)| PMID |
17997855 ( ) |
|---|---|
| Title | Expression of a Cu,Zn superoxide dismutase typical for familial amyotrophic lateral sclerosis increases the vulnerability of neuroblastoma cells to infectious injury. |
| Abstract | BACKGROUND: Infections can aggravate the course of neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). Mutations in the anti-oxidant enzyme Cu,Zn superoxide dismutase (EC 1.15.1.1, SOD1) are associated with familial ALS. Streptococcus pneumoniae, the most frequent respiratory pathogen, causes damage by the action of the cholesterol-binding virulence factor pneumolysin and by stimulation of the innate immune system, particularly via Toll-like-receptor 2. METHODS: SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SOD1 typical for familial ALS (G93A-SOD1) and SH-SY5Y neuroblastoma cells transfected with wildtype SOD1 were both exposed to pneumolysin and in co-cultures with cultured human macrophages treated with the Toll like receptor 2 agonist N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-[R]-cysteinyl-[S]-seryl -[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine x 3 HCl (Pam3CSK4). Cell viability and apoptotic cell death were compared morphologically and by in-situ tailing. With the help of the WST-1 test, cell viability was quantified, and by measurement of neuron-specific enolase in the culture supernatant neuronal damage in co-cultures was investigated. Intracellular calcium levels were measured by fluorescence analysis using fura-2 AM. RESULTS: SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SOD1 typical for familial ALS (G93A-SOD1) were more vulnerable to the neurotoxic action of pneumolysin and to the attack of monocytes stimulated by Pam3CSK4 than SH-SY5Y cells transfected with wild-type human SOD1. The enhanced pneumolysin toxicity in G93A-SOD1 neuronal cells depended on the inability of these cells to cope with an increased calcium influx caused by pores formed by pneumolysin. This inability was caused by an impaired capacity of the mitochondria to remove cytoplasmic calcium. Treatment of G93A-SOD1 SH-SY5Y neuroblastoma cells with the antioxidant N-acetylcysteine reduced the toxicity of pneumolysin. CONCLUSION: The particular vulnerability of G93A-SOD1 neuronal cells to hemolysins and inflammation may be partly responsible for the clinical deterioration of ALS patients during infections. These findings link infection and motor neuron disease and suggest early treatment of respiratory infections in ALS patients. Germany. mlotz@gwdg.de |
NOTE: Color highlight is limited to the abstract and SciMiner text-mining mode. If you see much more identified targets below from "Targets by SciMiner Summary" and "Targets by SciMiner Full list", they may have been identified from the full text.
Targets by SciMiner Summary
| HUGO ID | Symbol | Target Name | #Occur | ActualStr |
|---|---|---|---|---|
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | 24 | SOD1 | SOD | superoxide dismutase | |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | 8 | caspase 3 | |
| 11848 | TLR2 | toll-like receptor 2 | 8 | toll like receptor 2 | TLR2 | |
| 1693 | CD68 | CD68 molecule | 6 | CD68 | CD-68 | |
| 3353 | ENO2 | enolase 2 (gamma, neuronal) | 4 | NSE | |
| 620 | APP | amyloid beta (A4) precursor protein (peptidase nexin-II, Alzheimer disease) | 2 | amyloid | |
| 6871 | MAPK1 | mitogen-activated protein kinase 1 | 2 | MAPK | p38 | |
| 391 | AKT1 | v-akt murine thymoma viral oncogene homolog 1 | 1 | Rac | |
| 437 | ALPI | alkaline phosphatase, intestinal | 1 | alkaline phosphatase | |
| 15633 | TLR9 | toll-like receptor 9 | 1 | TLR9 | |
| 11850 | TLR4 | toll-like receptor 4 | 1 | TLR4 | |
| 670 | RHOD | ras homolog gene family, member D | 1 | Rho | |
| 2983 | DNTT | deoxynucleotidyltransferase, terminal | 1 | terminal transferase | |
| 399 | ALB | albumin | 1 | serum albumin | |
Targets by SciMiner Full list
| HUGO ID | Symbol | Name | ActualStr | Score | FlankingText |
|---|---|---|---|---|---|
