【目的】克隆黄鳝csf1r基因,并对其时空表达特性进行分析,为探明csf1r基因在黄鳝不同体色形成中的作用奠定基础。【方法】采用RACEs(Rapid-amplification of cDNA ends)技术从黄鳝皮肤cDNAs中克隆得到csf1r基因的全长cDNA序列,对其编码...【目的】克隆黄鳝csf1r基因,并对其时空表达特性进行分析,为探明csf1r基因在黄鳝不同体色形成中的作用奠定基础。【方法】采用RACEs(Rapid-amplification of cDNA ends)技术从黄鳝皮肤cDNAs中克隆得到csf1r基因的全长cDNA序列,对其编码蛋白进行生物信息学分析。采用实时荧光定量PCR(qRT-PCR)方法检测csf1r基因在黄鳝不同组织、不同发育时期胚胎或个体及3种体色黄鳝(黄黑斑鳝、碎花斑鳝和隐花斑鳝)皮肤和肾脏中的相对表达量,分析该基因的表达特征。测定3种体色黄鳝肝脏中的碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)活性和总抗氧化能力(T-AOC)。【结果】黄鳝csf1r cDNA序列全长为4430 bp(GenBank收录号:OP589303),其编码区长度为2937 bp,编码978个氨基酸,存在免疫球蛋白结构域和蛋白激酶催化结构域2个保守结构域。荧光定量PCR结果表明,csf1r基因在黄鳝脑、精巢、卵巢、肠、心脏、肾脏、肝脏、肌肉、皮肤和脾脏等组织中均有表达,在脾脏和心脏中表达量较高,其次是肾脏、皮肤和肌肉,卵巢中表达量最低;csf1r在胚胎眼晶体形成期开始大量表达,显微观察发现该时期胚胎的躯干上开始有色素颗粒出现。在3种体色黄鳝皮肤和肾脏中,csf1r基因在黄黑斑鳝的皮肤中表达量最低,而在其肾脏中表达量最高。3种体色黄鳝肝脏氧化应激指标测定结果发现,黄黑斑鳝肝脏中的碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)活性和总抗氧化能力比其他2种体色黄鳝高,但是差异未达显著水平。【结论】csf1r基因可能不仅参与了黄鳝体色的形成,还与黄鳝非特异性免疫相关。展开更多
Background:CSF1R-related leukoencephalopathy,also known as hereditary diffuse leukoencephalopathy with spheroids(HDLS),is a rare white-matter encephalopathy characterized by motor and neuropsychiatric symptoms due to ...Background:CSF1R-related leukoencephalopathy,also known as hereditary diffuse leukoencephalopathy with spheroids(HDLS),is a rare white-matter encephalopathy characterized by motor and neuropsychiatric symptoms due to colony-stimulating factor 1 receptor(CSF1R)gene mutation.Few of CSF1R mutations have been functionally testified and the pathogenesis remains unknown.Methods:In order to investigate clinical and pathological characteristics of patients with CSF1R-related leukoencephalopathy and explore the potential impact of CSF1R mutations,we analyzed clinical manifestations of 15 patients from 10 unrelated families and performed brain biopsy in 2 cases.Next generation sequencing was conducted for 10 probands to confirm the diagnosis.Sanger sequencing,segregation analysis and phenotypic reevaluation were utilized to substantiate findings.Functional examination of identified mutations was further explored.Results:Clinical and neuroimaging characteristics were summarized.The average age at onset was 35.9±6.4 years(range 24–46 years old).Younger age of onset was observed in female than male(34.2 vs.39.2 years).The most common initial symptoms were speech dysfunction,cognitive decline and parkinsonian symptoms.One patient also had marked peripheral neuropathy.Brain biopsy of two cases showed typical pathological changes,including myelin loss,axonal spheroids,phosphorylated neurofilament and activated macrophages.Electron microscopy disclosed increased mitochondrial vacuolation and disorganized neurofilaments in ballooned axons.A total of 7 pathogenic variants(4 novel,3 documented)were identified with autophosphorylation deficiency,among which c.2342C>T remained partial function of autophosphorylation.Western blotting disclosed the significantly lower level of c.2026C>T(p.R676*)than wild type.The level of microtubule associated protein 1 light chain 3-II(LC3-II),a classical marker of autophagy,was significantly lower in mutants expressed cells than wild type group by western blotting and immunofluorescence staining.Conclusions:Our findings support the loss-of-function and haploinsufficiency hypothesis in pathogenesis.Autophagy abnormality may play a role in the disease.Repairing or promoting the phosphorylation level of mutant CSF1R may shed light on therapeutic targets in the future.However,whether peripheral polyneuropathy potentially belongs to CSF1R-related spectrum deserves further study with longer follow-up and more patients enrolled.展开更多
