RESUMEN
Gene silencing by either small-interference RNAs (siRNA) or antisense oligodeoxynucleotides (aODN) is widely used in biomedical research. However, their use as therapeutic agents is hindered by two important limitations: their low stability and the activation of the innate immune response. Recently, we developed a new type of molecule to decrease gene expression named polypurine reverse Hoogsteen hairpins (PPRHs) that bind to polypyrimidine targets in the DNA. Herein, stability experiments performed in mouse, human, and fetal calf serum and in PC3 cells revealed that the half-life of PPRHs is much longer than that of siRNAs in all cases. Usage of PPRHs with a nicked-circular structure increased the binding affinity to their target sequence and their half-life in FCS when bound to the target. Regarding the innate immune response, we determined that the levels of the transcription factors IRF3 and its phosphorylated form, as well as NF-κB were increased by siRNAs and not by PPRHs; that the expression levels of several proinflammatory cytokines including IL-6, TNF-α, IFN-α, IFN-ß, IL-1ß, and IL-18 were not significantly increased by PPRHs; and that the cleavage and activation of the proteolytic enzyme caspase-1 was not triggered by PPRHs. These determinations indicated that PPRHs, unlike siRNAs, do not activate the innate inflammatory response.
Asunto(s)
Neoplasias de la Mama/inmunología , Silenciador del Gen/inmunología , Neoplasias de la Próstata/inmunología , Nucleótidos de Purina/química , Nucleótidos de Purina/inmunología , Animales , Western Blotting , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Proliferación Celular , Citocinas/genética , Citocinas/metabolismo , Ensayo de Cambio de Movilidad Electroforética , Femenino , Semivida , Humanos , Factor 3 Regulador del Interferón/genética , Factor 3 Regulador del Interferón/metabolismo , Masculino , Ratones , FN-kappa B/genética , FN-kappa B/metabolismo , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Nucleótidos de Purina/farmacología , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
BACKGROUND: In France, the coordinated healthcare circuit means that patients should be referred to specialists, for example nephrologists, by another physician. However, there are no recommendations concerning the reasons justifying the referral to a nephrologist. The main purpose of our study is to describe the motif of first consultations in nephrology in the health area 5 of Brittany. METHODS: We retrospectively collected medical reports of first consultations by 17 nephrologists in the 4 centers of the study area, during the year 2014. In these letters, we noted the consultation motif, the specialty of the physician who refers the patient, and main characteristics of patients. RESULTS: We included 662 first consultations. The main reason for consultations was chronic kidney disease (68.7%), including chronic renal insufficiency (56.9%), proteinuria (7.3%), microscopic hematuria (3.3%) and searching for chronic kidney disease in the presence of risk factor (1.2%). Other frequent reasons were the follow-up consultation after a pregnancy complicated by preeclampsia (9.5%), urinary lithiasis (5.7%), hypertension (3.8%) and hydroelectrolytic disorder (3.5%). Non-nephrology reasons represent 3.2% of first consultations. Almost all patients have been referred by a physician (99.7%), mainly a general practitioner (71.9%). CONCLUSION: Nephrology first consultations are realized according to the coordinated healthcare circuit since almost all are requested by another physician. The reasons are adapted to the specialty. The main reason is chronic kidney disease, often already associated with renal insufficiency chronic.