Microarray and proteomics expression profiling identifies several candidates, including the valosin-containing protein (VCP), involved in regulating high cellular growth rate in production CHO cell lines.
A high rate of cell growth (micro) leading to rapid accumulation of viable biomass is a desirable phenotype during scale up operations and the early stages of production cultures. In order to identify genes and proteins that contribute to higher growth rates in Chinese hamster ovary (CHO) cells, a combined approach using microarray and proteomic expression profiling analysis was carried out on two matched pairs of CHO production cell lines that displayed either fast or slow growth rates. Statistical analysis of the microarray and proteomic data separately resulted in the identification of 118 gene transcripts and 58 proteins that were differentially expressed between the fast- and slow-growing cells. Overlap comparison of both datasets identified a priority list of 21 candidates associated with a high growth rate phenotype in CHO. Functional analysis (by siRNA) of five of these candidates identified the valosin-containing protein (VCP) as having a substantial impact on CHO cell growth and viability. Knockdown of HSPB1 and ENO1 also had an effect on cell growth (negative and positive, respectively). Further functional validation in CHO using both gene knockdown (siRNA) and overexpression (cDNA) confirmed that altered VCP expression impacted CHO cell proliferation, indicating that VCP and other genes and proteins identified here may play an important role in the regulation of CHO cell growth during log phase culture and are potential candidates for CHO cell line engineering strategies.
Doolan
Padraig
P
National Institute for Cellular Biotechnology, Dublin City University, Dublin 9, Ireland. padraig.doolan@dcu.ie
Meleady
Paula
P
Barron
Niall
N
Henry
Michael
M
Gallagher
Ross
R
Gammell
Patrick
P
Melville
Mark
M
Sinacore
Martin
M
McCarthy
Kevin
K
Leonard
Mark
M
Charlebois
Timothy
T
Clynes
Martin
M
D000251:Adenosine Triphosphatases / Q000096:biosynthesis; D000818:Animals; D016466:CHO Cells; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000096:biosynthesis; D006224:Cricetinae; D003412:Cricetulus; D004847:Epithelial Cells / Q000502:physiology*; D015870:Gene Expression; D020869:Gene Expression Profiling; D020868:Gene Silencing; D020543:Proteome / Q000032:analysis*; D034741:RNA, Small Interfering / Q000378:metabolism; D015854:Up-Regulation; D000074405:Valosin Containing Protein
D018797:Cell Cycle Proteins; D020543:Proteome; D034741:RNA, Small Interfering; D000251:Adenosine Triphosphatases; D000074405:Valosin Containing Protein