项目来源
国家重点研发计划(NKRD)
项目主持人
曲静
项目受资助机构
中国科学院动物研究所
项目编号
2018YFC2000100
立项年度
2018
立项时间
未公开
研究期限
未知 / 未知
项目级别
国家级
受资助金额
3865.00万元
学科
主动健康和老龄化科技应对
学科代码
未公开
基金类别
“主动健康和老龄化科技应对”重点专项
关键词
灵长类动物 ; 早衰症 ; 衰老 ; 机制研究 ; non-human primate ; progeria ; aging ; mechanism study
参与者
毛志勇；慈维敏；初群
参与机构
同济大学；中国科学院北京基因组研究所
项目标书摘要：本年度项目团队围绕项目设置的研究目标，在建立衰老研究体系，尤其是优化和开发精准基因编辑工具方面取得了一定突破。一方面，团队建立了新一代基因编辑工具脱靶检测技术，并通过对单碱基编辑工具的改造，进一步完善了现有基因编辑工具。另一方面，团队通过对食蟹猴胚胎基因编辑，初步获得F0代WRN基因缺失的猴子模型。此外，通过进一步结合人胚胎干细胞技术，团队建立了人科凯恩氏综合征干细胞研究体系以及多种衰老相关疾病模型。基于上述研究体系，团队利用新型的DNA损伤修复报告工具初步发现干细胞衰老过程中特异的DNA损伤修复调控途径；成功获得了人源端粒酶HACA调节复合物5.9 分辨率的冷冻电镜电子密度数据；初步绘制了食蟹猴动物心脏、肝脏、肌肉增龄转录组图谱和m6A表观遗传修饰图谱，并构建衰老相关的生物学数据库。在衰老及衰老相关疾病的干预方面，团队通过靶向编辑单个长寿基因产生了首例遗传增强的人类血管细胞；揭示了CBX4,YAP-FOXD1通路和miRNA合成通路关键因子DGCR8在人干细胞衰老调控及骨关节炎基因治疗中的作用及分子机制；通过对小分子化合物库的通量筛选，发现长期低剂量槲皮素处理可以延长小鼠健康寿命。项目在本执行年度在包括Nature,Science以及Cell Stem Cell 等在内著名学术期刊发表共计23篇研究论文，申请或获授权了14项中国和国际PCT专利。
Application Abstract: Aiming to study the underlying mechanism of primate aging and identify aging biomarkers,we firstly developed a method named GOTI(Genome wide off target analysis by two cell embryo injection)to detect off-target mutations caused by either CRISPR-Cas9 or base editors,the most popular gene editing tools,and we identified that cytosine base editing induced SNVs at more than 20-fold higher frequencies,requiring a solution to address its fidelity.Subsequently,by engineering deaminases,we modified the both ABE and CBE gene editing tools with a significant reduction in off-target RNA SNVs.Meantime,utilizing gene editing tools,we generated F0 generation of“Werner Syndrome”cynomolgus monkey by knock out WRN gene.We also established the human Cockayne syndrome stem cell model revealing that defects in DNA repair account for CS pathologies.Based on the above research systems for primate aging study,we carried out the following mechanism studies:delineating the DNA damage repair pathways responsible for the senescence of human adipocyte stem cells;acquisition of the crystal structure of human telomerase complex with 5.9 resolution;profiling the N6-methyladenosine(m5A)epitranome of human progeria and multi organs of aged monkeys and their control counterparts,and further established the Agingbank providing profound and accurate resources and tools for aging research.In terms of the intervention of aging and aging associated diseases,we generated the first genetically enhanced human vascular cells through targeted editing of the longevity gene FOXO3A;revealed the role of key rejuvenation factors including CBX4,YAP,FOXD1 and DGCR8 for human stem cell aging and developed novel gene therapy of the aging-related disease such as osteoarthritis;revealed that the long-term low-dose quercetin treatment prolonged the healthy life of mice.All of the above work was accomplished by the end of year 2019,with 23 SCI papers published.
项目受资助省
北京市
