title: Identification of new genes involved in human adipogenesis and fat storage.
abstract: Since the worldwide increase in obesity represents a growing challenge for health care systems, new approaches are needed to effectively treat obesity and its associated diseases. One prerequisite for advances in this field is the identification of genes involved in adipogenesis and/or lipid storage. To provide a systematic analysis of genes that regulate adipose tissue biology and to establish a target-oriented compound screening, we performed a high throughput siRNA screen with primary (pre)adipocytes, using a druggable siRNA library targeting 7,784 human genes. The primary screen showed that 459 genes affected adipogenesis and/or lipid accumulation after knock-down. Out of these hits, 333 could be validated in a secondary screen using independent siRNAs and 110 genes were further regulated on the gene expression level during adipogenesis. Assuming that these genes are involved in neutral lipid storage and/or adipocyte differentiation, we performed InCell-Western analysis for the most striking hits to distinguish between the two phenotypes. Beside well known regulators of adipogenesis and neutral lipid storage (i.e. PPARγ, RXR, Perilipin A) the screening revealed a large number of genes which have not been previously described in the context of fatty tissue biology such as axonemal dyneins. Five out of ten axonemal dyneins were identified in our screen and quantitative RT-PCR-analysis revealed that these genes are expressed in preadipocytes and/or maturing adipocytes. Finally, to show that the genes identified in our screen are per se druggable we performed a proof of principle experiment using an antagonist for HTR2B. The results showed a very similar phenotype compared to knock-down experiments proofing the "druggability". Thus, we identified new adipogenesis-associated genes and those involved in neutral lipid storage. Moreover, by using a druggable siRNA library the screen data provides a very attractive starting point to identify anti-obesity compounds targeting the adipose tissue.
authors.affiliation: Beiersdorf AG, Research & Development, Hamburg, Germany.
authors.forename: Jörn
authors.initials: J
authors.lastname: Söhle
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Machuy
authors.forename: Elma
authors.initials: E
authors.lastname: Smailbegovic
authors.forename: Ursula
authors.initials: U
authors.lastname: Holtzmann
authors.forename: Elke
authors.initials: E
authors.lastname: Grönniger
authors.forename: Horst
authors.initials: H
authors.lastname: Wenck
authors.forename: Franz
authors.initials: F
authors.lastname: Stäb
authors.forename: Marc
authors.initials: M
authors.lastname: Winnefeld
mesh_terms: D017667:Adipocytes / Q000166:cytology; D050156:Adipogenesis; D000273:Adipose Tissue / Q000378:metabolism*; D002454:Cell Differentiation; D004247:DNA / Q000737:chemistry; D004398:Dyneins / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008954:Models, Biological; D009765:Obesity / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D011786:Quality Control; D034741:RNA, Small Interfering / Q000378:metabolism*; D058830:Serotonin 5-HT2 Receptor Antagonists / Q000378:metabolism
chemical_list: D008055:Lipids; D034741:RNA, Small Interfering; D058830:Serotonin 5-HT2 Receptor Antagonists; D004247:DNA; D004398:Dyneins