Our previous study has shown that the Autographa californica multiple nucleopolyhedrovirus(AcMNPV) p48(ac103)gene is essential for the nuclear egress of nucleocapsids and the formation of occlusion-derived virions(ODV...Our previous study has shown that the Autographa californica multiple nucleopolyhedrovirus(AcMNPV) p48(ac103)gene is essential for the nuclear egress of nucleocapsids and the formation of occlusion-derived virions(ODVs). However,the exact role of p48 in the morphogenesis of ODVs remains unknown. In this study, we demonstrated that p48 was required for the efficient formation of intranuclear microvesicles. To further understand its functional role in intranuclear microvesicle formation, we characterized the distribution of the P48 protein, which was found to be associated with the nucleocapsid and envelope fractions of both budded virions and ODVs. In Ac MNPV-infected cells, P48 was predominantly localized to nucleocapsids in the virogenic stroma and the nucleocapsids enveloped in ODVs, with a limited but discernible distribution in the plasma membrane, nuclear envelope, intranuclear microvesicles, and ODV envelope. Furthermore,coimmunoprecipitation assays showed that among the viral proteins required for intranuclear microvesicle formation, P48 associated with Ac93 in the absence of viral infection.展开更多
BACKGROUND Neuronal intranuclear inclusion disease(NIID)is a rare neurological degenerative disorder with diverse manifestations and inadequate awareness.Only a few cases of NIID have been reported,and typical imaging...BACKGROUND Neuronal intranuclear inclusion disease(NIID)is a rare neurological degenerative disorder with diverse manifestations and inadequate awareness.Only a few cases of NIID have been reported,and typical imaging findings can provide certain clues for the diagnosis of the disease.Furthermore,skin biopsy and genetic testing are important to confirm the diagnosis.CASE SUMMARY An 84-year-old man presented to the Neurology Department of our hospital complaining of a progressive course of cognitive impairment and unsteady gait for 2 years.The symptoms gradually progressed and affected his daily life.The patient was initially diagnosed with Parkinson’s disease and vascular dementia.The patient did not respond to conventional treatment,such as dopasehydrazine.Therefore,magnetic resonance imaging(MRI)was performed.Based on the imaging findings,we suspected an NIID diagnosis.During the 3-year follow-up in our hospital,his clinical symptoms gradually progressed,and imaging findings became more significant.A high signal intensity along the corticomedullary junction persisted on MRI.Gene testing and skin biopsy were recommended in our hospital;however,the patient refused these procedures.NIID was also considered when he went to a superior hospital in Shanghai.The patient eventually agreed to undergo gene testing.This revealed abnormal GGC repeat expansions in the NOTCH2NLC gene.CONCLUSION The clinical manifestations of NIID are diverse.Patients with clinical manifestations similar to Parkinson’s disease and dementia may have NIID.展开更多
BACKGROUND Neuronal intranuclear inclusion disease(NIID)is an unusual autosomal dominant,chronic progressive neurodegenerative disease.The clinical manifestations of NIID are complex and varied,complicating its clinic...BACKGROUND Neuronal intranuclear inclusion disease(NIID)is an unusual autosomal dominant,chronic progressive neurodegenerative disease.The clinical manifestations of NIID are complex and varied,complicating its clinical diagnosis.To the best of our knowledge,this report is the first to document sporadic adult-onset NIID mimicking acute cerebellitis(AC)that was finally diagnosed by imaging studies,skin biopsy,and genetic testing.CASE SUMMARY A 63-year-old man presented with fever,gait unsteadiness,dysarthria,and an episode of convulsion.His serum levels of white blood cells and C-reactive protein were significantly elevated.T2-weighted brain magnetic resonance imaging and fluid attenuation inversion recovery sequences showed bilateral high-intensity signals in the