RESUMEN
OBJETIVO: Evaluar el riesgo de progresión de la retinopatía diabética (RD) utilizando nuevas estrategias para obtener información genética en diabéticos tipo 2 (DT2) basadas en interferencia por ácido ribonucleico (ARN). MATERIAL Y MÉTODOS: Estudio multicéntrico, prospectivo de casos-controles en 132 participantes divididos en: grupo DT2 (GDT2) con RD (+RD) y sin RD (-RD) (n = 77) y grupo control (GC) (n = 55). Tras entrevista personal y examen oftalmológico, se extrajeron lágrimas para análisis molecular (expresión de micro-ARN [miARN] [miRCURY™ ARN Isolation Kit, Qiagen]). En 18 muestras (GDT2+RD = 6; GDT2-RD = 6; GC = 6) obtuvimos librerías de 137 vs. 140 pares de bases (GeneMapper, Applied Biosystems) y realizamos secuenciación de próxima generación (NGS). El programa SPSS 15.0 vehiculizó el análisis estadístico. RESULTADOS: Edad media: 67 ± 12 años en GDT2 vs. 55 ± 21 años en GC. Distribución hombres/mujeres: 51/28 en GDT2 vs. 25/30 en GC. Los antecedentes familiares de DM, cumplir dieta, fumar, beber y realizar ejercicio mostraron diferencias significativas entre grupos (p < 0,001). Con 20-25 μL de lágrimas extrajimos 9,42 ± 3,30 ng/mL de ARN purificado, con diferencias significativas entre GDT2/GC (p = 0,002) y GDT2+RD/GC (p = 0,004). La expresión lagrimal de miARN en GDT2 correlacionó directamente con: edad/obesidad/duración de DM (p < 0,05), e indirectamente con: agudeza visual (p < 0,05). Hemos identificado 14 miARN relacionados con la presencia, mecanismos patogénicos y factores de riesgo para la progresión de la RD. CONCLUSIONES: Proponemos utilizar lágrimas como fuente de información genética para la DM. Los miARN específicos implicados en desarrollo o progresión de la RD pueden utilizarse como biomarcadores moleculares y, a partir de ellos, desarrollar futuras bioterapias
OBJECTIVE: To evaluate the risk of progression of diabetic retinopathy (DR) using new strategies to obtain genetic information in type 2 diabetes (T2D) based on interfering ribonucleic acid (RNA). MATERIAL AND METHODS: A prospective multicentre case-control study of 132 participants was distributed into: T2D with (+DR) or without (-DR) (T2DG; n = 77), and a control group (CG; n = 55). After an eye examination and personal interview, tears were collected for molecular analysis (expression of microRNAs [miRNAs] (miRCURY(TM) ARN Isolation Kit, Qiagen)]. Libraries, 137 vs. 140 bp (GeneMapper, Applied Biosystems), were obtained in 18 samples (T2DG+DR = 6; T2DG-DR = 6; CG = 6) by performing next-generation sequencing (NGS). SPSS 15.0 statistical program was used to perform data analysis. RESULTS: The mean age was 67 ± 12 years in the T2DG vs. 55 ± 21 years in the CG. Distribution men/women: 25/30 in T2DG vs. 51/28 in CG. A family history of DM, diet compliance, smoking, drinking and exercise, showed significant differences between groups (P<.001). A 20-25 microlitre sample of tears contained a mean of 9.42 ± 3.30 ng/mL of purified ARN, with significant differences between T2DG/CG (P=.002) and T2DG+RD/CG (P=.004). Tear expression of miARNs in T2DG directly correlated with age/obesity/T2D duration (P<.05), and indirectly with visual acuity (P<.05). A total of 14 miRNAs related to the presence, pathogenic mechanisms and risk factors for the progression of diabetic retinopathy, were identified. CONCLUSIONS: We propose to use tears as a source of genetic information for DM. Specific miRNAs involved in DR development and/or progression can be used as molecular biomarkers, and based on these, for developing future biotherapies
Asunto(s)
Humanos , Masculino , Femenino , Adolescente , Adulto Joven , Adulto , Persona de Mediana Edad , Anciano , Interferencia de ARN/inmunología , Interferencia de ARN/fisiología , MicroARNs/genética , MicroARNs/uso terapéutico , Retinopatía Diabética/diagnóstico , Retinopatía Diabética/genética , Retinopatía Diabética/terapia , Biomarcadores/análisis , Biomarcadores/metabolismo , Ceguera/diagnóstico , Ceguera/prevención & control , Coagulación con Láser/instrumentación , Coagulación con Láser/métodos , Coagulación con Láser , Tomografía de Coherencia Óptica/instrumentación , Tomografía de Coherencia Óptica/métodos , Tomografía de Coherencia Óptica , Lágrimas/fisiología , Oftalmopatías/complicaciones , Estudios Multicéntricos como Asunto , Estudios Prospectivos , Estudios de Casos y Controles , EspañaRESUMEN
OBJECTIVE: To evaluate the risk of progression of diabetic retinopathy (DR) using new strategies to obtain genetic information in type 2 diabetes (T2D) based on interfering ribonucleic acid (RNA). MATERIAL AND METHODS: A prospective multicentre case-control study of 132 participants was distributed into: T2D with (+DR) or without (-DR) (T2DG; n=77), and a control group (CG; n=55). After an eye examination and personal interview, tears were collected for molecular analysis (expression of microRNAs [miRNAs] (miRCURY ™ ARN Isolation Kit, Qiagen)]. Libraries, 137 vs. 140bp (GeneMapper, Applied Biosystems), were obtained in 18 samples (T2DG+DR=6; T2DG-DR=6; CG=6) by performing next-generation sequencing (NGS). SPSS 15.0 statistical program was used to perform data analysis. RESULTS: The mean age was 67±12 years in the T2DG vs. 55±21 years in the CG. Distribution men/women: 25/30 in T2DG vs. 51/28 in CG. A family history of DM, diet compliance, smoking, drinking and exercise, showed significant differences between groups (P<.001). A 20-25 microlitre sample of tears contained a mean of 9.42±3.30 ng/mL of purified ARN, with significant differences between T2DG/CG (P=.002) and T2DG+RD/CG (P=.004). Tear expression of miARNs in T2DG directly correlated with age/obesity/T2D duration (P<.05), and indirectly with visual acuity (P<.05). A total of 14 miRNAs related to the presence, pathogenic mechanisms and risk factors for the progression of diabetic retinopathy, were identified. CONCLUSIONS: We propose to use tears as a source of genetic information for DM. Specific miRNAs involved in DR development and/or progression can be used as molecular biomarkers, and based on these, for developing future biotherapies.
