BAMBI (bone morphogenetic protein and activin membrane-bound inhibitor) reveals the involvement of the transforming growth factor-beta family in pain modulation

Transforming growth factors- (TGF- s) signal through type I and type II serine–threonine kinase receptor complexes. During ligand binding, type II receptors recruit and phosphorylate type I receptors, triggering downstream signaling. BAMBI [bone morphogenetic protein (BMP) and activin membrane-bound inhibitor] is a transmembrane pseudoreceptor structurally similar to type I receptors but lacks the intracellular kinase domain. BAMBI modulates negatively pan-TGF- family signaling; therefore, it can be used as an instrument for unraveling the roles of these cytokines in the adult CNS. BAMBI is expressed in regions of the CNS involved in pain transmission and modulation. The lack of BAMBI in mutant mice resulted in increased levels of TGF- signaling activity, which was associated with attenuation of acute pain behaviors, regardless of the modality of the stimuli (thermal, mechanical, chemical/inflammatory). The nociceptive hyposensitivity exhibited by BAMBI / mice was reversed by the opioid antagonist naloxone. Moreover, in a model of chronic neuropathic pain, the allodynic responses of BAMBI / mice also appeared attenuated through a mechanism involving -opioid receptor signaling. BasalmRNAand protein levels of precursor proteins of the endogenous opioid peptides proopiomelanocortin (POMC) and proenkephalin (PENK) appeared increased in the spinal cords of BAMBI / . Transcript levels of TGF- s and their intracellular effectors correlated directly with genes encoding opioid peptides, whereas BAMBI correlated inversely. Furthermore, incubation of spinal cord explants with activin A or BMP-7 increased POMC and/or PENK mRNA levels. Our findings identify TGF- family members as modulators of acute and chronic pain perception through the transcriptional regulation of genes encoding the endogenous opioids

Sex-Specific Regulation of miR-29b in the Myocardium Under Pressure Overload is Associated with Differential Molecular, Structural and Functional Remodeling Patterns in Mice and Patients with Aortic Stenosis

Pressure overload in patients with aortic stenosis (AS) induces an adverse remodeling of the left ventricle (LV) in a sex-specific manner. We assessed whether a sex-specific miR-29b dysregulation underlies this sex-biased remodeling pattern, as has been described in liver fibrosis. We studied mice with transverse aortic constriction (TAC) and patients with AS. miR-29b was determined in the LV (mice, patients) and plasma (patients). Expression of remodeling-related markers and histological fibrosis were determined in mouse LV. Echocardiographic morpho-functional parameters were evaluated at baseline and post-TAC in mice, and preoperatively and 1 year after aortic valve replacement (AVR) in patients with AS. In mice, miR-29b LV regulation was opposite in TAC-males (down-regulation) and TAC-females (up-regulation). The subsequent changes in miR-29b targets (collagens and GSK-3?) revealed a remodeling pattern that was more fibrotic in males but more hypertrophic in females. Both systolic and diastolic cardiac functions deteriorated more in TAC-females, thus suggesting a detrimental role of miR-29b in females, but was protective in the LV under pressure overload in males. Clinically, miR-29b in controls and patients with AS reproduced most of the sexually dimorphic features observed in mice. In women with AS, the preoperative plasma expression of miR-29b paralleled the severity of hypertrophy and was a significant negative predictor of reverse remodeling after AVR; therefore, it may have potential value as a prognostic biomarker

Identification of Epigenetic Interactions between MicroRNA-30c-5p and DNA Methyltransferases in Neuropathic Pain

Neuropathic pain is a prevalent and severe chronic syndrome, often refractory to treatment, whose development and maintenance may involve epigenetic mechanisms. We previously demonstrated a causal relationship between miR-30c-5p upregulation in nociception-related neural structures and neuropathic pain in rats subjected to sciatic nerve injury. Furthermore, a short course of an miR-30c-5p inhibitor administered into the cisterna magna exerts long-lasting antiallodynic effects via a TGF-?1-mediated mechanism. Herein, we show that miR-30c-5p inhibition leads to global DNA hyper-methylation of neurons in the lumbar dorsal root ganglia and spinal dorsal horn in rats subjected to sciatic nerve injury. Specifically, the inhibition of miR-30-5p significantly increased the expression of the novo DNA methyltransferases DNMT3a and DNMT3b in those structures. Furthermore, we identified the mechanism and found that miR-30c-5p targets the mRNAs of DNMT3a and DNMT3b. Quantitative methylation analysis revealed that the promoter region of the antiallodynic cytokine TGF-?1 was hypomethylated in the spinal dorsal horn of nerve-injured rats treated with the miR-30c-5p inhibitor, while the promoter of Nfyc, the host gene of miR-30c-5p, was hypermethylated. These results are consistent with long-term protection against neuropathic pain development after nerve injury. Altogether, our results highlight the key role of miR-30c-5p in the epigenetic mechanisms' underlying neuropathic pain and provide the basis for miR-30c-5p as a therapeutic target.Funding: This work was supported by: Grant SAF2016-77732-R funded by MCIN/AEI/10.13039/ 501100011033 and by “ERDF A way of making Europe”; Grant PID2019-104398RB-I00 funded by MCIN/AEI/ 10.13039/501100011033 and Instituto de Investigación Sanitaria Valdecilla (IDIVAL) (NVAL17/23). R.F. was the recipient of a pre-doctoral fellowship of the Foundation Tatiana Pérez de Guzmán el Bueno. Acknowledgments: We acknowledge the excellent technical assistance of Nieves García Iglesias