medial part of the cerebellar hemisphere beside the vermis.While we initially considered a diagnosis of AC,the patient’s symptoms improved significantly without special treatment,prompting our consideration of NIID.Diffusion-weighted imaging showed hyperintensity in the corticomedullary junction.Skin biopsy revealed eosinophilic inclusions positive for anti-p62 in epithelial sweat-gland cells.GGC repeat expansions in the Notch 2 N-terminal like C gene confirmed the diagnosis of NIID.CONCLUSION For patients with clinical manifestations mimicking AC,the possibility of underlying NIID should be considered along with prompt rigorous examinations.展开更多
ObjectiveeToinvestigate theclinical and neuroelectrophysiological characteristics of NOTCH2NLC gene-related neuronal intranuclear inclusion disease(NIID).Methods One hundred and forty patients with NOTCH2NLCC gene-rel...ObjectiveeToinvestigate theclinical and neuroelectrophysiological characteristics of NOTCH2NLC gene-related neuronal intranuclear inclusion disease(NIID).Methods One hundred and forty patients with NOTCH2NLCC gene-related NIID diagnosed in the Department of Neurology and Department of Geriatrics,Xiangya Hospital,Central South University from January 2018 to June 2024 were selected as the research subjects.Their clinical data as well as neuroelectrophysiological results were collected.Their clinical and neuroelectrophysiological characteristics were summarized.Results The onset age of 140 patients with NOTCH2NLC gene-related NIID was 56.00(45.25,62.75)years.Among them,55.0%(77/140)of patients with NIID presented with peripheral nerve symptoms,but up to 98.6%(138/140)of patients with NIID had peripheral nerve involvement.Out of the patients studied,97.1%(136/140)exhibited a reduction in motor nerve conduction velocity and 66.4%(93/140)showed a decreasee in sensory nerve conductionnvelocity.Furthermore,53.6%(75/140)of patients had mild decrease in compound muscle action potential,and 55.7%(78/140)of patients showed mild reduction in sensory nerve action potential.Motor nerve involvement was more severe than sensory nerve impairment,and lower limb involvement was more severe than upper limb involvement.The nerve conduction abnormalities in the muscle weakness type(n=32)of NIID patients were more severe than those in the non-muscle weakness type(cognitive impairment type,n=41;movement disorder type,n=43;paroxysmal symptom type,n=24),showing mixed demyelinating and axonal sensorimotor neuropathy,while the non-muscle weakness type of NID patients mostly showed mild demyelinating sensorimotor neuropathy.There was no significant difference in nerve conduction related electrophysiological results among the patientswith3 non-muscleweaknessphenotypes.Conclusion Peripheral neuropathy is common in NIID patients.The neuroelectrophysiological characteristics of NIID patients include slight demyelinating sensorimotor neuropathy,and some of NIID patients are also accompanied bymildaxonaldamage.Neuroelectrophysiological evaluation is helpful for the diagnosis of NIID.展开更多
Here we report a case of a 67-year-old female patient who presented with headache,limb tremors,and acute complete vision loss.Physical examination revealed bilateral miosis,and diffusion-weighted imaging sequences sho...Here we report a case of a 67-year-old female patient who presented with headache,limb tremors,and acute complete vision loss.Physical examination revealed bilateral miosis,and diffusion-weighted imaging sequences showed mild diffusion restriction in the subcortical regions of both occipital lobes.Genetic results revealed 85 GGC repeats in the 50-untranslated region of the NOTCH2NLC gene.The therapeutic effect of dexamethasone and acyclovir was minimal.NIID must be considered in patients with acute onset and various clinical manifestations and imagingfindings similar to encephalitis.We hope that our case presentation will enhance clinicians’awareness of NIID.展开更多
