Selective inhibitors of protein kinase A may be of therapeutic value in both sporadic and Familial Alzheimer’s disease, since they may decrease the production of A that is thought to be responsible for the neurodegenerative process

Selective inhibitors of protein kinase A may be of therapeutic value in both sporadic and Familial Alzheimer’s disease, since they may decrease the production of A that is thought to be responsible for the neurodegenerative process. Keywords: Alzheimer’s disease, amyloid peptides, APP, protein kinase A, PKI, H89, HEK293 cells, neurons, mutant APP, mutant presenilins Introduction Sporadic and familial forms of Alzheimer’s disease (FAD) are characterized by comparable extracellular proteinaceous deposits called senile plaques that invade the cortical and subcortical areas of affected brains (Hardy & Allsop, 1991). early-onset forms of Familial Alzheimer’s disease (FAD). By contrast, H89 and PKI do not significantly affect the recovery of the physiological -secretase-derived fragment APP. Our study indicates that protein kinase A inhibitors selectively lower the formation of A40 and A42 in human cells expressing normal and mutant APP and PS1 without affecting the physiological -secretase pathway in these cells. Selective inhibitors of protein kinase A may be of therapeutic value in both sporadic and Familial Alzheimer’s disease, since they may decrease the production of A that is thought to be responsible for the neurodegenerative process. Keywords: Alzheimer’s disease, amyloid peptides, APP, protein kinase A, PKI, H89, HEK293 cells, neurons, mutant APP, mutant presenilins Introduction Sporadic and familial forms of Alzheimer’s disease (FAD) are characterized by comparable extracellular proteinaceous deposits called senile plaques that invade the cortical and subcortical areas of affected brains (Hardy & Allsop, 1991). These neuropathological lesions are mainly composed of amyloid peptide (A), a 39C43 amino-acid poorly soluble peptide (Selkoe, 1991). The onset of genetic forms of Alzheimer’s disease generally precedes that of the sporadic cases by several decades. This is thought to be due to the drastic overproduction of A and, particularly that of the readily aggregable 42 aminoacid form of A (for review observe Checler, 1995). The acceleration of the A production has been demonstrated to be due to the presence of missense mutations in the amyloid precursor protein (APP, Citron et al., 1992; Cai et al., 1993; Cichoric Acid Felsenstein et al., 1994) and more recently, in two homologous proteins named presenilins 1 and 2 (PS1, PS2) (Borchelt et al., 1996; Duff et al., 1996; Citron et al., 1997; Tomita et al., 1997; Xia et al., 1997; Ancolio et al., 1997; Marambaud et al., 1998b). The fact that distinct proteins, all responsible for aggressive forms of Alzheimer’s disease, could trigger similar phenotypic overproduction of A argues in favour of a therapeutic strategy aimed at slowing down the production of this peptide. In this context, putative therapeutic targets could be – and -secretases, (the proteolytic activities responsible for the release of A from its precursor) or other mechanisms responsible for the regulation of APP processing. Effectors of the protein kinase C have been shown to decrease A production and increase secretion of the -secretase-derived physiological product APP in various cell lines (Caporaso et al., 1992; Gillespie et al., 1992; Buxbaum et al., 1993; Hung et al., 1993). Furthermore, in gene-targeted mice overproducing A, the administration of the PKC stimulator phorbol 12,13-dibutyrate (PDBu) led to drastic inhibition of the production of A (Savage et al., 1998). We recently showed that the maturation of APP appears to be under control of the protein kinase A (PKA) pathway in human cells and neurons overexpressing normal and FAD-linked APP (Marambaud et al., 1998a). However, unlike modulators of PKC, effectors of the PKA pathway stimulated production of both A and APP (Marambaud et al., 1998a) suggesting that the target of PKA was probably located upstream of both – and /- secretases cleavages. Here we show that two distinct PKA inhibitors drastically reduce the