Fibroblast Growth Factor Receptor-Mediated Activation of AKT-β-Catenin-CBP Pathway Regulates Survival and Proliferation of Murine Hepatoblasts and Hepatic Tumor Initiating Stem Cells
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{"title"=>"Fibroblast Growth Factor Receptor-Mediated Activation of AKT-β-Catenin-CBP Pathway Regulates Survival and Proliferation of Murine Hepatoblasts and Hepatic Tumor Initiating Stem Cells", "type"=>"journal", "authors"=>[{"first_name"=>"Nirmala", "last_name"=>"Mavila", "scopus_author_id"=>"6508175584"}, {"first_name"=>"David", "last_name"=>"James", "scopus_author_id"=>"57198797333"}, {"first_name"=>"Sarah", "last_name"=>"Utley", "scopus_author_id"=>"8301054500"}, {"first_name"=>"Nguyen", "last_name"=>"Cu", "scopus_author_id"=>"55512316700"}, {"first_name"=>"Orly", "last_name"=>"Coblens", "scopus_author_id"=>"57200596193"}, {"first_name"=>"Katrina", "last_name"=>"Mak", "scopus_author_id"=>"55511841100"}, {"first_name"=>"C. Bart", "last_name"=>"Rountree", "scopus_author_id"=>"16313819200"}, {"first_name"=>"Michael", "last_name"=>"Kahn", "scopus_author_id"=>"7402055370"}, {"first_name"=>"Kasper S.", "last_name"=>"Wang", "scopus_author_id"=>"7501399363"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-84870618348", "pui"=>"366207225", "doi"=>"10.1371/journal.pone.0050401", "issn"=>"19326203", "pmid"=>"23308088", "sgr"=>"84870618348"}, "id"=>"09a0236b-3052-38f1-ba58-ef62b9066a78", "abstract"=>"UNLABELLED: Fibroblast Growth Factor (FGF)-10 promotes the proliferation and survival of murine hepatoblasts during early stages of hepatogenesis through a Wnt-β-catenin dependent pathway. To determine the mechanism by which this occurs, we expanded primary culture of hepatoblasts enriched for progenitor markers CD133 and CD49f from embryonic day (E) 12.5 fetal liver and an established tumor initiating stem cell line from Mat1a(-/-) livers in media conditioned with recombinant (r) FGF10 or rFGF7. FGF Receptor (R) activation resulted in the downstream activation of MAPK, PI3K-AKT, and β-catenin pathways, as well as cellular proliferation. Additionally, increased levels of nuclear β-catenin phosphorylated at Serine-552 in cultured primary hepatoblasts, Mat1a(-/-) cells, and also in ex vivo embryonic liver explants indicate AKT-dependent activation of β-catenin downstream of FGFR activation; conversely, the addition of AKT inhibitor Ly294002 completely abrogated β-catenin activation. FGFR activation-induced cell proliferation and survival were also inhibited by the compound ICG-001, a small molecule inhibitor of β-catenin-CREB Binding Protein (CBP) in hepatoblasts, further indicating a CBP-dependent regulatory mechanism of β-catenin activity.\\n\\nCONCLUSION: FGF signaling regulates the proliferation and survival of embryonic and transformed progenitor cells in part through AKT-mediated activation of β-catenin and downstream interaction with the transcriptional co-activator CBP.", "link"=>"http://www.mendeley.com/research/fibroblast-growth-factor-receptormediated-activation-akt%CE%B2catenincbp-pathway-regulates-survival-proli", "reader_count"=>20, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>2, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>1, "Student > Master"=>4, "Student > Bachelor"=>2, "Lecturer"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>2, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>1, "Student > Master"=>4, "Student > Bachelor"=>2, "Lecturer"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Medicine and Dentistry"=>4, "Agricultural and Biological Sciences"=>13, "Biochemistry, Genetics and Molecular Biology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>13}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"United States"=>2, "Portugal"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/538692"], "description"=>"<p><i><sup>−/−</sup></i><b>cells are mediated in part through PI3K-AKT pathway.</b> Serum-starved <i>Mat1a<sup>−/−</sup></i> cells were treated with rFGF7/10 ± PI3K-AKT inhibitor LY294002 for detection and quantification of (A) pSer 552 β-CATENIN or (B) BrdU. Total and β-CATENIN/BrdU Positive cells were counted from 4–5 HPF images from 3 independent experiments and represented as relative % of control (n = 3, *p<0.001).