Pub Date : 2021-09-01DOI: 10.1136/jnnp-2021-ehdn.127
M. Geva, M. Shenkman, Luana Naia, Noga Gershoni Emek, P. Ly, S. Mota, Carla Lopes, M. Ankarcrona, A. Rego, Gerardo Z Lederkreme, M. Hayden
Background Pridopidine is a highly selective, potent Sigma-1 receptor (S1R) agonist in clinical development for HD and ALS. The S1R is a protein enriched at the endoplasmic reticulum (ER)-mitochondria interface and vital to multiple cellular mechanisms, including mitochondrial function and the ER stress response. By activating the S1R, pridopidine exerts neuroprotective effects in many models of neurodegenerative diseases, including HD. In HD neurons, abnormal ER and mitochondria function increase susceptibility to oxidative stress, causing cell death. Aims Assess the effects of pridopidine on mitochondrial function and ER stress in HD models. Methods In neurons from YAC128 HD model mice, mitochondria and ER structure were assessed., Mitochondrial function was assessed by measuring respiration, ATP production, membrane potential and reactive oxygen species. Motor and mitochondrial function was assessed in vivo in YAC128 mice. ER stress was assessed by measuring levels of proteins involved in the stress response in HEK293 cells expressing normal or mutant Htt. Results Pridopidine restores mitochondrial and ER structure and connectivity (p Pridopidine reduces levels of phosphorylated protein eIF2α (p Conclusion The protective effects of pridopidine are facilitated by S1R-mediated rescue of mitochondrial function and ER stress pathway, both disrupted in HD. These findings shed new light on pridopidine’s mechanism of action.
{"title":"I13 Pridopidine restores mitochondrial function and decreases er stress which is mediated through the S1R","authors":"M. Geva, M. Shenkman, Luana Naia, Noga Gershoni Emek, P. Ly, S. Mota, Carla Lopes, M. Ankarcrona, A. Rego, Gerardo Z Lederkreme, M. Hayden","doi":"10.1136/jnnp-2021-ehdn.127","DOIUrl":"https://doi.org/10.1136/jnnp-2021-ehdn.127","url":null,"abstract":"Background Pridopidine is a highly selective, potent Sigma-1 receptor (S1R) agonist in clinical development for HD and ALS. The S1R is a protein enriched at the endoplasmic reticulum (ER)-mitochondria interface and vital to multiple cellular mechanisms, including mitochondrial function and the ER stress response. By activating the S1R, pridopidine exerts neuroprotective effects in many models of neurodegenerative diseases, including HD. In HD neurons, abnormal ER and mitochondria function increase susceptibility to oxidative stress, causing cell death. Aims Assess the effects of pridopidine on mitochondrial function and ER stress in HD models. Methods In neurons from YAC128 HD model mice, mitochondria and ER structure were assessed., Mitochondrial function was assessed by measuring respiration, ATP production, membrane potential and reactive oxygen species. Motor and mitochondrial function was assessed in vivo in YAC128 mice. ER stress was assessed by measuring levels of proteins involved in the stress response in HEK293 cells expressing normal or mutant Htt. Results Pridopidine restores mitochondrial and ER structure and connectivity (p Pridopidine reduces levels of phosphorylated protein eIF2α (p Conclusion The protective effects of pridopidine are facilitated by S1R-mediated rescue of mitochondrial function and ER stress pathway, both disrupted in HD. These findings shed new light on pridopidine’s mechanism of action.","PeriodicalId":444837,"journal":{"name":"I: Experimental therapeutics – preclinical","volume":"133 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122806852","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-09-01DOI: 10.1136/jnnp-2021-ehdn.115
A. Bhattacharyya, Effenberger Kerstin, C. Trotta, J. Narasimhan, Wencheng Li, G. WollMatthew, B. JaniMinakshi, N. Risher, Shirley Yeh, Yaofeng Cheng, N. Sydorenko, M. Weetall, A. Southwell, Michael R. Hayden, J. Colacino, S. Peltz
{"title":"I01 Orally bioavailable small molecule splicing modifiers with systemic and even htt-lowering activity in vitro and in vivo","authors":"A. Bhattacharyya, Effenberger Kerstin, C. Trotta, J. Narasimhan, Wencheng Li, G. WollMatthew, B. JaniMinakshi, N. Risher, Shirley Yeh, Yaofeng Cheng, N. Sydorenko, M. Weetall, A. Southwell, Michael R. Hayden, J. Colacino, S. Peltz","doi":"10.1136/jnnp-2021-ehdn.115","DOIUrl":"https://doi.org/10.1136/jnnp-2021-ehdn.115","url":null,"abstract":"","PeriodicalId":444837,"journal":{"name":"I: Experimental therapeutics – preclinical","volume":"69 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132637667","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-09-01DOI: 10.1136/jnnp-2021-ehdn.118
