Hepatic Tissue Environment in NEMO-Deficient Mice Critically Regulates Positive Selection of Donor Cells after Hepatocyte Transplantation
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{"title"=>"Hepatic tissue environment in NEMO-deficient mice critically regulates positive selection of donor cells after hepatocyte transplantation", "type"=>"journal", "authors"=>[{"first_name"=>"Michaela", "last_name"=>"Kaldenbach", "scopus_author_id"=>"35774162900"}, {"first_name"=>"Francisco Javier", "last_name"=>"Cubero", "scopus_author_id"=>"8517959700"}, {"first_name"=>"Stephanie", "last_name"=>"Erschfeld", "scopus_author_id"=>"35338987300"}, {"first_name"=>"Christian", "last_name"=>"Liedtke", "scopus_author_id"=>"8041642800"}, {"first_name"=>"Christian", "last_name"=>"Trautwein", "scopus_author_id"=>"7005071218"}, {"first_name"=>"Konrad", "last_name"=>"Streetz", "scopus_author_id"=>"6603357715"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"24979756", "sgr"=>"84903648359", "doi"=>"10.1371/journal.pone.0100786", "scopus"=>"2-s2.0-84903648359", "pui"=>"373435012", "issn"=>"19326203"}, "id"=>"21a68970-a056-30dd-aab3-eef23d090218", "abstract"=>"BACKGROUND: Hepatocyte transplantation (HT) is a promising alternative treatment strategy for end-stage liver diseases compared with orthotopic liver transplantation. A limitation for this approach is the low engraftment of donor cells. The deletion of the I-kappa B kinase-regulatory subunit IKKγ/NEMO in hepatocytes prevents nuclear factor (NF)-kB activation and triggers spontaneous liver apoptosis, chronic hepatitis and the development of liver fibrosis and hepatocellular carcinoma. We hypothesized that NEMOΔhepa mice may therefore serve as an experimental model to study HT.\\n\\nMETHODS: Pre-conditioned NEMOΔhepa mice were transplanted with donor-hepatocytes from wildtype (WT) and mice deficient for the pro-apoptotic mediator Caspase-8 (Casp8Δhepa).\\n\\nRESULTS: Transplantation of isolated WT-hepatocytes into pre-conditioned NEMOΔhepa mice resulted in a 6-7 fold increase of donor cells 12 weeks after HT, while WT-recipients showed no liver repopulation. The use of apoptosis-resistant Casp8Δhepa-derived donor cells further enhanced the selection 3-fold after 12-weeks and up to 10-fold increase after 52 weeks compared with WT donors. While analysis of NEMOΔhepa mice revealed strong liver injury, HT-recipient NEMOΔhepa mice showed improved liver morphology and decrease in serum transaminases. Concomitant with these findings, the histological examination elicited an improved liver tissue architecture associated with significantly lower levels of apoptosis, decreased proliferation and a lesser amount of liver fibrogenesis. Altogether, our data clearly support the therapeutic benefit of the HT procedure into NEMOΔhepa mice.\\n\\nCONCLUSION: This study demonstrates the feasibility of the NEMOΔhepa mouse as an in vivo tool to study liver repopulation after HT. The improvement of the characteristic phenotype of chronic liver injury in NEMOΔhepa mice after HT suggests the therapeutic potential of HT in liver diseases with a chronic inflammatory phenotype and opens a new door for the applicability of this technique to combat liver disease in the human clinic.", "link"=>"http://www.mendeley.com/research/hepatic-tissue-environment-nemodeficient-mice-critically-regulates-positive-selection-donor-cells-af", "reader_count"=>4, "reader_count_by_academic_status"=>{"Student > Ph. D. Student"=>2, "Student > Bachelor"=>1, "Researcher"=>1}, "reader_count_by_user_role"=>{"Student > Ph. D. Student"=>2, "Student > Bachelor"=>1, "Researcher"=>1}, "reader_count_by_subject_area"=>{"Agricultural and Biological Sciences"=>1, "Medicine and Dentistry"=>2, "Biochemistry, Genetics and Molecular Biology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1575218"], "description"=>"<p>(<b>A</b>) NK cell staining was performed in order to address the role of the immune cell response to HT (blue: DAPI; green: NK1.1(+) cells). This shows, that NK-cells are present in a significantly lower number in HT-recipient mice as compared to controls and solely BM-treated animals. (<b>B</b>) Visualization of cell proliferation by Ki-67 and BrdU incorporation stainings. (blue: DAPI; green: BrdU(+); red: Ki-67(+)). (<b>C</b>) Determination of the total number of BrdU(+) cells by means of BrdU staining in a 100x magnification. (*p<0.05, **p<0.01). (<b>D</b>) Quantification of Ki-67 positive signals in immunofluorescent staining using a 200x magnification. (*p<0.05, **p<0.01). (<b>E</b>) For localization of proliferating cells, double immunofluorescence staining of BrdU and hAAT was performed (blue: DAPI; red: hAAT(+) cells; green: BrdU(+) cells). (<b>F</b>) DAB-staining of NEMO protein in NEMO<sup>Δhepa</sup> mice transplanted with Casp8<sup>loxP/loxP</sup>/hAAT(+) or Casp8<sup>Δhepa</sup>/hAAT(+) donor cells, respectively (DAB in brown: NEMO protein). This clearly shows the presence of NEMO(+) hepatocytes in the surrounding NEMO-deficient (NEMO<sup>Δhepa</sup>) liver tissue.