</p>", "links"=>[], "tags"=>["mediated", "proliferation"], "article_id"=>209184, "categories"=>["Chemistry", "Developmental Biology"], "users"=>["Nirmala Mavila", "David James", "Sarah Utley", "Nguyen Cu", "Orly Coblens", "Katrina Mak", "C. Bart Rountree", "Michael Kahn", "Kasper S. Wang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050401.g005", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FGFR_mediated_proliferation_of_Mat1a_/209184", "title"=>"FGFR mediated proliferation of <i>Mat1a</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-30 02:33:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/286408", "https://ndownloader.figshare.com/files/286450", "https://ndownloader.figshare.com/files/286470", "https://ndownloader.figshare.com/files/286557"], "description"=>"<div><p>Fibroblast Growth Factor (FGF)-10 promotes the proliferation and survival of murine hepatoblasts during early stages of hepatogenesis through a Wnt-β-catenin dependent pathway. To determine the mechanism by which this occurs, we expanded primary culture of hepatoblasts enriched for progenitor markers CD133 and CD49f from embryonic day (E) 12.5 fetal liver and an established tumor initiating stem cell line from <em>Mat1a<sup>−/−</sup></em> livers in media conditioned with recombinant (r) FGF10 or rFGF7. FGF Receptor (R) activation resulted in the downstream activation of MAPK, PI3K-AKT, and β-catenin pathways, as well as cellular proliferation. Additionally, increased levels of nuclear β-catenin phosphorylated at Serine-552 in cultured primary hepatoblasts, <em>Mat1a<sup>−/−</sup></em> cells, and also in <em>ex vivo</em> embryonic liver explants indicate AKT-dependent activation of β-catenin downstream of FGFR activation; conversely, the addition of AKT inhibitor Ly294002 completely abrogated β-catenin activation. FGFR activation-induced cell proliferation and survival were also inhibited by the compound ICG-001, a small molecule inhibitor of β-catenin-CREB Binding Protein (CBP) in hepatoblasts, further indicating a CBP-dependent regulatory mechanism of β-catenin activity.</p> <p><em>Conclusion:</em> FGF signaling regulates the proliferation and survival of embryonic and transformed progenitor cells in part through AKT-mediated activation of β-catenin and downstream interaction with the transcriptional co-activator CBP.</p> </div>", "links"=>[], "tags"=>["fibroblast", "receptor-mediated", "activation", "pathway", "regulates", "proliferation", "murine", "hepatoblasts", "hepatic", "initiating", "cells"], "article_id"=>116457, "categories"=>["Chemistry", "Developmental Biology"], "users"=>["Nirmala Mavila", "David James", "Sarah Utley", "Nguyen Cu", "Orly Coblens", "Katrina Mak", "C. Bart Rountree", "Michael Kahn", "Kasper S. Wang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0050401.s001", "https://dx.doi.org/10.1371/journal.pone.0050401.s002", "https://dx.doi.org/10.1371/journal.pone.0050401.s003", "https://dx.doi.org/10.1371/journal.pone.0050401.s004"], "stats"=>{"downloads"=>9, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Fibroblast_Growth_Factor_Receptor_Mediated_Activation_of_AKT_Catenin_CBP_Pathway_Regulates_Survival_and_Proliferation_of_Murine_Hepatoblasts_and_Hepatic_Tumor_Initiating_Stem_Cells__/116457", "title"=>"Fibroblast Growth Factor Receptor-Mediated Activation of AKT-β-Catenin-CBP Pathway Regulates Survival and Proliferation of Murine Hepatoblasts and Hepatic Tumor Initiating Stem Cells", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-11-30 01:47:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/538194"], "description"=>"<p>(A) Phase contrast pictures of <i>in vitro</i> expanded CD133<sup>pos</sup> CD49f<sup>pos</sup> CD45<sup>neg</sup> enriched cells from E12.5 embryonic liver at different time points. Co-immunofluorescence staining for (B) HNF4α and CK19 and (C) ALBUMIN and PCK. (D) Gene expression analysis by RTPCR using RNA isolated from the MACS enriched cells before and after 3-day culture. Negative control = water and positive control = E16.5 whole embryonic cDNA. Scale bar 25 µm. (E) Proliferation indices of CD133<sup>pos</sup> CD49f<sup>pos</sup> CD45<sup>neg</sup> cells treated with rFGF7/10 for 48 hrs and pulse labeled with BrdU (n = 4, *<i>p</i><0.001 compared to control). Data are representative of three or more independent experiments.