P. Bellosta, Stefania Santarelli, Chiara Londero
Autophagy is a fundamental cellular pathway involved in the clearance of protein aggregates, and it is particularly important in neurons. The toxic aggregates derived from the mutated Huntingtin have been shown to interfere with the physiological autophagic flux, resulting in neuronal death. Glutamate Dehydrogenase (GDH) is an evolutionary conserved enzyme that catalyses the conversion of glutamate and ammonia to α-ketoglutarate and vice versa and is also member of the Glutamate-Glutamine Cycle (GGC), a physiological process between glia and neurons that controls glutamate homeostasis. Through a genetic screen using a Drosophila model for Huntington’s disease (HD), we identified that reduction of GDH ameliorates animal motility and decreases the size of mutated Huntingtin’s (mHTT) aggregates in brains. The aim of our project is to analyze how GDH downregulation induces autophagy in neurons. We modeled HD phenotype in Drosophila by expressing mHTT with 93-CAG repetition (HTTQ93) in neurons. To investigate the effect of GDH we used motility and viability assay, while western blots and immunofluorescence analysis were used to investigate changes in mHTT aggregates. We found the reduction of GDH inhibits the accumulation of p62/Ref(2)P, an autophagic adaptor that abnormally increases in mHTT-expressing neurons. Reduction of GDH also leads to a substantial decrease in essential aminoacids in heads of adult flies. In particular we focused on Leucine and Glutamine, two major activators of TOR pathway. Leucine binds to its sensor Sestrin, while Glutamine enters the cell through specific receptors including LAT1/SCLA7/Minidisc, a Glutamine/Leucine antiporter. We are currently exploiting whether these sensors modulate TOR activity in the mechanism through which GDH downregulation induces autophagy. The goal of our work is also to design pharmacological inhibitors of GDH to be tested in vivo in flies to ameliorate HD pathology in humans.
{"title":"I04 Reduction of glutamate dehydrogenase increases autophagy and ameliorate motility and survival in a drosophila model for huntington’s disease","authors":"P. Bellosta, Stefania Santarelli, Chiara Londero","doi":"10.1136/jnnp-2021-ehdn.118","DOIUrl":"https://doi.org/10.1136/jnnp-2021-ehdn.118","url":null,"abstract":"Autophagy is a fundamental cellular pathway involved in the clearance of protein aggregates, and it is particularly important in neurons. The toxic aggregates derived from the mutated Huntingtin have been shown to interfere with the physiological autophagic flux, resulting in neuronal death. Glutamate Dehydrogenase (GDH) is an evolutionary conserved enzyme that catalyses the conversion of glutamate and ammonia to α-ketoglutarate and vice versa and is also member of the Glutamate-Glutamine Cycle (GGC), a physiological process between glia and neurons that controls glutamate homeostasis. Through a genetic screen using a Drosophila model for Huntington’s disease (HD), we identified that reduction of GDH ameliorates animal motility and decreases the size of mutated Huntingtin’s (mHTT) aggregates in brains. The aim of our project is to analyze how GDH downregulation induces autophagy in neurons. We modeled HD phenotype in Drosophila by expressing mHTT with 93-CAG repetition (HTTQ93) in neurons. To investigate the effect of GDH we used motility and viability assay, while western blots and immunofluorescence analysis were used to investigate changes in mHTT aggregates. We found the reduction of GDH inhibits the accumulation of p62/Ref(2)P, an autophagic adaptor that abnormally increases in mHTT-expressing neurons. Reduction of GDH also leads to a substantial decrease in essential aminoacids in heads of adult flies. In particular we focused on Leucine and Glutamine, two major activators of TOR pathway. Leucine binds to its sensor Sestrin, while Glutamine enters the cell through specific receptors including LAT1/SCLA7/Minidisc, a Glutamine/Leucine antiporter. We are currently exploiting whether these sensors modulate TOR activity in the mechanism through which GDH downregulation induces autophagy. The goal of our work is also to design pharmacological inhibitors of GDH to be tested in vivo in flies to ameliorate HD pathology in humans.","PeriodicalId":444837,"journal":{"name":"I: Experimental therapeutics – preclinical","volume":"6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126152617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-09-01DOI: 10.1136/jnnp-2021-ehdn.126
