829-836.
    Résumé : The rapid occurrence of emerging infectious diseases demonstrates an urgent need for a new preclinical experimental model that reliably replicates human immune responses. Here, a new homozygous humanized human leukocyte antigen (HLA)-A11/DR1 transgenic mouse (HLA-A11(+/+)/DR01(+/+)/H-2-β2m(-/-)/IAβ(-/-)) was generated by crossing HLA-A11 transgenic (Tg) mice with HLA-A2(+/+)/DR01(+/+)/H-2-β2m(-/-)/IAβ(-/-) mice. The HLA-A11-restricted immune response of this mouse model was then examined. HLA-A11 Tg mice expressing a chimeric major histocompatibility complex (MHC) molecule comprising the α1, α2, and β2m domains of human HLA-A11 and the α3 transmembrane and cytoplasmic domains of murine H-2D(b) were generated. The correct integration of HLA-A11 and HLA-DR1 into the genome of the HLA-A11/DR1 Tg mice (which lacked the expression of endogenous H-2-I/II molecules) was then confirmed. Immunizing mice with a recombinant HBV vaccine or a recombinant HIV-1 protein resulted in the generation of IFN-γ-producing cytotoxic T lymphocyte (CTL) and antigen-specific antibodies. The HLA-A11-restricted CTL response was directed at HLA immunodominant epitopes. These mice represent a versatile animal model for studying the immunogenicity of HLA CTL epitopes in the absence of a murine MHC response. The established animal model will also be useful for evaluating and optimizing T cell-based vaccines and for studying differences in antigen processing between mice and humans.
    Mots-clés : HLA, HLA-A11/DR1 transgenic mice, MHC, MHC-restricted epitopes, Vaccine.

2015

  • Ambroise, G, Portier, A, Roders, N, Arnoult, D & Vazquez, A 2015, « Subcellular localization of PUMA regulates its pro-apoptotic activity in Burkitt's lymphoma B cells », Oncotarget.
    Résumé : The BH3-only protein PUMA (p53-upregulated modulator of apoptosis) is a major regulator of apoptosis. It belongs to the Bcl-2 family of proteins responsible for maintaining mitochondrial outer membrane integrity by controlling the intrinsic (mitochondrial) apoptotic pathway. We describe here a new pathway regulating PUMA activation through the control of its subcellular distribution. Surprisingly, neither PUMA upregulation in normal activated human B lymphocytes nor high levels of PUMA in Burkitt's lymphoma (BL) were associated with cell death. We show that PUMA is localized to the cytosol in these cells. By contrast, various apoptosis-triggering signals were found to promote the translocation of PUMA to the mitochondria in these cells, leading to their death by apoptosis. This apoptosis was associated with the binding of mitochondrial PUMA to anti-apoptotic members of the Bcl-2 family, such as Bcl-2 and Mcl-1. This translocation was caspase-independent but was prevented by inhibiting or knocking down the expression of the MAPK kinase p38. Our data suggest that the accumulation of PUMA in the cytosol may be important for the participation of this protein in apoptosis without the need for prior transcription. This regulatory pathway may be an important feature of differentiation and tumorigenic processes.
    Mots-clés : Apoptosis, Burkitt’s Lymphoma, Mitochondria, PUMA, translocation.

  • Beaudreuil, S, Lorenzo, HK & Durrbach, A 2015, « The anti-CD40 auto-antibody: a biomarker or a factor for the permeability of recurrent focal segmental glomerulosclerosis? », Annals of Translational Medicine, vol. 3, no. 9, p. 115.

  • Benzoubir, N, Mussini, C, Lejamtel, C, Dos Santos, A, Guillaume, C, Desterke, C, Samuel, D, Bréchot, C, Bourgeade, M-F & Guettier, C 2015, « Gamma-Smooth Muscle Actin Expression Is Associated with Epithelial-Mesenchymal Transition and Stem-Like Properties in Hepatocellular Carcinoma », PloS One, vol. 10, no. 6, p. e0130559.
    Résumé : BACKGROUND AND AIMS: The prognosis of hepatocellular carcinoma (HCC) is hampered by frequent tumour recurrence and metastases. Epithelial-Mesenchymal Transition (EMT) is now recognized as a key process in tumour invasion, metastasis and the generation of cancer initiating cells. The morphological identification of EMT in tumour samples from the expression of novel mesenchymal markers could provide relevant prognostic information and aid in understanding the metastatic process. METHODS: The expression of Smooth Muscle Actins was studied using immunofluorescence and immunohistochemistry assays in cultured liver cells during an induced EMT process and in liver specimens from adult and paediatric HCC series. RESULTS: We report here that in HCC cell lines treated with TGF-β and in HCC specimens, the expression of αSMA, a known mesenchymal marker of EMT, could never be detected. In addition, our in vitro studies identified the enteric form of SMA, γSMA, as being a marker of EMT. Moreover, this SMA isoform was expressed in 46% of 58 tumours from 42 adult HCC patients and in 90% of 16 tumours from 12 paediatric HCC patients. Interestingly, this expression was significantly correlated with poor tumour differentiation and progenitor cell features characterized by the expression of EpCAM and K19. CONCLUSION: Taken together, our results support the conclusion that γSMA expression in HCC is strongly correlated with the EMT process, HCC aggressiveness and the identification of cancer stem cells. This correlation suggests that γSMA represents a novel and powerful marker to predict HCC progression.

  • Blasimme, A, Anegon, I, Concordet, J-P, De Vos, J, Dubart-Kupperschmitt, A, Fellous, M, Fouchet, P, Frydman, N, Giovannangeli, C, Jouannet, P, Serre, J-L, Steffann, J, Rial-Sebbag, E, Thomsen, M & Cambon-Thomsen, A 2015, « Genome Editing and Dialogic Responsibility: "What's in a Name?" », The American journal of bioethics: AJOB, vol. 15, no. 12, p. 54-57.
    Mots-clés : Humans, Social Behavior, Terminology as Topic.

  • Candelier, J-J 2015, « [Complete hydatidiform mole] », Medecine Sciences: M/S, vol. 31, no. 10, p. 861-868.
    Résumé : "The battle of the sexes begins in the zygote" W. Reik and J. Walter. Complete hydatidiform mole (CHM) is a pathology of the placenta with androgenetic diploid origin (chromosomes only from paternal origin). Placental villi present an abnormal hyperproliferation and hydropic degeneration associated with the absence of embryo. Three mechanisms can be envisaged at its origin: (1) destruction/expulsion of the female pronucleus at the time of fertilization by one or two spermatozoa, the former being followed by an endoreplication of the male pronucleus (homozygous mole), (2) a triploid zygote (fertilization by two spermatozoa) leading to a haploid and a diploid clones. The diploid clone may produce a normal fetus while the haploid clone, after endoreplication, generates a complete hydatidiform mole, (3) a nutritional defect during the differentiation of the oocytes of the female embryo that will affect the integrity and maturity of her oocytes during her adult life and lead to hydatidiform mole. In countries with a poor medical health care system, moles can be invasive or, in rare cases, lead to gestational choriocarcinomas.
    Mots-clés : Adult, Epigenesis, Genetic, Female, Humans, Hydatidiform Mole, Incidence, Male, Placenta, Pregnancy, Uterine Neoplasms.

  • Coullin, P, Diatta, AL, Boufettal, H, Feingold, J, Leguern, E & Candelier, JJ 2015, « The involvement of the trans-generational effect in the high incidence of the hydatidiform mole in Africa », Placenta, vol. 36, no. 1, p. 48-51.
    Résumé : INTRODUCTION: While the incidence of various chromosomal anomalies observed, including triploid partial moles is independent of the socio-economic level, higher incidences of complete hydatidiform mole "CHM" is generally associated with under developed areas. Moreover, studies have shown that some nutritional deficiencies are related to the abnormal development of oocytes and placenta. In Senegal and Morocco, the annual seasonal cycle contains one period with food shortages and the incidence of complete moles is significant. Accordingly, accurate statistical analyses have been performed in these two countries. METHODS: Each month during a one year period, we investigated the occurrence of normal conceptions, molar conceptions and the conception of the future patients in Senegal and Morocco. The comparisons of the conception dates for these three types of conception were analyzed using the Chi-squared test. RESULTS: 94% of the patients were conceived just prior to the period in the year with food shortages. Consequently, the development of the female embryos occurred under nutritional constraints, which negatively affect the recruitment of the vital factors required for the normal synthesis of DNA, proteins and placental differentiation. DISCUSSIONS: A nutritional deficiency in the mother at conception of their daughter (future patient) is implicated in the higher incidence of CHM in their daughters' filiation. These nutritional deficiencies during the first weeks of pregnancy will have repercussions on the normal development of the oocytes. Accordingly, these developmental impairments take place during the embryonic life of the future mothers of complete moles and not during the conception of the moles themselves.
    Mots-clés : Female, Humans, Hydatidiform Mole, Incidence, Maternal Nutritional Physiological Phenomena, Morocco, Nutrition, Nutritional Status, Oocyte, Placental development, Pregnancy, Senegal, trophoblast, Uterine Neoplasms.

