Mechanisms controlling the clinical response genes to antipsychotics: role of D1 receptor- cAMP-CREB signalling in the regulation of ADAMTS2

RESUMEN: El conocimiento de los mecanismos moleculares implicados en el desarrollo y las manifestaciones clínicas de la esquizofrenia pueden mejorar nuestra capacidad para diagnosticarlo y tratarlo. Datos previos asociaron la desregulación de ADAMTS2 en PBMCs de pacientes en FEP, así como en respondedores clínicos a AP. En esta tesis, realizamos una validación independiente de dichos datos y estudiamos los mecanismos implicados en el control de ADAMTS2. Verificamos nuestros resultados previos utilizando una nueva cohorte de pacientes con esquizofrenia con seguimiento clínico tras el tratamiento. Además, ADAMTS2 se activó mediante señalización de receptores de clase D1 y señalización CREB / ERK, que fue derogada in vitro mediante AP e inhibidores de PKA/NEK. En conclusión, receptores D1 y activación de CREB podrían participar en el inicio y respuesta clínicas a la terapia, controlando la expresión y actividad de ADAMTS2, y proporcionando un nuevo panorama de mejorar de su eficacia clínica.ABSTRACT: Molecular mechanisms in the development and clinical manifestations of schizophrenia can improve our ability to diagnose and treat it. Previous data associated ADAMTS2 expression deregulation in PBMCs from patients at FEP (up) as well as in clinical responders to APs (down). In this thesis, we performed an independent validation of such data and studied the mechanisms implicated in the control of ADAMTS2. Using a new cohort of drug-naïve schizophrenia patients with clinical follow-up, we confirmed our previous data. Also, ADAMTS2 was activated by D1-class receptors signalling and downstream by CREB/ERK signalling. APs and selective PKA/MEK inhibitors abrogated D1-mediated activation of ADAMTS2 in vitro. Thus, D1 receptors signalling towards CREB activation might participate in the onset and clinical responses to therapy in schizophrenia, by controlling ADAMTS2 expression and activity. The unbiased investigation of molecular mechanisms triggered by APs may provide a new landscape of novel targets potentially associated with clinical efficacy.La financiación necesaria para la realización de esta Tesis doctoral ha sido aportada por el MINECO mediante los proyectos SAF2013-46292-R y SAF2016-76046-R, y por el ISCII mediante el Proyecto PI16/00156 y la financiación específica al grupo de investigación CIBERSAM-G26. El autor de esta tesis ha disfrutado de contrato predoctoral de Formación de Personal Investigador (FPI), referencia BES-2014-070615, concedida por el MINECO, y de un contrato de 6 meses mediante la ayuda de Luchamos por la Vida 2019 en el Instituto de Investigación Marqués de Valdecilla (IDIVAL)

Time dependent expression of the blood biomarkers EIF2D and TOX in patients with schizophrenia

Background During last years, there has been an intensive search for blood biomarkers in schizophrenia to assist in diagnosis, prognosis and clinical management of the disease. Methods In this study, we first conducted a weighted gene coexpression network analysis to address differentially expressed genes in peripheral blood from patients with chronic schizophrenia (n?=?30) and healthy controls (n?=?15). The discriminating performance of the candidate genes was further tested in an independent cohort of patients with first-episode schizophrenia (n?=?124) and healthy controls (n?=?54), and in postmortem brain samples (cingulate and prefrontal cortices) from patients with schizophrenia (n?=?34) and healthy controls (n?=?35). Results The expression of the Eukaryotic Translation Initiation Factor 2D (EIF2D) gene, which is involved in protein synthesis regulation, was increased in the chronic patients of schizophrenia. On the contrary, the expression of the Thymocyte Selection-Associated High Mobility Group Box (TOX) gene, involved in immune function, was reduced. EIF2D expression was also altered in first-episode schizophrenia patients, but showing reduced levels. Any of the postmortem brain areas studied did not show differences of expression of both genes. Conclusions EIF2D and TOX are putative blood markers of chronic patients of schizophrenia, which expression change from the onset to the chronic disease, unraveling new biological pathways that can be used for the development of new intervention strategies in the diagnosis and prognosis of schizophrenia disease.Acknowledgments: This work was supported by Fondo de Investigación Sanitaria, Ministerio de Economía y Competitividad, Spain (PI10/01399, PI13/00447; PI17/00402, co-financed by FEDER) to J. Sanjuan and M.D. Moltó; Generalitat Valenciana PROMETEO Excellence Program, Spain (PROMETEO2016/082) to J Sanjuán. J Gilabert-Juan and N. Sebastiá-Ortega were recipients of research contracts from CIBERSAM, Spain. The RNA samples donated bythe Stanley Medical Research Institute Brain Collection were courtesy of Drs. Michael B. Knable, E. Fuller Torrey, Maree J. Webster, and Robert H. Yolken. The authors also thank the collaboration of the staff members of the hospitals

