Background:Chimeric antigen receptor T(CAR-T)cell therapy has achieved marked therapeutic success in ameliorating hematological malignancies.However,there is an extant void in the clinical guidelines concerning the mo...Background:Chimeric antigen receptor T(CAR-T)cell therapy has achieved marked therapeutic success in ameliorating hematological malignancies.However,there is an extant void in the clinical guidelines concerning the most effective chemotherapy regimen prior to chimeric antigen receptor T(CAR-T)cell therapy,as well as the optimal timing for CAR-T cell infusion post-chemotherapy.Materials and Methods:We employed cell-derived tumor xenograft(CDX)murine models to delineate the optimal pre-conditioning chemotherapy regimen and timing for CAR-T cell treatment.Furthermore,transcriptome sequencing was implemented to identify the therapeutic targets and elucidate the underlying mechanisms governing the treatment regimen.Results:Our preclinical in vivo evaluation determined that a combination of cyclophosphamide and fludarabine,followed by the infusion of CD19 CAR-T cells five days subsequent to the chemotherapy,exerts the most efficacious therapeutic effect in B-cell hematological malignancies.Concurrently,RNA-seq data indicated that the therapeutic efficacy predominantly perturbs tumor cell metabolism,primarily through the inhibition of key mitochondrial targets,such as C-Jun Kinase enzyme(C-JUN).Conclusion:In summary,the present study offers critical clinical guidance and serves as an authoritative reference for the deployment of CD19 CAR-T cell therapy in the treatment of B-cell hematological malignancies.展开更多
Hearing loss and deafness,as a worldwide disability disease,have been troubling human beings.However,the auditory organ of the inner ear is highly heterogeneous and has a very limited number of cells,which are largely...Hearing loss and deafness,as a worldwide disability disease,have been troubling human beings.However,the auditory organ of the inner ear is highly heterogeneous and has a very limited number of cells,which are largely uncharacterized in depth.Recently,with the development and utilization of single-cell RNA sequencing(scRNA-seq),researchers have been able to unveil the complex and sophisticated biological mechanisms of various types of cells in the auditory organ at the single-cell level and address the challenges of cellular heterogeneity that are not resolved through by conventional bulk RNA sequencing(bulk RNAseq).Herein,we reviewed the application of scRNA-seq technology in auditory research,with the aim of providing a reference for the development of auditory organs,the pathogenesis of hearing loss,and regenerative therapy.Prospects about spatial transcriptomic scRNA-seq,single-cell based genome,and Live-seq technology will also be discussed.展开更多
Sensory hair cells(HCs)in the cochlea cannot regenerate spontaneously in adult mammals after being damaged by external or genetic factors.However,several genes and signaling pathways are reported to induce cochlear HC...Sensory hair cells(HCs)in the cochlea cannot regenerate spontaneously in adult mammals after being damaged by external or genetic factors.However,several genes and signaling pathways are reported to induce cochlear HC regeneration at the early neonatal stage.Rps14 encodes a ribosomal protein that is involved in the regulation of cell differentiation and proliferation in mammals.However,its roles in the cochlea have not been reported in vivo.Here,we specifically overexpressed Rps14 in Lgr5+progenitor cells in the newborn mouse cochlea and found that Rps14 conditional overexpression(cOE)mice had significantly increased the ectopic HCs,including inner and outer HCs.We further explored the source of these ectopic HCs and found no EdU+supporting cells observed in the Rps14 cOE mice.The lineage tracing results,on the other hand,revealed that Rps14 cOE mice had significantly more tdTomato+HCs in their cochleae than control mice.These results indicated that regenerated HCs by cOE of Rps14 are most likely derived from inducing the direct trans-differentiation of Lgr5+progenitor cells into HCs.Moreover,real-time qPCR results suggested that the transcription factor genes Atoh1 and Gfi1,which are important in regulating HC differentiation,were upregulated in the cochlear basilar membrane of Rps14 cOE mice.In summary,this study provides in vivo evidence that,in the postnatal mouse cochlea,Rps14 is a potential gene that can promote the spontaneous trans-differentiation of Lgr5+progenitor cells into HCs.This gene may one day be exploited as a therapeutic target for treating hearing loss.展开更多
基金National Natural Science Foundation of China(No.82370164)Sanming Project of Medicine in Shenzhen(No.SZSM202011004)Shenzhen Science and Technology Innovation Commission(JCYJ20180307150419435 and JCYJ20210324123004011).