  • Desterke, C, Martinaud, C, Guerton, B, Pieri, L, Bogani, C, Clay, D, Torossian, F, Lataillade, J-J, Hasselbach, HC, Gisslinger, H, Demory, J-L, Dupriez, B, Boucheix, C, Rubinstein, E, Amsellem, S, Vannucchi, AM & Le Bousse-Kerdilès, M-C 2015, « Tetraspanin CD9 participates in dysmegakaryopoiesis and stromal interactions in primary myelofibrosis », Haematologica, vol. 100, no. 6, p. 757-767.
    Résumé : Primary myelofibrosis is characterized by clonal myeloproliferation, dysmegakaryopoiesis, extramedullary hematopoiesis associated with myelofibrosis and altered stroma in the bone marrow and spleen. The expression of CD9, a tetraspanin known to participate in megakaryopoiesis, platelet formation, cell migration and interaction with stroma, is deregulated in patients with primary myelofibrosis and is correlated with stage of myelofibrosis. We investigated whether CD9 participates in the dysmegakaryopoiesis observed in patients and whether it is involved in the altered interplay between megakaryocytes and stromal cells. We found that CD9 expression was modulated during megakaryocyte differentiation in primary myelofibrosis and that cell surface CD9 engagement by antibody ligation improved the dysmegakaryopoiesis by restoring the balance of MAPK and PI3K signaling. When co-cultured on bone marrow mesenchymal stromal cells from patients, megakaryocytes from patients with primary myelofibrosis displayed modified behaviors in terms of adhesion, cell survival and proliferation as compared to megakaryocytes from healthy donors. These modifications were reversed after antibody ligation of cell surface CD9, suggesting the participation of CD9 in the abnormal interplay between primary myelofibrosis megakaryocytes and stroma. Furthermore, silencing of CD9 reduced CXCL12 and CXCR4 expression in primary myelofibrosis megakaryocytes as well as their CXCL12-dependent migration. Collectively, our results indicate that CD9 plays a role in the dysmegakaryopoiesis that occurs in primary myelofibrosis and affects interactions between megakaryocytes and bone marrow stromal cells. These results strengthen the "bad seed in bad soil" hypothesis that we have previously proposed, in which alterations of reciprocal interactions between hematopoietic and stromal cells participate in the pathogenesis of primary myelofibrosis.

  • Desterke, C, Martinaud, C, Ruzehaji, N & Le Bousse-Kerdilès, M-C 2015, « Inflammation as a Keystone of Bone Marrow Stroma Alterations in Primary Myelofibrosis », Mediators of Inflammation, vol. 2015, p. 415024.
    Résumé : Primary myelofibrosis (PMF) is a clonal myeloproliferative neoplasm where severity as well as treatment complexity is mainly attributed to a long lasting disease and presence of bone marrow stroma alterations as evidenced by myelofibrosis, neoangiogenesis, and osteosclerosis. While recent understanding of mutations role in hematopoietic cells provides an explanation for pathological myeloproliferation, functional involvement of stromal cells in the disease pathogenesis remains poorly understood. The current dogma is that stromal changes are secondary to the cytokine "storm" produced by the hematopoietic clone cells. However, despite therapies targeting the myeloproliferation-sustaining clones, PMF is still regarded as an incurable disease except for patients, who are successful recipients of allogeneic stem cell transplantation. Although the clinical benefits of these inhibitors have been correlated with a marked reduction in serum proinflammatory cytokines produced by the hematopoietic clones, further demonstrating the importance of inflammation in the pathological process, these treatments do not address the role of the altered bone marrow stroma in the pathological process. In this review, we propose hypotheses suggesting that the stroma is inflammatory-imprinted by clonal hematopoietic cells up to a point where it becomes "independent" of hematopoietic cell stimulation, resulting in an inflammatory vicious circle requiring combined stroma targeted therapies.
    Mots-clés : Bone Marrow, Data Mining, DNA Methylation, Hematopoietic Stem Cells, Humans, Inflammation, primary myelofibrosis, Stromal Cells, Transforming Growth Factor beta.
  • García-Cano, J, Ambroise, G, Pascual-Serra, R, Carrión, MC, Serrano-Oviedo, L, Ortega-Muelas, M, Cimas, FJ, Sabater, S, Ruiz-Hidalgo, MJ, Sanchez Perez, I, Mas, A, Jalón, FA, Vazquez, A & Sánchez-Prieto, R 2015, « Exploiting the potential of autophagy in cisplatin therapy: A new strategy to overcome resistance », Oncotarget, vol. 6, no. 17, p. 15551-15565.
    Résumé : Resistance to cisplatin is a major challenge in the current cancer therapy. In order to explore new therapeutic strategies to cisplatin resistance, we evaluated, in a model of lung cancer (H1299 and H460 cell lines), the nature of the pathways leading to cell death. We observed that H1299 displayed a natural resistance to cisplatin due to an inability to trigger an apoptotic response that correlates with the induction of autophagy. However, pharmacological and genetic approaches showed how autophagy was a mechanism associated to cell death rather than to resistance. Indeed, pro-autophagic stimuli such as mTOR or Akt inhibition mediate cell death in both cell lines to a similar extent. We next evaluated the response to a novel platinum compound, monoplatin, able to promote cell death in an exclusive autophagy-dependent manner. In this case, no differences were observed between both cell lines. Furthermore, in response to monoplatin, two molecular hallmarks of cisplatin response (p53 and MAPKs) were not implicated, indicating the ability of this pro-autophagic compound to overcome cisplatin resistance. In summary, our data highlight how induction of autophagy could be used in cisplatin resistant tumours and an alternative treatment for p53 mutated patient in a synthetic lethally approach.
    Mots-clés : Apoptosis, autophagy, cisplatin, monoplatin, synthetic lethality.

  • Genêt, F, Kulina, I, Vaquette, C, Torossian, F, Millard, S, Pettit, AR, Sims, NA, Anginot, A, Guerton, B, Winkler, IG, Barbier, V, Lataillade, J-J, Le Bousse-Kerdilès, M-C, Hutmacher, DW & Levesque, J-P 2015, « Neurological heterotopic ossification following spinal cord injury is triggered by macrophage-mediated inflammation in muscle », The Journal of Pathology, vol. 236, no. 2, p. 229-240.
    Résumé : Neurological heterotopic ossification (NHO) is the abnormal formation of bone in soft tissues as a consequence of spinal cord or traumatic brain injury. NHO causes pain, ankyloses, vascular and nerve compression and delays rehabilitation in this high-morbidity patient group. The pathological mechanisms leading to NHO remain unknown and consequently there are no therapeutic options to prevent or reduce NHO. Genetically modified mouse models of rare genetic forms of heterotopic ossification (HO) exist, but their relevance to NHO is questionable. Consequently, we developed the first model of spinal cord injury (SCI)-induced NHO in genetically unmodified mice. Formation of NHO, measured by micro-computed tomography, required the combination of both SCI and localized muscular inflammation. Our NHO model faithfully reproduced many clinical features of NHO in SCI patients and both human and mouse NHO tissues contained macrophages. Muscle-derived mesenchymal progenitors underwent osteoblast differentiation in vitro in response to serum from NHO mice without additional exogenous osteogenic stimuli. Substance P was identified as a candidate NHO systemic neuropeptide, as it was significantly elevated in the serum of NHO patients. However, antagonism of substance P receptor in our NHO model only modestly reduced the volume of NHO. In contrast, ablation of phagocytic macrophages with clodronate-loaded liposomes reduced the size of NHO by 90%, supporting the conclusion that NHO is highly dependent on inflammation and phagocytic macrophages in soft tissues. Overall, we have developed the first clinically relevant model of NHO and demonstrated that a combined insult of neurological injury and soft tissue inflammation drives NHO pathophysiology.
    Mots-clés : Animals, Cardiotoxins, Disease Models, Animal, Female, heterotopic ossification, Humans, Inflammation, macrophage, Macrophages, Mice, Inbred C57BL, Muscle, Skeletal, Myositis, Ossification, Heterotopic, Paraplegia, Spinal Cord Injuries, spinal cord injury, Stem Cells.

  • Hasmim, M, Bruno, S, Azzi, S, Gallerne, C, Michel, JG, Chiabotto, G, Lecoz, V, Romei, C, Spaggiari, GM, Pezzolo, A, Pistoia, V, Angevin, E, Gad, S, Ferlicot, S, Messai, Y, Kieda, C, Clay, D, Sabatini, F, Escudier, B, Camussi, G, Eid, P, Azzarone, B & Chouaib, S 2015, « Isolation and characterization of renal cancer stem cells from patient-derived xenografts », Oncotarget.
    Résumé : As rapidly developing patient-derived xenografts (PDX) could represent potential sources of cancer stem cells (CSC), we selected and characterized non-cultured PDX cell suspensions from four different renal carcinomas (RCC). Only the cell suspensions from the serial xenografts (PDX-1 and PDX-2) of an undifferentiated RCC (RCC-41) adapted to the selective CSC medium. The cell suspension derived from the original tumor specimen (RCC-41-P-0) did not adapt to the selective medium and strongly expressed CSC-like markers (CD133 and CD105) together with the non-CSC tumor marker E-cadherin. In comparison, PDX-1 and PDX-2 cells exhibited evolution in their phenotype since PDX-1 cells were CD133high/CD105-/Ecadlow and PDX-2 cells were CD133low/CD105-/Ecad-. Both PDX subsets expressed additional stem cell markers (CD146/CD29/OCT4/NANOG/Nestin) but still contained non-CSC tumor cells. Therefore, using different cell sorting strategies, we characterized 3 different putative CSC subsets (RCC-41-PDX-1/CD132+, RCC-41-PDX-2/CD133-/EpCAMlow and RCC-41-PDX-2/CD133+/EpCAMbright). In addition, transcriptomic analysis showed that RCC-41-PDX-2/CD133- over-expressed the pluripotency gene ERBB4, while RCC-41-PDX-2/CD133+ over-expressed several tumor suppressor genes. These three CSC subsets displayed ALDH activity, formed serial spheroids and developed serial tumors in SCID mice, although RCC-41-PDX-1/CD132+ and RCC-41-PDX-2/CD133+ displayed less efficiently the above CSC properties. RCC-41-PDX-1/CD132+ tumors showed vessels of human origin with CSC displaying peri-vascular distribution. By contrast, RCC-41-PDX-2 originated tumors exhibiting only vessels of mouse origin without CSC peri-vascular distribution.Altogether, our results indicate that PDX murine microenvironment promotes a continuous redesign of CSC phenotype, unmasking CSC subsets potentially present in a single RCC or generating ex novo different CSC-like subsets.
    Mots-clés : cancer stem cells, CD133, clear cell renal cell carcinoma, EpCAM, patient-derived xenografts.