constitutive production of both A40 and A42 in stably transfected HEK293 cells expressing wild type (wt) and Swedish mutated (Sw) APP751. We also establish that PKA inhibitors almost completely prevent the formation of As by HEK293 cells overexpressing wt- and M146V-PS1. Interestingly, the inhibitors do not significantly affect the recoveries of APP or its -secretase-derived C-terminal stub, p10. Our data indicate that PKA inhibitors.Our data indicate that PKA inhibitors selectively affect the /-secretase pathway in human cells and are potential pharmacological which may be able to reduce A formation in both sporadic and FAD-linked Alzheimer’s disease. Methods Antibodies FCA3340 and FCA3542 specifically recognize the C- termini of A40 and A42, respectively (Barelli et al., 1997). early-onset forms of Familial Alzheimer’s disease (FAD). By contrast, H89 and PKI do not significantly affect the recovery of the physiological -secretase-derived fragment APP. Our study indicates that protein kinase A inhibitors selectively lower the formation of A40 and A42 in human cells expressing normal and mutant APP and PS1 without affecting the physiological -secretase pathway in these cells. Selective inhibitors of protein kinase A may be of therapeutic value in both sporadic and Familial Alzheimer’s disease, since they may decrease the production of A that is thought to be responsible for the neurodegenerative process. Keywords: Alzheimer’s disease, amyloid peptides, APP, protein kinase A, PKI, H89, HEK293 cells, neurons, mutant APP, mutant presenilins Introduction Sporadic Cichoric Acid and familial forms of Alzheimer’s disease (FAD) are characterized by similar extracellular proteinaceous deposits called senile plaques that invade the cortical and subcortical areas of affected brains (Hardy & Allsop, 1991). These neuropathological lesions are mainly composed of amyloid peptide (A), a 39C43 amino-acid poorly soluble peptide (Selkoe, 1991). The onset of genetic forms of Alzheimer’s disease generally precedes that of the sporadic cases by several decades. This is thought to be due to the drastic overproduction of A and, particularly that of the readily aggregable 42 aminoacid form of A (for review see Checler, 1995). The acceleration of the A production has been demonstrated to be due to the presence of missense mutations in the amyloid precursor protein (APP, Citron et al., 1992; Cai et al., 1993; Felsenstein et al., 1994) and more recently, in two homologous proteins named presenilins 1 and 2 (PS1, PS2) (Borchelt et al., 1996; Duff et al., 1996; Citron et al., 1997; Tomita et al., 1997; Xia et al., 1997; Ancolio et al., 1997; Marambaud et al., 1998b). The fact that distinct proteins, all responsible for aggressive forms of Alzheimer’s disease, could trigger similar phenotypic overproduction of A argues in favour of a therapeutic strategy aimed at slowing down the production of this peptide. In this context, putative restorative targets could be – and -secretases, (the proteolytic activities responsible for the release of A from its precursor) or additional mechanisms responsible for the rules of APP control. Effectors of the protein kinase C have been shown to decrease A production and increase secretion of the -secretase-derived physiological product APP in various cell lines (Caporaso et al., 1992; Gillespie et al., 1992; Buxbaum et al., 1993; Hung et al., 1993). Furthermore, in gene-targeted mice overproducing A, the administration of the PKC stimulator phorbol 12,13-dibutyrate (PDBu) led to drastic inhibition of the production of A (Savage et al., 1998). We recently showed the maturation of APP appears to be under control of the protein kinase A (PKA) pathway in human being cells and neurons overexpressing normal and FAD-linked APP (Marambaud et al., 1998a). However, unlike modulators of PKC, effectors of the PKA pathway stimulated production of both A and APP (Marambaud et al., 1998a) suggesting that the prospective of PKA was probably located upstream of both – and /- secretases cleavages. Here we display that two unique PKA inhibitors drastically reduce the constitutive production of both A40 and A42 in stably transfected HEK293 cells