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | the anti-oxidant enzyme Cu Zn superoxide dismutase (EC EC 1.15.1.1 SOD1 are associated with familial ALS |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | Methods SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SOD1 typical for familial ALS (G93A-SOD1) G93A-SOD1 and SH-SY5Y neuroblastoma cells |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | ALS (G93A-SOD1) G93A-SOD1 and SH-SY5Y neuroblastoma cells transfected with wildtype SOD1 were both exposed to pneumolysin and in co-cultures with cultured |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | Results SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SOD1 typical for familial ALS (G93A-SOD1) G93A-SOD1 were more vulnerable to |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | 3 CSK 4 than SH-SY5Y cells transfected with wild-type human SOD1 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | the gene encoding Cu Zn superoxide dismutase (EC EC 1.15.1.1 SOD1 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | cases are caused by a mutation in the gene encoding SOD1 4 5 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | the amino acid glycine is replaced by alanine in the SOD1 enzyme |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | model to study the cellular alterations associated with mutations of SOD1 was constructed by transfection of the human neuroblastoma cell line |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | was chosen because it does not affect the activity of SOD1 |
| 11848 | TLR2 | toll-like receptor 2 | TLR2 | 3.7 | of the innate immune system particularly Toll-like receptor 2 (TLR2) TLR2 18 19 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | neuroblastoma cell lines constitutively expressing either wild-type (Wt) Wt human SOD1 or the G93A mutant of this enzyme associated with familial |
| 1693 | CD68 | CD68 molecule | CD-68 | 0.3 | CD-68 staining showed 98_amp_#x02013 99% purity of the macrophage cultures |
| 1693 | CD68 | CD68 molecule | CD68 | 0.3 | Methods Hemalum staining CD68 and activated caspase-3 immunocytochemistry and light green staining Pneumolysin-stimulated SH-SY5Y |
| 1693 | CD68 | CD68 molecule | CD68 | 0.3 | CD68 and light green staining was used to distinguish between human |
| 1693 | CD68 | CD68 molecule | CD68 | 0.3 | 0.1% in PBS for 30 minutes and then incubated with CD68 antibody (clone clone KP1 dilution 1 50 DAKO Glostrup Denmark |
| 1693 | CD68 | CD68 molecule | CD68 | 0.3 | in a brown staining of the somata of macrophages (CD68)/apoptotic CD68 apoptotic G93A SOD1 cells (Caspase Caspase 3 |
| 1693 | CD68 | CD68 molecule | CD68 | 0.3 | yellowish solution (Chroma-Gesellschaft Chroma-Gesellschaft Schmidt _amp_#x00026 Co Stuttgart Germany after CD68 immunocytochemistry and with hemalum after caspase-3 immunocytochemistry for 1_amp_#x02013 2 |
| 11848 | TLR2 | toll-like receptor 2 | TLR2 | 3.7 | After stimulation of human neuroblastoma and macrophage co-cultures with the TLR2 agonist Pam 3 CSK 4 for a period of 72 |
| 3353 | ENO2 | enolase 2 (gamma, neuronal) | NSE | 0.3 | period of 72 hours the release of neuron-specific enolase (NSE) NSE was measured in the culture supernatants and expressed as per |
| 3353 | ENO2 | enolase 2 (gamma, neuronal) | NSE | 0.3 | the culture supernatants and expressed as per cent of the NSE release induced by cell lysis |
| 3353 | ENO2 | enolase 2 (gamma, neuronal) | NSE | 0.3 | with G93A-SOD1 SH-SY5Y cells showed a significantly higher release of NSE compared to co-cultures with Wt-SOD1 SH-SY5Y cells after stimulation with |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | that SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SOD1 typical for familial ALS (G93A-SOD1) G93A-SOD1 are more vulnerable to |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | more vulnerable to infectious stimuli than neuroblastoma cells overexpressing normal SOD1 |
| 670 | RHOD | ras homolog gene family, member D | Rho | 0.3 | At sublytic concentrations pneumolysin rapidly activates Rho and Rac GTPases and leads to the formation of actin |
| 391 | AKT1 | v-akt murine thymoma viral oncogene homolog 1 | Rac | 0.1 | At sublytic concentrations pneumolysin rapidly activates Rho and Rac GTPases and leads to the formation of actin stress fibers |