  • Jouannet, S, Saint-Pol, J, Fernandez, L, Nguyen, V, Charrin, S, Boucheix, C, Brou, C, Milhiet, P-E & Rubinstein, E 2015, « TspanC8 tetraspanins differentially regulate the cleavage of ADAM10 substrates, Notch activation and ADAM10 membrane compartmentalization », Cellular and molecular life sciences: CMLS.
    Résumé : The metalloprotease ADAM10 mediates the shedding of the ectodomain of various cell membrane proteins, including APP, the precursor of the amyloid peptide Aβ, and Notch receptors following ligand binding. ADAM10 associates with the members of an evolutionary conserved subgroup of tetraspanins, referred to as TspanC8, which regulate its exit from the endoplasmic reticulum. Here we show that 4 of these TspanC8 (Tspan5, Tspan14, Tspan15 and Tspan33) which positively regulate ADAM10 surface expression levels differentially impact ADAM10-dependent Notch activation and the cleavage of several ADAM10 substrates, including APP, N-cadherin and CD44. Sucrose gradient fractionation, single molecule tracking and quantitative mass-spectrometry analysis of the repertoire of molecules co-immunoprecipitated with Tspan5, Tspan15 and ADAM10 show that these two tetraspanins differentially regulate ADAM10 membrane compartmentalization. These data represent a unique example where several tetraspanins differentially regulate the function of a common partner protein through a distinct membrane compartmentalization.
    Mots-clés : ADAM10, Ectodomain shedding, Membrane compartmentalization, Microdomain, Notch, Tetraspanin.

  • Lecru, L, Desterke, C, Grassin-Delyle, S, Chatziantoniou, C, Vandermeersch, S, Devocelle, A, Vernochet, A, Ivanovski, N, Ledent, C, Ferlicot, S, Dalia, M, Saïd, M, Beaudreuil, S, Charpentier, B, Vazquez, A, Giron-Michel, J, Azzarone, B, Durrbach, A & François, H 2015, « Cannabinoid receptor 1 is a major mediator of renal fibrosis », Kidney International.
    Résumé : Chronic kidney disease, secondary to renal fibrogenesis, is a burden on public health. There is a need to explore new therapeutic pathways to reduce renal fibrogenesis. To study this, we used unilateral ureteral obstruction (UUO) in mice as an experimental model of renal fibrosis and microarray analysis to compare gene expression in fibrotic and normal kidneys. The cannabinoid receptor 1 (CB1) was among the most upregulated genes in mice, and the main endogenous CB1 ligand (2-arachidonoylglycerol) was significantly increased in the fibrotic kidney. Interestingly, CB1 expression was highly increased in kidney biopsies of patients with IgA nephropathy, diabetes, and acute interstitial nephritis. Both genetic and pharmacological knockout of CB1 induced a profound reduction in renal fibrosis during UUO. While CB2 is also involved in renal fibrogenesis, it did not potentiate the role of CB1. CB1 expression was significantly increased in myofibroblasts, the main effector cells in renal fibrogenesis, upon TGF-β1 stimulation. The decrease in renal fibrosis during CB1 blockade could be explained by a direct action on myofibroblasts. CB1 blockade reduced collagen expression in vitro. Rimonabant, a selective CB1 endocannabinoid receptor antagonist, modulated the macrophage infiltrate responsible for renal fibrosis in UUO through a decrease in monocyte chemoattractant protein-1 synthesis. Thus, CB1 has a major role in the activation of myofibroblasts and may be a new target for treating chronic kidney disease.Kidney International advance online publication, 11 March 2015; doi:10.1038/ki.2015.63.

  • Martinaud, C, Desterke, C, Konopacki, J, Pieri, L, Torossian, F, Golub, R, Schmutz, S, Anginot, A, Guerton, B, Rochet, N, Albanese, P, Henault, E, Pierre-Louis, O, Souraud, J-B, de Revel, T, Dupriez, B, Ianotto, J-C, Bourgeade, M-F, Vannucchi, AM, Lataillade, J-J & Le Bousse-Kerdilès, M-C 2015, « Osteogenic Potential of Mesenchymal Stromal Cells Contributes to Primary Myelofibrosis », Cancer Research, vol. 75, no. 22, p. 4753-4765.
    Résumé : Primary myelofibrosis is a myeloproliferative neoplasm that is a precursor to myeloid leukemia. Dysmegakaryopoiesis and extramedullary hematopoiesis characterize primary myelofibrosis, which is also associated with bone marrow stromal alterations marked by fibrosis, neoangiogenesis, and osteomyelosclerosis. In particular, contributions to primary myelofibrosis from mesenchymal stromal cells (MSC) have been suggested by mouse studies, but evidence in humans remains lacking. In this study, we show that bone marrow MSCs from primary myelofibrosis patients exhibit unique molecular and functional abnormalities distinct from other myeloproliferative neoplasms and these abnormalities are maintained stably ex vivo in the absence of leukemic cells. Primary myelofibrosis-MSC overexpressed heparin-binding cytokines, including proinflammatory TGFβ1 and osteogenic BMP-2, as well as glycosaminoglycans such as heparan sulfate and chondroitin sulfate. Transcriptome and functional analyses revealed alterations in MSC differentiation characterized by an increased osteogenic potential and a TGFβ1 signaling signature. Accordingly, phospho-Smad2 levels were intrinsically increased in primary myelofibrosis-MSC along with enhanced expression of the master bone regulator RUNX2, while inhibition of the endogenous TGFβ1 receptor TGFβR1 impaired osteogenic differentiation in these MSCs. Taken together, our results define the source of a critical osteogenic function in primary myelofibrosis that supports its pathophysiology, suggesting that combined targeting of both the hematopoietic and stromal cell compartments in primary myelofibrosis patients may heighten therapeutic efficacy.
    Mots-clés : Adult, Aged, Aged, 80 and over, Animals, Cell Differentiation, Cells, Cultured, Female, Heterografts, Humans, Male, Mesenchymal Stromal Cells, Mice, Mice, Nude, Middle Aged, Ossification, Heterotopic, Polymerase Chain Reaction, Primary myelofibrosis.

  • Martinaud, C, Desterke, C, Konopacki, J, Vannucchi, AM, Pieri, L, Guglielmelli, P, Dupriez, B, Ianotto, J-C, Boutin, L, Lataillade, J-J & Le Bousse-Kerdilès, M-C 2015, « Transcriptome analysis of bone marrow mesenchymal stromal cells from patients with primary myelofibrosis », Genomics Data, vol. 5, p. 1-2.
    Résumé : Primary myelofibrosis (PMF) is a clonal myeloproliferative neoplasm whose severity and treatment complexity are attributed to the presence of bone marrow (BM) fibrosis and alterations of stroma impairing the production of normal blood cells. Despite the recently discovered mutations including the JAK2V617F mutation in about half of patients, the primitive event responsible for the clonal proliferation is still unknown. In the highly inflammatory context of PMF, the presence of fibrosis associated with a neoangiogenesis and an osteosclerosis concomitant to the myeloproliferation and to the increase number of circulating hematopoietic progenitors suggests that the crosstalk between hematopoietic and stromal cells is deregulated in the PMF BM microenvironmental niches. Within these niches, mesenchymal stromal cells (BM-MSC) play a hematopoietic supportive role in the production of growth factors and extracellular matrix which regulate the proliferation, differentiation, adhesion and migration of hematopoietic stem/progenitor cells. A transcriptome analysis of BM-MSC in PMF patients will help to characterize their molecular alterations and to understand their involvement in the hematopoietic stem/progenitor cell deregulation that features PMF.
    Mots-clés : Bone Marrow, Mesenchymal stroma cells, Myeloproliferative disorders, Primary myelofibrosis.

  • Martínez del Hoyo, G, Ramírez-Huesca, M, Levy, S, Boucheix, C, Rubinstein, E, Minguito de la Escalera, M, González-Cintado, L, Ardavín, C, Veiga, E, Yáñez-Mó, M & Sánchez-Madrid, F 2015, « CD81 controls immunity to Listeria infection through rac-dependent inhibition of proinflammatory mediator release and activation of cytotoxic T cells », Journal of Immunology (Baltimore, Md.: 1950), vol. 194, no. 12, p. 6090-6101.
    Résumé : Despite recent evidence on the involvement of CD81 in pathogen binding and Ag presentation by dendritic cells (DCs), the molecular mechanism of how CD81 regulates immunity during infection remains to be elucidated. To investigate the role of CD81 in the regulation of defense mechanisms against microbial infections, we have used the Listeria monocytogenes infection model to explore the impact of CD81 deficiency in the innate and adaptive immune response against this pathogenic bacteria. We show that CD81(-/-) mice are less susceptible than wild-type mice to systemic Listeria infection, which correlates with increased numbers of inflammatory monocytes and DCs in CD81(-/-) spleens, the main subsets controlling early bacterial burden. Additionally, our data reveal that CD81 inhibits Rac/STAT-1 activation, leading to a negative regulation of the production of TNF-α and NO by inflammatory DCs and the activation of cytotoxic T cells by splenic CD8α(+) DCs. In conclusion, this study demonstrates that CD81-Rac interaction exerts an important regulatory role on the innate and adaptive immunity against bacterial infection and suggests a role for CD81 in the development of novel therapeutic targets during infectious diseases.
    Mots-clés : Animals, Antigens, CD81, Cell Differentiation, Cell Survival, Dendritic Cells, Disease Models, Animal, Disease Resistance, Inflammation Mediators, Listeria, Listeriosis, Lymphocyte Activation, Mice, Mice, Knockout, Nitric Oxide, Phagocytosis, Phosphorylation, Protein Binding, rac GTP-Binding Proteins, Receptor, Interferon alpha-beta, Signal Transduction, STAT1 Transcription Factor, T-Lymphocytes, Cytotoxic, Tumor Necrosis Factor-alpha.

