オプトジェネティクスを用いたPDGFRb活性化システムの構築

抄録

Platelet-derived growth factor receptor β (PDGFRβ) is often found in cerebrovascular pericytes. PDGFRβ is suggested to be involved in the pathogenic mechanism of neurodegenerative diseases. Thus, investigation of the function of PDGFRβ is essential for the drug discovery of neurodegenerative diseases. However, the function of PDGFRβ is partially unknown since there is no specific agonist as target of PDGFRβ. Then, we took advantage of an optogenetic approach to analysis the function of PDGFRβ. It was reported that Light-Oxygen-Voltage-sensing (LOV) domain of aureochrome1 from Vaucheria frigida (AuLOV), a blue-light-induced receptor, induces dimerized the receptor by irradiating blue light. Hence, we developed a system that can specifically control the activities of PDGFRβ by blue light using the LOV domain. AuLOV-PDGFRβ-mCherry was constructed by inserting AuLOV into mCherry-N1 vector and subcloning intracellular domain of PDGFRβ (ICD) using In-Fusion cloning. The vector was transfection into HEK293 cells. Phosphorylation of PDGFRβ during dark and blue light irradiation were analyzed by western blot. We successfully developed an optogenetic PDGFRβ system to phosphorylation of PDGFRβ in HRK293 cells by the photoactivation. The utilization of the system for specifically activating PDGFRβ is expected to be useful for the development of therapeutic agents for neurodegenerative diseases.