- Email: [email protected]
CEREBROSPINAL FLUID LEVELS OF ANGIOTENSIN-CONVERTING ENZYME ARE ASSOCIATED WITH AMYLOID-β IN ALZHEIMER’S DISEASE
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Poster Presentations: Sunday, July 16, 2017
have investigated the role of trans-acting factors; transcription factor SOX2 and miRNA-140, in ADAM10 gene regulation. The entire sequence of ADAM10 gene is analyzed by computational bioinformatics and screened for probable trans-actors with high regulatory importance. Results: Our study shows that miRNA-140-5p has enhanced expression in AD post-mortem brain hippocampus using high throughput micro-arrays and qRT-PCR. Interestingly we have also seen that miRNA-140 seed sequence is present on both ADAM10 and SOX2 3’UTR and thus reporter constructs containing regulatory elements were transfected into human cell lines along with miRNA-140 mimics and inhibitors to evaluate their interaction by dual luciferase reporter assays. The specific interaction of miRNA-140 with both ADAM10 and its transcription factor SOX2 signifies high regulatory importance of this miRNA in controlling ADAM10 expression. Conclusions: Thus our proposed investigation unravels the mechanisms underlying ADAM10 downregulation by miR-140 that exacerbates Alzheimer’s disease related neurotoxic effects. Our findings could reflect a strong basis for future research aimed at understanding the potential contribution of trans-acting factors as a biomarker or modulator of AD pathophysiology.
P1-212
CEREBROSPINAL FLUID LEVELS OF ANGIOTENSIN-CONVERTING ENZYME ARE ASSOCIATED WITH AMYLOID-b IN ALZHEIMER’S DISEASE
Natalia P. Rocha1,2, Andre ASF. Toledo2, Laiane TS. Corgosinho2, Leonardo Cruz de Souza2, Elisa de Paula Franc¸a Resende2, Nayara FT. Braz2, Ana C. Simoes e Silva2, Paulo Caramelli2, Antonio L. Teixeira2,3, 1The University of Texas Health Science Center at Houston, Houston, TX, USA; 2Universidade Federal de Minas Gerais, Belo Horizonte, Brazil; 3The University of Texas Health Science Center at Houston, Houston, TX, USA. Contact e-mail: [email protected] Background: Although the renin-angiotensin system (RAS) is traditionally regarded as a system that maintains blood pressure and fluid homeostasis, a range of evidence points to its role in different cerebral functions. For instance, all RAS components are found in the central nervous system. Experimental and epidemiological studies indicate that both angiotensin receptor type 1 (AT1) antagonists and angiotensin converting enzyme
(ACE) inhibitors may have beneficial effects for Alzheimer’s disease (AD). In the current study, we aimed to determine cerebrospinal (CSF) levels of RAS components in patients with AD and controls. We also investigated potential associations
Table 1 Clinical characteristics and cerebrospinal fluid levels of angiotensin-converting enzyme (ACE), amyloid-b, phosphorylated Tau (pTau) and Tau protein (hTau) in Alzheimer’s disease (AD) patients and controls.
Age in years, mean 6 SD (median) Female sex, N (%) Educational level MMSE, mean 6 SD (median) Ab40 (pg/mL), mean 6 SD (median) Ab42 (pg/mL), mean 6 SD (median) hTau (pg/mL), mean 6 SD (median) pTau (pg/mL), mean 6 SD (median) ACE (pg/mL), mean 6 SD (median) ACE2 (pg/mL), mean 6 SD (median) IATI, mean 6 SD (median) hTau/Ab42, mean 6 SD (median) pTau/Ab42, mean 6 SD (median)
Controls (n ¼ 10)
AD (n ¼ 18)
P value
69.0069.45 (65.50) 5 (50%) 3.4461.81 (3.00) 25.4463.13 (27.00) 1208.846732.42 (914.61) 1183.256248.30 (1275.70) 275.27676.44 (263.16) 52.77615.36 (48.41) 53.5564.01 (53.81) 12.4565.56 (53.81) 12.4565.56 (11.75) 0.2460.06 (0.24) 0.0560.01 (0.05)
66.6968.75 (67.50) 9 (50%) 8.9265.06 (8.00) 20.1567.82 (23.00) 638.046508.93 (549.78) 513.856225.22 (504.99) 1096.056957.24 (809.84) 106.73649.09 (104.67) 49.6965.01 (49.55) 13.9064.92 (13.67) 0.4260.17 (0.45) 2.1261.67 (1.53) 0.2160.07 (0.19)
0.531a 1.000b 0.003c 0.071c 0.020a <0.0001c <0.0001c 0.002a 0.046a 0.477a <0.0001a <0.0001c <0.0001c
MMSE ¼ mini-mental state examination, IATI ¼ Innotest Amyloid Tau Index a Student’s t test; bFisher’s exact test; cMann-Whitney test. Significant values are highlighted in bold.