Utilización de la simulación clínica en un Máster Interuniversitario en Estudio y Tratamiento del Dolor

Objectives: To describe our experience integrating simulation based training in an Interuniversitary Master's degree in Pain Study and Management from the University Rey Juan Carlos of Madrid and University of Cantabria, Spain. Material and methods: After taking the online and on-site modules, students participated in 6 scenarios of simulated clinical immersion, in order to train patient management, clinical diagnosis, and patient safety competencies, in a realistic environment reproducing the actual clinical work. Results: Five physiotherapists, 1 psychologist, 10 medical doctors, and 9 nurses participated in the simulations during two consecutive editions of the Master. They found the scenarios very realistic and simulation a valuable teaching tool, reporting a global satisfaction of 4.5 over 5. It was also an opportunity to experiment adverse events of the analgesic that are uncommon to observe during clinical rotations. Conclusions: Participants were satisfied with the use of clinical simulation as part of the training program of an Interuniversitary Master in Pain Study and Management.Objetivos: En los sistemas de formación tradicional el entrenamiento se realiza practicando directamente sobre los pacientes durante las rotaciones clínicas. Esta organización tiene como consecuencia que los contenidos docentes dependan de los diferentes lugares de rotación, el tiempo disponible para cada uno de ellos y de la casualidad de encontrar determinadas patologías. Además, la variabilidad de los clínicos puede dificultar la estandarización de contenidos. Nuestro objetivo es describir la experiencia con la integración de la simulación con otros métodos de formación en el marco del Máster Oficial Interuniversitario para el Estudio y Tratamiento del Dolor de la Universidad Rey Juan Carlos de Madrid y la Universidad de Cantabria, con enfoque interprofesional. Material y métodos: Se integró el aprendizaje online y las sesiones presenciales, con una práctica de 7 horas utilizando la simulación clínica. Se realizaron 4 escenarios con 6 casos clínicos con los objetivos de reconocer y comprender las principales entidades patológicas que cursan con dolor en la práctica clínica. Se utilizaron pacientes estandarizados y simuladores de paciente de alta fidelidad, dependiendo del escenario clínico. Resultados: Los participantes (5 fisioterapeutas, 1 psicólogo, 10 médicos y 9 enfermeras) consideraron la simulación como herramienta docente muy útil. Encontraron los escenarios realistas y útiles para entrenar el enfoque diagnóstico y terapéutico en pacientes con dolor. Además, fue una oportunidad para experimentar los efectos adversos de los analgésicos que son difíciles de observar durante las rotaciones clínicas, y para que, aquellos alumnos sin competencias profesionales para tratar pacientes, pudieran tomar decisiones clínicas en un ambiente realista y sin riesgo. Conclusiones: La integración de la simulación clínica dentro de un Máster para el Estudio y Tratamiento del Dolor es percibida de modo muy satisfactorio por los participantes. Proporciona oportunidades para integrar y aplicar de modo práctico los conocimientos adquiridos durante el curso, independientemente de la titulación de origen, de un modo que evoca la realidad y es seguro. Además, facilita la formación específica en la comunicación y el trabajo en equipo interprofesional, cuando se toman decisiones para diagnosticar y tratar pacientes con dolor. Es una alternativa a las actuaciones directas sobre los pacientes que están limitadas por el riesgo de comprometer su seguridad clínica

Mecanismos implicados en el efecto protector de citoquinas pertenecientes a la familia Factores de Crecimiento Transformante-beta frente al desarrollo de dolor neuropático: XXXII Congreso Nacional de la SEF: premio Almirall