Melanoma is a malignant neoplasm with a high propensity to metastasize,arising from melanocytes and contributing significantly to global morbidity and mortality.Despite the demonstrated efficacy of many immunotherapy ap...Melanoma is a malignant neoplasm with a high propensity to metastasize,arising from melanocytes and contributing significantly to global morbidity and mortality.Despite the demonstrated efficacy of many immunotherapy approaches,these methods rely on direct destruction of tumor cells with minimal impact on the aggregate of nearby non-tumor cells,the extracellular matrix,and blood vessels that form the tumor microenvironment(TME).The TME is known to be heterogeneous and dynamic,exerting both antitumor and pro-tumor effects depending on the specific features and stage of carcinogenesis.TME has been shown in several studies to promote malignancy,angiogenesis,and metastasis in tumors in general and melanoma in particular.Consequently,a significant number of studies in thefield of melanoma therapy have been redirected to investigate the effects of individual TME constituents,their prognostic significance for patients,and the potential of therapeutic intervention to improve overall patient survival.This review highlights novel therapeutic approaches targeting two key resident cell types in the melanoma microenvironment:tumor-associated macrophages(TAMs)and cancer-associatedfibroblasts(CAFs).The review discusses their role in disease progression and summarizes the results of preclinical and clinical trials of targeted therapies against these cell types in the melanoma TME.展开更多
目的探讨遗传性弥漫性脑白质病变合并球状轴索(hereditary diffuseleukoencephalopathy with spher⁃oids,HDLS)临床及影像学特点,为临床正确诊断HDLS提供思路。方法回顾性分析2012年12月至2021年11月基因检测确诊的3例HDLS临床表现、体...目的探讨遗传性弥漫性脑白质病变合并球状轴索(hereditary diffuseleukoencephalopathy with spher⁃oids,HDLS)临床及影像学特点,为临床正确诊断HDLS提供思路。方法回顾性分析2012年12月至2021年11月基因检测确诊的3例HDLS临床表现、体格检查、神经影像、治疗、预后等资料,结合文献分析HDLS患者的临床特点。结果3例HDLS患者中,男1例,女2例。发病年龄分别为41岁、47岁、46岁。2例患者以反应迟钝、记忆力下降起病,1例以行动迟缓起病。2例病程中出现锥体系受累症状。3例患者头颅MRI均提示双侧侧脑室旁、半卵圆中心脑白质病变。1例DWI序列持续高信号。2例出现脑萎缩,脑室扩大。3例患者均完善脑白质病相关基因检测,结果显示3例患者集落刺激因子1受体(colony⁃stimulating factor 1 receptor,CSF1R)基因突变。结论HDLS临床表现具有异质性。对以神经精神症状和运动障碍起病且头颅MRI示双侧脑白质病变的中青年患者,需考虑CSF1R基因突变所致HDLS的可能。展开更多
Colony-stimulating factor 1 receptor(CSF1R)is highly expressed in mononuclear phagocytes and in the central nervous system.It has emerged as a promising target for tumor therapy and neuroinflammation imaging.Although ...Colony-stimulating factor 1 receptor(CSF1R)is highly expressed in mononuclear phagocytes and in the central nervous system.It has emerged as a promising target for tumor therapy and neuroinflammation imaging.Although therapeutic agents targeting CSF1R have shown great success,the development of diagnostic radiotracers for CSF1R has faced numerous challenges.Consequently,there is an urgent need to overcome these obstacles for the development of CSF1R radiotracers,particularly positron emission tomography tracers,not only for diagnostic purposes but also to aid the development of more effective therapeutic drugs.Here,we provide a comprehensive overview of the development of CSF1R radiotracers,presenting detailed profiles of each tracer's ability to image CSF1R.Additionally,we discuss reported CSF1R small-molecule inhibitors and antibodies,highlighting their relevance to the further development of CSF1R radiotracers.We aim to shed light on the current state of CSF1R radiotracer research and development,provide an insight into the challenges in this field,and offer guidance for future exploration.展开更多
In the central nervous system,immunologic surveillance and response are carried out,in large part,by microglia.These resident macrophages derive from myeloid precursors in the embryonic yolk sac,migrating to the brain...In the central nervous system,immunologic surveillance and response are carried out,in large part,by microglia.These resident macrophages derive from myeloid precursors in the embryonic yolk sac,migrating to the brain and eventually populating local tissue prior to blood-brain barrier formation.Preserved for the duration of lifespan,microglia serve the host as more than just a central arm