In the past studies,it is shown that cardiac troponin I(cTnI,encoded by TNNI3),as a cytoplasmic protein,is an inhibitory subunit in troponin complex,and involves in cardiomyocyte diastolic regulation.Here,we assessed ...In the past studies,it is shown that cardiac troponin I(cTnI,encoded by TNNI3),as a cytoplasmic protein,is an inhibitory subunit in troponin complex,and involves in cardiomyocyte diastolic regulation.Here,we assessed a novel role of cTnI as a nucleoprotein.Firstly,the nuclear translocation of cTnI was found in mouse,human fetuses and rat heart tissues.In addition,there were differences in percentage of intranuclear cTnI in different conditions.Based on weighted gene co-expression network analyses(WGCNA)and verification in cell experiments,a strong expression correlation was found between TNNI3 and Atp2a2,which encodes sarco-endoplasmic reticulum Ca2t ATPase isoform 2a(SERCA2a),and involves in ATP hydrolysis and Ca2t transient.TNNI3 gain and loss caused Atpa2a2 increase/decrease in a dosedependent manner both in mRNA and protein levels,in vivo and in vitro.By using ChIP-sequence we demonstrated specific binding DNA sequences of cTnI were enriched in ATP2a2 promoter239we889 region and the specific binding sequence motif of cTnI was analyzed by software as"CCAT",which has been reported to be required for YY1 binding to the promoter region of YY1-related genes.Moreover,it was further verified that pcDNA3.1()-TNNI3 could express cTnI proteins and increase the promoter activity of Atp2a2 through luciferase report assay.In the end,we evaluated beat frequencies,total ATP contents,Ca2t transients in TNNI3-siRNA myocardial cells.These findings indicated,for the first time,cTnI may regulate Atp2a2 in cardiomyocytes as a co-regulatory factor and participate in the regulation of intracellular Ca ions.展开更多
The nucleolus,the locus of ribosome biogenesis,was found to be the predominant intracellular target of a new fluorescent probe,V-P1.In solution,the probe demonstrated both a selectivity to RNA G-quadruplexes and a sen...The nucleolus,the locus of ribosome biogenesis,was found to be the predominant intracellular target of a new fluorescent probe,V-P1.In solution,the probe demonstrated both a selectivity to RNA G-quadruplexes and a sensitivity to the viscosity,while G-quadruplex binding did not disturb the viscosity sensing.In cells,confocal and fluorescence lifetime imaging,combined with digestion and competition experiments,lent support to the hypothesis of an RNA-based G-quadruplex as the intracellular target,postulated to be nucleolar ribosomal RNA(rRNA).The probe demonstrated a high sensitivity to viscosity in both the cytoplasm and the nuclear compartment and was used to precisely interrogate the viscosity changes resulting from diverse stimuli,such as temperature,monensin treatment,and etoposide-induced apoptosis.Owing to the putative rRNA G-quadruplex binding in vitro and in vivo,and further combined with a relatively low degree of toxicity,the dye enabled the interrogation of cytoplasm and intranuclear viscosity changes under diverse conditions and found applications in studying the influence and significance of cytoplasm and intranuclear viscosity as well as in gaining insight into the native secondary structure of rRNA in nucleoli.展开更多
Lack of the fragile X mental retardation protein leads to Fragile X syndrome(FXS)while increased levels of FMR1 mRNA,as those observed in premutation carriers can lead to Fragile X-associated tremor ataxia syndrome(FX...Lack of the fragile X mental retardation protein leads to Fragile X syndrome(FXS)while increased levels of FMR1 mRNA,as those observed in premutation carriers can lead to Fragile X-associated tremor ataxia syndrome(FXTAS).Until recently,FXTAS had been observed only in carriers of an FMR1 premutation(55–200 CGG repeats);however the disorder has now been described in individuals carriers of an intermediate allele(45–54 CGG repeats)as well as in a subject with a full mutation with mosaicism.Here,we report on molecular and clinical data of a male FMR1 mosaic individual with full and premutation alleles.Molecular analysis of FMR1 and FMRP expression in this subject is consistent with a FXS phenotype.We observed reduced expression