目的:研究CLN3基因在卵巢浆液性癌及子宫内膜癌中的表达特点及与其发生发展的关系。方法:采用RT-PCR和W estern B lot方法检测卵巢浆液性肿瘤(良性浆液性肿瘤、交界性肿瘤、浆液性癌)组织、子宫内膜癌及癌旁组织、正常卵巢组织和正常子...目的:研究CLN3基因在卵巢浆液性癌及子宫内膜癌中的表达特点及与其发生发展的关系。方法:采用RT-PCR和W estern B lot方法检测卵巢浆液性肿瘤(良性浆液性肿瘤、交界性肿瘤、浆液性癌)组织、子宫内膜癌及癌旁组织、正常卵巢组织和正常子宫内膜组织中CLN3基因表达情况。结果:卵巢浆液性癌组织中CLN3 mRNA表达为3.04±0.58,交界性卵巢浆液性肿瘤组织中表达为1.63±0.43,均明显高于正常卵巢组织和良性卵巢浆液性肿瘤,差异显著(P<0.01);卵巢浆液性癌组织CLN3表达高于交界性卵巢浆液性肿瘤(P<0.01);卵巢浆液性癌患者不同临床分期(Ⅰ/Ⅱ期与Ⅲ/Ⅳ期)、病理分级及有无腹水间比较差异显著(P<0.05)。正常子宫内膜组织中CLN3呈低表达,在所有的子宫内膜癌及癌旁组织中发现CLN3 mRNA及蛋白表达上调(P<0.01)。结论:①CLN3基因与卵巢浆液性癌的发生发展、生物学行为和预后可能有关,可作为卵巢浆液性癌的重要的生物学标记物,是卵巢浆液癌防治的一个新靶点;②CLN3基因过表达可能与子宫内膜癌发生有关。展开更多
目的探讨CLN3(青少年神经元腊样质脂褐质沉积症Juvenile,form of neuronalceroid-lipofuscinosis,JNCL,Batten病)基因在卵巢浆液性囊腺癌中的表达特点及其在卵巢癌化疗耐药中的作用。方法采用半定量RT-PCR、Western blot以及免疫组化的...目的探讨CLN3(青少年神经元腊样质脂褐质沉积症Juvenile,form of neuronalceroid-lipofuscinosis,JNCL,Batten病)基因在卵巢浆液性囊腺癌中的表达特点及其在卵巢癌化疗耐药中的作用。方法采用半定量RT-PCR、Western blot以及免疫组化的方法检测CLN3在卵巢浆液性囊腺癌中的表达,分析其表达与临床病理特征及化疗耐药的相关性。结果 CLN3在卵巢浆液性囊腺癌中的表达高于正常卵巢组织,且临床分期越晚其表达越高(P<0.05),而与肿瘤患者年龄无显著相关性。CLN3在卵巢癌化疗耐药组表达阳性率明显高于化疗敏感组(P<0.05)。结论卵巢癌中CLN3高表达与卵巢癌的发生、发展、浸润和转移有关,与卵巢癌化疗耐药密切相关。CLN3可成为卵巢癌化疗新的基因治疗靶点。展开更多
Lysosomes have many roles,including degrading macromolecules and signalling to the nucleus1.Lysosomal dysfunction occurs in various human conditions,such as common neurodegenerative diseases and monogenic lysosomal st...Lysosomes have many roles,including degrading macromolecules and signalling to the nucleus1.Lysosomal dysfunction occurs in various human conditions,such as common neurodegenerative diseases and monogenic lysosomal storage disorders(LSDs)2-4.For most LSDs,the causal genes have been identified but,in some,the function of the implicated gene is unknown,in part because lysosomes occupy a small fraction of the cellular volume so that changes in lysosomal contents are difficult to detect.Here we develop the LysoTag mouse for the tissue-specific isolation of intact lysosomes that are compatible with the multimodal profiling of their contents.We used the LysoTag mouse to study CLN3,a lysosomal transmembrane protein with an unknown function.In children,the loss of CLN3 causes juvenile neuronal ceroid lipofuscinosis(Batten disease),a lethal neurodegenerative LSD.Untargeted metabolite profiling of lysosomes from the brains of mice lacking CLN3 revealed a massive accumulation of glycerophosphodiesters(GPDs)-the end products of glycerophospholipid catabolism.GPDs also accumulate in the lysosomes of CLN3-deficient cultured cells and we show that CLN3 is required for their lysosomal egress.Loss of CLN3 also disrupts glycerophospholipid catabolism in the lysosome.Finally,we found elevated levels of glycerophosphoinositol in the cerebrospinal fluid of patients with Batten disease,suggesting the potential use of glycerophosphoinositol as a disease biomarker.Our results show that CLN3 is required for the lysosomal clearance of GPDs and reveal Batten disease as a neurodegenerative LSD with a defect in glycerophospholipid metabolism.展开更多
CLN3 is a lysosomal transmembrane protein and loss of CLN3 is known to cause a juvenile lethal neurodegenerative lysosomal storage disorder(LSD),called Batten disease.In a recent study published in Nature,Laqtom et al...CLN3 is a lysosomal transmembrane protein and loss of CLN3 is known to cause a juvenile lethal neurodegenerative lysosomal storage disorder(LSD),called Batten disease.In a recent study published in Nature,Laqtom et al.reported a novel function of CLN3 in the clearance of glycerophospholipid from lysosomes via lysosomal efflux of glycerophosphodiesters(GPDs),not only establishing a deeper mechanistic understanding of Batten disease,but also suggesting both the diagnostic and therapeutic potential of CLN3-GPDs in this type of neurodegenerative LSD.展开更多
