The BioInvestigationIndex (BII)

Title:

Human adipocytes induced to differentiate with indomethacin. [original title: Identification of new genes involved in human adipogenesis and fat storage]

Organism(s):

Homo sapiens

Description:

Three donors, three time points each (0d, 3,d, 7d). [original description: Since the worldwide increase in obesity represents a growing challenge for the health care systems, new approaches are needed to treat obesity and associated disease effectively. The food intake is primarily stored in the adipose tissue and therefore this organ is in focus to develop new anti-obesity treatments. 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. Beside well known regulators of adipogenesis and neutral lipid storage (like PPAR?, RXR, Perilipin A) the screening revealed a large number of genes which were not previously described in the context of fatty tissue biology. An enrichment of genes was observed for axonemal dyneins. Five out of ten axonemal dyneins were identified in our primary screen and retested positive with independent siRNAs and cell-donors. Quantitative RT-PCR- and immunoblot analysis revealed that axonemal dyneins are expressed in preadipocytes and maturing adipocytes. Using microarray analysis, we further characterize the remaining genes identified in our primary screen to determine their expression pattern during adipogenesis. In the course of fat cell differentiation, 149 among the 459 positive genes were regulated on the gene expression level. Assuming that an adipogenesis-specific expression pattern is another independent hind [hint? -ed.], that these genes are involved in neutral lipid storage and/or adipocyte differentiation, we retested this gene-pool with independent siRNAs and cell donors. Finally, to show that the genes identified are per se druggable we performed a proof of principle experiment using an antagonist for one randomly chosen gene. 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. For microarray analysis, RNA was isolated at 3 different points in time during adipogenesis (day 0, day 3 and day 7 after induction of differentiation). This experiment was carried out with cells obtained from three different donors. No replicates are included for each point in time.]

Design(s):

transcription profiling by array

Experimental factor(s):

time 3 recorded
3 days of differentiation 0 days of differentiation 7 days of differentiation
individual 3 recorded
donor 3 donor 1 donor 2

Publication(s):

Shle J, Machuy N, Smailbegovic E, Holtzmann U, Grnniger E, Wenck H, Stb F, Winnefeld M
Identification of new genes involved in human adipogenesis and fat storage. PubMed:22384002

Sample attribute(s):

Material Type 1 recorded
total RNA
sex 1 recorded
female organism
biopsy site tissue 1 recorded
subcutaneous adipose tissue
individual 3 recorded
donor 3 donor 1 donor 2
drug treatment 4 1 recorded
3-isobutyl-1-methylxanthine
biopsy site 2 recorded
abdomen thigh
drug treatment 3 1 recorded
indometacin
age 3 recorded
34 years 33 years 37 years
drug treatment 2 1 recorded
dexamethasone
BMI 3 recorded
22.24 kg/m^2 21.56 kg/m^2 21.67 kg/m^2
Label 1 recorded
Cy3
Organism 1 recorded
Homo sapiens
cell type 1 recorded
preadipocyte
drug treatment 1 recorded
insulin (human)
cell type2 1 recorded
preadipocyte