| 6871 | MAPK1 | mitogen-activated protein kinase 1 | p38 | 0.5 | The massive Ca 2 influx causes activation of p38 MAPK opening of the mitochondrial permeability transition (MPT) MPT pore |
| 6871 | MAPK1 | mitogen-activated protein kinase 1 | MAPK | 0.5 | The massive Ca 2 influx causes activation of p38 MAPK opening of the mitochondrial permeability transition (MPT) MPT pore and |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | than the capacity of neuroblastoma cells not transfected with mutant SOD1 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | production of reactive oxygen species in neuroblastoma cells expressing mutant SOD1 34 |
| 11850 | TLR4 | toll-like receptor 4 | TLR4 | 2.2 | microglia has been observed in vitro following stimulation with the TLR4 agonist lipopolysaccharide (LPS) LPS 44 45 and analogues of bacterial |
| 15633 | TLR9 | toll-like receptor 9 | TLR9 | 1.2 | (LPS) LPS 44 45 and analogues of bacterial DNA (TLR9 TLR9 agonist 13 |
| 11848 | TLR2 | toll-like receptor 2 | TLR2 | 3.7 | Here we demonstrate that activation of macrophages by the TLR2 agonist Pam 3 CSK 4 can also cause neuronal death |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | the attack of activated immune cells than those expressing wild-type SOD1 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD | 1.9 | the disease predominated by motor neuron damage caused by mutant SOD in a later phase of disease progression is linked to |
| 620 | APP | amyloid beta (A4) precursor protein (peptidase nexin-II, Alzheimer disease) | amyloid | 1.0 | Co-stimulation of microglia with host-derived compounds (_amp_#x003b2;-amyloid, _amp_#x003b2 -amyloid fibronectin advanced glycation end products and bacterial products can lead |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | increased susceptibility of neurons expressing the G93A mutant in their SOD1 to the attack of activated immune cells may be the |
| 620 | APP | amyloid beta (A4) precursor protein (peptidase nexin-II, Alzheimer disease) | amyloid | 1.0 | Endogenous compounds deposited in the extracellular space (e.g., e.g. _amp_#x003b2 -amyloid 17 entering the brain through the leaky blood-brain barrier (e.g., |
| 11848 | TLR2 | toll-like receptor 2 | TLR2 | 3.7 | of the monocyte/macrophage/microglia monocyte macrophage microglia type activated by a TLR2 agonist can kill neurons |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | Neuronal cells expressing a SOD1 mutant frequently encountered in familial cases of amyotrophic lateral sclerosis |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | attack of activated immune cells than neuronal cells expressing wild-type SOD1 |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | Figure 11 Vulnerability of G93A-SOD1 and wild-type SOD1 neuroblastoma cells to the attack of monocytes stimulated with Pam |
| 3353 | ENO2 | enolase 2 (gamma, neuronal) | NSE | 0.3 | (values values expressed in per cent _amp_#x000b1 SD of the NSE release induced by cell lysis |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | SOD1 | 3.2 | Figure 12 Vulnerability of G93A-SOD1 and wild-type SOD1 neuroblastoma cells to the attack of monocytes stimulated with Pam |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | superoxide dismutase | 1.0 | mutations in the anti oxidant enzyme cu zn superoxide dismutase ec 1.15.1.1 sod1 are associated with familial als. |
| 11848 | TLR2 | toll-like receptor 2 | toll like receptor 2 | 1.0 | s pneumoniae the most frequent respiratory pathogen causes damage by the action of the cholesterol binding virulence factor pneumolysin and by stimulation of the innate immune system particularly via toll like receptor 2. |
| 11848 | TLR2 | toll-like receptor 2 | toll like receptor 2 | 1.0 | typical for familial als g93a sod1 and sh sy5y neuroblastoma cells transfected with wildtype sod1 were both exposed to pneumolysin and in co cultures with cultured human macrophages treated with the toll like receptor 2 agonist n palmitoyl s [2 3 bis palmitoyloxy 2rs propyl] [r] cysteinyl [s] seryl [s] lysyl [s] lysyl [s] lysyl [s] lysyl [s] lysine _amp_#x000d7; 3 hcl pam 3 csk 4 . |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | superoxide dismutase | 1.0 | mitochondrial function can be disturbed by mutations in the gene encoding cu zn superoxide dismutase ec 1.15.1.1 sod1 . |