Poster Presentations: Sunday, July 16, 2017
between CSF levels of RAS components and AD biomarkers. Methods: This study included 18 patients with probable AD recruited from the Behavioral and Cognitive Neurology Clinic of the Universidade Federal de Minas Gerais (UFMG), Brazil. A control group composed by 10 subjects undergoing lumbar puncture for anesthetic purpose was also enrolled. These subjects had no history of neurological and/or psychiatric disorders. After collection, CSF samples were centrifuged at 1,800 g for 10 minutes (4⁰C) and stored at -80⁰C until biomarker analysis. Ab40, Ab42, total tau (hTau), and phosphorylated-tau (pTau) were measured by enzyme-linked immunosorbent assay (ELISA) with commercially available kits (Innogenetics/Fujirebio, Gent, Belgium). Angiotensin (Ang) II, Ang-(1-7), ACE and ACE2 levels were also measured by ELISA according to the procedures supplied by the manufacturer (MyBioSource, San Diego, CA, USA). Results: All patients with AD met the CSF criteria for biomarker-based diagnosis, (i.e., IATI < 0.8; hTau/ Ab42 ratio > 0.52 and pTau/Ab42 ratio >0.08). Accordingly, we found decreased levels of Ab40 and Ab42, and increased levels of hTau and pTau in the CSF of patients with AD in comparison with controls (Table 1). Ang II and Ang-(1-7) were not detected in the CSF samples. Patients with AD presented decreased levels of ACE, but similar levels of ACE2 when compared with controls (Table 1; Fig.1A). Among patients with AD, ACE levels correlated positively with Ab42 levels (Fig.1B). Conclusions: CSF levels of ACE are reduced in AD, correlating with Ab42 levels. Our results strengthen the hypothesis that ACE is associated with amyloid- b pathology in AD.
P1-213
THE EFFECT OF SERUM FACTORS FROM PATIENTS WITH/WITHOUT ESSENTIAL HYPERTENSIOIN ON MEMBRANE RECEPTORS FROM RAT CEREBRAL CORTEX VASCULAR AND CARDIAC MUSCLE CELLS
Cristian Romeo S Revnic1, Floarea Revnic2, Silviu Voinea3, 1UMF Carol Davila, Bucharest, Romania; 2NIGG Ana Aslan, Bucharest, Romania; 3 UMF Carol Davila, Bucharest, Romania. Contact e-mail: kityrom@yahoo. com Background: Essential Hypertension (EHT)is one of the main risk factors for Alzheimer’s disease(AD). The aim of our study was to point out experimental data on the potential role of some serum factors from blood of patients with (EHT) in determining some changes in the ionic flow and neuromediators uptake by the rat cerebral cortex, cardiac and arterial smooth muscle cell receptors with a view to estimate the relevance of these factors in the development of cardiovascular pathology and (AD) and the possible therapeutic management of these agents under study. Methods: Our study has been conducted on 34 patients :17 aged 35-86 with EHT (10 men and 7 women) and 17 normotensive subjects (aged 30-84 years old) hospitalized in NIGG Ana Aslan. The radioactive incubation experiments of rat cerebral cortex, heart and aorta have been done according with the published technique. The uptake of 45CaCl2,3H Serotonine,3H Cortisol and 3H Norepinephrine by animal tissues in the presence of serum from (EHT) and normotensive patients has been assayed using a Beta Berthold Scintilator Counter. Results: Our data have pointed out a a significant increase in 45Ca uptake by cerebral cortex incubated
P325
with serum from (EHT) patients while in rat heart and aorta there is a decrease in 45Ca uptake. When incubated with serum from (EHT) patients in the presence of 3H Norepinephrine, the uptake is increased in cerebral cortex and heart and decreased in aorta. In samples incubated with serum from hypertensive patients, the uptake of 3H Serotonine was significantly decreased in rat brain cortex, heart and aorta. in comparison with biological samples incubated with serum from normothensive patients. Regarding 3H Cortisol uptake, a significant increase in brain cortex and aorta has been recorded in the presence of serum from hypertensive patients while in rat heart there was a decrease in 3H Cortisol uptake. Conclusions: According to our experimental results serum factors present in the blood of (EHT) patients are responsible for changes in receptor state behaviour concerning the uptake of 45Ca,3H SErotonine,3H Noradrenaline and 3H Cortisol in rat cerebral cortex as well as in heart and aorta which may have a significance in the onset of (AD) in (EHT) patients.