Gianmarco Pascarella, T. Battista, G. Colotti, Jessica Rosati, A. Fiorillo, Daniele Narzi, L. Guidoni, F. Squitieri, V. Morea, A. Ilari
Background Huntington’s Disease (HD) is a devastating and presently untreatable neurodegenerative disease, characterized by progressively disabling motor and mental manifestations. Sigma-1 receptor (σ1R), whose 3D structure has been recently determined by X-ray crystallography, is expressed in the central nervous system, and σ1R agonists have been shown to possess neuroprotective activity in neurodegenerative diseases. Aims Our overall aim is to exploit σ1R neuroprotective activity for HD therapy. Methods We are integrating computational and experimental methods: i) Virtual Screening (VS) of compound libraries available through the ZINC database towards σ1R; ii) Experimental ligand binding to purified σ1R in vitro by Surface Plasmon Resonance; iii) Assessment of ligand ability to improve the growth of fibroblasts obtained from HD patients, which is significantly impaired with respect to control cells. Additionally, we are performing Molecular Dynamics (MD) simulations to elucidate the mechanism of ligand entrance to σ1R binding site. Results i) Six known drugs have been demonstrated to be able to bind purified σ1R in vitro and improve survival and growth of HD fibroblasts; ii) Several human metabolites have been predicted by VS to bind σ1R. iii) Predictions on routes of ligand entrance have been provided by MD studies. Conclusions Our results support the validity of σ1R as a molecular target for HD therapy, and of the drug repositioning procedure implemented herein for the identification of new therapeutic agents against HD. Experimental validation of metabolites selected by VS and results of MD simulations will contribute to identify endogenous σ1R ligand(s) and mechanism of σ1R entrance.
{"title":"I12 Deciphering the neuroprotective role of sigma1 receptor, an important function to overcome the symptoms of neurodegenerative disorders","authors":"Gianmarco Pascarella, T. Battista, G. Colotti, Jessica Rosati, A. Fiorillo, Daniele Narzi, L. Guidoni, F. Squitieri, V. Morea, A. Ilari","doi":"10.1136/jnnp-2021-ehdn.126","DOIUrl":"https://doi.org/10.1136/jnnp-2021-ehdn.126","url":null,"abstract":"Background Huntington’s Disease (HD) is a devastating and presently untreatable neurodegenerative disease, characterized by progressively disabling motor and mental manifestations. Sigma-1 receptor (σ1R), whose 3D structure has been recently determined by X-ray crystallography, is expressed in the central nervous system, and σ1R agonists have been shown to possess neuroprotective activity in neurodegenerative diseases. Aims Our overall aim is to exploit σ1R neuroprotective activity for HD therapy. Methods We are integrating computational and experimental methods: i) Virtual Screening (VS) of compound libraries available through the ZINC database towards σ1R; ii) Experimental ligand binding to purified σ1R in vitro by Surface Plasmon Resonance; iii) Assessment of ligand ability to improve the growth of fibroblasts obtained from HD patients, which is significantly impaired with respect to control cells. Additionally, we are performing Molecular Dynamics (MD) simulations to elucidate the mechanism of ligand entrance to σ1R binding site. Results i) Six known drugs have been demonstrated to be able to bind purified σ1R in vitro and improve survival and growth of HD fibroblasts; ii) Several human metabolites have been predicted by VS to bind σ1R. iii) Predictions on routes of ligand entrance have been provided by MD studies. Conclusions Our results support the validity of σ1R as a molecular target for HD therapy, and of the drug repositioning procedure implemented herein for the identification of new therapeutic agents against HD. Experimental validation of metabolites selected by VS and results of MD simulations will contribute to identify endogenous σ1R ligand(s) and mechanism of σ1R entrance.","PeriodicalId":444837,"journal":{"name":"I: Experimental therapeutics – preclinical","volume":"11 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131028506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-09-01DOI: 10.1136/jnnp-2021-ehdn.122
C. Mounier, Maxime Brilland, P. Vanhoutte, J. Caboche, S. Betuing
