Development of a methodology for in vivo follow-up of hepatocellular carcinoma in hepatocyte specific Trim24-null mice treated with myo-inositol trispyrophosphate.
BACKGROUND
Genetically induced hepatocellular carcinoma (HCC) models are generally used to investigate carcinogenesis pathways, but very few attempts were made to valorize them for pharmacological testing. This study describes a micro-computed tomography (micro-CT) - based methodology for the diagnostic and lifelong follow-up of HCC in the hepatocyte-specific Trim24-null mouse line. Myo-inositol trispyrophosphate (ITPP) was tested as anti-cancer drug.


METHODS
Partial hepatectomy was performed in 2 months-old Trim24-null mice, in order to accelerate the carcinogenesis process. HCC diagnosis was obtained by micro-CT scan with double contrast agent: 10 μl/g Fenestra™ LC was injected intraperitoneally 6 h prior to imaging and 10 μl/g Fenestra™ VC was injected intravenously 15 min prior to imaging. Twenty three hepatocyte-specific Trim24-null mice were considered for ITPP testing (3 mg/g/week intraperitoneally during 10 months in 12 mice, versus 11 controls). Lifelong follow-up was performed using micro-CT. Comparative analysis was performed using unpaired t test with Welch correction and survival curves were compared by log-rank test. Gene expression analysis was performed using the RT q-PCR technique.


RESULTS
Double contrast micro-CT scan allowed HCC diagnosis as hypodense, isodense or hyperdense nodules. Positive predictive value was 81.3 %. Negative predictive value was 83.3 %. Tumor growth could be objectified by micro-CT scan before the ITPP treatment was started, and at 3 and 9 months follow-up. Significant progression of tumor volume was demonstrated in the both groups, with no difference between groups (p > 0.05). In the ITPP group, a mild decrease in tumor doubling time was first observed (31.9 +/- 12 days, p > 0.05) followed by a significant increase (59.8 +/- 18.3 days, p = 0.008). However, tumor doubling time was not different between groups (p > 0.05). Median survival after treatment initiation was 223 days (controls) versus 296 days (ITPP group, p = 0.0027). HIF1α, VEGF, glutamine synthase, osteopontin expression levels were not significantly modified at the end of follow-up. In the ITPP group, the p53 expression profile was inversed as compared to the control group, higher in non-tumor livers than in tumors.


CONCLUSIONS
ITPP treatment allowed for a two-month survival improvement, with better tolerance of tumor burden and apoptosis increase in non-tumor, pathological livers.
IRCAD, 1 place de l'hôpital, 67091, Strasbourg, France. dana-mihaela.ignat@chru-strasbourg.fr.|Department of Digestive and Endocrine Surgery, University Hospital of Strasbourg, 1 place de l'Hôpital, 67091, Strasbourg, France. dana-mihaela.ignat@chru-strasbourg.fr.
Mihaela
http://orcid.org/0000-0002-0250-3280
M
Ignat
Department of Digestive and Endocrine Surgery, University Hospital of Strasbourg, 1 place de l'Hôpital, 67091, Strasbourg, France.
Cherif Youssef
CY
Akladios
Department of Digestive and Endocrine Surgery, University Hospital of Strasbourg, 1 place de l'Hôpital, 67091, Strasbourg, France.
Véronique
V
Lindner
Institute of Genetics and Molecular and Cellular Biology, F-67404, Illkirch, France.|Laboratoire de génétique moléculaire, Institut de recherches cliniques de Montréal (IRCM), Montréal, QC, H2W 1R7, Canada.
Konstantin
K
Khetchoumian
Department of Digestive and Endocrine Surgery, University Hospital of Strasbourg, 1 place de l'Hôpital, 67091, Strasbourg, France.|Institute of Genetics and Molecular and Cellular Biology, F-67404, Illkirch, France.
Marius
M
Teletin
IRCAD, 1 place de l'hôpital, 67091, Strasbourg, France.|Department of Digestive and Endocrine Surgery, University Hospital of Strasbourg, 1 place de l'Hôpital, 67091, Strasbourg, France.
Didier
D
Muttter
IRCAD, 1 place de l'hôpital, 67091, Strasbourg, France.
Pierre Marc
PM
Aprahamian
IRCAD, 1 place de l'hôpital, 67091, Strasbourg, France.
Jacques
J
Marescaux
Genetically induced hepatocellular carcinoma; Hepatocyte-specific Trim24-null mouse; High-resolution micro-computed tomography (micro-CT-scan); Myo-inositol trispyrophosphate (ITPP); Transgenic mouse
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