  • Rostaing, L, Hertig, A, Albano, L, Anglicheau, D, Durrbach, A, Vuiblet, V, Moulin, B, Merville, P, Hazzan, M, Lang, P, Touchard, G, Hurault deLigny, B, Quéré, S, Di Giambattista, F, Dubois, Y-C, Rondeau, E & CERTITEM Study Group, 2015, « Fibrosis progression according to epithelial-mesenchymal transition profile: a randomized trial of everolimus versus CsA », American Journal of Transplantation: Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons, vol. 15, no. 5, p. 1303-1312.
    Résumé : Markers of epithelial-mesenchymal transition (EMT) may identify patients at high risk of graft fibrogenesis who could benefit from early calcineurin inhibitor (CNI) withdrawal. In a randomized, open-label, 12-month trial, de novo kidney transplant patients received cyclosporine, enteric-coated mycophenolate sodium (EC-MPS) and steroids to month 3. Patients were stratified as EMT+ or EMT- based on month 3 biopsy, then randomized to start everolimus with half-dose EC-MPS (720 mg/day) and cyclosporine withdrawal (CNI-free) or continue cyclosporine with standard EC-MPS (CNI). The primary endpoint was progression of graft fibrosis (interstitial fibrosis/tubular atrophy [IF/TA] grade increase ≥1 between months 3-12) in EMT+ patients. 194 patients were randomized (96 CNI-free, 98 CNI); 153 (69 CNI-free, 84 CNI) were included in histological analyses. Fibrosis progression occurred in 46.2% (12/26) CNI-free EMT+ patients versus 51.6% (16/31) CNI EMT+ patients (p = 0.68). Biopsy-proven acute rejection (BPAR, including subclinical events) occurred in 25.0% and 5.1% of CNI-free and CNI patients, respectively (p < 0.001). In conclusion, early CNI withdrawal with everolimus initiation does not prevent interstitial fibrosis. Using this CNI-free protocol, in which everolimus exposure was relatively low and administered with half-dose EC-MPS, CNI-free patients were overwhelmingly under-immunosuppressed and experienced an increased risk of BPAR.

  • Steichen, C, Maluenda, J, Tosca, L, Luce, E, Pineau, D, Dianat, N, Hannoun, Z, Tachdjian, G, Melki, J & Dubart-Kupperschmitt, A 2015, « An atypical human induced pluripotent stem cell line with a complex, stable, and balanced genomic rearrangement including a large de novo 1q uniparental disomy », Stem Cells Translational Medicine, vol. 4, no. 3, p. 224-229.
    Résumé : Human induced pluripotent stem cells (hiPSCs) hold great promise for cell therapy through their use as vital tools for regenerative and personalized medicine. However, the genomic integrity of hiPSCs still raises some concern and is one of the barriers limiting their use in clinical applications. Numerous articles have reported the occurrence of aneuploidies, copy number variations, or single point mutations in hiPSCs, and nonintegrative reprogramming strategies have been developed to minimize the impact of the reprogramming process on the hiPSC genome. Here, we report the characterization of an hiPSC line generated by daily transfections of modified messenger RNAs, displaying several genomic abnormalities. Karyotype analysis showed a complex genomic rearrangement, which remained stable during long-term culture. Fluorescent in situ hybridization analyses were performed on the hiPSC line showing that this karyotype is balanced. Interestingly, single-nucleotide polymorphism analysis revealed the presence of a large 1q region of uniparental disomy (UPD), demonstrating for the first time that UPD can occur in a noncompensatory context during nonintegrative reprogramming of normal fibroblasts.
    Mots-clés : Abnormal karyotype, Aneuploidy, Cell Line, Cellular Reprogramming, Chromosomes, Human, Pair 1, Directed differentiation, Fibroblasts, Genome, Human, Genomic integrity, Humans, Induced Pluripotent Stem Cells, Teratoma formation, Uniparental Disomy.

  • Tolosa, L, Caron, J, Hannoun, Z, Antoni, M, López, S, Burks, D, Castell, JV, Weber, A, Gomez-Lechon, M-J & Dubart-Kupperschmitt, A 2015, « Transplantation of hESC-derived hepatocytes protects mice from liver injury », Stem Cell Research & Therapy, vol. 6, p. 246.
    Résumé : BACKGROUND: Hepatic cell therapy has become a viable alternative to liver transplantation for life-threatening liver diseases. However, the supply of human hepatocytes is limited due to the shortage of suitable donor organs required to isolate high-quality cells. Human pluripotent stem cells reflect a potential renewable source for generating functional hepatocytes. However, most differentiation protocols use undefined matrices or factors of animal origin; as such, the resulting hepatocytes are not Good Manufacturing Practice compliant. Moreover, the preclinical studies employed to assess safety and function of human embryonic stem cell (hESC)-derived hepatocytes are generally limited to immunodeficient mice. In the present study, we evaluate the generation of hepatocytes under defined conditions using a European hESC line (VAL9) which was derived under animal-free conditions. The function capacity of VAL9-derived hepatocytes was assessed by transplantation into mice with acetaminophen-induced acute liver failure, a clinically relevant model. METHODS: We developed a protocol that successfully differentiates hESCs into bipotent hepatic progenitors under defined conditions, without the use of chromatin modifiers such as dimethyl sulphoxide. These progenitors can be cryopreserved and are able to generate both committed precursors of cholangiocytes and neonate-like hepatocytes. RESULTS: Thirty days post-differentiation, hESCs expressed hepatocyte-specific markers such as asialoglycoprotein receptor and hepatic nuclear factors including HNF4α. The cells exhibited properties of mature hepatocytes such as urea secretion and UGT1A1 and cytochrome P450 activities. When transplanted into mice with acetaminophen-induced acute liver failure, a model of liver damage, the VAL9-derived hepatocytes efficiently engrafted and proliferated, repopulating up to 10 % of the liver. In these transplanted livers, we observed a significant decrease of liver transaminases and found no evidence of tumourigenicity. Thus, VAL9-derived hepatocytes were able to rescue hepatic function in acetaminophen-treated animals. CONCLUSIONS: Our study reveals an efficient protocol for differentiating VAL9 hESCs to neonatal hepatocytes which are then able to repopulate livers in vivo without tumour induction. The human hepatocytes are able to rescue liver function in mice with acetaminophen-induced acute toxicity. These results provide proof-of-concept that replacement therapies using hESC-derived hepatocytes are effective for treating liver diseases.
    Mots-clés : Acetaminophen, Animals, Biliary Tract, Cell Differentiation, Cell Line, Chemical and Drug Induced Liver Injury, Embryonic Stem Cells, Hepatocytes, Heterografts, Humans, Male, Mice, Mice, Inbred NOD, Mice, SCID.

  • Touzot, M, Poisson, J, Faguer, S, Ribes, D, Cohen, P, Geffray, L, Anguel, N, François, H, Karras, A, Cacoub, P, Durrbach, A & Saadoun, D 2015, « Rituximab in anti-GBM disease: A retrospective study of 8 patients », Journal of Autoimmunity, vol. 60, p. 74-79.
    Résumé : OBJECTIVES: Anti-glomerular basement membrane (GBM) disease is a rare autoantibody-mediated disorder presenting as rapidly progressive glomerulonephritis, and often with pulmonary hemorraghe. Antibody removal with plasmapheresis and immunosuppressive drugs are the cornerstones of the treatment. Data regarding the use of specific B-cell depleting therapy such as rituximab are lacking. METHODS: We conducted a retrospective observational study of 8 patients with severe and/or refractory GBM disease that received rituximab therapy. RESULTS: Eight patients (2 men, 6 women) with a mean age of 26 ± 13.1 years old were included. Seven had severe renal involvement [median creatinin level was 282 μmol/l, range (65-423)] requiring high immunosuppressive or plasmapheresis dependent, and two had relapse of pulmonary hemorraghe including one with renal failure. Patients received an initial immunosuppressive treatment including steroid and cyclosphosphamide (n = 8) and plasmapheresis (n = 5). Except one late relapse, rituximab therapy was started within two months after diagnosis. All patients except one received 4 weekly dose of rituximab (375 mg(2)). Anti-GBM antibodies were still present in 6/8 patients, at rituximab initiation. Complete remission was observed in 7 out of 8 patients, mostly 3 months after rituximab therapy. After a mean follow-up of 25.6 months (range 4-93), patient and renal survival were 100% and 75% respectively, but rituximab use did not improve GFR. Anti-GBM antibodies remained negative for all patients during follow-up. Only one patient developed a severe bacterial infection but no opportunistic or viral infections were reported. CONCLUSION: Rituximab may represent an additional and/or alternative therapy in the induction treatment of anti-GBM disease.

  • Zhu, C, Trabado, S, Fan, Y, Trojan, J, Lone, Y-C, Giron-Michel, J & Duc, H-T 2015, « Characterization of effector components from the humoral and cellular immune response stimulated by melanoma cells exhibiting modified IGF-1 expression », Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie, vol. 70, p. 53-57.
    Résumé : Modified melanoma B16 cells inhibited in their IGF-1 expression (B16MOD), on the contrary to the IGF-1 fully expressed parental wild-type (B16WT) counterpart, were shown to stimulate humoral as well as cellular immune responses. Among humoral components, the neutralizing and complement-fixing antibodies of IgM and essentially IgG2 (a+b) isotypes exhibited in vitro and in vivo effects upon tumour growth, while the IgG1 antibody isotype promoted enhanced tumour proliferation. As for the cellular immunity, it was found that the T CD8(+) lymphocyte subpopulation remained the main potent and long lasting immune active effector regulating tumour growth.

2014


  • Achour, A, Baychelier, F, Marty, M, Debré, P, Samuel, D & Vieillard, V 2014, « Transplantation-induced cancers: Emerging evidence that clonal CMV-specific NK cells are causal immunogenic factors », Oncoimmunology, vol. 3, p. e28782.
    Résumé : Solid cancers are a major adverse outcome of liver transplantation. Recent reassessments have revealed insights into causal factors, primarily centering on modulations of the natural killer (NK) cell compartment in liver transplant recipients. In the presence of cytomegalovirus, the clonal expansion of differentiated NK cells could restrict the diversity of the NK repertoire favoring the development of certain tumors.

  • Adam, R 2014, « Surrogate markers of overall survival in metastatic colorectal cancer: an evolving challenge still more complex with repeat surgery », Annals of Surgical Oncology, vol. 21, no. 6, p. 1763-1764.

  • Adam, R & Bhangui, P 2014, « Reply to Letter: "Resection or Transplantation for Early Hepatocellular Carcinoma in a Cirrhotic Liver: Does Size Define the Best Oncological Strategy?" », Annals of Surgery.