of innate immunity,also contributing significantly to the development and maintenance of neurons and neural networks,as well as neuroregeneration.The critical nature of these varied functions makes the characterization of key roles played by microglia in neurodegenerative disorders,especially Alzheimer’s disease,of paramount importance.While genetic models and rudimentary pharmacologic approaches for microglial manipulation have greatly improved our understanding of central nervous system health and disease,significant advances in the selective and near complete in vitro and in vivo depletion of microglia for neuroscience application continue to push the boundaries of research.Here we discuss the research efficacy and utility of various microglial depletion strategies,including the highly effective CSF1R inhibitor models,noteworthy insights into the relationship between microglia and neurodegeneration,and the potential for therapeutic repurposing of microglial depletion and repopulation.展开更多
We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,inc...We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,including microglia.To examine whether this therapeutic effect is due to the regulation of activated microglia,we analyzed mic roglial gene expression profiles and the existence of microglia in the brain in this study.5xFAD mice at the age of 1.5 months were assigned to sham-and radiofrequency electromagnetic fields-exposed groups and then animals were exposed to 1950 MHz radiofrequency electromagnetic fields at a specific absorption rate of 5 W/kg for 2 hours/day and 5 days/week for 6 months.We conducted behavioral tests including the object recognition and Y-maze tests and molecular and histopathological analysis of amyloid precursor protein/a myloid-beta metabolism in brain tissue.We confirmed that radiofrequency electromagnetic field exposure for 6 months ameliorated cognitive impairment and amyloid-βdeposition.The expression levels of Iba1(pan-microglial marker)and colony-stimulating factor 1 receptor(CSF1R;regulates microglial prolife ration)in the hippocampus in 5xFAD mice treated with radiofrequency electromagnetic fields were significantly reduced compared with those of the sham-exposed group.Subsequently,we analyzed the expression levels of genes related to mic rogliosis and microglial function in the radiofrequency electromagnetic fields-exposed group compared to those of a CSF1R inhibitor(PLX3397)-treated group.Both radiofrequency electromagnetic fields and PLX3397 suppressed the levels of genes related to microgliosis(Csf1r,CD68,and Ccl6)and pro-inflammatory cytokine interleukin-1β.N otably,the expression levels of genes related to mic roglial function,including Trem2,Fcgr1α,Ctss,and Spi1,were decreased after long-term radiofrequency electromagnetic field exposure,which was also observed in response to microglial suppression by PLX3397.These results showed that radiofrequency electromagnetic fields ameliorated amyloid-βpathology and cognitive impairment by suppressing amyloid-βdeposition-induced microgliosis and their key regulator,CSF1R.展开更多
基金This work was partially supported by the Cohort Study of Cerebral White Matter Change(SWATCH)Multicenter Network(ChiCTR1800015295)This study was supported by the grants from the National Natural Science Foundation of China(No.81571086,81870889,81600978,81200965 and 81430022)+6 种基金National Key R&D Program of China(No.2016YFC1305804 and 2017YFC1310200)Doctoral Innovation Fund of Shanghai Jiao Tong University School of Medicine(No.BXJ201913)Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant Support(No.20161401)Interdisciplinary Project of Shanghai Jiao Tong University(No.YG2016MS64)the Research Fund for the Doctoral Program of Higher Education(No.20110073120088)Natural Science Foundation of Science and Technology of Shanghai(No.15ZR1426700)Guang Ci Qing Nian Grant(No.GCQN-2017-A03).
