Diffuse midline glioma(DMG),H3K27-altered,is lethal pediatric-type,high-grade,localized to the midline region of the central nervous system.Effective treatment guidelines are absent,and clinical trials are preferred f...Diffuse midline glioma(DMG),H3K27-altered,is lethal pediatric-type,high-grade,localized to the midline region of the central nervous system.Effective treatment guidelines are absent,and clinical trials are preferred for primary or recur-rent DMG patients.Recently,epigenetic agent-based immunotherapy has exhibited promising therapeutic effects in the clinical setting.However,the underlying mechanisms remain a mystery.The rare DMG tumor samples from biopsy or resection largely impede basic research,by using patient-derived tumor cells which better recapitulate the parental tu-mor’s heterogeneity compared to established cell lines.As an epigenetic reprogramming disease,DMG exhibits a global loss of H3K27 trimethylation(H3K27me3)and a gain of H3K27 acetylation(H3K27ac).Analysis of multiple epige-netic marks is fundamentally necessary.However,traditional techniques cannot allow ultra-low input and high-throughput.Herein we have developed a new method called high-throughput in situ tagged immunoprecipitation sequencing(HiTIP-seq),which uses an integrated superhydrophobic microwell array technology(InSMART).We were able to perform 100 parallel assays from as few as 100 cells per microwell on a single chip.We applied the tech-nology to profile epigenetic alterations of three-dimensional(3D)cell cultures derived from DMG patients.Our HiTIP-seq integrated with RNA sequencing(RNA-seq)analysis revealed that the combination of epigenetic agents(panobino-stat and tazemetostat),reprogrammed histone modifications and drove transcriptome changes.Among them,Wnt inhibitory factor 1(WIF1)has a gain of H3K27ac and a loss of H3K27me3,which leads to the upregulated expression.Altogether,HiTIP-seq is a versatile method for high-throughput analysis of histone modifications,suitable for both DMG research and studying rare 3D models.展开更多
Brainstem gliomas comprise both slow-growing and highly aggressive tumors,the latter carrying a dismal prognosis of approximately 10 months in children.Given their common locations along the brainstem,they are often n...Brainstem gliomas comprise both slow-growing and highly aggressive tumors,the latter carrying a dismal prognosis of approximately 10 months in children.Given their common locations along the brainstem,they are often not amenable to surgical resection.There are currently a host of exploratory therapies under investigation ranging from immunotherapy,small molecular inhibitors,epigenetic-modifying agents,and radiation protocols to combat these difficult-to-treat tumors.Recent discoveries highlighting the role of H3 histone mutations in diffuse midline glioma oncogenesis have yielded a variety of new targetable antigens and aberrant signaling pathways.Although many of these studies have shown promise in terms of inhibiting tumor growth and disease progression,results to date have been modest in their ability to translate into meaningful clinical benefit.This review will serve as an updated report on the current state of literature concerning pre-clinical and clinical therapies being investigated for brainstem glioma.In addition,this review will serve as a guide for clinicians as we review the evolving nomenclature of brainstem gliomas,commonly presenting symptoms,diagnostic tools,and standard therapies.展开更多
基金supported by grants from the National Science Foundation of China(No.22127804)the National Natural Science Foundation of China(No.81771931 and 31971325)+1 种基金Institute of Biomedicine,the Tsinghua-Peking Joint Center for Life Sciences,and the Clinical Medicine Development Fund of Tsinghua University(No.10001020510)the National Key Research and Development Program of China(No.2016YFC0900200).
文摘Diffuse midline glioma(DMG),H3K27-altered,is lethal pediatric-type,high-grade,localized to the midline region of the central nervous system.Effective treatment guidelines are absent,and clinical trials are preferred for primary or recur-rent DMG patients.Recently,epigenetic agent-based immunotherapy has exhibited promising therapeutic effects in the clinical setting.However,the underlying mechanisms remain a mystery.The rare DMG tumor samples from biopsy or resection largely impede basic research,by using patient-derived tumor cells which better recapitulate the parental tu-mor’s heterogeneity compared to established cell lines.As an epigenetic reprogramming disease,DMG exhibits a global loss of H3K27 trimethylation(H3K27me3)and a gain of H3K27 acetylation(H3K27ac).Analysis of multiple epige-netic marks is fundamentally necessary.However,traditional techniques cannot allow ultra-low input and high-throughput.Herein we have developed a new method called high-throughput in situ tagged immunoprecipitation sequencing(HiTIP-seq),which uses an integrated superhydrophobic microwell array technology(InSMART).We were able to perform 100 parallel assays from as few as 100 cells per microwell on a single chip.We applied the tech-nology to profile epigenetic alterations of three-dimensional(3D)cell cultures derived from DMG patients.Our HiTIP-seq integrated with RNA sequencing(RNA-seq)analysis revealed that the combination of epigenetic agents(panobino-stat and tazemetostat),reprogrammed histone modifications and drove transcriptome changes.Among them,Wnt inhibitory factor 1(WIF1)has a gain of H3K27ac and a loss of H3K27me3,which leads to the upregulated expression.Altogether,HiTIP-seq is a versatile method for high-throughput analysis of histone modifications,suitable for both DMG research and studying rare 3D models.
文摘Brainstem gliomas comprise both slow-growing and highly aggressive tumors,the latter carrying a dismal prognosis of approximately 10 months in children.Given their common locations along the brainstem,they are often not amenable to surgical resection.There are currently a host of exploratory therapies under investigation ranging from immunotherapy,small molecular inhibitors,epigenetic-modifying agents,and radiation protocols to combat these difficult-to-treat tumors.Recent discoveries highlighting the role of H3 histone mutations in diffuse midline glioma oncogenesis have yielded a variety of new targetable antigens and aberrant signaling pathways.Although many of these studies have shown promise in terms of inhibiting tumor growth and disease progression,results to date have been modest in their ability to translate into meaningful clinical benefit.This review will serve as an updated report on the current state of literature concerning pre-clinical and clinical therapies being investigated for brainstem glioma.In addition,this review will serve as a guide for clinicians as we review the evolving nomenclature of brainstem gliomas,commonly presenting symptoms,diagnostic tools,and standard therapies.