| 11848 | TLR2 | toll-like receptor 2 | toll like receptor 2 | 1.0 | s damage by the direct action of the cholesterol binding pore forming hemolysin pneumolysin and through microglia/monocyte activation by agonists of receptors of the innate immune system particularly toll like receptor 2 tlr2 [ 18 19 ]. |
| 399 | ALB | albumin | serum albumin | 1.0 | bathing solutions were either rpmi 1640 or in mm nacl 140 kcl 2 cacl 2 2.5 mgcl 2 1 hepes 10 glucose 40 and bovine serum albumin 0.05% at ph 7.3. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | methods hemalum staining cd68 and activated caspase 3 immunocytochemistry and light green staining pneumolysin stimulated sh sy5y cell mono cultures wt sod1 and g93a sod1 were plated on glass coverslips and were fixated with 4% formaldehyde dehydrated t |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | cd68 and light green staining was used to distinguish between human macrophages and the sh sy5y cells in co culture and to visualize the morphology of the cells; immunostaining for activated caspase 3 was used to detect apoptosis in pneumolysin treated g93a sod1 cells: fixated cells were permeabilised with triton x 0.1% in pbs for 30 minutes and then incubated with cd68 antibody clone kp1 dilution |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | sin treated g93a sod1 cells: fixated cells were permeabilised with triton x 0.1% in pbs for 30 minutes and then incubated with cd68 antibody clone kp1 dilution 1:50 dako glostrup denmark or activated caspase 3 antibody [rabbit anti caspase 3 cleaved dilution 1:100 kindly donated by zytomed systems berlin germany] for 90 minutes. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | antibody [rabbit anti caspase 3 cleaved dilution 1:100 kindly donated by zytomed systems berlin germany] for 90 minutes. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | biotin complex abc vector burlingame ca for 30 min and diaminobenzidine was used for visualisation 5 minutes resulting in a brown staining of the somata of macrophages cd68 /apoptotic g93a sod1 cells caspase 3 . |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | sh sy5y cells were counterstained by light green sf yellowish solution chroma gesellschaft schmidt _amp_#x00026; co stuttgart germany after cd68 immunocytochemistry and with hemalum after caspase 3 immunocytochemistry for 1_amp_#x02013;2 minutes rinsed in water dehydrated and mounted with depex serva heidelberg germany . |
| 2983 | DNTT | deoxynucleotidyltransferase, terminal | terminal transferase | 1.0 | 37_amp_#x000b0;c in a reaction mixture that contained 10 _amp_#x003bc;l of 5_amp_#x000d7; tailing buffer 1 _amp_#x003bc;l of digoxigenin dna labeling mix 2 _amp_#x003bc;l of cobalt chloride 12.5 u of terminal transferase and the amount of distilled water necessary to give a volume of 50 _amp_#x003bc;l. |
| 437 | ALPI | alkaline phosphatase, intestinal | alkaline phosphatase | 1.0 | a solution of alkaline phosphatase labeled anti digoxigenin antibody in 10% fcs 1:250 was placed on the sections for 60 min at 37_amp_#x000b0;c. |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | by in situ tailing morphology arrows fig 3 and by immunocytochemistry for activated caspase 3 fig 4 of g93a sod1 neuroblastoma cells it became apparent that a large proportion of these cells died by apoptosis. |
| 11848 | TLR2 | toll-like receptor 2 | toll like receptor 2 | 1.0 | results increased vulnerability of neuroblastoma cells transfected with g93a sod1 to the attack of monocytes stimulated with the toll like receptor 2 agonist pam 3 csk 4 after stimulation of human neuroblastoma and macrophage co cultures with the tlr2 agonist pam 3 csk 4 for a period of 72 hours the release of neuron specific enolase nse was measu |
| 1504 | CASP3 | caspase 3, apoptosis-related cysteine peptidase | caspase 3 | 1.0 | apoptotic g93a sod1 sh sy5y cells after incubation with 0.5 _amp_#x003bc;g/ml ply for three hours detected by immunocytochemistry for activated caspase 3. |