P1-214
ANALYSIS OF THE EXPRESSION OF TASTE AND OLFACTORY RECEPTORS IN CHOROID PLEXUS AND ORBITOFRONTAL CORTEX OF ALZHEIMER’S DISEASE PATIENTS
Victoria Cunha Alves1, Joana Figueiro Silva1, Isabel Gonc¸alves2, Cecılia Santos2, Isidre Ferrer3, Eva Carro4, 1Instituto de Investigacion Hospital 12 de Octubre, Madrid, Spain; 2CICS - Centro de Investigac¸~ao em Ci^encias da Saude, Covilha, Portugal; 3Institut d’Investigacio Biomedica de Bellvitge-Hospital Universitari de Bellvitge, Barcelona, Spain; 4Research Institute Hospital 12 de Octubre (i+12), Madrid, Spain. Contact e-mail: [email protected] Background: Alzheimer’s disease (AD) is a chronic neurode-
generative disease characterized by progressive memory loss and cognitive deterioration, attributed to neuropathological lesions within specific regions in the brain. However, other areas of the brain such as the choroid plexus and the orbitofrontal cortex, have gained much attention since the choroid plexus is a multifunctional tissue responsible for a wide range of functions crucial to the central nervous system and the orbitofrontal cortex is considered among the most polymodal regions of the brain. Previous studies have shown the expression of taste and olfactory transduction pathways in rat choroid plexus as well as taste and olfactory regulation and activation of orbitofrontal cortex. Methods: Transcriptomic analysis of these chemoreceptors were performed by means of Real-time quantitative Polymerase Chain Reaction (RT-qPCR), and compared between AD patients (ad different Braak stages of the disease) and age-matched controls. Results: Transcriptomic analysis indicated that orbitofrontal cortical olfactory receptors (ORs) and taste receptors (TASRs) are expressed and regulated at different stages of AD in male and female patients. Moreover, these receptors were differentially regulated at Braak stages I, V, VI, compared to age-matched controls in the orbitofrontal cortex of AD patients. The strongest differences were found at Braak stage I of AD. These receptors we also found to be expressed in choroid plexus, however, their expression is very low. Conclusions: Taken together, these results suggest that dysregulation of ORs and TASRs is an early event in the pathogenesis of AD. Nevertheless further studies are needed
Poster Presentations: Sunday, July 16, 2017
have investigated the role of trans-acting factors; transcription factor SOX2 and miRNA-140, in ADAM10 gene regulation. The entire sequence of ADAM10 gene is analyzed by computational bioinformatics and screened for probable trans-actors with high regulatory importance. Results: Our study shows that miRNA-140-5p has enhanced expression in AD post-mortem brain hippocampus using high throughput micro-arrays and qRT-PCR. Interestingly we have also seen that miRNA-140 seed sequence is present on both ADAM10 and SOX2 3’UTR and thus reporter constructs containing regulatory elements were transfected into human cell lines along with miRNA-140 mimics and inhibitors to evaluate their interaction by dual luciferase reporter assays. The specific interaction of miRNA-140 with both ADAM10 and its transcription factor SOX2 signifies high regulatory importance of this miRNA in controlling ADAM10 expression. Conclusions: Thus our proposed investigation unravels the mechanisms underlying ADAM10 downregulation by miR-140 that exacerbates Alzheimer’s disease related neurotoxic effects. Our findings could reflect a strong basis for future research aimed at understanding the potential contribution of trans-acting factors as a biomarker or modulator of AD pathophysiology.
P1-212
CEREBROSPINAL FLUID LEVELS OF ANGIOTENSIN-CONVERTING ENZYME ARE ASSOCIATED WITH AMYLOID-b IN ALZHEIMER’S DISEASE
Natalia P. Rocha1,2, Andre ASF. Toledo2, Laiane TS. Corgosinho2, Leonardo Cruz de Souza2, Elisa de Paula Franc¸a Resende2, Nayara FT. Braz2, Ana C. Simoes e Silva2, Paulo Caramelli2, Antonio L. Teixeira2,3, 1The University of Texas Health Science Center at Houston, Houston, TX, USA; 2Universidade Federal de Minas Gerais, Belo Horizonte, Brazil; 3The University of Texas Health Science Center at Houston, Houston, TX, USA. Contact e-mail: [email protected] Background: Although the renin-angiotensin system (RAS) is traditionally regarded as a system that maintains blood pressure and fluid homeostasis, a range of evidence points to its role in different cerebral functions. For instance, all RAS components are found in the central nervous system. Experimental and epidemiological studies indicate that both angiotensin receptor type 1 (AT1) antagonists and angiotensin converting enzyme
(ACE) inhibitors may have beneficial effects for Alzheimer’s disease (AD). In the current study, we aimed to determine cerebrospinal (CSF) levels of RAS components in patients with AD and controls. We also investigated potential associations
Table 1 Clinical characteristics and cerebrospinal fluid levels of angiotensin-converting enzyme (ACE), amyloid-b, phosphorylated Tau (pTau) and Tau protein (hTau) in Alzheimer’s disease (AD) patients and controls.