RB-101 fue generosamente facilitado por el Dr. Bernard Roques. El trabajo está financiado por: Sociedad Española de Farmacología y Laboratorios Almirall; Instituto de Salud Carlos III (RTICS: RD06/001/1016); Ministerio de Ciencia e Innovación (SAF2010-16894); Fundació La Marató de TV3 (Grant 072131)

TGF-β and opioid receptor signaling crosstalk results in improvement of endogenous and exogenous opioid analgesia under pathological pain conditions

Transforming growth factor-β1 (TGF-β1) protects against neuroinflammatory events underlying neuropathic pain. TGF-β signaling enhancement is a phenotypic characteristic of mice lacking the TGF-β pseudoreceptor BAMBI (BMP and activin membrane-bound inhibitor), which leads to an increased synaptic release of opioid peptides and to a naloxone-reversible hypoalgesic/antiallodynic phenotype. Herein, we investigated the following: (1) the effects of BAMBI deficiency on opioid receptor expression, functional efficacy, and analgesic responses to endogenous and exogenous opioids; and (2) the involvement of the opioid system in the antiallodynic effect of TGF-β1. BAMBI-KO mice were subjected to neuropathic pain by sciatic nerve crash injury (SNI). Gene (PCR) and protein (Western blot) expressions of μ- and δ-opioid receptors were determined in the spinal cord. The inhibitory effects of agonists on the adenylyl cyclase pathway were investigated. Two weeks after SNI, wild-type mice developed mechanical allodynia and the functionality of μ-opioid receptors was reduced. By this time, BAMBI-KO mice were protected against allodynia and exhibited increased expression and function of opioid receptors. Four weeks after SNI, when mice of both genotypes had developed neuropathic pain, the analgesic responses induced by morphine and RB101 (an inhibitor of enkephalin-degrading enzymes, which increases the synaptic levels of enkephalins) were enhanced in BAMBI-KO mice. Similar results were obtained in the formalin-induced chemical-inflammatory pain model. Subcutaneous TGF-β1 infusion prevented pain development after SNI. The antiallodynic effect of TGF-β1 was naloxone-sensitive. In conclusion, modulation of the endogenous opioid system by TGF-β signaling improves the analgesic effectiveness of exogenous and endogenous opioids under pathological pain conditions.This work was supported by Ministerio de Ciencia e Innovacio´n Grant SAF2010-16894, Fundacio´ La Marato´ de TV3 Grant 072131, Instituto de Salud Carlos III Grant RTICS:RD06/001/1016, and Sociedad Espan˜ola de Farmacología and Laboratorios Almirall-Prodesfarm

BAMBI (bone morphogenetic protein and activin membrane-bound inhibitor) reveals the involvement of the transforming growth factor-ß family in pain modulation

Transforming growth factors-beta (TGF-betas) signal through type I and type II serine-threonine kinase receptor complexes. During ligand binding, type II receptors recruit and phosphorylate type I receptors, triggering downstream signaling. BAMBI [bone morphogenetic protein (BMP) and activin membrane-bound inhibitor] is a transmembrane pseudoreceptor structurally similar to type I receptors but lacks the intracellular kinase domain. BAMBI modulates negatively pan-TGF-beta family signaling; therefore, it can be used as an instrument for unraveling the roles of these cytokines in the adult CNS. BAMBI is expressed in regions of the CNS involved in pain transmission and modulation. The lack of BAMBI in mutant mice resulted in increased levels of TGF-beta signaling activity, which was associated with attenuation of acute pain behaviors, regardless of the modality of the stimuli (thermal, mechanical, chemical/inflammatory). The nociceptive hyposensitivity exhibited by BAMBI(-/-) mice was reversed by the opioid antagonist naloxone. Moreover, in a model of chronic neuropathic pain, the allodynic responses of BAMBI(-/-) mice also appeared attenuated through a mechanism involving delta-opioid receptor signaling. Basal mRNA and protein levels of precursor proteins of the endogenous opioid peptides proopiomelanocortin (POMC) and proenkephalin (PENK) appeared increased in the spinal cords of BAMBI(-/-). Transcript levels of TGF-betas and their intracellular effectors correlated directly with genes encoding opioid peptides, whereas BAMBI correlated inversely. Furthermore, incubation of spinal cord explants with activin A or BMP-7 increased POMC and/or PENK mRNA levels. Our findings identify TGF-beta family members as modulators of acute and chronic pain perception through the transcriptional regulation of genes encoding the endogenous opioids.This work was supported by Ministerio de Ciencia e Innovación Grant SAF2007-65451, Instituto de Salud Carlos III Grant RD06/0001/1016, and Fundación La Marató de TV3 Grant 072131 (M.A.H.), by Instituto de Salud Carlos III Grant FIS-PI 060240, by Ministerio de Ciencia e Innovación Grant SAF2005-00811 (R.M.), and by the National Institutes of Health and the G. Harold and Leila Y. Mathers Charitable Foundation (J.C.I.-B.). We thank N. García, S. Pérez, M. F. Calderón, and C. Badía for their technical assistance