  • Ailane, N, Greco, C, Zhu, Y, Sala-Valdés, M, Billard, M, Casal, I, Bawa, O, Opolon, P, Rubinstein, E & Boucheix, C 2014, « Effect of an anti-human Co-029/tspan8 mouse monoclonal antibody on tumor growth in a nude mouse model », Frontiers in Physiology, vol. 5, p. 364.
    Résumé : New therapeutic agents are needed in digestive tract tumors. Co-029/tspan8 is a tetraspanin frequently expressed on human colorectal tumors, In this work, we report the effects of the monoclonal antibody Ts29.2, targeting Co-029/tspan8, on colorectal tumor cells in vitro and after implantation in nude mice. HT29, Isreco1 and SW480 colorectal tumor cell lines were used for this study. HT29 has a strong endogenous expression of Co-029/tspan8, whereas Isreco1 cells don't express Co-029/tspan8 and SW480 has only a weak expression. Isreco1 and SW480 were transduced to express Co-029/tspan8 at the same level as HT29. In order to check the specificity of the effect of monoclonal antibody Ts29.2, low Co-029/tspan8 expressing SW480 cells were injected simultaneously with transduced cells in the back, on the left and right sides of the mice. With an early treatment, Ts29.2 mAb inhibited growth of tumors expressing Co-029/tspan8 up to 70%, whereas a delayed treatment was less efficient. No effect of the antibody on cell proliferation or apoptosis induction was detected in vitro. No increase of activated caspase 3 labeling was observed in vivo and areas occupied by vessels were not significantly different between treated mice and controls. This suggests that the action of Ts29.2 is linked neither to cellular toxicity nor to the inhibition of the previously reported angiogenic properties of Co-029/tspan8. An inhibition of cell proliferation in vivo is demonstrated by a reduction of the mitotic index in HT29 tumors of Ts29.2 treated mice. The discrepancy between in vitro and in vivo data on cell proliferation suggests that the binding of Ts29.2 to tumor cells may modify their response to signals issued from the microenvironment. Given the restricted pattern of tissue expression of the tetraspanin Co-029/tspan8, these preliminary results put forth for consideration the antibody targeting of this tetraspanin in further investigations for therapeutic applications.

  • Allard, MA & Adam, R 2014, « Reply letter to: Simultaneous surgical treatment for both colorectal liver metastases and peritoneal carcinomatosis », European Journal of Surgical Oncology: The Journal of the European Society of Surgical Oncology and the British Association of Surgical Oncology, vol. 40, no. 3, p. 368-369.
    Résumé : PMID: 24370282
    Mots-clés : Carcinoma, Colorectal Neoplasms, Female, Hepatectomy, Humans, Liver Neoplasms, Male, Peritoneal Neoplasms.

  • Allard, M-A, Sa Cunha, A, Ruiz, A, Vibert, E, Sebagh, M, Castaing, D & Adam, R 2014, « The postresection alpha-fetoprotein in cirrhotic patients with hepatocellular carcinoma. An independent predictor of outcome », Journal of Gastrointestinal Surgery: Official Journal of the Society for Surgery of the Alimentary Tract, vol. 18, no. 4, p. 701-708.
    Résumé : BACKGROUND: The postresection alpha-fetoprotein (AFP) in cirrhotic patients with hepatocellular carcinoma (HCC) may predict overall survival (OS) and recurrence beyond Milan criteria (MC) among the subgroup of initially transplantable patients. METHODS: All patients with cirrhosis resected for HCC between January 1990 and December 2010 in a single institution and presenting a serum AFP value > 15 ng/ml at diagnosis were included. The postresection AFP was analyzed as a dichotomized variable: normalization (norm + group) or not (norm - group) within the 90-day postresection period. RESULTS: Among 271 resected patients, 141 patients (52%) had a level of serum AFP ≥ 15 ng/ml at diagnosis. Five-year OS and median survival were 42% and 52 months in group norm + versus 20% and 23 months in the group norm - (P = 0.009). On multivariate analysis, the absence of AFP normalization was an independent factor of poor OS as well as microvascular invasion, and satellites nodules. Among theoretically transplantable patients, independent predictors of recurrence beyond MC were the absence of AFP normalization (risk ratio (RR) 5.02 [1.53-16.34]) and microvascular invasion (RR 4.76 [1.42-15.34]). CONCLUSION: The postresection AFP has an independent prognostic value. Transplantable patients resected for HCC without 90-day AFP normalization should be discussed for early liver transplantation.

  • Balbous, A, Cortes, U, Guilloteau, K, Villalva, C, Flamant, S, Gaillard, A, Milin, S, Wager, M, Sorel, N, Guilhot, J, Bennaceur-Griscelli, A, Turhan, A, Chomel, J-C & Karayan-Tapon, L 2014, « A mesenchymal glioma stem cell profile is related to clinical outcome », Oncogenesis, vol. 3, p. e91.
    Résumé : Recent studies have demonstrated a relationship between the expression of stem cell-associated genes and relapses in glioblastoma (GBM), suggesting a key role for tumor stem cells in this process. Although there is increasing interest in this field, glioma stem cells (GSCs) are still poorly characterized, their 'stemness' state and factors maintaining these properties remain largely unknown. We performed an expression profiling analysis of pluripotency in gliomaspheres derived from 11 patients. Comparative analysis between GSCs and H1 and H9 human embryonic stem cells as well as H9-derived neural stem cells indicates major variations in gene expression of pluripotency factors Nanog and OCT4, but a stable pattern for SOX2 suggesting its important function in maintaining pluripotency in GSCs. Our results also showed that all GSC lines have the capacity to commit to neural differentiation and express mesenchymal or endothelial differentiation markers. In addition, hierarchical clustering analysis revealed two groups of GSCs reflecting their heterogeneity and identified COL1A1 and IFITM1 as the most discriminating genes. Similar patterns have been observed in tumors from which gliomaspheres have been established. To determine whether this heterogeneity could be clinically relevant, the expression of both genes was further analyzed in an independent cohort of 30 patients with GBM and revealed strong correlation with overall survival. In vitro silencing of COL1A1 and IFTM1 confirmed the effect of these mesenchymal-associated genes on cell invasion and gliomasphere initiation. Our results indicate that COL1A1 and IFITM1 genes could be considered for use in stratifying patients with GBM into subgroups for risk of recurrence at diagnosis, as well as for prognostic and therapeutic evolution.

  • Boyer-Di Ponio, J, El-Ayoubi, F, Glacial, F, Ganeshamoorthy, K, Driancourt, C, Godet, M, Perrière, N, Guillevic, O, Couraud, PO & Uzan, G 2014, « Instruction of circulating endothelial progenitors in vitro towards specialized blood-brain barrier and arterial phenotypes », PloS One, vol. 9, no. 1, p. e84179.
    Résumé : OBJECTIVE: The vascular system is adapted to specific functions in different tissues and organs. Vascular endothelial cells are important elements of this adaptation, leading to the concept of 'specialized endothelial cells'. The phenotype of these cells is highly dependent on their specific microenvironment and when isolated and cultured, they lose their specific features after few passages, making models using such cells poorly predictive and irreproducible. We propose a new source of specialized endothelial cells based on cord blood circulating endothelial progenitors (EPCs). As prototype examples, we evaluated the capacity of EPCs to acquire properties characteristic of cerebral microvascular endothelial cells (blood-brain barrier (BBB)) or of arterial endothelial cells, in specific inducing culture conditions. APPROACH AND RESULTS: First, we demonstrated that EPC-derived endothelial cells (EPDCs) co-cultured with astrocytes acquired several BBB phenotypic characteristics, such as restricted paracellular diffusion of hydrophilic solutes and the expression of tight junction proteins. Second, we observed that culture of the same EPDCs in a high concentration of VEGF resulted, through activation of Notch signaling, in an increase of expression of most arterial endothelial markers. CONCLUSIONS: We have thus demonstrated that in vitro culture of early passage human cord blood EPDCs under specific conditions can induce phenotypic changes towards BBB or arterial phenotypes, indicating that these EPDCs maintain enough plasticity to acquire characteristics of a variety of specialized phenotypes. We propose that this property of EPDCs might be exploited for producing specialized endothelial cells in culture to be used for drug testing and predictive in vitro assays.

  • Charbord, P, Pouget, C, Binder, H, Dumont, F, Stik, G, Levy, P, Allain, F, Marchal, C, Richter, J, Uzan, B, Pflumio, F, Letourneur, F, Wirth, H, Dzierzak, E, Traver, D, Jaffredo, T & Durand, C 2014, « A Systems Biology Approach for Defining the Molecular Framework of the Hematopoietic Stem Cell Niche », Cell Stem Cell.
    Résumé : Despite progress in identifying the cellular composition of hematopoietic stem/progenitor cell (HSPC) niches, little is known about the molecular requirements of HSPC support. To address this issue, we used a panel of six recognized HSPC-supportive stromal lines and less-supportive counterparts originating from embryonic and adult hematopoietic sites. Through comprehensive transcriptomic meta-analyses, we identified 481 mRNAs and 17 microRNAs organized in a modular network implicated in paracrine signaling. Further inclusion of 18 additional cell strains demonstrated that this mRNA subset was predictive of HSPC support. Our gene set contains most known HSPC regulators as well as a number of unexpected ones, such as Pax9 and Ccdc80, as validated by functional studies in zebrafish embryos. In sum, our approach has identified the core molecular network required for HSPC support. These cues, along with a searchable web resource, will inform ongoing efforts to instruct HSPC ex vivo amplification and formation from pluripotent precursors.

  • Chevalier, F, Lavergne, M, Negroni, E, Ferratge, S, Carpentier, G, Gilbert-Sirieix, M, Siñeriz, F, Uzan, G & Albanese, P 2014, « Glycosaminoglycan mimetic improves enrichment and cell functions of human endothelial progenitor cell colonies », Stem Cell Research, vol. 12, no. 3, p. 703-715.
    Résumé : Human circulating endothelial progenitor cells isolated from peripheral blood generate in culture cells with features of endothelial cells named late-outgrowth endothelial colony-forming cells (ECFC). In adult blood, ECFC display a constant quantitative and qualitative decline during life span. Even after expansion, it is difficult to reach the cell dose required for cell therapy of vascular diseases, thus limiting the clinical use of these cells. Glycosaminoglycans (GAG) are components from the extracellular matrix (ECM) that are able to interact and potentiate heparin binding growth factor (HBGF) activities. According to these relevant biological properties of GAG, we designed a GAG mimetic having the capacity to increase the yield of ECFC production from blood and to improve functionality of their endothelial outgrowth. We demonstrate that the addition of [OTR(4131)] mimetic during the isolation process of ECFC from Cord Blood induces a 3 fold increase in the number of colonies. Moreover, addition of [OTR(4131)] to cell culture media improves adhesion, proliferation, migration and self-renewal of ECFC. We provide evidence showing that GAG mimetics may have great interest for cell therapy applied to vascular regeneration therapy and represent an alternative to exogenous growth factor treatments to optimize potential therapeutic properties of ECFC.