of FMRP in both peripheral blood and brain leading to the FXS diagnosis.In addition,a dramatic 90%depletion of both FMR1 mRNA and FMRP levels was observed in the blood,as normally observed in FXS cases,and an even greater depletion in the brain.A clinical report of this patient,at age 71,described neurodegenerative signs of parkinsonism that were likely,in retrospect,part of a FXTAS scenario as post-mortem examination shows the presence of intranuclear inclusions,the hallmark pathology of FXTAS.The findings presented in this study indicate co-morbidity for both FXS and FXTAS in this individual carrying both full and premutation FMR1 alleles.In addition,based on symptoms and pathological and molecular evidence,this report suggests the need to redefine the diagnostic criteria of FXTAS.展开更多
Aim:β-catenin activation is known to promote liver regeneration and play a role in the pathogenesis of liver cancer.Recently,we detected intranuclear inclusions(NI)in hepatocellular carcinoma(HCC)containing degenerat...Aim:β-catenin activation is known to promote liver regeneration and play a role in the pathogenesis of liver cancer.Recently,we detected intranuclear inclusions(NI)in hepatocellular carcinoma(HCC)containing degenerated cell organelles and lysosomal proteins and delimited by a completely closed nuclear membrane.The presence of NI was positively associated with patient survival.The aim of the current study was to investigate a possible association between proteins of the Wnt/β-catenin pathway with NI morphology and survival.Methods:We examined NI in 72 paraffin-embedded specimens of HCC.Immunohistochemistry(IHC)and immunofluorescence(IF)were performed to investigate the content and shape of NI.β-catenin gene(CTNNB1)mutations were analyzed by next generation sequencing.Results:We detected the accumulation ofβ-catenin and glutamine synthetase(a target gene ofβ-catenin)proteins within NI.Further,we found immunopositivity for the lysine demethylase KDM2A in NI.KDM2A is known to be involved inβ-catenin degradation.We detected significant associations between the presence ofβ-catenin and autophagy-associated proteins in NI.Double-IF revealed co-localization ofβ-catenin and p62 in the same NI.Kaplan-Meier survival analysis showed that the presence of NI containing KDM2A protein accumulations displayed a significant benefit in overall survival.Conclusion:We detected accumulations ofβ-catenin and proteins associated with the Wnt/β-catenin pathway partly together with autophagy-associated proteins in the same inclusion.Our finding that KDM2A immunopositivity within NIs was associated with favorable clinical outcomes and suggests a biological significance of NI.展开更多
基金supported by the National Natural Science Foundation of China (31572056 and 31872025)the Key Project of Natural Science Foundation of Guangdong Province (2018B030311018)+1 种基金the National Key R&D Program of China (2017YFD0200404)the Guangzhou Science and Technology Project (201707020003)
文摘Our previous study has shown that the Autographa californica multiple nucleopolyhedrovirus(AcMNPV) p48(ac103)gene is essential for the nuclear egress of nucleocapsids and the formation of occlusion-derived virions(ODVs). However,the exact role of p48 in the morphogenesis of ODVs remains unknown. In this study, we demonstrated that p48 was required for the efficient formation of intranuclear microvesicles. To further understand its functional role in intranuclear microvesicle formation, we characterized the distribution of the P48 protein, which was found to be associated with the nucleocapsid and envelope fractions of both budded virions and ODVs. In Ac MNPV-infected cells, P48 was predominantly localized to nucleocapsids in the virogenic stroma and the nucleocapsids enveloped in ODVs, with a limited but discernible distribution in the plasma membrane, nuclear envelope, intranuclear microvesicles, and ODV envelope. Furthermore,coimmunoprecipitation assays showed that among the viral proteins required for intranuclear microvesicle formation, P48 associated with Ac93 in the absence of viral infection.