Circulating levels of miR-133a predict the regression potential of left ventricular hypertrophy after valve replacement surgery in patients with aortic stenosis

Background: Myocardial microRNA-133a (miR-133a) is directly related to reverse remodeling after pressure overload release in aortic stenosis patients. Herein, we assessed the significance of plasma miR-133a as an accessible biomarker with prognostic value in predicting the reversibility potential of LV hypertrophy after aortic valve replacement (AVR) in these patients. Methods and results: The expressions of miR-133a and its targets were measured in LV biopsies from 74 aortic stenosis patients. Circulating miR-133a was measured in peripheral and coronary sinus blood. LV mass reduction was determined echocardiographically. Myocardial and plasma levels of miR-133a correlated directly (r=0.46, P<0.001) supporting the myocardium as a relevant source of plasma miR-133a. Accordingly, a significant gradient of miR-133a was found between coronary and systemic venous blood. The preoperative plasma level of miR-133a was higher in the patients who normalized LV mass 1 year after AVR than in those exhibiting residual hypertrophy. Logistic regression analysis identified plasma miR-133a as a positive predictor of the hypertrophy reversibility after surgery. The discrimination of the model yielded an area under the receiver operator characteristic curve of 0.89 (P<0.001). Multiple linear regression analysis revealed plasma miR-133a and its myocardial target Wolf-Hirschhorn syndrome candidate 2/Negative elongation factor A as opposite predictors of the LV mass loss (g) after AVR. Conclusions: Preoperative plasma levels of miR-133a reflect their myocardial expression and predict the regression potential of LV hypertrophy after AVR. The value of this bedside information for the surgical timing, particularly in asymptomatic aortic stenosis patients, deserves confirmation in further clinical studies.This work was supported by: Instituto de Salud Carlos III (PS09/01097 and PI12/00999); Fundación Marqués de Valdecilla-Universidad de Cantabria (FMV-UC 09/01); Instituto de Formación e Investigación Marqués de Valdecilla (FMV-API 10/20).Aortic stenosisMicroRNAMiR-133aMyocardial hypertrophyMyocardial reverse remodelin

BMP7-based peptide agonists of BMPR1A protect the left ventricle against pathological remodeling induced by pressure overload

Aortic stenosis (AS) exposes the left ventricle (LV) to pressure overload leading to detrimental LV remodeling and heart failure. In animal models of cardiac injury or hemodynamic stress, bone morphogenetic protein-7 (BMP7) protects LV against remodeling by counteracting TGF-β effects. BMP receptor 1A (BMPR1A) might mediate BMP7 antifibrotic effects. Herein we evaluated BMP7-based peptides, THR123 and THR184, agonists of BMPR1A, as cardioprotective drugs in a pressure overload model. We studied patients with AS, mice subjected to four-week transverse aortic constriction (TAC) and TAC release (de-TAC). The LV of AS patients and TAC mice featured Bmpr1a downregulation. Also, pSMAD1/5/(8)9 was reduced in TAC mice. Pre-emptive treatment of mice with THR123 and THR184, during the four-week TAC period, normalized pSMAD1/5/(8)9 levels in the LV, attenuated overexpression of remodeling-related genes (Col 1α1, β-MHC, BNP), palliated structural damage (hypertrophy and fibrosis) and alleviated LV dysfunction (systolic and diastolic). THR184 administration, starting fifteen days after TAC, halted the ongoing remodeling and partially reversed LV dysfunction. The reverse remodeling after pressure overload release was facilitated by THR184. Both peptides diminished the TGF-β1-induced hypertrophic gene program in cardiomyocytes, collagen transcriptional activation in fibroblasts, and differentiation of cardiac fibroblasts to myofibroblasts. Molecular docking suggests that both peptides bind with similar binding energies to the BMP7 binding domain at the BMPR1A. The present study results provide a preclinical proof-of-concept of potential therapeutic benefits of BMP7-based small peptides, which function as agonists of BMPR1A, against the pathological LV remodeling in the context of aortic stenosis.Acknowledgments. We acknowledge the excellent technical assistance of Nieves García-Iglesias, Ana Cayón, Roberto Moreta, and Abraham Velasco