title: Effects of TGF-beta signalling inhibition with galunisertib (LY2157299) in hepatocellular carcinoma models and in ex vivo whole tumor tissue samples from patients.
abstract: Galunisertib (LY2157299) is a selective ATP-mimetic inhibitor of TGF-β receptor (TβR)-I activation currently under clinical investigation in hepatocellular carcinoma (HCC) patients. Our study explored the effects of galunisertib in vitro in HCC cell lines and ex vivo on patient samples. Galunisertib was evaluated in HepG2, Hep3B, Huh7, JHH6 and SK-HEP1 cells as well as in SK-HEP1-derived cells tolerant to sorafenib (SK-Sora) and sunitinib (SK-Suni). Exogenous stimulation of all HCC cell lines with TGF-β yielded downstream activation of p-Smad2 and p-Smad3 that was potently inhibited with galunisertib treatment at micromolar concentrations. Despite limited antiproliferative effects, galunisertib yielded potent anti-invasive properties. Tumor slices from 13 patients with HCC surgically resected were exposed ex vivo to 1 µM and 10 µM galunisertib, 5 µM sorafenib or a combination of both drugs for 48 hours. Galunisertib but not sorafenib decreased p-Smad2/3 downstream TGF-β signaling. Immunohistochemistry analysis of galunisertib and sorafenib-exposed samples showed a significant decrease of the proliferative marker Ki67 and increase of the apoptotic marker caspase-3. In combination, galunisertib potentiated the effect of sorafenib efficiently by inhibiting proliferation and increasing apoptosis. Our data suggest that galunisertib may be active in patients with HCC and could potentiate the effects of sorafenib.
authors.affiliation: AAREC Filia Research, Boulogne-Billancourt, France.|INSERM U728, Department of Medical Oncology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Maria
authors.initials: M
authors.lastname: Serova
authors.affiliation: AAREC Filia Research, Boulogne-Billancourt, France.|INSERM U728, Department of Medical Oncology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Annemilaï
authors.initials: A
authors.lastname: Tijeras-Raballand
authors.affiliation: AAREC Filia Research, Boulogne-Billancourt, France.|INSERM U728, Department of Medical Oncology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Célia
authors.initials: C
authors.lastname: Dos Santos
authors.affiliation: INSERM U773, Department of Pathology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Miguel
authors.initials: M
authors.lastname: Albuquerque
authors.affiliation: INSERM U773, Department of Pathology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Valerie
authors.initials: V
authors.lastname: Paradis
authors.affiliation: INSERM U728, Department of Medical Oncology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Cindy
authors.initials: C
authors.lastname: Neuzillet
authors.affiliation: Eli Lilly and Co., Neuilly-sur-Seine, France.
authors.forename: Karim A
authors.initials: KA
authors.lastname: Benhadji
authors.affiliation: INSERM U728, Department of Medical Oncology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Raymond
authors.affiliation: INSERM U728, Department of Medical Oncology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.|INSERM U773, Department of Pathology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Faivre
authors.affiliation: AAREC Filia Research, Boulogne-Billancourt, France.|INSERM U728, Department of Medical Oncology, Beaujon University Hospital (AP-HP - PRES Paris 7 Diderot), Clichy, France.
authors.forename: Armand
authors.initials: A
authors.lastname: de Gramont
mesh_terms: D000328:Adult; D000368:Aged; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D056945:Hep G2 Cells; D006801:Humans; D007150:Immunohistochemistry; D007211:Indoles / Q000494:pharmacology; D019394:Ki-67 Antigen / Q000378:metabolism; D008113:Liver Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D009536:Niacinamide / Q000031:analogs & derivatives / Q000494:pharmacology; D010671:Phenylurea Compounds / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology*; D011758:Pyrroles / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology*; D015398:Signal Transduction; D000077157:Sorafenib; D000077210:Sunitinib; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D007211:Indoles; D019394:Ki-67 Antigen; D010671:Phenylurea Compounds; D011720:Pyrazoles; D011758:Pyrroles; D011804:Quinolines; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; D009536:Niacinamide; C557799:LY-2157299; D000077157:Sorafenib; D053148:Caspase 3; D000077210:Sunitinib
keywords: SMAD; TGF-β inhibition; galunisertib; hepatocellular carcinoma; sorafenib