  • Coilly, A, Roche, B, Duclos-Vallée, J-C & Samuel, D 2014, « Management of HCV transplant patients with triple therapy », Liver International: Official Journal of the International Association for the Study of the Liver, vol. 34 Suppl 1, p. 46-52.
    Résumé : Hepatitis C virus (HCV) infection is one of the leading causes of end-stage liver disease and the main indication for liver transplantation (LT) in most countries. All patients who undergo LT with detectable serum HCV RNA experience graft reinfection. Between 20 and 30% of patients have developed cirrhosis at 5 years post-LT. The outcome of transplant patients with cirrhosis on the graft is severe, with a rate of decompensation at 1 year of approximately 40%. To date, retransplantation is the only option in patients with decompensated liver disease. Until 2011, standard antiviral therapy with pegylated interferon (PEG-IFN) and ribavirin (RBV), was the only effective therapy. Obtaining a sustained virological response (SVR) in patients with LT greatly improves overall and graft survival but this only occurs in 30% of transplanted patients. Direct acting antivirals (DAAs) such as protease inhibitors (PI), polymerase or other non-structural proteins inhibitors represent a new era in HCV associated liver disease. Although their use in the field of LT will certainly be essential there are some limitations because of safety and tolerance. One limitation is the potential interaction with calcineurin inhibitors. We describe the results of triple therapy with boceprevir (BOC) or telaprevir (TVR) for efficacy and safety and comment on future therapeutic strategies in liver transplant recipients.

  • Coilly, A, Roche, B, Dumortier, J, Leroy, V, Botta-Fridlund, D, Radenne, S, Pageaux, G-P, Si-Ahmed, S-N, Guillaud, O, Antonini, TM, Haïm-Boukobza, S, Roque-Afonso, A-M, Samuel, D & Duclos-Vallée, J-C 2014, « Safety and efficacy of protease inhibitors to treat hepatitis C after liver transplantation: a multicenter experience », Journal of Hepatology, vol. 60, no. 1, p. 78-86.
    Résumé : BACKGROUND & AIMS: Protease inhibitors (PI) with peginterferon/ribavirin have significantly improved SVR rates in HCV G1 patients. Their use to treat HCV recurrence after liver transplantation (LT) is a challenge. METHODS: This cohort study included 37 liver transplant recipients (male, 92%, age 57 ± 11 years), treated with boceprevir (n=18) or telaprevir (n=19). The indication for therapy was HCV recurrence (fibrosis stage ≥F2 (n=31, 83%) or fibrosing cholestatic hepatitis (n=6, 16%). RESULTS: Eighteen patients were treatment-naive, five were relapsers and fourteen were non-responders to dual therapy after LT. Twenty-two patients received cyclosporine and fifteen tacrolimus. After 12 weeks of PI therapy, a complete virological response was obtained in 89% of patients treated with boceprevir, and 58% with telaprevir (p=0.06). The end of treatment virological response rate was 72% (13/18) in the boceprevir group and 40% (4/10) in the telaprevir group (p=0.125). A sustained virological response 12 weeks after treatment discontinuation was observed in 20% (1/5) and 71% (5/7) of patients in the telaprevir and boceprevir groups, respectively (p=0.24). Treatment was discontinued in sixteen patients (treatment failures (n=11), adverse events (n=5)). Infections occurred in ten patients (27%), with three fatal outcomes (8%). The most common adverse effect was anemia (n=34, 92%), treated with erythropoietin and/or a ribavirin dose reduction; thirteen patients (35%) received red blood cell transfusions. The cyclosporine dose was reduced by 1.8 ± 1.1-fold and 3.4 ± 1.0-fold with boceprevir and telaprevir, respectively. The tacrolimus dose was reduced by 5.2 ± 1.5-fold with boceprevir and 23.8±18.2-fold with telaprevir. CONCLUSIONS: Our results suggest that triple therapy is effective in LT recipients, particularly those experiencing a severe recurrence. The occurrence of anemia and drug-drug interactions, and the risk of infections require close monitoring.

  • Coilly, A, Sebagh, M & Duclos-Vallée, J-C 2014, « Reply to: "Frequent plasma cell hepatitis during telaprevir-based triple therapy for hepatitis C after liver transplantation" », Journal of Hepatology, vol. 60, no. 4, p. 896-897.

  • Dianat, N, Dubois-Pot-Schneider, H, Steichen, C, Desterke, C, Leclerc, P, Raveux, A, Combettes, L, Weber, A, Corlu, A & Dubart-Kupperschmitt, A 2014, « Generation of functional cholangiocyte-like cells from human pluripotent stem cells and HepaRG cells », Hepatology (Baltimore, Md.).
    Résumé : Cholangiocytes are biliary epithelial cells, which, like hepatocytes, originate from hepatoblasts during embryonic development. In this study, we investigated the potential of human embryonic stem cells (hESCs) to differentiate into cholangiocytes and we report a new approach, which drives differentiation of hESCs toward the cholangiocytic lineage using feeder-free and defined culture conditions. After differentiation into hepatic progenitors, hESCs were differentiated further into cholangiocytes using growth hormone, epidermal growth factor, Interleukin-6 and then sodium taurocholate. These conditions also allowed us to generate cholangiocytes from HepaRG-derived hepatoblasts. hESC- and HepaRG-derived cholangiocyte-like cells expressed markers of cholangiocytes including Cytokeratin 7 and Osteopontin, and transcription factors SOX9 and Hepatocyte nuclear factor 6. The cells also displayed specific proteins important for cholangiocyte functions including cystic fibrosis transmembrane conductance regulator, secretin receptor and nuclear receptors. They formed primary cilia and also responded to hormonal stimulation by increase of intracellular Ca(2+) . We demonstrated by integrative genomics that the expression of genes, which signed hESC- or HepaRG-cholangiocytes separates hepatocytic lineage from cholangiocyte lineage. When grown in a 3D matrix, cholangiocytes developed epithelial/apicobasal polarity and formed functional cysts and biliary ducts. In addition, we showed that cholangiocyte-like cells could also be generated from human induced pluripotent stem cells, demonstrating the efficacy of our approach with stem/progenitor cells of diverse origins. Conclusion: We have developed a robust and efficient method for differentiating pluripotent stem cells into cholangiocyte-like cells, which display structural and functional similarities with bile duct cells in normal liver. These cells will be useful for the in vitro study of the molecular mechanisms of bile duct development and have important potential for therapeutic strategies, including bioengineered liver approaches. (Hepatology 2014).

  • Dubois, SM, Alexia, C, Wu, Y, Leclair, HM, Leveau, C, Schol, E, Fest, T, Tarte, K, Chen, ZJ, Gavard, J & Bidère, N 2014, « A catalytic-independent role for the LUBAC in NF-κB activation upon antigen receptor engagement and in lymphoma cells », Blood, vol. 123, no. 14, p. 2199-2203.
    Résumé : Antigen receptor-mediated nuclear factor κB (NF-κB) activation relies on the formation of a large multi-protein complex that contains CARMA1, BCL10, and MALT1 (CBM complex). This signalosome is pirated in the activated B-cell-like subgroup of diffuse large B-cell lymphoma (ABC DLBCL) to drive aberrant NF-κB activation, thereby promoting cell survival and propagation. Using an unbiased proteomic approach, we screened for additional components of the CBM in lymphocytes. We found that the linear ubiquitin chain assembly complex (LUBAC), which was previously linked to cytokine-mediated NF-κB activation, dynamically integrates the CBM and marshals NF-κB optimal activation following antigen receptor ligation independently of its catalytic activity. The LUBAC also participates in preassembled CBM complex in cells derived from ABC DLBCL. Silencing the LUBAC reduced NF-κB activation and was toxic in ABC DLBCL cell lines. Thus, our findings reveal a role for the LUBAC during lymphocyte activation and in B-cell malignancy.
    Mots-clés : Catalysis, Cell Line, Tumor, Humans, Jurkat Cells, Lymphocyte Activation, Lymphoma, NF-kappa B, Protein Binding, Protein Interaction Domains and Motifs, Receptors, Antigen, T-Cell, Ubiquitin, Ubiquitination.

  • Dumortier, J, Salamé, E, Roche, B, Hurtova, M, Conti, F, Radenne, S, Vanlemmens, C, Pageaux, G-P, Saliba, F, Samuel, D, Compagnon, P, Neau-Cransac, M, Calmus, Y, Guillaud, O, Gugenheim, J, Altieri, M, Durand, F, Hardwigsen, J, Lorho, R, Dharancy, S, Leroy, V, Di Giambattista, F & Duvoux, C 2014, « Severe fibrosis in patients with recurrent hepatitis C after liver transplantation: A French experience on 250 patients over 15 years (the Orfèvre study) », Clinics and Research in Hepatology and Gastroenterology, vol. 38, no. 3, p. 292-299.
    Résumé : BACKGROUND AND AIMS: Recurrent hepatitis C after liver transplantation (LT) is associated with rapid fibrosis progression. The aim of this study was to evaluate the cumulative risk for severe fibrosis and the factors influencing it. PATIENTS AND METHODS: Two hundred and fifty LT patients were included 1 to 15years after LT. Recurrence of chronic hepatitis C on liver graft was classified according to Metavir score. RESULTS: Kaplan-Meyer estimates for actuarial progression to severe fibrosis (Metavir>F3) showed a probability of 15.2% and 44.5% at 5 and 10years, respectively. Predictive factors for progression to severe fibrosis were: use of tacrolimus as main CNI, recipient age at time of biopsy<55, donor age ≥45, graft HCV re-infection<3months, biologically suspected graft re-infection and lack of response to antiviral treatment after LT. Multivariate analysis disclosed that only donor age ≥45 (hazard ratio 2.243, 95%CI 1.264-3.983, P=0.0058) and lack of response to antiviral treatment (hazard ratio 2.816, 95%CI 1.227-6.464, P=0.0146) were associated to severe fibrosis. CONCLUSIONS: Our study confirms that donor age ≥45 and lack of response to antiviral treatment after LT are major predictive factors of progression of HCV recurrence on liver graft.

