Vertebrate axis patterning requires precise control of the differentiation of neuromesodermal progenitors(NMPs),which generate spinal cord(SC)and presomitic mesoderm(PSM).Previously,we identified a gastrula-premarked ...Vertebrate axis patterning requires precise control of the differentiation of neuromesodermal progenitors(NMPs),which generate spinal cord(SC)and presomitic mesoderm(PSM).Previously,we identified a gastrula-premarked posterior enhancer(p-Enh)that is essential for posterior tissue development by regulating somite and SC in organogenetic embryos,while its role in early NMPs cells remains elusive.Here,using a highly efficient in vitro differentiation system,we found that the genetic removal of p-Enh leads to the aberrantly up-regulated PSM-related genes during both PSM and SC differentiation.Time-resolved transcriptomic analysis and experimental characterization revealed the activated PSM transcriptomic signature arose from disorganized NMPs composition,with an over-representation of the T^(high)SOX2^(low)NMPs subtype.Besides,through a newly developed bioinformatic tool,ST-Pheno,which effectively bridges the in vitro samples to in vivo embryonic phenotypes within spatiotemporal context,we determined that the over-produced T^(high)SOX2^(low)NMPs subtype is predominantly enriched in the anterior primitive streak and adjacent mesoderm region at E7.5,which may disrupt the proper development of NMPs towards prospective PSM and SC,ultimately leading to the posterior development failure.In summary,this study demonstrates a critical role of p-Enh in regulating NMPs subtype composition,which will broaden the molecular understanding of mammalian embryogenesis.展开更多
Spinal motor neurons deficiency results in a series of devastating disorders such as amyotrophic lateral sclerosis(ALS),spinal muscular atrophy(SMA)and spinal cord injury(SCI).These disorders are currently incurable,w...Spinal motor neurons deficiency results in a series of devastating disorders such as amyotrophic lateral sclerosis(ALS),spinal muscular atrophy(SMA)and spinal cord injury(SCI).These disorders are currently incurable,while human pluripotent stem cells(hPSCs)-derived spinal motor neurons are promising but suffered from inappropriate regional identity and functional immaturity for the study and treatment of posterior spinal cord related injuries.In this study,we have established human spinal cord neural progenitor cells(hSCNPCs)via hPSCs differentiated neuromesodermal progenitors(NMPs)and demonstrated the hSCNPCs can be continuously expanded up to 40 passages.hSCNPCs can be rapidly differentiated into posterior spinal motor neurons with high efficiency.The functional maturity has been examined in detail.Moreover,a co-culture scheme which is compatible for both neural and muscular differentiation is developed to mimic the neuromuscular junction(NMJ)formation in vitro.Together,these studies highlight the potential avenues for generating clinically relevant spinal motor neurons and modeling neuromuscular diseases through our defined hSCNPCs.展开更多
基金supported in part by the National Key Basic Research and Development Program of China(2025YFE0200600,2018YFA0800100,2019YFA0801402)the Major Project of Guangzhou National Laboratory(GZNL2025C02014,GZNL2023A02005)+1 种基金the National Natural Science Foundation of China(32130030,32470866,31900454)the Union Project by Guangzhou National Laboratory and State Key Laboratory of Respiratory Disease,Guangzhou Medical University(GZNL2024B01007).
文摘Vertebrate axis patterning requires precise control of the differentiation of neuromesodermal progenitors(NMPs),which generate spinal cord(SC)and presomitic mesoderm(PSM).Previously,we identified a gastrula-premarked posterior enhancer(p-Enh)that is essential for posterior tissue development by regulating somite and SC in organogenetic embryos,while its role in early NMPs cells remains elusive.Here,using a highly efficient in vitro differentiation system,we found that the genetic removal of p-Enh leads to the aberrantly up-regulated PSM-related genes during both PSM and SC differentiation.Time-resolved transcriptomic analysis and experimental characterization revealed the activated PSM transcriptomic signature arose from disorganized NMPs composition,with an over-representation of the T^(high)SOX2^(low)NMPs subtype.Besides,through a newly developed bioinformatic tool,ST-Pheno,which effectively bridges the in vitro samples to in vivo embryonic phenotypes within spatiotemporal context,we determined that the over-produced T^(high)SOX2^(low)NMPs subtype is predominantly enriched in the anterior primitive streak and adjacent mesoderm region at E7.5,which may disrupt the proper development of NMPs towards prospective PSM and SC,ultimately leading to the posterior development failure.In summary,this study demonstrates a critical role of p-Enh in regulating NMPs subtype composition,which will broaden the molecular understanding of mammalian embryogenesis.
基金supported in part by the National Key Basic Research and Development Program of China(2019YFA0801402,2018YFA0800100,2018YFA0108000,2018YFA0107200)“Strategic Priority Research Program”of the Chinese Academy of Sciences,Grant No.(XDA16020501,XDA16020404)National Natural Science Foundation of China(32130030,31630043,31871456,31900454).
文摘Spinal motor neurons deficiency results in a series of devastating disorders such as amyotrophic lateral sclerosis(ALS),spinal muscular atrophy(SMA)and spinal cord injury(SCI).These disorders are currently incurable,while human pluripotent stem cells(hPSCs)-derived spinal motor neurons are promising but suffered from inappropriate regional identity and functional immaturity for the study and treatment of posterior spinal cord related injuries.In this study,we have established human spinal cord neural progenitor cells(hSCNPCs)via hPSCs differentiated neuromesodermal progenitors(NMPs)and demonstrated the hSCNPCs can be continuously expanded up to 40 passages.hSCNPCs can be rapidly differentiated into posterior spinal motor neurons with high efficiency.The functional maturity has been examined in detail.Moreover,a co-culture scheme which is compatible for both neural and muscular differentiation is developed to mimic the neuromuscular junction(NMJ)formation in vitro.Together,these studies highlight the potential avenues for generating clinically relevant spinal motor neurons and modeling neuromuscular diseases through our defined hSCNPCs.