</p>", "links"=>[], "tags"=>["cells", "assisted", "enriches", "hepatoblast", "progenitor", "proliferate", "fgfr"], "article_id"=>208679, "categories"=>["Chemistry", "Developmental Biology"], "users"=>["Nirmala Mavila", "David James", "Sarah Utley", "Nguyen Cu", "Orly Coblens", "Katrina Mak", "C. Bart Rountree", "Michael Kahn", "Kasper S. Wang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050401.g002", "stats"=>{"downloads"=>1, "page_views"=>34, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Isolation_and_in_vitro_expansion_of_CD133_pos_CD49f_pos_CD45_neg_cells_by_Magnetic_Assisted_Cells_Separation_MACS_from_E12_5_liver_enriches_for_hepatoblast_progenitor_cells_that_proliferate_in_response_to_FGFR_activation_/208679", "title"=>"Isolation and <i>in vitro</i> expansion of CD133<sup>pos</sup>CD49f<sup>pos</sup>CD45<sup>neg</sup> cells by Magnetic Assisted Cells Separation (MACS) from E12.5 liver enriches for hepatoblast progenitor cells that proliferate in response to FGFR activation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-30 02:24:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/538006"], "description"=>"<p>(A) Immunofluorescence staining for progenitor cell markers CD133 and CD49f on E12.5 embryonic liver. Yellow cells are double positive cells. DAPI staining (blue) identifies nuclei. (B) Immunostaining of CD133 and Cytokeratin (CK19). (C) Immunostaining for CD133, CD49f and FGFR 1 and (D) FGFR2. White arrows mark triple positive cells and yellow arrows mark positive cells that do not express receptors. Data are representative of three or more independent experiments. Scale bar represents 25 µm.</p>", "links"=>[], "tags"=>["murine", "populated", "hepatoblast", "progenitor", "cells", "expressing", "fgfr1"], "article_id"=>208494, "categories"=>["Chemistry", "Developmental Biology"], "users"=>["Nirmala Mavila", "David James", "Sarah Utley", "Nguyen Cu", "Orly Coblens", "Katrina Mak", "C. Bart Rountree", "Michael Kahn", "Kasper S. Wang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050401.g001", "stats"=>{"downloads"=>1, "page_views"=>66, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_murine_E12_5_liver_is_populated_with_CD133_pos_CD49f_pos_hepatoblast_progenitor_cells_expressing_FGFR1_and_2_/208494", "title"=>"The murine E12.5 liver is populated with CD133<sup>pos</sup>CD49f<sup>pos</sup> hepatoblast progenitor cells expressing FGFR1 and 2.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-30 02:21:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/538369"], "description"=>"<p><i><sup>−/−</sup></i><b>cells.</b> (A) Co-localization of ALBUMIN and pCK in <i>Mat1a<sup>−/−</sup></i> cells. Scale bar 25 µm. (B) RTPCR analysis shows <i>Mat1a<sup>−/−</sup></i> cells express putative stem gene markers and <i>Fgfr</i>’s. Positive (+) control E16.5 = embryonic cDNA. Data are representative of three or more independent experiments. (C) qPCR analysis shows that rFGF7/10 upregulates expression of <i>Fgfr2IIIb</i> (n = 3, *<i>p</i><0.01). (D) Western blot analysis of time course rFGF7 treatment on <i>Mat1a<sup>−/−</sup></i> cells results in the downstream phosphorylation and activation of FGF receptor, AKT, ERK and β-CATENIN (ctrl (control) = untreated cells, +ctrl = E12.5 embryonic lung). Data are representative of two independent experiments. (E) Proliferation indices for <i>Mat1a<sup>−/−</sup></i> cells treated with rFGF7/10 for 48 hrs and pulse labeled with BrdU (n = 4, *<i>p</i><0.001 compared to control).</p>", "links"=>[], "tags"=>["activation", "downstream", "pathways", "proliferation"], "article_id"=>208857, "categories"=>["Chemistry", "Developmental Biology"], "users"=>["Nirmala Mavila", "David James", "Sarah Utley", "Nguyen Cu", "Orly Coblens", "Katrina Mak", "C. Bart Rountree", "Michael Kahn", "Kasper S. Wang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050401.g003", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FGFR_activation_promotes_downstream_activation_of_AKT_ERK_and_catenin_pathways_as_well_as_proliferation_of_Mat1a_/208857", "title"=>"FGFR activation promotes downstream activation of AKT, ERK, and β-catenin pathways as well as proliferation of <i>Mat1a</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-30 02:27:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/538502"], "description"=>"<p><i><sup>−/− </sup></i><b><i>cells</i></b>. Immunofluorescence staining and quantification for pSer-552 β-CATENIN in rFGF7/10-treated MACS-enriched day-4 CD133<sup>pos</sup> CD49f<sup>pos</sup> CD45<sup>neg</sup> (A, B) and <i>Mat1a<sup>−/−</sup></i> cells (C, D). Total number of pSer-552 β-CATENIN positive cells were counted in 3–4 HPF from three independent experiments each and represented as % of total cells (n = 3, *p<0.001). Scale bar 25 µm. (E, F) Western blot analysis of nuclear extracts from <i>Mat1a<sup>−/−</sup></i> cells treated with rFGF10. HISTONE-H3 was used to normalize for relative nuclear protein expression levels (n = 3, *p< 0.005).