  • Durand, M, Collombet, J-M, Frasca, S, Begot, L, Lataillade, J-J, Le Bousse-Kerdilès, M-C & Holy, X 2014, « In Vivo Hypobaric Hypoxia Performed During the Remodeling Process Accelerates Bone Healing in Mice », Stem Cells Translational Medicine.
    Résumé : We investigated the effects of respiratory hypobaric hypoxia on femoral bone-defect repair in mice because hypoxia is believed to influence both mesenchymal stromal cell (MSC) and hematopoietic stem cell mobilization, a process involved in the bone-healing mechanism. To mimic conditions of non-weight-bearing limb immobilization in patients suffering from bone trauma, our hypoxic mouse model was further subjected to hind-limb unloading. A hole was drilled in the right femur of adult male C57/BL6J mice. Four days after surgery, mice were subjected to hind-limb unloading for 1 week. Seven days after surgery, mice were either housed for 4 days in a hypobaric room (FiO2 at 10%) or kept under normoxic conditions. Unsuspended control mice were housed in either hypobaric or normoxic conditions. Animals were sacrificed on postsurgery day 11 to allow for collection of both contralateral and lesioned femurs, blood, and spleen. As assessed by microtomography, delayed hypoxia enhanced bone-healing efficiency by increasing the closing of the cortical defect and the newly synthesized bone volume in the cavity by +55% and +35%, respectively. Proteome analysis and histomorphometric data suggested that bone-repair improvement likely results from the acceleration of the natural bone-healing process rather than from extended mobilization of MSC-derived osteoprogenitors. Hind-limb unloading had hardly any effect beyond delayed hypoxia-enhanced bone-healing efficiency.
    Mots-clés : Animals, Anoxia, Biological Markers, Bone Remodeling, Disease Models, Animal, Femoral Fractures, Femur, Fracture Healing, Hematopoietic Stem Cells, Hindlimb Suspension, Male, Mesenchymal Stromal Cells, Mice, Mice, Inbred C57BL, Proteomics, Time Factors, X-Ray Microtomography.

  • Feillet, C, Krusche, P, Tamanini, F, Janssens, RC, Downey, MJ, Martin, P, Teboul, M, Saito, S, Lévi, FA, Bretschneider, T, van der Horst, GTJ, Delaunay, F & Rand, DA 2014, « Phase locking and multiple oscillating attractors for the coupled mammalian clock and cell cycle », Proceedings of the National Academy of Sciences of the United States of America, vol. 111, no. 27, p. 9828-9833.
    Résumé : Daily synchronous rhythms of cell division at the tissue or organism level are observed in many species and suggest that the circadian clock and cell cycle oscillators are coupled. For mammals, despite known mechanistic interactions, the effect of such coupling on clock and cell cycle progression, and hence its biological relevance, is not understood. In particular, we do not know how the temporal organization of cell division at the single-cell level produces this daily rhythm at the tissue level. Here we use multispectral imaging of single live cells, computational methods, and mathematical modeling to address this question in proliferating mouse fibroblasts. We show that in unsynchronized cells the cell cycle and circadian clock robustly phase lock each other in a 1:1 fashion so that in an expanding cell population the two oscillators oscillate in a synchronized way with a common frequency. Dexamethasone-induced synchronization reveals additional clock states. As well as the low-period phase-locked state there are distinct coexisting states with a significantly higher period clock. Cells transition to these states after dexamethasone synchronization. The temporal coordination of cell division by phase locking to the clock at a single-cell level has significant implications because disordered circadian function is increasingly being linked to the pathogenesis of many diseases, including cancer.

  • Féray, C, Pawlotsky, J-M, Roque-Afonso, A-M, Samuel, D & Dhumeaux, D 2014, « Should we screen blood products for hepatitis E virus RNA ? », Lancet, vol. 383, no. 9913, p. 218.
    Résumé : PMID: 24439737
    Mots-clés : Blood Donors, Blood Safety, Flaviviridae Infections, GB virus C, Hepatitis, Viral, Human, Humans, Mass Screening, RNA, Viral.

  • Filipski, E, Berland, E, Ozturk, N, Guettier, C, van der Horst, GTJ, Lévi, F & Okyar, A 2014, « Optimization of irinotecan chronotherapy with P-glycoprotein inhibition », Toxicology and Applied Pharmacology, vol. 274, no. 3, p. 471-479.
    Résumé : The relevance of P-glycoprotein (P-gp) for irinotecan chronopharmacology was investigated in female B6D2F1 mice. A three-fold 24h change in the mRNA expression of Abcb1b was demonstrated in ileum mucosa, with a maximum at Zeitgeber Time (ZT) 15 (p<0.001). No rhythm was found for abcb1a in ileum mucosa, or for Abcb1a/b in Glasgow osteosarcoma (GOS), a mouse tumor cell line moderately sensitive to irinotecan. Non-tumor-bearing mice received irinotecan (50mg/kg/day i.v.×4days) as a single agent or combined with P-gp inhibitor PSC833 (6.25mg/kg/day i.p.×4 days) at ZT3 or ZT15, respectively corresponding to the worst or the best irinotecan tolerability. Endpoints involved survival, body weight change and hematologic toxicity. Antitumor efficacy was studied in GOS-bearing mice receiving irinotecan (25, 30 or 40mg/kg/day×4days) and +/-PSC833 at ZT3 or ZT15, with survival, body weight change, and tumor growth inhibition as endpoints. Non-tumor bearing mice lost an average of 17% or 9% of their body weight according to irinotecan administration at ZT3 or ZT15 respectively (p<0.001). Dosing at ZT15 rather than ZT3 reduced mean leucopenia (9% vs 53%; p<0.001). PSC833 aggravated irinotecan lethal toxicity from 4 to ~60%. In tumor-bearing mice, body weight loss was ~halved in the mice on irinotecan or irinotecan-PSC833 combination at ZT15 as compared to ZT3 (p<0.001). PSC833-irinotecan at ZT15 increased tumor inhibition by ~40% as compared to irinotecan only at ZT15. In conclusion, P-gp was an important determinant of the circadian balance between toxicity and efficacy of irinotecan.
    Mots-clés : Animals, Body Weight, Camptothecin, Cell Line, Tumor, Chronotherapy, Circadian Rhythm, Cyclosporins, Endpoint Determination, Female, Ileum, Mice, Mucous Membrane, Osteosarcoma, P-Glycoprotein, RNA, Messenger, Time Factors.

  • Gane, EJ, Deray, G, Liaw, Y-F, Lim, SG, Lai, C-L, Rasenack, J, Wang, Y, Papatheodoridis, G, Di Bisceglie, A, Buti, M, Samuel, D, Uddin, A, Bosset, S & Trylesinski, A 2014, « Telbivudine improves renal function in patients with chronic hepatitis B », Gastroenterology, vol. 146, no. 1, p. 138-146.e5.
    Résumé : BACKGROUND & AIMS: There is a close relationship between chronic hepatitis B virus infection and chronic renal disease. We analyzed changes in renal function using different markers of glomerular filtration rate (GFR) in multiple studies of telbivudine treatment of patients with chronic hepatitis B virus infection. METHODS: We used serum creatinine-based equations (ie, Cockcroft-Gault, Modification of Diet in Renal Disease, and Chronic Kidney Disease Epidemiology Collaboration) to estimate GFR (eGFR) in adults with chronic hepatitis B virus infection and compensated liver disease who participated in a phase III, randomized, double-blind study comparing the efficacy and safety of telbivudine (600 mg/d) and lamivudine (100 mg/d) for 2 years (the GLOBE study) and in long-term extension studies (4-6 years), as well as in patients with decompensated cirrhosis (2 years). RESULTS: eGFRs calculated using the Cockcroft-Gault, Modification of Diet in Renal Disease, and Chronic Kidney Disease Epidemiology Collaboration equations were concordant, indicating improved renal function in telbivudine-treated patients during the 2-year GLOBE study (there was an 8.5% increase in mean eGFR, based on the Modification of Diet in Renal Disease equation). Improved renal function was maintained for 4-6 years. Increased eGFR with telbivudine treatment was also observed in patients at increased risk for renal impairment: patients with baseline eGFRs of 60-89 mL/min/1.73 m(2) (+17.2%), older than 50 years (+11.4%), and with liver fibrosis/cirrhosis (+7.2% for patients with Ishak fibrosis score at 5-6). In decompensated patients with high renal risk, eGFR was also improved on telbivudine (+2.0%). CONCLUSIONS: In global trials of patients with compensated and decompensated cirrhosis, long-term telbivudine therapy was associated with a sustained improvement of renal function-particularly among patients with increased risk of renal impairment. The mechanisms of this renal protective effect remain to be determined.
    Mots-clés : Adult, Antiviral Agents, Creatinine, Double-Blind Method, Female, Glomerular Filtration Rate, Hepatitis B, Chronic, Humans, Male, Renal Insufficiency, Chronic, Thymidine, Treatment Outcome.

  • Giuliani, M, Bennaceur-Griscelli, A, Nanbakhsh, A, Oudrhiri, N, Chouaib, S, Azzarone, B, Durrbach, A & Lataillade, J-J 2014, « TLR ligands stimulation protects MSC from NK killing », Stem cells (Dayton, Ohio), vol. 32, no. 1, p. 290-300.
    Résumé : Mesenchymal stem cells (MSCs) play a fundamental role in allograft rejection and graft-versus-host disease through their immunosuppressive abilities. Recently, Toll-like receptors (TLR) have been shown to modulate MSC functions. The aim of this study was to investigate the effects of several TLR ligands on the interaction between MSC and natural killer (NK) cells. Our results show that TLR-primed adult bone marrow and embryonic MSC are more resistant than unprimed MSC to IL-2-activated NK-induced killing. Such protection can be explained by the modulation of Natural Killer group 2D ligands major histocompatibility complex class I chain A and ULBP3 and DNAM-1 ligands by TLR-primed MSC. These results indicate that MSCs are able to adapt their immuno-behavior in an inflammatory context, decreasing their susceptibility to NK killing. In addition, TLR3 but not TLR4-primed MSC enhance their suppressive functions against NK cells. However, the efficiency of this response is heterogeneous, even if the phenotypes of different analyzed MSC are rather homogeneous. The consequences could be important in MSC-mediated cell therapy, since the heterogeneity of adult MSC responders may be explored in order to select the more efficient responders.

