Phosphodiesterase 7B regulates prostanoid- but not ß-adrenergic-stimulated cAMP levels in primary human airway smooth muscle cells.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology(2022)

引用 0|浏览18
暂无评分
摘要
Human airway smooth muscle (HASM) cells contain G-protein coupled receptor (GPCR) signal transduction pathways that are organized in the plasma membrane and form cAMP signaling compartments. One such compartment contains ß -adrenergic receptors (ß AR) coupled to adenylyl cyclase (AC)6 and is principally localized in lipid rafts. Another compartment expresses E prostanoid receptors 2 and 4 (EP ) coupled to AC2 and is restricted to non-raft regions of the plasma membrane. Various phosphodiesterase (PDE) isozymes modulate cyclic AMP (cAMP) levels in response to these pathways. Non-specific or PDE-family specific PDE inhibitors have been used to treat asthma and chronic obstructive pulmonary disease with limited success, in part due to expression of these PDE isoforms throughout the body. Drugs that inhibit specific PDE isozymes might produce therapeutic effects in airways with fewer off-target effects. RNA-sequencing data in HASM cells indicates that PDE7B is the third most highly expressed PDE isoform and includes expression of both the PDE7B-201 and PDE7B-203 splice variants. We sought to characterize the role of PDE7B in the lipid raft and non-raft cAMP signaling compartments. We used siRNA to knock down both PDE7B splice variants, confirmed protein reduction via Western Blot analysis, and then observed live cells' real-time cAMP levels in response to either isoproterenol (ßAR agonist) or PGE2 (EP agonist). The cADDis-FMP biosensor, which localizes in lipid raft regions, did not report altered cAMP dynamics in response to ß AR or EPR agonists following PDE7B knockdown. However, the cADDis-FS15 biosensor, which reports cAMP levels in non-raft plasma membrane regions, reported significant increases in cAMP levels stimulated by EPR agonists but no change in cAMP levels stimulated by ß AR agonist. PDE7B may play a unique role in regulating non-raft associated EP signaling without altering cAMP levels in other areas of the cell, thereby presenting a novel therapeutic target for pulmonary diseases.
更多
查看译文
AI 理解论文
溯源树
样例
生成溯源树,研究论文发展脉络
Chat Paper
正在生成论文摘要