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Long-term in vivo chimeric cells tracking in non-human primate

Protein & Cell 2024年第3期15卷 207-222页

作者： Junmo Wu Yu Kang Xiang Luo Shaoxing Dai Yuxi Shi Zhuoyao Li Zengli Tang Zhenzhen Chen Ran Zhu Pengpeng Yang Zifan Li Hong Wang Xinglong Chen Ziyi Zhao weizhi ji Yuyu Niu State Key Laboratory of Primate Biomedical Research Institute of Primate Translational MedicineKunming University of Science and TechnologyKunming 650500China Yunnan Key Laboratory of Primate Biomedical Research Kunming 650500China Faculty of Life Science and Technology Kunming University of Science and TechnologyKunming 650500China

Non-human primates(NHPs)are increasingly used in preclinical trials to test the safety and efficacy of biotech-nology therapies.Nonetheless,given the ethical issues and costs associated with this model,it would be highly advantageous to use NHP cellular models in clinical studies.However,developing and maintaining the naive state of primate pluripotent stem cells(PSCs)remains difficult as does in vivo detection of PSCs,thus limiting biotech-nology application in the cynomolgus monkey.Here,we report a chemically defined,xeno-free culture system for culturing and deriving monkey PSCs in vitro.The cells display global gene expression and genome-wide hypometh-ylation patterns distinct from monkey-primed cells.We also found expression of signaling pathways components that may increase the potential for chimera formation.Crucially for biomedical applications,we were also able to integrate bioluminescent reporter genes into monkey PsCs and track them in chimeric embryos in vivo and in vitro.The engineered cells retained embryonic and extra-embryonic developmental potential.Meanwhile,we generated a chimeric monkey carrying bioluminescent cells,which were able to track chimeric cells for more than 2 years in living animals.Our study could have broad utility in primate stem cell engineering and in utilizing chimeric monkey models forclinical studies.

关键词： pluripotent stem cells bioluminescence imaging non-human primates

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Genome editing in large animals: current status and future prospects

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National Science Review 2019年第3期6卷 402-420页

作者： jianguo Zhao Liangxue Lai weizhi ji Qi Zhou State Key Laboratory of Stem Cell and Reproductive Biology Institute of ZoologyChinese Academy of Sciences Savaid Medical School University of Chinese Academy of Sciences Institute of Stem Cell and Regeneration Chinese Academy of Sciences South China Institute for Stem Cell Biology and Regenerative Medicine Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Yunnan Key Laboratory of Primate Biomedicine Research Institute of Primate Translational Medicine Kunming University of Science and Technology CAS Center for Excellence in Brain Science and Intelligence Technology (CEBSIT)

Large animals(non-human primates, livestock and dogs) are playing important roles in biomedical research, and large livestock animals serve as important sources of meat and milk. The recently developed programmable DNA nucleases have revolutionized the generation of gene-modified large animals that are used for biological and biomedical research. In this review, we briefly introduce the recent advances in nuclease-meditated gene editing tools, and we outline these editing tools’ applications in human disease modeling, regenerative medicine and agriculture. Additionally, we provide perspectives regarding the challenges and prospects of the new genome editing technology.

关键词： genome editing large animal disease model agriculture xenotransplantation

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Long-read sequencing and de novo assembly of the cynomolgus macaque genome

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Journal of Genetics and Genomics 2022年第10期49卷 975-978页

作者： Bing Bai Yi Wang Ran Zhu Yaolei Zhang Hong Wang Guangyi Fan Xin Liu Hong Shi Yuyu Niu weizhi ji State Key Laboratory of Primate Biomedical Research Institute of Primate Translational MedicineKunming University of Science and TechnologyKunmingYunnan 650500China Yunnan Key Laboratory of Primate Biomedical Research KunmingYunnan 650500China BGI-Qingdao BGI-ShenzhenQingdaoShandong 266555China Translational Immunology Group Department of Biotechnology and BiomedicineTechnical University of DenmarkLyngbyDenmark State Key Laboratory of Primate Biomedical Research Institute of Primate Translational MedicineKunming University of Science and TechnologyKunmingYunnnan 650500China BGI-Shenzhen ShenzhenGuangdong 518083China China National GeneBank BGI-ShenzhenShenzhenGuangdong 518120China Faculty of Life Science and Technology Kunming University of Science and TechnologyKunmingYunnan 650500China ji1 2

Cynomolgus macaques(Macaca fascicularis)belong to the genus Macaca.They are the most widespread nonhuman primate(NHP)and share a common ancestor with humans from about 25 million years ago(Kumar and Hedges,1998).Because of their phylogenetic proximity to humans,macaques are an attractive NHP research model for a wide range of biomedical research(Yan et al.,2011).