Age in years, mean 6 SD (median) Female sex, N (%) Educational level MMSE, mean 6 SD (median) Ab40 (pg/mL), mean 6 SD (median) Ab42 (pg/mL), mean 6 SD (median) hTau (pg/mL), mean 6 SD (median) pTau (pg/mL), mean 6 SD (median) ACE (pg/mL), mean 6 SD (median) ACE2 (pg/mL), mean 6 SD (median) IATI, mean 6 SD (median) hTau/Ab42, mean 6 SD (median) pTau/Ab42, mean 6 SD (median)
Controls (n ¼ 10)
AD (n ¼ 18)
P value
69.0069.45 (65.50) 5 (50%) 3.4461.81 (3.00) 25.4463.13 (27.00) 1208.846732.42 (914.61) 1183.256248.30 (1275.70) 275.27676.44 (263.16) 52.77615.36 (48.41) 53.5564.01 (53.81) 12.4565.56 (53.81) 12.4565.56 (11.75) 0.2460.06 (0.24) 0.0560.01 (0.05)
66.6968.75 (67.50) 9 (50%) 8.9265.06 (8.00) 20.1567.82 (23.00) 638.046508.93 (549.78) 513.856225.22 (504.99) 1096.056957.24 (809.84) 106.73649.09 (104.67) 49.6965.01 (49.55) 13.9064.92 (13.67) 0.4260.17 (0.45) 2.1261.67 (1.53) 0.2160.07 (0.19)
0.531a 1.000b 0.003c 0.071c 0.020a <0.0001c <0.0001c 0.002a 0.046a 0.477a <0.0001a <0.0001c <0.0001c
MMSE ¼ mini-mental state examination, IATI ¼ Innotest Amyloid Tau Index a Student’s t test; bFisher’s exact test; cMann-Whitney test. Significant values are highlighted in bold.
Poster Presentations: Sunday, July 16, 2017
between CSF levels of RAS components and AD biomarkers. Methods: This study included 18 patients with probable AD recruited from the Behavioral and Cognitive Neurology Clinic of the Universidade Federal de Minas Gerais (UFMG), Brazil. A control group composed by 10 subjects undergoing lumbar puncture for anesthetic purpose was also enrolled. These subjects had no history of neurological and/or psychiatric disorders. After collection, CSF samples were centrifuged at 1,800 g for 10 minutes (4⁰C) and stored at -80⁰C until biomarker analysis. Ab40, Ab42, total tau (hTau), and phosphorylated-tau (pTau) were measured by enzyme-linked immunosorbent assay (ELISA) with commercially available kits (Innogenetics/Fujirebio, Gent, Belgium). Angiotensin (Ang) II, Ang-(1-7), ACE and ACE2 levels were also measured by ELISA according to the procedures supplied by the manufacturer (MyBioSource, San Diego, CA, USA). Results: All patients with AD met the CSF criteria for biomarker-based diagnosis, (i.e., IATI < 0.8; hTau/ Ab42 ratio > 0.52 and pTau/Ab42 ratio >0.08). Accordingly, we found decreased levels of Ab40 and Ab42, and increased levels of hTau and pTau in the CSF of patients with AD in comparison with controls (Table 1). Ang II and Ang-(1-7) were not detected in the CSF samples. Patients with AD presented decreased levels of ACE, but similar levels of ACE2 when compared with controls (Table 1; Fig.1A). Among patients with AD, ACE levels correlated positively with Ab42 levels (Fig.1B). Conclusions: CSF levels of ACE are reduced in AD, correlating with Ab42 levels. Our results strengthen the hypothesis that ACE is associated with amyloid- b pathology in AD.