Huntington’s disease (HD) is caused by expanded poly-glutamine in Huntingtin (mHTT), inducing many cellular dysfunctions, including cholesterol metabolism deregulation. The main pathway of cholesterol elimination is its catabolization by the neuronal 24-hydroxylase enzyme (CYP46A1) into 24S-hydroxycholesterol (24S-OHC), a ligand of Liver X Receptor (LXR). CYP46A1 level is decreased in HD, and its restoration induces a neuroprotection with an upregulation of LXR target genes. A therapeutic interest was raised for the LXR in several neurodegenerative diseases. We hypothesized the involvement of LXR in CYP46A1 neuroprotection. There are two LXR isoforms, LXRalpha mainly expressed in liver and LXRbeta enriched in the brain for cholesterol metabolism regulation. Commercialized LXR agonists suffer from side effects on lipogenesis due to the activation of LXRalpha in the liver. The aim of the project is to take advantage of new LXRbeta agonists to investigate the role of LXR activation in HD. Primary cultures of striatal neurons and astrocytes were treated with LXR agonists to validate their bioactivity and study their neuroprotective role in a HD cellular model. Wild Type mice were treated with LXR commercial agonist to determine the more efficient administration root and protocol. In neurons and astrocytes culture, LXRalpha, LXRbeta and commercial agonists induce an increase of mRNA level of LXR target genes, involved in cholesterol metabolism and known to be downregulated in HD. The LXR agonists induce a neuroprotection in HD striatal neurons in culture, with a decrease of mHTT aggregates and an increase of cell survival. When treated with inhibitor of proteasome or autophagy machinery, the neuroprotective role induced by LXR agonists is reversed. These results support the biological efficacy of these new LXR compounds and their neuroprotective role in HD striatal neurons. The next step will be to explore their effect in HD mice model.
{"title":"I08 LXR signaling in the striatum and neuroprotection in huntington’s disease","authors":"C. Mounier, Maxime Brilland, P. Vanhoutte, J. Caboche, S. Betuing","doi":"10.1136/jnnp-2021-ehdn.122","DOIUrl":"https://doi.org/10.1136/jnnp-2021-ehdn.122","url":null,"abstract":"Huntington’s disease (HD) is caused by expanded poly-glutamine in Huntingtin (mHTT), inducing many cellular dysfunctions, including cholesterol metabolism deregulation. The main pathway of cholesterol elimination is its catabolization by the neuronal 24-hydroxylase enzyme (CYP46A1) into 24S-hydroxycholesterol (24S-OHC), a ligand of Liver X Receptor (LXR). CYP46A1 level is decreased in HD, and its restoration induces a neuroprotection with an upregulation of LXR target genes. A therapeutic interest was raised for the LXR in several neurodegenerative diseases. We hypothesized the involvement of LXR in CYP46A1 neuroprotection. There are two LXR isoforms, LXRalpha mainly expressed in liver and LXRbeta enriched in the brain for cholesterol metabolism regulation. Commercialized LXR agonists suffer from side effects on lipogenesis due to the activation of LXRalpha in the liver. The aim of the project is to take advantage of new LXRbeta agonists to investigate the role of LXR activation in HD. Primary cultures of striatal neurons and astrocytes were treated with LXR agonists to validate their bioactivity and study their neuroprotective role in a HD cellular model. Wild Type mice were treated with LXR commercial agonist to determine the more efficient administration root and protocol. In neurons and astrocytes culture, LXRalpha, LXRbeta and commercial agonists induce an increase of mRNA level of LXR target genes, involved in cholesterol metabolism and known to be downregulated in HD. The LXR agonists induce a neuroprotection in HD striatal neurons in culture, with a decrease of mHTT aggregates and an increase of cell survival. When treated with inhibitor of proteasome or autophagy machinery, the neuroprotective role induced by LXR agonists is reversed. These results support the biological efficacy of these new LXR compounds and their neuroprotective role in HD striatal neurons. The next step will be to explore their effect in HD mice model.","PeriodicalId":444837,"journal":{"name":"I: Experimental therapeutics – preclinical","volume":"30 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128786208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-09-01DOI: 10.1136/jnnp-2021-ehdn.120
Giulia Birolini, G. Verlengia, F. Talpo, Claudia Maniezzi, L. Zentilin, M. Giacca, P. Conforti, C. Cordiglieri, C. Caccia, V. Leoni, F. Taroni, G. Biella, M. Simonato, E. Cattaneo, M. Valenza