关键词： widespread attractive proximity

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Primate stem cells: bridge the translation from basic research to clinic application

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Science China(Life Sciences) 2019年第1期62卷 12-21页

作者： Tianqing Li Zongyong Ai weizhi ji Yunnan Key Laboratory of Primate Biomedical Research Institute of Primate Translational Medicine Kunming University of Science and Technology

A growing body of literature has shown that stem cells are very effective for the treatment of degenerative diseases in rodents but these exciting results have not translated to clinical practice. The difference results from the divergence in genetic, metabolic, and physiological phenotypes between rodents and humans. The high degree of similarity between non-human primates(NHPs) and humans provides the most accurate models for preclinical studies of stem cell therapy. Using a NHP model to understand the following key issues, which cannot be addressed in humans or rodents, will be helpful for extending stem cell applications in the basic science and the clinic. These issues include pluripotency of primate stem cells, the safety and efficiency of stem cell therapy, and transplantation procedures of stem cells suitable for clinical translation. Here we review studies of the above issues in NHPs and current challenges of stem cell applications in both basic science and clinical therapies. We propose that the use of NHP models, in particular combining the serial production and transplantation procedures of stem cells is the most useful for preclinical studies designed to overcome these challenges.

关键词： non-human primate pluripotent stem cells monkey disease model stem cell therapy pluripotency state transition transplantation procedures

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Current state of stem cell research in non-human primates:an overview

Medical Review

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Medical Review 2023年第4期3卷 277-304页

作者： Junmo Wu Yuxi Shi Shanshan Yang Zengli Tang Zifan Li Zhuoyao Li jiawei Zuo weizhi ji Yuyu Niu Kunming University of Science and Technology KunmingYunnan ProvinceChina Yunnan Key Laboratory of Primate Biomedical Research KunmingYunnan ProvinceChina

The remarkable similarity between non-human primates(NHPs)and humans establishes them as essential models for understanding human biology and diseases,as well as for developing novel therapeutic strategies,thereby providing more comprehensive reference data for clinical treatment.Pluripotent stem cells such as embryonic stem cells and induced pluripotent stem cells provide unprecedented opportunities for cell therapies against intractable diseases and injuries.As continue to harness the potential of these biotechnological therapies,NHPs are increasingly being employed in preclinical trials,serving as a pivotal tool to evaluate the safety and efficacy of these interventions.Here,we review the recent advancements in the fundamental research of stem cells and the progress made in studies involving NHPs.

关键词： non-human primates cell therapy stem cell

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Cross-species single-cell transcriptomic analysis reveals divergence of cell composition and functions in mammalian ileum epithelium

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Cell Regeneration 2022年第1期11卷 172-187页

作者： Haonan Li Xiaodan Wang Yalong Wang Mengxian Zhang Fan Hong Hong Wang Along Cui jianguo Zhao weizhi ji Ye-Guang Chen The State Key Laboratory of Membrane Biology Tsinghua-Peking Center for Life SciencesSchool of Life SciencesTsinghua UniversityBeijing 100084China Guangzhou Institutes of Biomedicine and Health Chinese Academy of SciencesGuangzhou Science ParkGuangzhou 510530China Guangzhou Laboratory Guangzhou 510005China State Key Laboratory of Primate Biomedical Research Institute of Primate Translational MedicineKunming University of Science and TechnologyKunming 650500YunnanChina State Key Laboratory of Stem cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China

Animal models are widely used for biomedical studies and drug evaluation.The small intestine plays key roles in nutrient absorption,hormone secretion,microbiota defense and drug absorption and metabolism.Although the intestinal structure of mammals is conserved,the differences on epithelial cell composition,functional assignments and drug absorption among mammals are largely unknown.Here,cross-species analysis of single-cell transcriptomic atlas of the ileum epithelium from mouse,rat,pig,macaque and human reveals the conserved and differential cell types and functions among species,identifies a new CA7+cell type in pig,macaque and human ileum,uncovers the distinct expression pattern in enterocytes,enteroendocrine cells and Paneth cells,and defines the conserved and species-specific intestinal stem cell signature genes.The examination of drug absorption across species suggests that drug metabolism in mouse ileum is closer to human while drug transport in macaque ileum is more similar to human.Together,our data provide the comprehensive information about cell composition and functional assignments in five species,and offer the valuable guidance for animal model selection and drug testing.

关键词： scRNA-seq Transcriptome Ileum Monkey Pig Rat CA7 cells

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Paving the road for biomedicine: genome editing and stem cells in primates

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National Science Review 2017年第4期4卷 543-549页

作者： Yuyu Niu Tianqing Li weizhi ji Yunnan Key Laboratory of Primate Biomedicine Research Institute of Primate Translational Medicine Kunming University of Science and Technology

Non-human primates(NHPs) are similar with humans in their anatomy, physiology, genetics and neural functions, and in their cognition, emotions and social behavior. The NHP model has played an important role in the biomedical research, development of vaccines and drugs and new therapies for many diseases.Recent advances in genome editing have generated NHP models for human diseases. Genome editing integrated with derivation of NHP pluripotent stem cells now provides an optimal resource to regenerate medicine.