文摘BACKGROUND Neuronal intranuclear inclusion disease(NIID)is a rare neurological degenerative disorder with diverse manifestations and inadequate awareness.Only a few cases of NIID have been reported,and typical imaging findings can provide certain clues for the diagnosis of the disease.Furthermore,skin biopsy and genetic testing are important to confirm the diagnosis.CASE SUMMARY An 84-year-old man presented to the Neurology Department of our hospital complaining of a progressive course of cognitive impairment and unsteady gait for 2 years.The symptoms gradually progressed and affected his daily life.The patient was initially diagnosed with Parkinson’s disease and vascular dementia.The patient did not respond to conventional treatment,such as dopasehydrazine.Therefore,magnetic resonance imaging(MRI)was performed.Based on the imaging findings,we suspected an NIID diagnosis.During the 3-year follow-up in our hospital,his clinical symptoms gradually progressed,and imaging findings became more significant.A high signal intensity along the corticomedullary junction persisted on MRI.Gene testing and skin biopsy were recommended in our hospital;however,the patient refused these procedures.NIID was also considered when he went to a superior hospital in Shanghai.The patient eventually agreed to undergo gene testing.This revealed abnormal GGC repeat expansions in the NOTCH2NLC gene.CONCLUSION The clinical manifestations of NIID are diverse.Patients with clinical manifestations similar to Parkinson’s disease and dementia may have NIID.
基金Supported by Project of Science and Technology Department of Jilin Province,China,No.20190303181 SF.
文摘BACKGROUND Neuronal intranuclear inclusion disease(NIID)is an unusual autosomal dominant,chronic progressive neurodegenerative disease.The clinical manifestations of NIID are complex and varied,complicating its clinical diagnosis.To the best of our knowledge,this report is the first to document sporadic adult-onset NIID mimicking acute cerebellitis(AC)that was finally diagnosed by imaging studies,skin biopsy,and genetic testing.CASE SUMMARY A 63-year-old man presented with fever,gait unsteadiness,dysarthria,and an episode of convulsion.His serum levels of white blood cells and C-reactive protein were significantly elevated.T2-weighted brain magnetic resonance imaging and fluid attenuation inversion recovery sequences showed bilateral high-intensity signals in the medial part of the cerebellar hemisphere beside the vermis.While we initially considered a diagnosis of AC,the patient’s symptoms improved significantly without special treatment,prompting our consideration of NIID.Diffusion-weighted imaging showed hyperintensity in the corticomedullary junction.Skin biopsy revealed eosinophilic inclusions positive for anti-p62 in epithelial sweat-gland cells.GGC repeat expansions in the Notch 2 N-terminal like C gene confirmed the diagnosis of NIID.CONCLUSION For patients with clinical manifestations mimicking AC,the possibility of underlying NIID should be considered along with prompt rigorous examinations.
文摘ObjectiveeToinvestigate theclinical and neuroelectrophysiological characteristics of NOTCH2NLC gene-related neuronal intranuclear inclusion disease(NIID).Methods One hundred and forty patients with NOTCH2NLCC gene-related NIID diagnosed in the Department of Neurology and Department of Geriatrics,Xiangya Hospital,Central South University from January 2018 to June 2024 were selected as the research subjects.Their clinical data as well as neuroelectrophysiological results were collected.Their clinical and neuroelectrophysiological characteristics were summarized.Results The onset age of 140 patients with NOTCH2NLC gene-related NIID was 56.00(45.25,62.75)years.Among them,55.0%(77/140)of patients with NIID presented with peripheral nerve symptoms,but up to 98.6%(138/140)of patients with NIID had peripheral nerve involvement.Out of the patients studied,97.1%(136/140)exhibited a reduction in motor nerve conduction velocity and 66.4%(93/140)showed a decreasee in sensory nerve conductionnvelocity.Furthermore,53.6%(75/140)of patients had mild decrease in compound muscle action potential,and 55.7%(78/140)of patients showed mild reduction in sensory nerve action potential.Motor nerve involvement was more severe than sensory nerve impairment,and lower limb involvement was more severe than upper limb involvement.The nerve conduction abnormalities in the muscle weakness type(n=32)of NIID patients were more severe than those in the non-muscle weakness type(cognitive impairment type,n=41;movement disorder type,n=43;paroxysmal symptom type,n=24),showing mixed demyelinating and axonal sensorimotor neuropathy,while the non-muscle weakness type of NID patients mostly showed mild demyelinating sensorimotor neuropathy.There was no significant difference in nerve conduction related electrophysiological results among the patientswith3 non-muscleweaknessphenotypes.Conclusion Peripheral neuropathy is common in NIID patients.The neuroelectrophysiological characteristics of NIID patients include slight demyelinating sensorimotor neuropathy,and some of NIID patients are also accompanied bymildaxonaldamage.Neuroelectrophysiological evaluation is helpful for the diagnosis of NIID.