文摘Background:Chimeric antigen receptor T(CAR-T)cell therapy has achieved marked therapeutic success in ameliorating hematological malignancies.However,there is an extant void in the clinical guidelines concerning the most effective chemotherapy regimen prior to chimeric antigen receptor T(CAR-T)cell therapy,as well as the optimal timing for CAR-T cell infusion post-chemotherapy.Materials and Methods:We employed cell-derived tumor xenograft(CDX)murine models to delineate the optimal pre-conditioning chemotherapy regimen and timing for CAR-T cell treatment.Furthermore,transcriptome sequencing was implemented to identify the therapeutic targets and elucidate the underlying mechanisms governing the treatment regimen.Results:Our preclinical in vivo evaluation determined that a combination of cyclophosphamide and fludarabine,followed by the infusion of CD19 CAR-T cells five days subsequent to the chemotherapy,exerts the most efficacious therapeutic effect in B-cell hematological malignancies.Concurrently,RNA-seq data indicated that the therapeutic efficacy predominantly perturbs tumor cell metabolism,primarily through the inhibition of key mitochondrial targets,such as C-Jun Kinase enzyme(C-JUN).Conclusion:In summary,the present study offers critical clinical guidance and serves as an authoritative reference for the deployment of CD19 CAR-T cell therapy in the treatment of B-cell hematological malignancies.
基金supported by grants from National Key R&D Program of China(2021YFA1101300,2021YFA1101800,2020YFA0112503)Strategic Priority Research Program of the Chinese Academy of Science(XDA16010303)+3 种基金National Natural Science Foundation of China(82030029,81970882,and 92149304)Science and Technology Department of Sichuan Province(2021YFS0371)Shenzhen Fundamental Research Program(JCYJ20190814093401920,JCYJ20210324125608022)Open Research Fund of State Key Laboratory of Genetic Engineering,Fudan University(SKLGE-2104).
文摘Hearing loss and deafness,as a worldwide disability disease,have been troubling human beings.However,the auditory organ of the inner ear is highly heterogeneous and has a very limited number of cells,which are largely uncharacterized in depth.Recently,with the development and utilization of single-cell RNA sequencing(scRNA-seq),researchers have been able to unveil the complex and sophisticated biological mechanisms of various types of cells in the auditory organ at the single-cell level and address the challenges of cellular heterogeneity that are not resolved through by conventional bulk RNA sequencing(bulk RNAseq).Herein,we reviewed the application of scRNA-seq technology in auditory research,with the aim of providing a reference for the development of auditory organs,the pathogenesis of hearing loss,and regenerative therapy.Prospects about spatial transcriptomic scRNA-seq,single-cell based genome,and Live-seq technology will also be discussed.
基金supported by the National Key R&D Program of China (No.2021YFA1101300,2021YFA1101800,2020YFA0112503,2022YFA0807000)the Strategic Priority Research Program of the Chinese Academy of Science (XDA16010303)+3 种基金the National Natural Science Foundation of China (Nos.81970892,82171149,82030029,81970882,92149304)the Science and Technology Department of Sichuan Province (No.2021YFS0371)the Shenzhen Fundamental Research Program (JCYJ20190814093401920,JCYJ20210324125608022)the Open Research Fund of the State Key Laboratory of Genetic Engineering,Fudan University (No.SKLGE-2104).
文摘Sensory hair cells(HCs)in the cochlea cannot regenerate spontaneously in adult mammals after being damaged by external or genetic factors.However,several genes and signaling pathways are reported to induce cochlear HC regeneration at the early neonatal stage.Rps14 encodes a ribosomal protein that is involved in the regulation of cell differentiation and proliferation in mammals.However,its roles in the cochlea have not been reported in vivo.Here,we specifically overexpressed Rps14 in Lgr5+progenitor cells in the newborn mouse cochlea and found that Rps14 conditional overexpression(cOE)mice had significantly increased the ectopic HCs,including inner and outer HCs.We further explored the source of these ectopic HCs and found no EdU+supporting cells observed in the Rps14 cOE mice.The lineage tracing results,on the other hand,revealed that Rps14 cOE mice had significantly more tdTomato+HCs in their cochleae than control mice.These results indicated that regenerated HCs by cOE of Rps14 are most likely derived from inducing the direct trans-differentiation of Lgr5+progenitor cells into HCs.Moreover,real-time qPCR results suggested that the transcription factor genes Atoh1 and Gfi1,which are important in regulating HC differentiation,were upregulated in the cochlear basilar membrane of Rps14 cOE mice.In summary,this study provides in vivo evidence that,in the postnatal mouse cochlea,Rps14 is a potential gene that can promote the spontaneous trans-differentiation of Lgr5+progenitor cells into HCs.This gene may one day be exploited as a therapeutic target for treating hearing loss.