expressing crazy type (wt) and Swedish mutated (Sw) APP751. We also set up that PKA inhibitors almost completely prevent the formation of As by HEK293 cells overexpressing wt- and M146V-PS1. Interestingly, the inhibitors do not significantly impact the recoveries of APP or its -secretase-derived C-terminal stub, p10. Our data show that PKA inhibitors selectively impact the /-secretase pathway in human being cells and are potential pharmacological which may be able to reduce A formation in both sporadic and FAD-linked Alzheimer’s disease. Methods Antibodies FCA3340 and FCA3542 specifically identify the C- termini of A40 and A42, respectively (Barelli et al., 1997). FCA18 (Barelli et.Densitometric analyses of A40 and A42 recoveries were quantified as with the Number 2 and are the meanss.e.mean of 4C5 indie experiments. HEK293 cells expressing crazy type and M146V-PS1 display low endogenous amounts of APP, leading to poorly detectable A42 secretion. Selective inhibitors of protein kinase A may be of restorative value in both sporadic and Familial Alzheimer’s disease, since they may decrease the production of A that is thought to be responsible for the neurodegenerative process. Keywords: Alzheimer’s disease, amyloid peptides, APP, protein kinase A, PKI, H89, HEK293 cells, neurons, mutant APP, mutant presenilins Intro Sporadic and familial forms of Alzheimer’s disease (FAD) are characterized by related extracellular proteinaceous deposits called senile plaques that invade the cortical and subcortical areas of affected brains (Hardy & Allsop, 1991). These neuropathological lesions are primarily composed of amyloid peptide (A), a 39C43 amino-acid poorly soluble peptide (Selkoe, 1991). The onset of genetic forms of Alzheimer’s disease generally precedes that of the sporadic instances by several decades. This is thought to be due to the drastic overproduction of A and, particularly that of the readily aggregable 42 aminoacid form of A (for review observe Checler, 1995). The acceleration of the A production has been demonstrated to be due to the presence of missense mutations in the amyloid precursor protein (APP, Citron et al., 1992; Cai et al., 1993; Felsenstein et al., 1994) and more recently, in two homologous proteins named presenilins 1 and 2 (PS1, PS2) (Borchelt et al., 1996; Duff et al., 1996; Citron et al., 1997; Tomita et al., 1997; Xia et al., 1997; Ancolio et al., 1997; Marambaud et al., 1998b). The fact that unique proteins, all responsible for aggressive forms of Alzheimer’s disease, could result in related phenotypic overproduction of A argues in favour of a restorative strategy aimed at slowing down the production of this peptide. With this context, putative restorative targets could be – and -secretases, (the proteolytic activities responsible for the release of A from its precursor) or additional mechanisms responsible for the rules of APP control. Effectors of the protein kinase C have been shown to decrease A production and increase secretion of the -secretase-derived physiological product APP in various cell lines (Caporaso et al., 1992; Gillespie et al., 1992; Buxbaum et al., 1993; Hung et al., 1993). Furthermore, in gene-targeted mice overproducing A, the administration of the PKC stimulator phorbol 12,13-dibutyrate (PDBu) led to drastic inhibition of the production of A (Savage et al., 1998). We recently showed the maturation of APP appears to be under control of the protein kinase A (PKA) pathway in human being cells and neurons overexpressing normal and FAD-linked APP (Marambaud et al., 1998a). However, unlike modulators of PKC, effectors of the PKA pathway stimulated production of both A and APP (Marambaud et al., 1998a) suggesting that the target of PKA was probably located upstream of both – and /- secretases cleavages. Here we show that two unique PKA inhibitors drastically reduce the constitutive production of both A40 and A42 in stably transfected HEK293 cells expressing wild type (wt) and Swedish mutated (Sw) APP751. We also establish that PKA inhibitors almost completely prevent the formation of As by HEK293 cells overexpressing wt- and M146V-PS1. Interestingly, the inhibitors do not significantly