文摘Here we report a case of a 67-year-old female patient who presented with headache,limb tremors,and acute complete vision loss.Physical examination revealed bilateral miosis,and diffusion-weighted imaging sequences showed mild diffusion restriction in the subcortical regions of both occipital lobes.Genetic results revealed 85 GGC repeats in the 50-untranslated region of the NOTCH2NLC gene.The therapeutic effect of dexamethasone and acyclovir was minimal.NIID must be considered in patients with acute onset and various clinical manifestations and imagingfindings similar to encephalitis.We hope that our case presentation will enhance clinicians’awareness of NIID.
基金This study was supported by research grants from Natural Science Foundation of China(No.81700214,and 81670212).
文摘In the past studies,it is shown that cardiac troponin I(cTnI,encoded by TNNI3),as a cytoplasmic protein,is an inhibitory subunit in troponin complex,and involves in cardiomyocyte diastolic regulation.Here,we assessed a novel role of cTnI as a nucleoprotein.Firstly,the nuclear translocation of cTnI was found in mouse,human fetuses and rat heart tissues.In addition,there were differences in percentage of intranuclear cTnI in different conditions.Based on weighted gene co-expression network analyses(WGCNA)and verification in cell experiments,a strong expression correlation was found between TNNI3 and Atp2a2,which encodes sarco-endoplasmic reticulum Ca2t ATPase isoform 2a(SERCA2a),and involves in ATP hydrolysis and Ca2t transient.TNNI3 gain and loss caused Atpa2a2 increase/decrease in a dosedependent manner both in mRNA and protein levels,in vivo and in vitro.By using ChIP-sequence we demonstrated specific binding DNA sequences of cTnI were enriched in ATP2a2 promoter239we889 region and the specific binding sequence motif of cTnI was analyzed by software as"CCAT",which has been reported to be required for YY1 binding to the promoter region of YY1-related genes.Moreover,it was further verified that pcDNA3.1()-TNNI3 could express cTnI proteins and increase the promoter activity of Atp2a2 through luciferase report assay.In the end,we evaluated beat frequencies,total ATP contents,Ca2t transients in TNNI3-siRNA myocardial cells.These findings indicated,for the first time,cTnI may regulate Atp2a2 in cardiomyocytes as a co-regulatory factor and participate in the regulation of intracellular Ca ions.
基金supported by CRI project(no.2018R1A3B1052702J.S.K.)from the National Research Foundation of Korea(NRF)+3 种基金by the China Scholarship Fund(CSC no.201907030009L.Y.)by the Interne Fondsen KU Leuven/Internal Funds KU Leuven(STG/19/029P.V.).