</p>", "links"=>[], "tags"=>["anatomy", "histology", "genetics", "systems biology", "Gastroenterology and hepatology", "nk", "activation", "hepatic"], "article_id"=>1089260, "categories"=>["Biological Sciences"], "users"=>["Michaela Kaldenbach", "Francisco Javier Cubero", "Stephanie Erschfeld", "Christian Liedtke", "Christian Trautwein", "Konrad Streetz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100786.g005", "stats"=>{"downloads"=>4, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Assessment_of_NK_cell_activation_and_hepatic_proliferation_/1089260", "title"=>"Assessment of NK cell activation and hepatic proliferation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-30 03:44:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1575210"], "description"=>"<p>(<b>A</b>) Analysis of Collagen accumulation in transplanted mice via Sirius red staining under polarized light. NEMO<sup>Δhepa</sup> mice that underwent transplantation with either Casp8<sup>loxP/loxP</sup>/hAAT(+) or Casp8<sup>Δhepa</sup>/hAAT(+) donor cells 52 weeks after HT were compared to age-matched completely untreated control mice as well as to bone marrow-transplanted mice 56 weeks (age matched) after BMT. (<b>B</b>) Collagen-1α staining for visualisation of a specific fibrotic collagen subtype (blue: DAPI: red/yellow: Collagen1-α). (<b>C</b>) Assessment of Collagen-1α mRNA expression via quantitative realtime PCR. (**p<0.01, ***p<0.001)</p>", "links"=>[], "tags"=>["anatomy", "histology", "genetics", "systems biology", "Gastroenterology and hepatology", "fibrogenesis", "hepatocyte", "transplanted"], "article_id"=>1089256, "categories"=>["Biological Sciences"], "users"=>["Michaela Kaldenbach", "Francisco Javier Cubero", "Stephanie Erschfeld", "Christian Liedtke", "Christian Trautwein", "Konrad Streetz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100786.g003", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reduced_fibrogenesis_in_hepatocyte_transplanted_NEMO_916_hepa_mice_/1089256", "title"=>"Reduced fibrogenesis in hepatocyte transplanted NEMO<sup>Δhepa</sup> mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-30 03:44:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1575208"], "description"=>"<p>(<b>A</b>) Macroscopical and histological analysis of livers from NEMO<sup>loxP/loxP</sup> and NEMO<sup>Δhepa</sup> mice that underwent HT compared to untreated and solely bone marrow transplanted livers. Livers were analysed 52 weeks after HT with either Casp8<sup>loxP/loxP</sup> (WT) or Casp8<sup>Δhepa</sup> donor mice in comparison to either naïve or solely BM-transplanted control mice of the appropriate age respectively. Control NEMO<sup>Δhepa</sup> mice display a progressive fatty degeneration and fibrosis development of their livers over time. This is illustrated by mononuclear cell infiltration, tissue necrosis and hepatocyte ballooning (left side, lower panel). Hepatocyte transplanted NEMO<sup>Δhepa</sup> mice displayed in this direct comparison less intrahepatic fat accumulation and tissue destruction, thus indicating a beneficial effect of HT in recipient NEMO<sup>Δhepa</sup> mice. (<b>B</b>) Liver <i>versus</i> bodyweight ratio, 52 weeks after HT. (**p<0.01, ***p<0.001). (<b>C, D</b>) Serum transaminases were measured as an indicator of liver function, showing a clear enhancement in transplanted compared to control mice. (**p<0.01)</p>", "links"=>[], "tags"=>["anatomy", "histology", "genetics", "systems biology", "Gastroenterology and hepatology", "transplantation", "mice"], "article_id"=>1089254, "categories"=>["Biological Sciences"], "users"=>["Michaela Kaldenbach", "Francisco Javier Cubero", "Stephanie Erschfeld", "Christian Liedtke", "Christian Trautwein", "Konrad Streetz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100786.g002", "stats"=>{"downloads"=>2, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Hepatocyte_transplantation_in_NEMO_916_hepa_mice_results_in_improved_liver_function_/1089254", "title"=>"Hepatocyte transplantation in NEMO<sup>Δhepa</sup> mice results in improved liver function.