Dopaminergic control of ADAMTS2 expression through cAMP/CREB and ERK: molecular effects of antipsychotics

A better understanding of the molecular mechanisms that participate in the development and clinical manifestations of schizophrenia can lead to improve our ability to diagnose and treat this disease. Previous data strongly associated the levels of deregulated ADAMTS2 expression in peripheral blood mononuclear cells (PBMCs) from patients at first episode of psychosis (up) as well as in clinical responders to treatment with antipsychotic drugs (down). In this current work, we performed an independent validation of such data and studied the mechanisms implicated in the control of ADAMTS2 gene expression. Using a new cohort of drug-naïve schizophrenia patients with clinical follow-up, we confirmed that the expression of ADAMTS2 was highly upregulated in PBMCs at the onset (drug-naïve patients) and downregulated, in clinical responders, after treatment with antipsychotics. Mechanistically, ADAMTS2 expression was activated by dopaminergic signalling (D1-class receptors) and downstream by cAMP/CREB and mitogen-activated protein kinase (MAPK)/ERK signalling. Incubation with antipsychotic drugs and selective PKA and MEK inhibitors abrogated D1-mediated activation of ADAMTS2 in neuronal-like cells. Thus, D1 receptors signalling towards CREB activation might participate in the onset and clinical responses to therapy in schizophrenia patients, by controlling ADAMTS2 expression and activity. The unbiased investigation of molecular mechanisms triggered by antipsychotic drugs may provide a new landscape of novel targets potentially associated with clinical efficacy.Acknowledgements: We are highly indebted to the participants and their families for their cooperation in this study. We also thank IDIVAL biobank (Inés Santiuste and Jana Arozamena) for clinical samples and data as well as the PAFIP members (Marga Corredera) for the data collection. This work was supported by: SAF2016-76046-R and SAF2013-46292-R (MINECO and FEDER) to B.C.F., PI16/00156 (isciii and FEDER) to J.P.V., LUCHAMOS POR LA VIDA project to F.R.J. and J.P.V., SAF2017-83702-R (MINECO and FEDER), Red TERCEL RD12/0019/0024 (ISCIII) and GVA-PROMETEO 2018/041 (Generalitat Valenciana) to S.M. J.P.V. is supported by the RyC research programme (RYC-2013-14097) and F.R.J. by the predoctoral research programme (BES-2014-070615), from MINECO and FEDER

Dopaminergic control of ADAMTS2 expression through cAMP/CREB and ERK: molecular effects of antipsychotics

© The Author(s) 2019.A better understanding of the molecular mechanisms that participate in the development and clinical manifestations of schizophrenia can lead to improve our ability to diagnose and treat this disease. Previous data strongly associated the levels of deregulated ADAMTS2 expression in peripheral blood mononuclear cells (PBMCs) from patients at first episode of psychosis (up) as well as in clinical responders to treatment with antipsychotic drugs (down). In this current work, we performed an independent validation of such data and studied the mechanisms implicated in the control of ADAMTS2 gene expression. Using a new cohort of drug-naïve schizophrenia patients with clinical follow-up, we confirmed that the expression of ADAMTS2 was highly upregulated in PBMCs at the onset (drug-naïve patients) and downregulated, in clinical responders, after treatment with antipsychotics. Mechanistically, ADAMTS2 expression was activated by dopaminergic signalling (D1-class receptors) and downstream by cAMP/CREB and mitogen-activated protein kinase (MAPK)/ERK signalling. Incubation with antipsychotic drugs and selective PKA and MEK inhibitors abrogated D1-mediated activation of ADAMTS2 in neuronal-like cells. Thus, D1 receptors signalling towards CREB activation might participate in the onset and clinical responses to therapy in schizophrenia patients, by controlling ADAMTS2 expression and activity. The unbiased investigation of molecular mechanisms triggered by antipsychotic drugs may provide a new landscape of novel targets potentially associated with clinical efficacy.This work was supported by: SAF2016-76046-R and SAF2013-46292-R (MINECO and FEDER) to B.C.F., PI16/00156 (isciii and FEDER) to J.P.V., LUCHAMOS POR LA VIDA project to F.R.J. and J.P.V., SAF2017-83702-R (MINECO and FEDER), Red TERCEL RD12/0019/0024 (ISCIII) and GVA-PROMETEO 2018/041 (Generalitat Valenciana) to S.M. J.P.V. is supported by the RyC research programme (RYC-2013-14097) and F.R.J. by the predoctoral research programme (BES-2014-070615), from MINECO and FEDER