  • Guillaud, O, Dumortier, J, Sobesky, R, Debray, D, Wolf, P, Vanlemmens, C, Durand, F, Calmus, Y, Duvoux, C, Dharancy, S, Kamar, N, Boudjema, K, Bernard, PH, Pageaux, G-P, Salamé, E, Gugenheim, J, Lachaux, A, Habes, D, Radenne, S, Hardwigsen, J, Chazouillères, O, Trocello, J-M, Woimant, F, Ichai, P, Branchereau, S, Soubrane, O, Castaing, D, Jacquemin, E, Samuel, D & Duclos-Vallée, J-C 2014, « Long term results of liver transplantation for Wilson's disease: experience in France », Journal of Hepatology, vol. 60, no. 3, p. 579-589.
    Résumé : BACKGROUND & AIMS: Liver transplantation (LT) is the therapeutic option for severe complications of Wilson's disease (WD). We aimed to report on the long-term outcome of WD patients following LT. METHODS: The medical records of 121 French patients transplanted for WD between 1985 and 2009 were reviewed retrospectively. Seventy-five patients were adults (median age: 29 years, (18-66)) and 46 were children (median age: 14 years, (7-17)). The indication for LT was (1) fulminant/subfulminant hepatitis (n = 64, 53%), median age = 16 years (7-53), (2) decompensated cirrhosis (n = 50, 41%), median age = 31.5 years (12-66) or (3) severe neurological disease (n = 7, 6%), median age = 21.5 years (14.5-42). Median post-transplant follow-up was 72 months (0-23.5). RESULTS: Actuarial patient survival rates were 87% at 5, 10, and 15 years. Male gender, pre-transplant renal insufficiency, non elective procedure, and neurological indication were significantly associated with poorer survival rate. None of these factors remained statistically significant under multivariate analysis. In patients transplanted for hepatic indications, the prognosis was poorer in case of fulminant or subfulminant course, non elective procedure, pretransplant renal insufficiency and in patients transplanted before 2000. Multivariate analysis disclosed that only recent period of LT was associated with better prognosis. At last visit, the median calculated glomerular filtration rate was 93 ml/min (33-180); 11/93 patients (12%) had stage II renal insufficiency and none had stage III. CONCLUSIONS: Liver failure associated with WD is a rare indication for LT (<1%), which achieves an excellent long-term outcome, including renal function.

  • Hézode, C, Fontaine, H, Dorival, C, Zoulim, F, Larrey, D, Canva, V, De Ledinghen, V, Poynard, T, Samuel, D, Bourliere, M, Alric, L, Raabe, J-J, Zarski, J-P, Marcellin, P, Riachi, G, Bernard, P-H, Loustaud-Ratti, V, Chazouilleres, O, Abergel, A, Guyader, D, Metivier, S, Tran, A, Di Martino, V, Causse, X, Dao, T, Lucidarme, D, Portal, I, Cacoub, P, Gournay, J, Grando-Lemaire, V, Hillon, P, Attali, P, Fontanges, T, Rosa, I, Petrov-Sanchez, V, Barthe, Y, Pawlotsky, J-M, Pol, S, Carrat, F, Bronowicki, J-P & CUPIC Study Group, 2014, « Effectiveness of telaprevir or boceprevir in treatment-experienced patients with HCV genotype 1 infection and cirrhosis », Gastroenterology, vol. 147, no. 1, p. 132-142.e4.
    Résumé : BACKGROUND & AIMS: We investigated the effectiveness of the protease inhibitors peginterferon and ribavirin in treatment-experienced patients with hepatitis C virus (HCV) genotype 1 infection and cirrhosis. METHODS: In the Compassionate Use of Protease Inhibitors in Viral C Cirrhosis study, 511 patients with HCV genotype 1 infection and compensated cirrhosis who did not respond to a prior course of peginterferon and ribavirin (44.3% relapsers or patients with viral breakthrough, 44.8% partial responders, and 8.0% null responders) were given either telaprevir (n = 299) or boceprevir (n = 212) for 48 weeks. We assessed percentages of patients with sustained viral responses 12 weeks after therapy and safety. This observational study did not allow for direct comparison of the 2 regimens. RESULTS: Among patients given telaprevir, 74.2% of relapsers, 40.0% of partial responders, and 19.4% of null responders achieved SVR12. Among those given boceprevir, 53.9% of relapsers, 38.3% of partial responders, and none of the null responders achieved SVR12. In multivariate analysis, factors associated with SVR12 included prior response to treatment response, no lead-in phase, HCV subtype 1b (vs 1a), and baseline platelet count greater than 100,000/mm(3). Severe adverse events occurred in 49.9% of cases, including liver decompensation, severe infections in 10.4%, and death in 2.2%. In multivariate analysis, baseline serum albumin level less than 35 g/L and baseline platelet counts of 100,000/mm(3) or less predicted severe side effects or death. CONCLUSIONS: Relatively high percentages of real-life, treatment-experienced patients with HCV genotype 1 infection and cirrhosis respond to the combination of peginterferon and ribavirin with telaprevir or boceprevir. However, side effects are frequent and often severe. Baseline levels of albumin and platelet counts can be used to guide treatment decisions. ClinicalTrials.gov number: NCT01514890.

  • Jalan, R, Saliba, F, Pavesi, M, Amoros, A, Moreau, R, Ginès, P, Levesque, E, Durand, F, Angeli, P, Caraceni, P, Hopf, C, Alessandria, C, Rodriguez, E, Solis-Muñoz, P, Laleman, W, Trebicka, J, Zeuzem, S, Gustot, T, Mookerjee, R, Elkrief, L, Soriano, G, Cordoba, J, Morando, F, Gerbes, A, Agarwal, B, Samuel, D, Bernardi, M, Arroyo, V & for the CANONIC study investigators of the EASL-CLIF Consortium, 2014, « Development and Validation of a Prognostic Score to Predict Mortality in Patients with Acute on Chronic Liver Failure », Journal of Hepatology.
    Résumé : BACKGROUND & AIMS: Acute-on Chronic Liver Failure (ACLF) is a frequent syndrome (30% prevalence) characterized by acute decompensation of cirrhosis, organ failure(s) and high short-term mortality. This study develops and validates a specific prognostic score for ACLF patients. METHODS: Data from 1,349 patients included in the CANONIC study were used. First, a simplified organ function scoring system (CLIF-Consortium Organ Failure score, CLIF-C OFs) to diagnose ACLF was developed using data from all patients. Subsequently, in 275 patients with ACLF, CLIF-C OFs and two other independent predictors of mortality (age and white-cell count) were combined to develop a specific prognostic score for ACLF (CLIF CONSORTIUM score for ACLF, CLIF-C ACLFs). Concordance index (C-index) was used to compare the discrimination abilities of CLIF-C ACLFs, MELD (MELDs), MELD-Sodium (MELD-Nas) and Child-Pugh (CPs) scores. CLIF-C ACLFs was validated in an external cohort and assessed for sequential use. RESULTS: CLIF-C ACLFs showed a significantly higher predictive accuracy than MELDs, MELD-Nas and CPs, reducing (19-28%) the corresponding prediction error rates at all the main time-points after ACLF diagnosis (28, 90, 180 and 365 days) in both the CANONIC and the external validation cohort. CLIF-C ACLFs computed at 48 hours, 3-7 days and 8-15 days after ACLF diagnosis predicted 28-day mortality significantly better than at diagnosis. CONCLUSIONS: CLIF-C ACLFs at ACLF diagnosis is superior to MELDs and MELD-Nas in predicting mortality. CLIF-C ACLFs is a clinically relevant, validated scoring system that can be used sequentially to stratify the risk of mortality in ACLF patients.

  • Larbi, A, Mitjavila-Garcia, MT, Flamant, S, Valogne, Y, Clay, D, Usunier, B, l'Homme, B, Féraud, O, Casal, I, Gobbo, E, Divers, D, Chapel, A, Turhan, AG, Bennaceur-Griscelli, A & Haddad, R 2014, « Generation of Multipotent Early Lymphoid Progenitors from Human Embryonic Stem Cells », Stem Cells and Development.
    Résumé : During human embryonic stem cell (ESC) hematopoietic differentiation, the description of the initial steps of lymphopoiesis remains elusive. Using a two-step culture procedure, we identified two original populations of ESC-derived hematopoietic progenitor cells (HPCs) with CD34(+)CD45RA(+)CD7(-) and CD34(+)CD45RA(+)CD7(+) phenotypes. Bulk cultures and limiting dilution assays, culture with MS5 cells in the presence of Notch ligand Delta-like-1 (DL-1), and ex vivo colonization tests using fetal thymic organ cultures showed that although CD34(+)CD45RA(+)CD7(-) HPCs could generate cells of the three lymphoid lineages, their potential was skewed toward the B cell lineages. In contrast, CD34(+)CD45RA(+)CD7(+) HPCs predominantly exhibited a T/natural killer (NK) cell differentiation potential. Furthermore these cells could differentiate equivalently into cells of the granulo-macrophagic lineage and dendritic cells and lacked erythroid potential. Expression profiling of 18 markers by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) revealed that CD34(+)CD45RA(+)CD7(-) and CD34(+)CD45RA(+)CD7(+) HPCs express genes of the lymphoid specification and that CD34(+)CD45RA(+)CD7(-) cells express B-cell-associated genes, while CD34(+)CD45RA(+)CD7(+) HPCs display a T-cell molecular profile. Altogether, these findings indicate that CD34(+)CD45RA(+)CD7(-) and CD34(+)CD45RA(+)CD7(+) HPCs correspond to candidate multipotent early lymphoid progenitors polarized toward either the B or T/NK lineage, respectively. This work should improve our understanding of the early steps of lymphopoiesis from pluripotent stem cells and pave the way for the production of lymphocytes for cell-based immunotherapy and lymphoid development studies.

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