Evidence from genetics and from analyzing cellular and animal models have converged to suggest links between neurodegenerative disorders of early and late life.Here,we summarize emerging links between the most common ...Evidence from genetics and from analyzing cellular and animal models have converged to suggest links between neurodegenerative disorders of early and late life.Here,we summarize emerging links between the most common late life neurodegenerative disease,Alzheimer’s disease,and the most common early life neurodegenerative diseases,neuronal ceroid lipofuscinoses.Genetic studies reported an overlap of clinically diagnosed Alzheimer’s disease and mutations in genes known to cause neuronal ceroid lipofuscinoses.Accumulating data strongly suggest dysfunction of intracellular trafficking mechanisms and the autophagy-endolysosome system in both types of neurodegenerative disorders.This suggests shared cytopathological processes underlying these different types of neurodegenerative diseases.A better understanding of the common mechanisms underlying the different diseases is important as this might lead to the identification of novel targets for therapeutic concepts,the transfer of therapeutic strategies from one disease to the other and therapeutic approaches tailored to patients with specific mutations.Here,we review dysfunctions of the endolysosomal autophagy pathway in Alzheimer’s disease and neuronal ceroid lipofuscinoses and summarize emerging etiologic and genetic overlaps.展开更多
Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural pro...Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural profiles. The gene underlying BD is designated CLN3 and encodes a protein, Battenin, of unknown function that localizes in lysosomes and/or mitochondria. Previously, we hypothesized that Battenin associates with other membrane protein（s） to form a membrane complex. Dysfunction of this complex could result in the pathological changes of BD, and possibly in other NCLs. Two such membranous proteins, the slow and fast Battenin-interactive proteins （BIPs and BIPf） of unknown functions, have been identified. In this study, we have characterized the functional domains of Battenin that interact with both BIP proteins. Methods: Protein-protein interactions with a yeast two-hybrid system were employed. A “deletion assay” was employed to localize the interactive segment（s）. Different lengths of cDNA sequences lacking exon 1-5 were used to express CLN3-encoded proteins lacking N-terminal segments in the yeast two-hybrid system. N-terminal exons of CLN3 were deleted with PCR-cloning strategies.Results: We eliminated the possibility of interacting domains from the exon 7-encoded region because both Battenin and mBattenin interact with the BIP proteins. We have shown that peptide sequences encoded by exons 2 and 4 of CLN3 gene include the functional domains by which Battenin interacts with the BIP proteins. Conclusion: Our studies provide evidence that the N-terminus of Battenin is the functional domain for these protein interactions.展开更多
文摘目的探讨CLN3(青少年神经元腊样质脂褐质沉积症Juvenile,form of neuronalceroid-lipofuscinosis,JNCL,Batten病)基因在卵巢浆液性囊腺癌中的表达特点及其在卵巢癌化疗耐药中的作用。方法采用半定量RT-PCR、Western blot以及免疫组化的方法检测CLN3在卵巢浆液性囊腺癌中的表达,分析其表达与临床病理特征及化疗耐药的相关性。结果 CLN3在卵巢浆液性囊腺癌中的表达高于正常卵巢组织,且临床分期越晚其表达越高(P<0.05),而与肿瘤患者年龄无显著相关性。CLN3在卵巢癌化疗耐药组表达阳性率明显高于化疗敏感组(P<0.05)。结论卵巢癌中CLN3高表达与卵巢癌的发生、发展、浸润和转移有关,与卵巢癌化疗耐药密切相关。CLN3可成为卵巢癌化疗新的基因治疗靶点。
文摘Lysosomes have many roles,including degrading macromolecules and signalling to the nucleus1.Lysosomal dysfunction occurs in various human conditions,such as common neurodegenerative diseases and monogenic lysosomal storage disorders(LSDs)2-4.For most LSDs,the causal genes have been identified but,in some,the function of the implicated gene is unknown,in part because lysosomes occupy a small fraction of the cellular volume so that changes in lysosomal contents are difficult to detect.Here we develop the LysoTag mouse for the tissue-specific isolation of intact lysosomes that are compatible with the multimodal profiling of their contents.We used the LysoTag mouse to study CLN3,a lysosomal transmembrane protein with an unknown function.In children,the loss of CLN3 causes juvenile neuronal ceroid lipofuscinosis(Batten disease),a lethal neurodegenerative LSD.Untargeted metabolite profiling of lysosomes from the brains of mice lacking CLN3 revealed a massive accumulation of