关键词： genome editing stem cells non-human primate

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Efficient Modulation of Exon Skipping via Antisense Circular RNAs

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Research 2023年第3期6卷 485-490页

作者： Shuaiwei Ren Mei Huang Raoxian Bai Lijiao Chen jiao Yang Junyu Zhang Wenting Guo weizhi ji Yongchang Chen State Key Laboratory of Primate Biomedical Research Institute of Primate Translational MedicineKunming University of Science and TechnologyKunming 650500China Yunnan Key Laboratory of Primate Biomedical Research Kunming 650500China

Splice-switching antisense oligonucleotides(ASOs)and engineered U7 small nuclear ribonucleoprotein(U7 Sm OPT)are the most commonly used methods for exon skipping.However,challenges remain,such as limited organ delivery and repeated dosing for ASOs and unknown risks of by-products produced by U7 Sm OPT.Here,we showed that antisense circular RNAs(AS-circRNAs)can effectively mediate exon skipping in both minigene and endogenous transcripts.We also showed a relatively higher exon skipping efficiency at the tested Dmd minigene than U7 Sm OPT.AS-circRNA specifically targets the precursor mRNA splicing without off-target effects.Moreover,AS-circRNAs with adeno-associated virus(AAV)delivery corrected the open reading frame and restored the dystrophin expression in a mouse model of Duchenne muscular dystrophy.In conclusion,we develop an alternative method for regulating RNA splicing,which might be served as a novel tool for genetic disease treatment.

关键词： endogenous dystrophy Duchenne

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Establishment of porcine and monkey colonic organoids for drug toxicity study

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Cell Regeneration 2021年第1期10卷 342-351页

作者： Haonan Li Yalong Wang Mengxian Zhang Hong Wang Along Cui jianguo Zhao weizhi ji Ye-Guang Chen The State Key Laboratory of Membrane Biology Tsinghua-Peking Center for Life SciencesSchool of Life SciencesTsinghua UniversityBeijing 100084China State Key Laboratory of Primate Biomedical Research Institute of Primate Translational MedicineKunming University of Science and TechnologyKunming 650500YunnanChina School of Life Sciences University of Science and Technology of ChinaHefei 230027China State Key Laboratory of Stem Cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China Max-Planck Center for Tissue Stem Cell Research and Regenerative Medicine Guangzhou Regenerative Medicine and Health Guangdong LaboratoryGuangzhou 510700China

Pig and monkey are widely used models for exploration of human diseases and evaluation of drug efficiency and toxicity,but high cost limits their uses.Organoids have been shown to be promising models for drug test as they reasonably preserve tissue structure and functions.However,colonic organoids of pig and monkey are not yet established.Here,we report a culture medium to support the growth of porcine and monkey colonic organoids.Wnt signaling and PGE2 are important for long-term expansion of the organoids,and their withdrawal results in lineage differentiation to mature cells.Furthermore,we observe that porcine colonic organoids are closer to human colonic organoids in terms of drug toxicity response.Successful establishment of porcine and monkey colonic organoids would facilitate the mechanistic investigation of the homeostatic regulation of the intestine of these animals and is useful for drug development and toxicity studies.

关键词： Pig Monkey Colonic organoids Culture Drug toxicity

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Cu_(2)O/Cu-垂直石墨烯微电极的制备及尿酸/脑电的检测

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无机化学学报 2024年第7期40卷 1309-1319页

作者：白浩冀伟志陈瑾妍李红姬李明吉天津理工大学化学化工学院天津300384 天津理工大学集成电路科学与工程学院天津300384

结合Cu(Ⅱ)离子浸渍吸附方法及直流电弧等离子体喷射化学气相沉积技术制备了一种电化学/电生理双模Cu_(2)O/Cu-垂直石墨烯微电极,并研究了电化学方法检测尿酸以及记录脑电信号的双响应性能。使用扫描电子显微镜、透射电子显微镜、X射线... 详细信息

结合Cu(Ⅱ)离子浸渍吸附方法及直流电弧等离子体喷射化学气相沉积技术制备了一种电化学/电生理双模Cu_(2)O/Cu-垂直石墨烯微电极,并研究了电化学方法检测尿酸以及记录脑电信号的双响应性能。使用扫描电子显微镜、透射电子显微镜、X射线衍射仪表征了形貌、微结构及晶体成分,并测试了电化学及脑电记录能力。结果表明,该微电极直径仅为200μm,大量镶嵌Cu_(2)O/Cu纳米粒子的石墨烯纳米片垂直生长在基片上,排列成了一种三维的多孔结构,使其具有了高的电化学催化活性、短程离子扩散路径、以及长程导电网络。由此,以10μL的饱和NaCl溶液为介质记录脑电信号时,该微电极的皮肤接触电阻低至约7.05 kΩ,生理电采集性能接近涂导电膏的商用湿电极。此外,该微电极还灵敏响应尿酸的氧化电流,检测浓度范围在0.5~500μmol·L^(-1),检测限低至0.024μmol·L^(-1),且具有良好的抗干扰能力及长期稳定性。

关键词：垂直石墨烯电化学传感器脑电尿酸检测

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