Small-interfering RNA-mediated identification and regulation of the ternary SNARE complex mediating RBL-2H3 mast cell...

Dysregulation of mast cell function contributes to allergic and autoimmune disease that affects more than 70 million persons in the United States alone. Identifying novel mast cell targets that mediate disease or disease progression is required for the development of innovative therapeutics for the treatment of allergy/asthma and autoimmune disease. RNA interference technologies offer hope both as basic research tools for target identification and as potential, novel, specific therapeutics. Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are a family of evolutionarily conserved proteins that have been postulated to mediate the transport and fusion of inflammatory mediator-laden vesicles to the membrane in mast cells leading to their subsequent exocytosis. The functional role(s) of specific SNARE family member complexes in mast cell degranulation has not been fully elucidated. Here, we characterize the functional importance of SNARE complexes in FcεRI receptor-mediated degranulation of RBL-2H3 cells utilizing RNA interference. We demonstrate that ternary SNARE complexes of synaptosomal-associated protein-23, Syntaxin 4 and vesicle-associated membrane protein-7 (VAMP-7) or VAMP-8 are directly involved in mast cell degranulation. Additionally, we evaluate the siRNAs directed against these molecules as potential therapeutic agents for disease intervention. These studies have identified specific SNARE proteins and complexes that serve as novel targets for the development of siRNA therapies to treat allergic and autoimmune disease.