Phosphoproteomics identifies potential downstream targets of the integrin alpha 2 beta 1 inhibitor BTT-3033 in prostate stromal cells

Li, Bingsheng; Li, Pan; Xia, Weiping; You, Baiyang; Yu, Qingfeng; Zhang, Bo; Huang, Ru; Wang, Ruixiao; Liu, Yuhan; Chen, Zhi; Gan, Yu; He, Yao; Hennenberg, Martin; Stief, Christian G. und Chen, Xiang (2021): Phosphoproteomics identifies potential downstream targets of the integrin alpha 2 beta 1 inhibitor BTT-3033 in prostate stromal cells. In: Annals of Translational Medicine, Bd. 9, Nr. 17, 1380

Abstract

Background: Integrin alpha 2 beta 1 inhibitor BTT-3033 (1-(4-fluorophenyl)-N-methyl-N-[4[[(phenylamino) carbonyl]amino]phenyl]-1H-pyrazole-4-sulfonamide) was recently reported to inhibit neurogenic and thromboxane A2-induced human prostate smooth muscle contraction, and thus represents a target with a different inhibition spectrum than that of alpha 1-blockers in benign prostate hyperplasia (BPH) treatments. Clarifying the underlying mechanisms of the inhibition effects will provide insights into the role of integrin alpha 2 beta 1 in prostate contraction and enable new intracellular targets for smooth muscle contraction to be explored. Methods: ProteomeHD was used to predict and enrich the top co-regulated proteins of integrin alpha 2 (ITGA2). A phosphoproteomic analysis was conducted on human prostate stromal cells (WPMY-1) treated with 1 or 10 mu M of BTT-3033 or solvent for controls. A clustering analysis was conducted to identify the intracellular targets that were inhibited in a dose-dependent manner. Gene ontology (GO) and annotation enrichments were conducted to examine any functional alterations and identify possible downstream targets. A Kinase-substrate enrichment analysis (KSEA) was conducted to identify kinases-substrate relationships. Results: Enrichments of the actin cytoskeleton and guanosine triphosphatases (GTPases) signaling were predicted from the co-regulated proteins with ITGA2. LIM domain kinases, including LIM domain and actin-binding 1 (LIMA1), zyxin (ZYX), and thyroid receptor-interacting protein 6 (TRIP6), which are functionally associated with focal adhesions and the cytoskeleton, were present in the clusters with dose-dependent phosphorylation inhibition pattern. 15 substrates were dose-dependently inhibited according to the KSEA, including polo-like kinase 1 (PLK1), and GTPases signaling proteins, such as disheveled segment polarity protein 2 (DVL2). Conclusions: In this study, we proposed that the mechanisms underlying the contractile and proliferative effects of integrin alpha 2 beta 1 are the LIM domain kinases, including the ZYX family, and substrates, including PLK1 and DVL2.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• ISSN: 2305-5839
• Sprache: Englisch