RESUMEN
Variants in the leucine-rich repeat kinase 2 (LRRK2) gene are associated with increased risk for familial and sporadic Parkinson's disease (PD). Pathogenic variants in LRRK2, including the common variant G2019S, result in increased LRRK2 kinase activity, supporting the therapeutic potential of LRRK2 kinase inhibitors for PD. To better understand the role of LRRK2 in disease and to support the clinical development of LRRK2 inhibitors, quantitative and high-throughput assays to measure LRRK2 levels and activity are needed. We developed and applied such assays to measure the levels of LRRK2 as well as the phosphorylation of LRRK2 itself or one of its substrates, Rab10 (pT73 Rab10). We observed increased LRRK2 activity in various cellular models of disease, including iPSC-derived microglia, as well as in human subjects carrying the disease-linked variant LRRK2 G2019S. Capitalizing on the high-throughput and sensitive nature of these assays, we detected a significant reduction in LRRK2 activity in subjects carrying missense variants in LRRK2 associated with reduced disease risk. Finally, we optimized these assays to enable analysis of LRRK2 activity following inhibition in human peripheral blood mononuclear cells (PBMCs) and whole blood, demonstrating their potential utility as biomarkers to assess changes in LRRK2 expression and activity in the clinic.
Asunto(s)
Proteína 2 Quinasa Serina-Treonina Rica en Repeticiones de Leucina/metabolismo , Proteínas de Unión al GTP rab/metabolismo , Animales , Biomarcadores , Activación Enzimática , Pruebas de Enzimas/métodos , Pruebas de Enzimas/normas , Expresión Génica , Ensayos Analíticos de Alto Rendimiento , Humanos , Proteína 2 Quinasa Serina-Treonina Rica en Repeticiones de Leucina/genética , Leucocitos Mononucleares/metabolismo , Ratones , Neuroglía/metabolismo , Proteínas de Unión al GTP rab/genéticaRESUMEN
OBJECTIVE: To identify markers of resistance to developing Parkinson disease (PD) among LRRK2 mutation carriers (LRRK2+), we carried out metabolomic profiling in individuals with PD and unaffected controls (UC), with and without the LRRK2 mutation. METHODS: Plasma from 368 patients with PD and UC in the LRRK2 Cohort Consortium (LCC), comprising 118 LRRK2+/PD+, 115 LRRK2+/UC, 70 LRRK2-/PD+, and 65 LRRK2-/UC, and CSF available from 68 of them, were analyzed by liquid chromatography with mass spectrometry. For 282 analytes quantified in plasma and CSF, we assessed differences among the 4 groups and interactions between LRRK2 and PD status, using analysis of covariance models adjusted by age, study site cohort, and sex, with p value corrections for multiple comparisons. RESULTS: Plasma caffeine concentration was lower in patients with PD vs UC (p < 0.001), more so among LRRK2+ carriers (by 76%) than among LRRK2- participants (by 31%), with significant interaction between LRRK2 and PD status (p = 0.005). Similar results were found for caffeine metabolites (paraxanthine, theophylline, 1-methylxanthine) and a nonxanthine marker of coffee consumption (trigonelline) in plasma, and in the subset of corresponding CSF samples. Dietary caffeine was also lower in LRRK2+/PD+ compared to LRRK2+/UC with significant interaction effect with the LRRK2+ mutation (p < 0.001). CONCLUSIONS: Metabolomic analyses of the LCC samples identified caffeine, its demethylation metabolites, and trigonelline as prominent markers of resistance to PD linked to pathogenic LRRK2 mutations, more so than to idiopathic PD. Because these analytes are known both as correlates of coffee consumption and as neuroprotectants in animal PD models, the findings may reflect their avoidance by those predisposed to develop PD or their protective effects among LRRK2 mutation carriers.
Asunto(s)
Alcaloides/sangre , Cafeína/sangre , Proteína 2 Quinasa Serina-Treonina Rica en Repeticiones de Leucina/genética , Fármacos Neuroprotectores/sangre , Enfermedad de Parkinson/sangre , Enfermedad de Parkinson/genética , Anciano , Alcaloides/líquido cefalorraquídeo , Cafeína/líquido cefalorraquídeo , Cromatografía Liquida , Estudios de Cohortes , Femenino , Heterocigoto , Humanos , Masculino , Espectrometría de Masas , Metabolómica , Persona de Mediana Edad , Fármacos Neuroprotectores/líquido cefalorraquídeo , Enfermedad de Parkinson/líquido cefalorraquídeo , Teofilina/sangre , Teofilina/líquido cefalorraquídeo , Xantinas/sangre , Xantinas/líquido cefalorraquídeoRESUMEN
El objetivo de este trabajo es estudiar los niveles y relaciones entre burnout, salud mental ocupacional y características del puesto en las distintas categorías profesionales. Los instrumentos utilizados han sido las Escalas Síndrome de Burnout, Salud Mental Ocupacional (SMO) y características del puesto (variedad, identidad, importancia y autonomía, retroalimentación y contacto social) JDS. La muestra se realizó entre 175 trabajadores: médicos, enfermeros/as, auxiliares de enfermería, celadores, administrativos, asistentes sociales y técnicos. Las conclusiones fueron que es necesario un mayor ajuste de las características del puesto, una mayor retroalimentación, y un mayor reconocimiento de los supervisores y compañeros y un ajuste en la autonomía y las tareas más importantes llevarían a una vivencia del puesto adecuada y evitarían despersonalización y agotamiento emocional (burnout) pues, los niveles indican padecimiento sostenido de estrés sin soluciones adecuadas (AU)
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