文摘Background:CSF1R-related leukoencephalopathy,also known as hereditary diffuse leukoencephalopathy with spheroids(HDLS),is a rare white-matter encephalopathy characterized by motor and neuropsychiatric symptoms due to colony-stimulating factor 1 receptor(CSF1R)gene mutation.Few of CSF1R mutations have been functionally testified and the pathogenesis remains unknown.Methods:In order to investigate clinical and pathological characteristics of patients with CSF1R-related leukoencephalopathy and explore the potential impact of CSF1R mutations,we analyzed clinical manifestations of 15 patients from 10 unrelated families and performed brain biopsy in 2 cases.Next generation sequencing was conducted for 10 probands to confirm the diagnosis.Sanger sequencing,segregation analysis and phenotypic reevaluation were utilized to substantiate findings.Functional examination of identified mutations was further explored.Results:Clinical and neuroimaging characteristics were summarized.The average age at onset was 35.9±6.4 years(range 24–46 years old).Younger age of onset was observed in female than male(34.2 vs.39.2 years).The most common initial symptoms were speech dysfunction,cognitive decline and parkinsonian symptoms.One patient also had marked peripheral neuropathy.Brain biopsy of two cases showed typical pathological changes,including myelin loss,axonal spheroids,phosphorylated neurofilament and activated macrophages.Electron microscopy disclosed increased mitochondrial vacuolation and disorganized neurofilaments in ballooned axons.A total of 7 pathogenic variants(4 novel,3 documented)were identified with autophosphorylation deficiency,among which c.2342C>T remained partial function of autophosphorylation.Western blotting disclosed the significantly lower level of c.2026C>T(p.R676*)than wild type.The level of microtubule associated protein 1 light chain 3-II(LC3-II),a classical marker of autophagy,was significantly lower in mutants expressed cells than wild type group by western blotting and immunofluorescence staining.Conclusions:Our findings support the loss-of-function and haploinsufficiency hypothesis in pathogenesis.Autophagy abnormality may play a role in the disease.Repairing or promoting the phosphorylation level of mutant CSF1R may shed light on therapeutic targets in the future.However,whether peripheral polyneuropathy potentially belongs to CSF1R-related spectrum deserves further study with longer follow-up and more patients enrolled.
基金performed at the expense of the subsidy allocated to Kazan Federal University for the fulfillment of the stated task in the field of scientific activity,No.FZSM-2023-0011.
文摘Melanoma is a malignant neoplasm with a high propensity to metastasize,arising from melanocytes and contributing significantly to global morbidity and mortality.Despite the demonstrated efficacy of many immunotherapy approaches,these methods rely on direct destruction of tumor cells with minimal impact on the aggregate of nearby non-tumor cells,the extracellular matrix,and blood vessels that form the tumor microenvironment(TME).The TME is known to be heterogeneous and dynamic,exerting both antitumor and pro-tumor effects depending on the specific features and stage of carcinogenesis.TME has been shown in several studies to promote malignancy,angiogenesis,and metastasis in tumors in general and melanoma in particular.Consequently,a significant number of studies in thefield of melanoma therapy have been redirected to investigate the effects of individual TME constituents,their prognostic significance for patients,and the potential of therapeutic intervention to improve overall patient survival.This review highlights novel therapeutic approaches targeting two key resident cell types in the melanoma microenvironment:tumor-associated macrophages(TAMs)and cancer-associatedfibroblasts(CAFs).The review discusses their role in disease progression and summarizes the results of preclinical and clinical trials of targeted therapies against these cell types in the melanoma TME.