</p>", "links"=>[], "tags"=>["signaling", "increases", "activation", "localization", "pser-552", "hepatoblast"], "article_id"=>208983, "categories"=>["Chemistry", "Developmental Biology"], "users"=>["Nirmala Mavila", "David James", "Sarah Utley", "Nguyen Cu", "Orly Coblens", "Katrina Mak", "C. Bart Rountree", "Michael Kahn", "Kasper S. Wang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050401.g004", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FGFR_signaling_increases_the_activation_and_nuclear_localization_pSer_552_catenin_in_hepatoblast_and_Mat1a_/208983", "title"=>"FGFR signaling increases the activation and nuclear localization pSer-552 β-catenin in hepatoblast and <i>Mat1a</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-30 02:29:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/539091"], "description"=>"<p>Immunodetection and quantification of BrdU positive cells in (A, B) CD133<sup>pos</sup> CD49f<sup>pos</sup> Cd45<sup>neg</sup> enriched embryonic hepatoblasts (3-day cultured) and (C, D) <i>Mat1a<sup>−/−</sup></i> cells after treating with rFGF7 or rFGF10 ± β-catenin-CBP inhibitor ICG-001. Total number of BrdU positive cells were counted from 3–4 HPF from three independent experiments and represented as % of total cells (n = 3, *p<0.0001 versus control). FGF-ICG-001 was compared to respective control FGF treated experiments. (E) qPCR analysis of <i>Survivin</i> mRNA expression level in <i>Mat1a<sup>−/−</sup></i> cells for control, DMSO control, and rFGF7/10 ± ICG-001 groups. Expression levels are normalized to <i>Actin</i> (n = 3, *<i>p</i>< 0.005, #<i>p</i><0.05).</p>", "links"=>[], "tags"=>["fgf", "signaling", "drives", "embryonic", "hepatoblasts", "cells", "cbp-dependent"], "article_id"=>209573, "categories"=>["Chemistry", "Developmental Biology"], "users"=>["Nirmala Mavila", "David James", "Sarah Utley", "Nguyen Cu", "Orly Coblens", "Katrina Mak", "C. Bart Rountree", "Michael Kahn", "Kasper S. Wang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050401.g007", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Activation_of_FGF_signaling_drives_embryonic_hepatoblasts_and_Mat1a_8722_8722_cells_into_the_cell_cycle_through_a_CBP_dependent_pathway_/209573", "title"=>"Activation of FGF signaling drives embryonic hepatoblasts and <i>Mat1a</i><i><sup>−/−</sup></i> cells into the cell cycle through a CBP-dependent pathway.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-30 02:39:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/538888"], "description"=>"<p>Co-staining for (A) pSer-552 β-CATENIN and PCNA, (B) pSer-552 β-CATENIN and CD133, (C) pSer-552 β-CATENIN and CD49f <i>in vivo</i>. (Di & ii) Co-staining for pSer-552 β-CATENIN and PCNA on E12.5 liver explant cultured for three days ± rFGF10. Data are representative of three or more independent experiments. White arrows represent single positive cells and yellow arrows show double positive cells. Scale bar = 25 µm.</p>", "links"=>[], "tags"=>["activation", "embryonic"], "article_id"=>209380, "categories"=>["Chemistry", "Developmental Biology"], "users"=>["Nirmala Mavila", "David James", "Sarah Utley", "Nguyen Cu", "Orly Coblens", "Katrina Mak", "C. Bart Rountree", "Michael Kahn", "Kasper S. Wang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0050401.g006", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_AKT_mediated_activation_of_946_catenin_in_E12_5_embryonic_liver_/209380", "title"=>"AKT-mediated activation of β-catenin in E12.5 embryonic liver.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-30 02:36:20"}

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Relative Metric

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