文摘The nucleolus,the locus of ribosome biogenesis,was found to be the predominant intracellular target of a new fluorescent probe,V-P1.In solution,the probe demonstrated both a selectivity to RNA G-quadruplexes and a sensitivity to the viscosity,while G-quadruplex binding did not disturb the viscosity sensing.In cells,confocal and fluorescence lifetime imaging,combined with digestion and competition experiments,lent support to the hypothesis of an RNA-based G-quadruplex as the intracellular target,postulated to be nucleolar ribosomal RNA(rRNA).The probe demonstrated a high sensitivity to viscosity in both the cytoplasm and the nuclear compartment and was used to precisely interrogate the viscosity changes resulting from diverse stimuli,such as temperature,monensin treatment,and etoposide-induced apoptosis.Owing to the putative rRNA G-quadruplex binding in vitro and in vivo,and further combined with a relatively low degree of toxicity,the dye enabled the interrogation of cytoplasm and intranuclear viscosity changes under diverse conditions and found applications in studying the influence and significance of cytoplasm and intranuclear viscosity as well as in gaining insight into the native secondary structure of rRNA in nucleoli.
基金This work was supported by NICH through individual research awards HD02274 and HD36071Brain tissue for this study was obtained from the UC Davis brain repository(NIH HD040661)Control tissues were obtained from the Brain Endowment Bank,Dept.Neurology University of Miami Miller School of Medicine.
文摘Lack of the fragile X mental retardation protein leads to Fragile X syndrome(FXS)while increased levels of FMR1 mRNA,as those observed in premutation carriers can lead to Fragile X-associated tremor ataxia syndrome(FXTAS).Until recently,FXTAS had been observed only in carriers of an FMR1 premutation(55–200 CGG repeats);however the disorder has now been described in individuals carriers of an intermediate allele(45–54 CGG repeats)as well as in a subject with a full mutation with mosaicism.Here,we report on molecular and clinical data of a male FMR1 mosaic individual with full and premutation alleles.Molecular analysis of FMR1 and FMRP expression in this subject is consistent with a FXS phenotype.We observed reduced expression of FMRP in both peripheral blood and brain leading to the FXS diagnosis.In addition,a dramatic 90%depletion of both FMR1 mRNA and FMRP levels was observed in the blood,as normally observed in FXS cases,and an even greater depletion in the brain.A clinical report of this patient,at age 71,described neurodegenerative signs of parkinsonism that were likely,in retrospect,part of a FXTAS scenario as post-mortem examination shows the presence of intranuclear inclusions,the hallmark pathology of FXTAS.The findings presented in this study indicate co-morbidity for both FXS and FXTAS in this individual carrying both full and premutation FMR1 alleles.In addition,based on symptoms and pathological and molecular evidence,this report suggests the need to redefine the diagnostic criteria of FXTAS.
文摘Aim:β-catenin activation is known to promote liver regeneration and play a role in the pathogenesis of liver cancer.Recently,we detected intranuclear inclusions(NI)in hepatocellular carcinoma(HCC)containing degenerated cell organelles and lysosomal proteins and delimited by a completely closed nuclear membrane.The presence of NI was positively associated with patient survival.The aim of the current study was to investigate a possible association between proteins of the Wnt/β-catenin pathway with NI morphology and survival.Methods:We examined NI in 72 paraffin-embedded specimens of HCC.Immunohistochemistry(IHC)and immunofluorescence(IF)were performed to investigate the content and shape of NI.β-catenin gene(CTNNB1)mutations were analyzed by next generation sequencing.Results:We detected the accumulation ofβ-catenin and glutamine synthetase(a target gene ofβ-catenin)proteins within NI.Further,we found immunopositivity for the lysine demethylase KDM2A in NI.KDM2A is known to be involved inβ-catenin degradation.We detected significant associations between the presence ofβ-catenin and autophagy-associated proteins in NI.Double-IF revealed co-localization ofβ-catenin and p62 in the same NI.Kaplan-Meier survival analysis showed that the presence of NI containing KDM2A protein accumulations displayed a significant benefit in overall survival.Conclusion:We detected accumulations ofβ-catenin and proteins associated with the Wnt/β-catenin pathway partly together with autophagy-associated proteins in the same inclusion.Our finding that KDM2A immunopositivity within NIs was associated with favorable clinical outcomes and suggests a biological significance of NI.