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-30 03:44:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1575205"], "description"=>"<p>(<b>A</b>) Hepatocyte transplantation was applied to NEMO<sup>Δhepa</sup> and WT recipient mice with preceding BMT. Donor cells were derived from WT (Casp8<sup>loxP/loxP</sup>/hAAT(+)) or Caspase-8-deficient (Casp8<sup>Δhepa</sup>/hAAT(+)) mice, respectively. (<b>B</b>) Serum hAAT levels were analysed via quantitative ELISA and displayed on a logarithmic scale. Basic values represent serum hAAT level 1d post transplantation and are used as a reference value for the calculation of the relative increase in the amount of donor derived hepatocytes over time. (<b>C</b>) Quantitative evaluation of hAAT (+) liver tissue areas 52 weeks after HT. (<b>D</b>) Visualization of engrafting donor cells using hAAT immunofluorescence staining. Clusters of donor derived hepatocytes are displayed 12 and 52 weeks after HT (blue: DAPI; red: hAAT(+) cells).</p>", "links"=>[], "tags"=>["anatomy", "histology", "genetics", "systems biology", "Gastroenterology and hepatology", "deficiency", "mice", "favours"], "article_id"=>1089251, "categories"=>["Biological Sciences"], "users"=>["Michaela Kaldenbach", "Francisco Javier Cubero", "Stephanie Erschfeld", "Christian Liedtke", "Christian Trautwein", "Konrad Streetz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100786.g001", "stats"=>{"downloads"=>2, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_NEMO_deficiency_in_recipient_mice_favours_donor_cell_selection_/1089251", "title"=>"NEMO deficiency in recipient mice favours donor cell selection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-30 03:44:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1575220"], "description"=>"<p>Liver histologies were graded for fibrosis levels according to an adapted METAVIR fibrosis score. F0 =  no fibrosis; F1 =  portal fibrosis without septa; F2 =  portal fibrosis with septa; F3 =  numerous septa without cirrhosis; F4 =  cirrhosis.</p>", "links"=>[], "tags"=>["anatomy", "histology", "genetics", "systems biology", "Gastroenterology and hepatology", "histological"], "article_id"=>1089262, "categories"=>["Biological Sciences"], "users"=>["Michaela Kaldenbach", "Francisco Javier Cubero", "Stephanie Erschfeld", "Christian Liedtke", "Christian Trautwein", "Konrad Streetz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100786.t001", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Quantitative_histological_assessment_of_mouse_livers_/1089262", "title"=>"Quantitative histological assessment of mouse livers.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-06-30 03:44:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1575212"], "description"=>"<p>(<b>A</b>) TUNEL staining was performed to mark apoptotic cells (blue: DAPI; green: TUNEL (+)). (<b>B</b>) Quantification of TUNEL(+) cells in recipients transplanted with Casp8<sup>Δhepa</sup>/hAAT(+) cells or Casp8<sup>Δhepa</sup>/hAAT(+) cells, respectively, compared to naïve control mice and mice subjected to BMT. The total number of apoptotic cells per view field was counted in a 100x magnification. (*p<0.01). (<b>C</b>) An analysis of the localization of TUNEL (+) cells was performed via serial sections and immunofluorescence staining of hAAT(+) areas and TUNEL-staining. (<b>D</b>) Displayed is a co-immunostaining of cleaved Caspase-3 and hAAT(+) donor cells, which shows an equal distribution of apoptotic cells as compared to figure (A). The use of double-staining reveals, that Caspase-3(+) and hAAT(+) cells are distinct and not overlapping.</p>", "links"=>[], "tags"=>["anatomy", "histology", "genetics", "systems biology", "Gastroenterology and hepatology", "hepatic"], "article_id"=>1089258, "categories"=>["Biological Sciences"], "users"=>["Michaela Kaldenbach", "Francisco Javier Cubero", "Stephanie Erschfeld", "Christian Liedtke", "Christian Trautwein", "Konrad Streetz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0100786.g004", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_hepatic_apoptosis_/1089258", "title"=>"Analysis of hepatic apoptosis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-30 03:44:32"}

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Relative Metric

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