Blood Gene Expression Profile Predicts Response to Antipsychotics

Antipsychotic drugs are one of the largest types of prescribed drugs and have large inter-individual differences in efficacy, but there is no methodology to predict their clinical effect. Here we show a four-gene blood expression profile to predict the response to antipsychotics in schizophrenia patients before treatment. We sequenced total mRNA from blood samples of antipsychotic naïve patients who, after 3 months of treatment, were in the top 40% with the best response (15 patients) and in the bottom 40% with the worst response (15 patients) according to the Brief Psychiatric Rating Scale (BPRS). We characterized the transcriptome before treatment of these 30 patients and found 130 genes with significant differential expression (Padj value < 0.01) associated with clinical response. Then, we used Random Forests, an ensemble learning method for classification and regression, to obtain a list of predictor genes. The expression of four genes can predict the response to antipsychotic medication with a cross-validation accuracy estimation of 0.83 and an area under the curve of 0.97 using a logistic regression. We anticipate that this approach is a gateway to select the specific antipsychotic that will produce the best response to treatment for each specific patient.FUNDING: This work was supported by the Ministerio de Ciencia e Innovación (MICINN) in a coordinated project (grant SAF2010-20840-C02-01/02) and by the Spanish Ministry of Economy, Industry and Competitiveness (grants SAF2013-46292-R; PTA2015-10483-I); and by the National Institutes of Health (grant 5R01HD056465-07. Sub-award #320793 from The Children’s Hospital of Philadelphia). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Papel de la proteína RhoE en la proliferación de células troncales neurales de la zona subventricular (SVZ) del cerebro

[EN] Rnd proteins are essential regulators of the cytoskeleton organization, playing a key role in several processes of the nervous system development. We have previously demonstrated that the absence of RhoE/Rnd3 affects the nervous system development, such as neuromotor and neuromuscular abnormalities, a decrease in the axonal and dendritic growth and a delayed neuronal polarization. Our goal was to investigate the role of RhoE in neurogenesis, specifically on the stem cell proliferation in the SVZ, by using neurosphere culture system. Our results show a decrease in the proliferation of progenitor cells, which correlates with a reduction in the levels of cyclin D1. This effect is essentially due to a decrease in DNA synthesis rather than an effect on survival. The lack of RhoE does not affect the stem cells (type B) self-renewal rate, suggesting that the decrease in proliferation would affect only the intermediate progenitor cells (type C). Finally, we have observed that the absence of RhoE alters the expression of certain proteins in neural progenitors, such as the GSK3 activation. This is a kinase related to the proliferation and differentiation of neural[ES] Las proteínas Rnd son reguladores esenciales de la organización del citoesqueleto celular, jugando un papel fundamental en diversos procesos del desarrollo del sistema nervioso. Nuestro grupo ha demostrado con anterioridad que la ausencia de RhoE/Rnd3 afecta al desarrollo sistema nervioso, observándose alteraciones neuromotoras y neuromusculares, una disminución en crecimiento dendrítico y axonal, y un retraso en la polarización neuronal. Nuestro objetivo ha sido investigar el papel de RhoE en la neurogénesis, concretamente en la proliferación de las células troncales de la SVZ, mediante su cultivo en forma de neuroesferas. Los resultados del presente trabajo indican una disminución en la proliferación de las células progenitoras, que se correlaciona con una reducción en los niveles de ciclina D1. Este efecto sobre la proliferación se debería fundamentalmente a una disminución en la síntesis de DNA, y no a un efecto sobre la supervivencia. La ausencia de RhoE no afecta a la tasa de autorrenovación de las células troncales (tipo B), lo que sugiere que la disminución en proliferación afectaría a las células progenitoras intermedias (tipo C). Finalmente, hemos observado que la ausencia de RhoE modifica la expresión de determinadas proteínas en los progenitores neurales, como son la activación de la GSK3, kinasa ampliamente relacionada con la proliferación y diferenciación de los progenitores neurales, que altera la expresión de las proteínas Tau y Ciclina D1.Ruso Julve, F. (2013). Papel de la proteína RhoE en la proliferación de células troncales neurales de la zona subventricular (SVZ) del cerebro. http://hdl.handle.net/10251/39182Archivo delegad

