Name: Dr. Emilio M. Casanova Hevia
Institution: Ludwig Boltzmann Institute for Cancer Research (LBI-CR)
Address: Währinger Str. 13a, A-1090, Vienna, Austria
Phone +43 1 40160 71210
Fax +43 1 40160 931300
Our group is interested in modelling human diseases in mice. We make use of mice genetically modified to study different pathological process such as liver fibrosis and cancer. Recently we have developed a new mouse model for liver fibrosis and we are currently analyzing. Also, by making use of mouse genetic technology, we have generated a mouse model for breast cancer.
1 Role of Stat5 in liver fibrosis
Liver fibrosis constitutes a considerable health problem in the human population. In order to further understand the molecular mechanisms underlying liver fibrogenesis it is mandatory to generate genetic mouse models closely resembling human disease development and progression.
Stat5 is a transcription factor that plays a central role in liver physiology. By making use of the Cre/loxP system we have generated mice lacking Stat5 in the liver. These mice show a strong liver fibrosis phenotype in model for cholestasis. Mice show a dramatic increase of collagen deposition, disturbed liver architecture, liver necrosis, bile duct infarcts and bile duct proliferation. Gene expression analysis in these animals shows impairment of the Growth hormone-Stat5-IGF-1 axis. Interestingly, also liver cirrhosis in human patients is associated to growth hormone resistance and low levels of IGF-1, thus making this mouse model and attractive tool to contribute to the understanding of liver fibrogenesis and progression.
Left panel: control liver. Right panel: fibrotic liver
2 AKT and breast tumors
AKT/PKB is a family of serine-threonine kinases activated in response to growth stimuli downstream of the PI3K /PTEN pathway. The AKT family consists of three members: AKT-1, AKT-2 and AKT-3, and has been implicated in different malignancies, such as glioblastoma, ovarian, gastric, colorectal, prostate and breast tumors. Gene amplification and/or over-activation of the three AKT members have been found in different tumors. However, it is not clear to what extent each member of the AKT family contributes to different tumors individually or in combination with other members. To explore how each individual or combination of the AKT isoforms contributes to the development of mammary gland tumors, we have generated a “multi-hit” mouse expressing each member of the AKT family (“AKT multi-hit”) in an inducible manner. We have crossed these mice to the Rosa26CreERT2 line and, upon Cre induction, mice develop breast tumors. Within this project, we expect to gain understanding of the molecular mechanisms underlying the contribution the AKT family to mammary gland tumors.
AKT dependent mammary gland tumors developed in our mouse genetic model.
3 Recombinant protein production using Bacterial Artificial Chromosomes as expression vectors in eukaryotic cells.
Recombinant protein production in eukaryotic cells is one of the main topics of biotechnology. One of the most critical steps is the development of appropriate vectors to express the protein of interest. Bacterial Artificial Chromosomes (BACs) containing the appropriate locus can be used as expression vectors in the protein production field. In collaboration with Dr. A. Bauer, we have developed a BAC-based vector system that improves the protein production by a factor of 10 when compared to a conventional vector in stable HEK293 cell lines. We are currently expanding our vector tool-box and exploring the use in different cell lines, such as CHO cells.
A) Schematic representation of the vectors used in this work. CAGGS Fc: conventional vector containing the CAGGS promoter and the human IgG1-Fc as protein of interest. Rosa26CAGGS Fc: CAGGS Fc vector recombined into a BAC containing the Rosa26 locus (BAC-based vector). B) Protein production in HEK293 stable bulk cultures from a conventional vector (CAGGS Fc) and a BAC based vector (Rosa26CAGGS Fc). The use of a BAC-based vector improves the protein production by a factor of 10.
Patricia Stiedl, DI
Beatrice Grabner, MSc
Mueller KM, Kornfeld JW, Friedbichler K, Blaas L, Egger G, Esterbauer H, Hasselblatt P, Schlederer M, Haindl S, Wagner KU, Engblom D, Haemmerle G, Kratky D, Sexl V, Kenner L, Kozlov AV, Terracciano L, Zechner R, Schuetz G, Casanova E, Pospisilik JA, Heim MH, Moriggl R.
Impairment of hepatic growth hormone and glucocorticoid receptor signaling causes steatosis and hepatocellular carcinoma in mice
Hepatology. (in press)
Blaas L, Musteanu M, Grabner B, Eferl R, Bauer A and Casanova E.
The Use of Bacterial Artificial Chromosomes for Recombinant Protein Production in Mammalian Cell Lines
Methods in Molecular Biology (in press)
Mair M, Blaas L, Osterreicher CH, Casanova E, and Eferl R.
JAK-STAT signaling in hepatic fibrosis
Front Biosci. 2011 Jun 1;17:2794-811
Eckelhart E, Warsch W, Zebedin E, Simma O, Stoiber D, Kolbe T, Rülicke T, Mueller M, Casanova E*, Sexl V*.
A novel Ncr1-Cre mouse reveals the essential role of STAT5 for NK cell survival and development.
Blood. 2011 Feb 3;117(5):1565-73. *Equally contributed
Grabner B, Blaas L, Musteanu M, Hoffmann T, Birbach A, Eferl R and Casanova E.
A mouse tool for conditional mutagenesis in ovarian granulosa cells
Genesis 2010 Oct 1;48(10):612-7.
Guetg N, Aziz S, Holbro N, Turecek R, Riad S, Gassmann M, Moes S, Jenoe P, Oertner T, Casanova E and Bettler B
NMDA Receptor-Dependent GABAB Receptor Internalization via CaMKII Phosphorylation of Serine 867 in GABAB1
PNAS, 2010 Aug 3;107(31)
Mair M; Zollner G; Schneller D; Musteanu M; Fickert P; Gumhold J; Schuster C; Fuchsbichler A; Bilban M; Tauber S; Esterbauer H; Kenner L; Poli V; Blaas L; Kornfeld JW; Casanova E; Mikulits W; Trauner M; Eferl R.
Stat3 protects from liver injury and fibrosis in a mouse model of sclerosing cholangitis
Gastroenterology. 2010 Feb 26
Musteanu M, Blaas L, Mair M, Schlederer M, Bilban M, Tauber S, Esterbauer H, Mueller M, Casanova E, Kenner L, Poli V and Eferl R1
Stat3 is a negative regulator of intestinal tumor progression in ApcMin mice
Gastroenterology. 2010 Mar;138(3):1003-11
Blaas L, Kornfeld JW, Schramek D, Musteanu M, Zollner G, Gumhold J, Schneller D, Esterbauer H, Mair M, Kenner L, Mikulits W, Eferl R, Moriggl R, Penninger J, Trauner M and Casanova E.
Disruption of the GH-STAT5-IGF-1 axis severely aggravates liver fibrosis in a mouse model of cholestasis
Hepatology. 2010 Apr;51(4):1319-260