基金National Natural Science Foundation of China,Grant/Award Number:82102120。
文摘Colony-stimulating factor 1 receptor(CSF1R)is highly expressed in mononuclear phagocytes and in the central nervous system.It has emerged as a promising target for tumor therapy and neuroinflammation imaging.Although therapeutic agents targeting CSF1R have shown great success,the development of diagnostic radiotracers for CSF1R has faced numerous challenges.Consequently,there is an urgent need to overcome these obstacles for the development of CSF1R radiotracers,particularly positron emission tomography tracers,not only for diagnostic purposes but also to aid the development of more effective therapeutic drugs.Here,we provide a comprehensive overview of the development of CSF1R radiotracers,presenting detailed profiles of each tracer's ability to image CSF1R.Additionally,we discuss reported CSF1R small-molecule inhibitors and antibodies,highlighting their relevance to the further development of CSF1R radiotracers.We aim to shed light on the current state of CSF1R radiotracer research and development,provide an insight into the challenges in this field,and offer guidance for future exploration.
基金This work was supported by DePaul University grant URC450622(to EC).
文摘In the central nervous system,immunologic surveillance and response are carried out,in large part,by microglia.These resident macrophages derive from myeloid precursors in the embryonic yolk sac,migrating to the brain and eventually populating local tissue prior to blood-brain barrier formation.Preserved for the duration of lifespan,microglia serve the host as more than just a central arm of innate immunity,also contributing significantly to the development and maintenance of neurons and neural networks,as well as neuroregeneration.The critical nature of these varied functions makes the characterization of key roles played by microglia in neurodegenerative disorders,especially Alzheimer’s disease,of paramount importance.While genetic models and rudimentary pharmacologic approaches for microglial manipulation have greatly improved our understanding of central nervous system health and disease,significant advances in the selective and near complete in vitro and in vivo depletion of microglia for neuroscience application continue to push the boundaries of research.Here we discuss the research efficacy and utility of various microglial depletion strategies,including the highly effective CSF1R inhibitor models,noteworthy insights into the relationship between microglia and neurodegeneration,and the potential for therapeutic repurposing of microglial depletion and repopulation.
基金Institute of Information&Communications Technology Planning&Evaluation(IITP)grant funded by Korea government(MSIT),Nos.2017-0-00961 and 2019-0-00102(to HDC)。
文摘We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,including microglia.To examine whether this therapeutic effect is due to the regulation of activated microglia,we analyzed mic roglial gene expression profiles and the existence of microglia in the brain in this study.5xFAD mice at the age of 1.5 months were assigned to sham-and radiofrequency electromagnetic fields-exposed groups and then animals were exposed to 1950 MHz radiofrequency electromagnetic fields at a specific absorption rate of 5 W/kg for 2 hours/day and 5 days/week for 6 months.We conducted behavioral tests including the object recognition and Y-maze tests and molecular and histopathological analysis of amyloid precursor protein/a myloid-beta metabolism in brain tissue.We confirmed that radiofrequency electromagnetic field exposure for 6 months ameliorated cognitive impairment and amyloid-βdeposition.The expression levels of Iba1(pan-microglial marker)and colony-stimulating factor 1 receptor(CSF1R;regulates microglial prolife ration)in the hippocampus in 5xFAD mice treated with radiofrequency electromagnetic fields were significantly reduced compared with those of the sham-exposed group.Subsequently,we analyzed the expression levels of genes related to mic rogliosis and microglial function in the radiofrequency electromagnetic fields-exposed group compared to those of a CSF1R inhibitor(PLX3397)-treated group.Both radiofrequency electromagnetic fields and PLX3397 suppressed the levels of genes related to microgliosis(Csf1r,CD68,and Ccl6)and pro-inflammatory cytokine interleukin-1β.N otably,the expression levels of genes related to mic roglial function,including Trem2,Fcgr1α,Ctss,and Spi1,were decreased after long-term radiofrequency electromagnetic field exposure,which was also observed in response to microglial suppression by PLX3397.These results showed that radiofrequency electromagnetic fields ameliorated amyloid-βpathology and cognitive impairment by suppressing amyloid-βdeposition-induced microgliosis and their key regulator,CSF1R.