glycerophosphodiesters(GPDs)-the end products of glycerophospholipid catabolism.GPDs also accumulate in the lysosomes of CLN3-deficient cultured cells and we show that CLN3 is required for their lysosomal egress.Loss of CLN3 also disrupts glycerophospholipid catabolism in the lysosome.Finally,we found elevated levels of glycerophosphoinositol in the cerebrospinal fluid of patients with Batten disease,suggesting the potential use of glycerophosphoinositol as a disease biomarker.Our results show that CLN3 is required for the lysosomal clearance of GPDs and reveal Batten disease as a neurodegenerative LSD with a defect in glycerophospholipid metabolism.
文摘CLN3 is a lysosomal transmembrane protein and loss of CLN3 is known to cause a juvenile lethal neurodegenerative lysosomal storage disorder(LSD),called Batten disease.In a recent study published in Nature,Laqtom et al.reported a novel function of CLN3 in the clearance of glycerophospholipid from lysosomes via lysosomal efflux of glycerophosphodiesters(GPDs),not only establishing a deeper mechanistic understanding of Batten disease,but also suggesting both the diagnostic and therapeutic potential of CLN3-GPDs in this type of neurodegenerative LSD.
基金supported by the Deutsche Forschungsgemeinschaft(DFG,425373668,HE 3220/4-1)(to GH).
文摘Evidence from genetics and from analyzing cellular and animal models have converged to suggest links between neurodegenerative disorders of early and late life.Here,we summarize emerging links between the most common late life neurodegenerative disease,Alzheimer’s disease,and the most common early life neurodegenerative diseases,neuronal ceroid lipofuscinoses.Genetic studies reported an overlap of clinically diagnosed Alzheimer’s disease and mutations in genes known to cause neuronal ceroid lipofuscinoses.Accumulating data strongly suggest dysfunction of intracellular trafficking mechanisms and the autophagy-endolysosome system in both types of neurodegenerative disorders.This suggests shared cytopathological processes underlying these different types of neurodegenerative diseases.A better understanding of the common mechanisms underlying the different diseases is important as this might lead to the identification of novel targets for therapeutic concepts,the transfer of therapeutic strategies from one disease to the other and therapeutic approaches tailored to patients with specific mutations.Here,we review dysfunctions of the endolysosomal autophagy pathway in Alzheimer’s disease and neuronal ceroid lipofuscinoses and summarize emerging etiologic and genetic overlaps.
文摘Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural profiles. The gene underlying BD is designated CLN3 and encodes a protein, Battenin, of unknown function that localizes in lysosomes and/or mitochondria. Previously, we hypothesized that Battenin associates with other membrane protein（s） to form a membrane complex. Dysfunction of this complex could result in the pathological changes of BD, and possibly in other NCLs. Two such membranous proteins, the slow and fast Battenin-interactive proteins （BIPs and BIPf） of unknown functions, have been identified. In this study, we have characterized the functional domains of Battenin that interact with both BIP proteins. Methods: Protein-protein interactions with a yeast two-hybrid system were employed. A “deletion assay” was employed to localize the interactive segment（s）. Different lengths of cDNA sequences lacking exon 1-5 were used to express CLN3-encoded proteins lacking N-terminal segments in the yeast two-hybrid system. N-terminal exons of CLN3 were deleted with PCR-cloning strategies.Results: We eliminated the possibility of interacting domains from the exon 7-encoded region because both Battenin and mBattenin interact with the BIP proteins. We have shown that peptide sequences encoded by exons 2 and 4 of CLN3 gene include the functional domains by which Battenin interacts with the BIP proteins. Conclusion: Our studies provide evidence that the N-terminus of Battenin is the functional domain for these protein interactions.