P1-213
THE EFFECT OF SERUM FACTORS FROM PATIENTS WITH/WITHOUT ESSENTIAL HYPERTENSIOIN ON MEMBRANE RECEPTORS FROM RAT CEREBRAL CORTEX VASCULAR AND CARDIAC MUSCLE CELLS
Cristian Romeo S Revnic1, Floarea Revnic2, Silviu Voinea3, 1UMF Carol Davila, Bucharest, Romania; 2NIGG Ana Aslan, Bucharest, Romania; 3 UMF Carol Davila, Bucharest, Romania. Contact e-mail: kityrom@yahoo. com Background: Essential Hypertension (EHT)is one of the main risk factors for Alzheimer’s disease(AD). The aim of our study was to point out experimental data on the potential role of some serum factors from blood of patients with (EHT) in determining some changes in the ionic flow and neuromediators uptake by the rat cerebral cortex, cardiac and arterial smooth muscle cell receptors with a view to estimate the relevance of these factors in the development of cardiovascular pathology and (AD) and the possible therapeutic management of these agents under study. Methods: Our study has been conducted on 34 patients :17 aged 35-86 with EHT (10 men and 7 women) and 17 normotensive subjects (aged 30-84 years old) hospitalized in NIGG Ana Aslan. The radioactive incubation experiments of rat cerebral cortex, heart and aorta have been done according with the published technique. The uptake of 45CaCl2,3H Serotonine,3H Cortisol and 3H Norepinephrine by animal tissues in the presence of serum from (EHT) and normotensive patients has been assayed using a Beta Berthold Scintilator Counter. Results: Our data have pointed out a a significant increase in 45Ca uptake by cerebral cortex incubated
P325
with serum from (EHT) patients while in rat heart and aorta there is a decrease in 45Ca uptake. When incubated with serum from (EHT) patients in the presence of 3H Norepinephrine, the uptake is increased in cerebral cortex and heart and decreased in aorta. In samples incubated with serum from hypertensive patients, the uptake of 3H Serotonine was significantly decreased in rat brain cortex, heart and aorta. in comparison with biological samples incubated with serum from normothensive patients. Regarding 3H Cortisol uptake, a significant increase in brain cortex and aorta has been recorded in the presence of serum from hypertensive patients while in rat heart there was a decrease in 3H Cortisol uptake. Conclusions: According to our experimental results serum factors present in the blood of (EHT) patients are responsible for changes in receptor state behaviour concerning the uptake of 45Ca,3H SErotonine,3H Noradrenaline and 3H Cortisol in rat cerebral cortex as well as in heart and aorta which may have a significance in the onset of (AD) in (EHT) patients.
P1-214
ANALYSIS OF THE EXPRESSION OF TASTE AND OLFACTORY RECEPTORS IN CHOROID PLEXUS AND ORBITOFRONTAL CORTEX OF ALZHEIMER’S DISEASE PATIENTS
Victoria Cunha Alves1, Joana Figueiro Silva1, Isabel Gonc¸alves2, Cecılia Santos2, Isidre Ferrer3, Eva Carro4, 1Instituto de Investigacion Hospital 12 de Octubre, Madrid, Spain; 2CICS - Centro de Investigac¸~ao em Ci^encias da Saude, Covilha, Portugal; 3Institut d’Investigacio Biomedica de Bellvitge-Hospital Universitari de Bellvitge, Barcelona, Spain; 4Research Institute Hospital 12 de Octubre (i+12), Madrid, Spain. Contact e-mail: [email protected] Background: Alzheimer’s disease (AD) is a chronic neurode-
generative disease characterized by progressive memory loss and cognitive deterioration, attributed to neuropathological lesions within specific regions in the brain. However, other areas of the brain such as the choroid plexus and the orbitofrontal cortex, have gained much attention since the choroid plexus is a multifunctional tissue responsible for a wide range of functions crucial to the central nervous system and the orbitofrontal cortex is considered among the most polymodal regions of the brain. Previous studies have shown the expression of taste and olfactory transduction pathways in rat choroid plexus as well as taste and olfactory regulation and activation of orbitofrontal cortex. Methods: Transcriptomic analysis of these chemoreceptors were performed by means of Real-time quantitative Polymerase Chain Reaction (RT-qPCR), and compared between AD patients (ad different Braak stages of the disease) and age-matched controls. Results: Transcriptomic analysis indicated that orbitofrontal cortical olfactory receptors (ORs) and taste receptors (TASRs) are expressed and regulated at different stages of AD in male and female patients. Moreover, these receptors were differentially regulated at Braak stages I, V, VI, compared to age-matched controls in the orbitofrontal cortex of AD patients. The strongest differences were found at Braak stage I of AD. These receptors we also found to be expressed in choroid plexus, however, their expression is very low. Conclusions: Taken together, these results suggest that dysregulation of ORs and TASRs is an early event in the pathogenesis of AD. Nevertheless further studies are needed