2 research outputs found
Proteome Analysis of a Hepatocyte-Specific BIRC5 (Survivin)-Knockout Mouse Model during Liver Regeneration
The
Baculoviral IAP repeat-containing protein 5 (BIRC5), also known
as inhibitor of apoptosis protein survivin, is a member of the chromosomal
passenger complex and a key player in mitosis. To investigate the
function of BIRC5 in liver regeneration, we analyzed a hepatocyte-specific
BIRC5-knockout mouse model using a quantitative label-free proteomics
approach. Here, we present the analyses of the proteome changes in
hepatocyte-specific BIRC5-knockout mice compared to wildtype mice,
as well as proteome changes during liver regeneration induced by partial
hepatectomy in wildtype mice and mice lacking hepatic BIRC5, respectively.
The BIRC5-knockout mice showed an extensive overexpression of proteins
related to cellular maintenance, organization and protein synthesis.
Key regulators of cell growth, transcription and translation MTOR
and STAT1/STAT2 were found to be overexpressed. During liver regeneration
proteome changes representing a response to the mitotic stimulus were
detected in wildtype mice. Mainly proteins corresponding to proliferation,
cell cycle and cytokinesis were up-regulated. The hepatocyte-specific
BIRC5-knockout mice showed impaired liver regeneration, which had
severe consequences on the proteome level. However, several proteins
with function in mitosis were found to be up-regulated upon the proliferative
stimulus. Our results show that the E3 ubiquitin-protein ligase UHRF1
is strongly up-regulated during liver regeneration independently of
BIRC5
Analysis of Disease-Associated Protein Expression Using Quantitative ProteomicsīøFibulinā5 Is Expressed in Association with Hepatic Fibrosis
Hepatic
fibrosis and cirrhosis are major health problems worldwide.
Until now, highly invasive biopsy remains the diagnostic gold standard
despite many disadvantages. To develop noninvasive diagnostic assays
for the assessment of liver fibrosis, it is urgently necessary to
identify molecules that are robustly expressed in association with
the disease. We analyzed biopsied tissue samples from 95 patients
with HBV/HCV-associated hepatic fibrosis using three different quantification
methods. We performed a label-free proteomics discovery study to identify
novel disease-associated proteins using a subset of the cohort (<i>n</i> = 27). Subsequently, gene expression data from all available
clinical samples were analyzed (<i>n</i> = 77). Finally,
we performed a targeted proteomics approach, multiple reaction monitoring
(MRM), to verify the disease-associated expression in samples independent
from the discovery approach (<i>n</i> = 68). We identified
fibulin-5 (FBLN5) as a novel protein expressed in relation to hepatic
fibrosis. Furthermore, we confirmed the altered expression of microfibril-associated
glycoprotein 4 (MFAP4), lumican (LUM), and collagen alpha-1Ā(XIV) chain
(COL14A1) in association to hepatic fibrosis. To our knowledge, no
tissue-based quantitative proteomics study for hepatic fibrosis has
been performed using a cohort of comparable size. By this means, we
add substantial evidence for the disease-related expression of the
proteins examined in this study