Background Cholesterol is a multifaceted molecule essential for brain function (Martin 2014). In the adult brain, cholesterol is produced locally by astrocytes and transferred to neurons through apoE-containing lipoproteins (Jurevics & Morell 1995; Mauch 2001). Disruption of brain cholesterol pathways has been linked to several neurological disorders, including Huntington’s disease (HD), a genetic, neurodegenerative disorder caused by a CAG expansion in the gene encoding the Huntingtin protein (Valenza & Cattaneo 2011). Brain cholesterol biosynthesis and content are reduced in several HD models (Valenza 2005; 2007; 2010; Shankaran 2017). The underlying molecular mechanism relies on reduced nuclear translocation of SREBP2, the transcription factor that controls the transcription of several genes involved in cholesterol biosynthesis (Valenza 2015; Di Pardo 2020). We have recently shown that cholesterol supplementation to the brain, with different delivery systems, ameliorates synaptic and behavioral defects in the R6/2 mouse model (Valenza 2015; Birolini 2020; Birolini 2021). Aims and Methods Here, we used recombinant adeno-associated virus 2/5 to deliver exogenous SREBP2 specifically in astrocytes in order to enhance the endogenous cholesterol biosynthesis in the striatum of R6/2 mice. Results We found that exogenous SREBP2 stimulates the transcription of key cholesterol biosynthesis genes resulting in fully restoration of synaptic transmission, reversal of Drd2 transcript levels, clearance of mutant Huntingtin (muHTT) aggregates and rescue of behavioral deficits. Conclusions These results demonstrate that stimulating cholesterol biosynthesis in striatal astrocytes has a positive effect on behavioral decline and disease-related phenotypes in HD mice. Furthermore, we have demonstrated that glial SREBP2 participates in HD pathogenesis in vivo, highlighting the translational potential of cholesterol-based strategies for this disease.
{"title":"I06 SREBP2 delivery to striatal astrocytes normalizes transcription of cholesterol biosynthesis genes and ameliorates pathological features in huntington’s disease","authors":"Giulia Birolini, G. Verlengia, F. Talpo, Claudia Maniezzi, L. Zentilin, M. Giacca, P. Conforti, C. Cordiglieri, C. Caccia, V. Leoni, F. Taroni, G. Biella, M. Simonato, E. Cattaneo, M. Valenza","doi":"10.1136/jnnp-2021-ehdn.120","DOIUrl":"https://doi.org/10.1136/jnnp-2021-ehdn.120","url":null,"abstract":"Background Cholesterol is a multifaceted molecule essential for brain function (Martin 2014). In the adult brain, cholesterol is produced locally by astrocytes and transferred to neurons through apoE-containing lipoproteins (Jurevics & Morell 1995; Mauch 2001). Disruption of brain cholesterol pathways has been linked to several neurological disorders, including Huntington’s disease (HD), a genetic, neurodegenerative disorder caused by a CAG expansion in the gene encoding the Huntingtin protein (Valenza & Cattaneo 2011). Brain cholesterol biosynthesis and content are reduced in several HD models (Valenza 2005; 2007; 2010; Shankaran 2017). The underlying molecular mechanism relies on reduced nuclear translocation of SREBP2, the transcription factor that controls the transcription of several genes involved in cholesterol biosynthesis (Valenza 2015; Di Pardo 2020). We have recently shown that cholesterol supplementation to the brain, with different delivery systems, ameliorates synaptic and behavioral defects in the R6/2 mouse model (Valenza 2015; Birolini 2020; Birolini 2021). Aims and Methods Here, we used recombinant adeno-associated virus 2/5 to deliver exogenous SREBP2 specifically in astrocytes in order to enhance the endogenous cholesterol biosynthesis in the striatum of R6/2 mice. Results We found that exogenous SREBP2 stimulates the transcription of key cholesterol biosynthesis genes resulting in fully restoration of synaptic transmission, reversal of Drd2 transcript levels, clearance of mutant Huntingtin (muHTT) aggregates and rescue of behavioral deficits. Conclusions These results demonstrate that stimulating cholesterol biosynthesis in striatal astrocytes has a positive effect on behavioral decline and disease-related phenotypes in HD mice. Furthermore, we have demonstrated that glial SREBP2 participates in HD pathogenesis in vivo, highlighting the translational potential of cholesterol-based strategies for this disease.","PeriodicalId":444837,"journal":{"name":"I: Experimental therapeutics – preclinical","volume":"169 2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116636557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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