impact the recoveries of APP or its -secretase-derived C-terminal stub, p10. Our data show that PKA inhibitors selectively impact the /-secretase pathway in human cells and are potential pharmacological which may be able to reduce A formation in both sporadic and FAD-linked Alzheimer’s disease. Methods Antibodies FCA3340 and FCA3542 specifically identify the C-.Here we show that two distinct PKA inhibitors drastically reduce the constitutive production of both A40 and A42 in stably transfected HEK293 cells expressing wild type (wt) and Swedish mutated (Sw) APP751. selectively lesser the formation of A40 and A42 in human cells expressing normal and mutant APP and PS1 without affecting the physiological -secretase pathway in these cells. Selective inhibitors of protein kinase A may be of therapeutic value in both sporadic and Familial Alzheimer’s disease, since they may decrease the production of A that is thought to be responsible for the neurodegenerative process. Keywords: Alzheimer’s disease, amyloid peptides, APP, protein kinase A, PKI, H89, HEK293 cells, neurons, mutant APP, mutant presenilins Introduction Sporadic and familial forms of Alzheimer’s disease (FAD) are characterized by comparable extracellular proteinaceous deposits called senile plaques that invade the cortical and subcortical areas of affected brains (Hardy & Allsop, 1991). These neuropathological lesions are mainly composed of amyloid peptide (A), a 39C43 amino-acid poorly soluble peptide (Selkoe, 1991). The onset of genetic forms of Alzheimer’s disease generally precedes that of the sporadic cases by several decades. This is thought to be due to the drastic overproduction of A and, particularly that of the readily aggregable 42 aminoacid form of A (for review observe Checler, 1995). The acceleration of the A production has been demonstrated to be due to the presence of missense mutations in the amyloid precursor protein (APP, Citron et al., 1992; Cai et al., 1993; Felsenstein et al., 1994) and more recently, in two homologous proteins named presenilins 1 and 2 (PS1, PS2) (Borchelt et al., 1996; Duff et al., 1996; Citron et al., 1997; Tomita et al., 1997; Xia et al., 1997; Ancolio et al., 1997; Marambaud et al., 1998b). The fact that unique proteins, all responsible for aggressive forms of Alzheimer’s disease, could trigger comparable phenotypic overproduction of A argues in favour of a therapeutic strategy aimed at slowing down the production of this peptide. In this context, putative therapeutic targets could be – and -secretases, (the proteolytic activities responsible for the release of A from its precursor) or other mechanisms responsible for the regulation of APP processing. Effectors of the protein kinase C have been shown to decrease A production and increase secretion of the -secretase-derived physiological product APP in various cell lines (Caporaso et al., 1992; Gillespie et al., 1992; Buxbaum et al., 1993; Hung et al., 1993). Furthermore, in gene-targeted mice overproducing A, the administration of the PKC stimulator phorbol 12,13-dibutyrate (PDBu) led to drastic inhibition of the production of A (Savage et al., 1998). We recently showed that this maturation of APP appears to be under control of the protein kinase A (PKA) pathway in human cells and neurons overexpressing normal and FAD-linked APP (Marambaud et al., 1998a). However, unlike modulators of PKC, effectors of the PKA pathway stimulated production of both A and APP (Marambaud et al., 1998a) suggesting that the target of PKA was probably located upstream of both – and /- secretases cleavages. Here we show that two unique PKA inhibitors drastically reduce the constitutive production of both A40 and A42 in stably transfected HEK293 cells expressing wild type (wt) and Swedish mutated (Sw) APP751. We also establish that PKA inhibitors almost completely prevent the formation of As by HEK293 cells overexpressing wt- and M146V-PS1. Interestingly, the inhibitors do not significantly impact the recoveries of APP or its -secretase-derived C-terminal stub, p10..Furthermore, in Cichoric Acid gene-targeted mice overproducing A, the administration of the PKC stimulator phorbol 12,13-dibutyrate (PDBu) led to drastic inhibition of the production of A (Savage et al., 1998). We recently showed that this maturation of APP appears to be under control of the protein kinase A (PKA) pathway in human cells and neurons overexpressing normal and FAD-linked APP (Marambaud et al., 1998a). not really affect the recovery from the physiological -secretase-derived fragment APP considerably. Our study shows that proteins kinase A inhibitors selectively lower the forming of A40 and A42 in human being cells expressing regular and mutant APP and PS1 without influencing the physiological -secretase pathway in these cells. Selective inhibitors of proteins kinase A could be of restorative worth in both sporadic and Familial Alzheimer’s disease, given that they may reduce the creation of the that is regarded as in charge of the neurodegenerative procedure. Keywords: Alzheimer’s disease, amyloid peptides, APP, proteins kinase A, PKI, H89, HEK293 cells, neurons, mutant APP, mutant presenilins Intro Sporadic and familial types of Alzheimer’s disease (Trend) are seen as a identical extracellular proteinaceous debris known as senile plaques that invade the cortical and subcortical regions of affected brains (Hardy & Allsop, 1991). These neuropathological lesions are primarily made up of amyloid peptide (A), a 39C43 amino-acid badly soluble peptide (Selkoe, 1991). The onset of hereditary types of Alzheimer’s disease generally precedes that Rabbit Polyclonal to PTGIS of the sporadic instances by several years. This is regarded as because of the extreme overproduction of the and, especially that of the easily aggregable 42 aminoacid type of A (for review discover Checler, 1995). The acceleration from the A creation has been proven because of the existence of missense mutations in the amyloid precursor proteins (APP, Citron et al., 1992; Cai et al., 1993; Felsenstein et al., 1994) and recently, in two homologous protein called presenilins 1 and 2 (PS1, PS2) (Borchelt et al., 1996; Duff et al., 1996; Citron et al., 1997; Tomita et al., 1997; Xia et al., 1997; Ancolio et al., 1997; Marambaud et al., 1998b). The actual fact that specific proteins, all in charge of aggressive types of Alzheimer’s disease, could result in identical phenotypic overproduction of the argues towards a restorative strategy targeted at slowing the creation of the peptide. With this framework, putative restorative targets could possibly be – and -secretases, (the proteolytic actions responsible for the discharge of the from its precursor) or additional mechanisms in charge of the rules of APP control. Effectors from the proteins kinase C have already been shown to reduce A creation and boost secretion from the -secretase-derived physiological item APP in a variety of cell lines (Caporaso et al., 1992; Gillespie et al., 1992; Buxbaum et al., 1993; Hung et al., 1993). Furthermore, in gene-targeted mice overproducing A, the administration from the PKC stimulator phorbol 12,13-dibutyrate (PDBu) resulted in extreme inhibition from the creation of the (Savage et al., 1998). We lately showed how the maturation of APP is apparently under control from the proteins kinase A (PKA) pathway in human being cells and neurons overexpressing regular and FAD-linked APP (Marambaud et al., 1998a). Nevertheless, unlike modulators of PKC, effectors from the PKA pathway activated creation of both A and APP (Marambaud et al., 1998a) recommending that the prospective of PKA was most likely located upstream of both – and /- secretases cleavages. Right here we display that two specific PKA inhibitors significantly decrease the constitutive creation of both A40 and A42 in stably transfected HEK293 cells expressing crazy type (wt) and Swedish mutated (Sw) APP751. We also set up that PKA inhibitors nearly completely avoid the development of As by HEK293 cells overexpressing wt- and M146V-PS1. Oddly enough, the inhibitors usually do not considerably influence the recoveries of APP or its -secretase-derived C-terminal stub, p10. Our data reveal that PKA inhibitors selectively influence the /-secretase pathway in human being cells and so are potential pharmacological which might be able to decrease.