Effects of BAMBI deficiency in the development of systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies and immune complexes deposition in various organs. Follicular CD4+ T cells play a crucial role in this autoimmune syndrome by activating autoreactive germinal center B cells. The transforming growth factor ß (TGBß) inhibitor BAMBI (Bone Morphogenetic Protein and Activin Membrane-Bound Inhibitor) is known to control the differentiation of CD4+ T lymphocytes into either tolerogenic Tregs or pathogenic Th17 cells. BAMBI absence in mutant mice or its pharmacological inhibition with the B101.37 monoclonal antibody promotes an increase in Treg differentiation and activity and at the same time, a reduction in Th17 cell differentiation, resulting in the inhibition of autoimmune disease development. Based on these findings, in the present study we have analysed the effects of BAMBI deficiency in the development of SLE. We have employed the chronic graft-versus-host disease lupus model induced after the transference of 40-60 × 106 lymphocytes from B6 or B6-BAMBI-KO mice into either Bm12 or Bm12-BAMBI-KO recipients. As reported previously, Bm12 mice receiving lymphocytes from B6 mice produce high levels of circulating IgG anti-DNA and anti-nucleosomal autoantibodies and exhibit linear glomerular IgG deposits in their kidneys. In contrast, both Bm12 mice injected with allogeneic B6-BAMBI-KO lymphocytes and Bm12-BAMBI-KO recipients receiving lymphocytes from B6 donors do not produce significant levels of circulating autoantibodies. Taken together, these results indicate that the absence of BAMBI in either donor or recipient lymphocytes protects mice against the development of SLE

Table_3.DOCX

<p>Antipsychotic drugs are one of the largest types of prescribed drugs and have large inter-individual differences in efficacy, but there is no methodology to predict their clinical effect. Here we show a four-gene blood expression profile to predict the response to antipsychotics in schizophrenia patients before treatment. We sequenced total mRNA from blood samples of antipsychotic naïve patients who, after 3 months of treatment, were in the top 40% with the best response (15 patients) and in the bottom 40% with the worst response (15 patients) according to the Brief Psychiatric Rating Scale (BPRS). We characterized the transcriptome before treatment of these 30 patients and found 130 genes with significant differential expression (P_adj value < 0.01) associated with clinical response. Then, we used Random Forests, an ensemble learning method for classification and regression, to obtain a list of predictor genes. The expression of four genes can predict the response to antipsychotic medication with a cross-validation accuracy estimation of 0.83 and an area under the curve of 0.97 using a logistic regression. We anticipate that this approach is a gateway to select the specific antipsychotic that will produce the best response to treatment for each specific patient.</p

Table_1.PDF

<p>Antipsychotic drugs are one of the largest types of prescribed drugs and have large inter-individual differences in efficacy, but there is no methodology to predict their clinical effect. Here we show a four-gene blood expression profile to predict the response to antipsychotics in schizophrenia patients before treatment. We sequenced total mRNA from blood samples of antipsychotic naïve patients who, after 3 months of treatment, were in the top 40% with the best response (15 patients) and in the bottom 40% with the worst response (15 patients) according to the Brief Psychiatric Rating Scale (BPRS). We characterized the transcriptome before treatment of these 30 patients and found 130 genes with significant differential expression (P_adj value < 0.01) associated with clinical response. Then, we used Random Forests, an ensemble learning method for classification and regression, to obtain a list of predictor genes. The expression of four genes can predict the response to antipsychotic medication with a cross-validation accuracy estimation of 0.83 and an area under the curve of 0.97 using a logistic regression. We anticipate that this approach is a gateway to select the specific antipsychotic that will produce the best response to treatment for each specific patient.</p

Table_5.DOCX

<p>Antipsychotic drugs are one of the largest types of prescribed drugs and have large inter-individual differences in efficacy, but there is no methodology to predict their clinical effect. Here we show a four-gene blood expression profile to predict the response to antipsychotics in schizophrenia patients before treatment. We sequenced total mRNA from blood samples of antipsychotic naïve patients who, after 3 months of treatment, were in the top 40% with the best response (15 patients) and in the bottom 40% with the worst response (15 patients) according to the Brief Psychiatric Rating Scale (BPRS). We characterized the transcriptome before treatment of these 30 patients and found 130 genes with significant differential expression (P_adj value < 0.01) associated with clinical response. Then, we used Random Forests, an ensemble learning method for classification and regression, to obtain a list of predictor genes. The expression of four genes can predict the response to antipsychotic medication with a cross-validation accuracy estimation of 0.83 and an area under the curve of 0.97 using a logistic regression. We anticipate that this approach is a gateway to select the specific antipsychotic that will produce the best response to treatment for each specific patient.</p