雷帕霉素荧光碳点诱导OPTN（E50K）突变鼠BMCs年轻化促靶向归巢对RGCs损伤修复的研究

OPTN是正常眼压性青光眼（NTG）相关致病基因，OPTN（E50K）突变能够干扰机体自噬水平引起视网膜神经节细胞（RGCs）凋亡。申请者研究发现OPTN（E50K）还通过抑制自噬引起骨髓干细胞（BMCs）功能性衰老，阻碍其修复受损RGCs。因此，本项目着力于改善OPTN（E50K）所致的自噬通量障碍，修复自体BMCs功能，拟利用雷帕霉素（RAPA）作为碳源提取新型功能性生物材料碳点（CDs），通过改善自噬和氧化应激水平逆转OPTN（E50K）突变导致的骨髓优势Sca-1+亚群细胞老化损伤，体外和体内靶向追踪观察RAPA-CDs对OPTN（E50K）突变BMCs年轻化作用及对RGCs损伤修复能力；探讨RAPA-CDs在调控突变机体BMCs自噬功能，恢复BMCs干性，强化干细胞靶向归巢促RGCs修复的多重信号机制，为OPTN（E50K）致NTG等神经退行性病变干细胞治疗提供切实有效的新策略。

One of the genes associated with normal-tension glaucoma (NTG) is OPTN. And the missense mutation of E50K in the coding region of OPTN, known to inhibit autophagy and cause the apoptosis of retinal ganglion cells (RGCs), seems to be associated with more progressive and severe disease with aging. Recent efforts have showed the possibilities of bone marrow-derived stem cells (BMCs) based repair of retinal RGCs injury. However, functional senescence associated with autophagy inhibition caused by OPTN(E50K) impaired endogenous stem cells, including BMCs, which may contribute to the limited regeneration of damaged RGCs. Our previous study identified the stem cell antigen 1 (Sca-1+) cells as the young BMCs type which had the greatest ability to home to the retina and protect the aged RGCs after injury. However, the allogenic stem cells may bring the chronic immune rejection. Recent researches have provided fundamental information about fluorescent carbon dots (CDs) as an emerging class of nanomaterials in the carbon family and the interactions with BMCs for tissue engineering. The amalgamation of stem cell therapy with nanotechnology brings new prospects to the stem cell research, as it improves the specificity of the treatment and controls the stem cell proliferation and differentiation. The present proposal is designed to explore the therapeutic potential for regulation of autophagy of rapamycin CDs (RAPA-CDs) against aged injury of BMC-Sca-1+ subpopulation caused by OPTN(E50K) for RGCs recovery in vitro and in vivo. We hypothesize the BMC-Sca-1+ subpopulation from OPTN(E50K) rejuvenated by RAPA-CDs exerts beneficial stemness and homing abilities actions on the NTG retinal neuroprotection. We believe that the current proposal will prospectively provide a new strategy of nano-biomaterials combined stem cell therapy for NTG and identify the most efficient therapy for cell application in ophthalmological treatment.