STAT3 phosphorylation at serine 727 activates specific genetic programs and promotes clear cell renal cell carcinoma (ccRCC) aggressiveness

Cancer models; Molecular medicineModelos de cáncer; Medicina molecularModels de càncer; Medicina molecularThe signal transducer and activator of transcription 3 (STAT3) is a transcription factor mainly activated by phosphorylation in either tyrosine 705 (Y705) or serine 727 (S727) residues that regulates essential processes such as cell differentiation, apoptosis inhibition, or cell survival. Aberrant activation of STAT3 has been related to development of nearly 50% of human cancers including clear cell renal cell carcinoma (ccRCC). In fact, phosho-S727 (pS727) levels correlate with overall survival of ccRCC patients. With the aim to elucidate the contribution of STAT3 phosphorylation in ccRCC development and progression, we have generated human-derived ccRCC cell lines carrying STAT3 Y705 and S727 phosphomutants. Our data show that the phosphomimetic substitution Ser727Asp facilitates a pro-tumoral phenotype in vitro, in a Y705-phosphorylation-independent manner. Moreover, we describe that STAT3 phosphorylation state determines the expression of different subsets of target genes associated with distinct biological processes, being pS727-dependent genes the most related to cellular hallmarks of cancer. In summary, the present study constitutes the first analysis on the role of overall STAT3 phosphorylation state in ccRCC and demonstrates that pS727 promotes the expression of a specific subset of target genes that might be clinically relevant as novel biomarkers and potential therapeutic targets for ccRCC.This work was supported by Ministerio de Ciencia e Innovación (Grant No. SAF201459945-R and SAF2017-89989-R) to A.M.; American Association for Cancer Research (AACR, Ref #419589) to A.M. and Red de Investigación Renal REDinREN (Grant No. 12/0021/0013) to. A.M. The group holds the Quality Mention from the Generalitat de Catalunya (Grant No. 2021 SGR 01,600). J.A. was a recipient of the Ph.D. Fellow Program from Consejo Nacional de Ciencia y Tecnología (CONACyT), México (Grant No. 549678)

Isolation and characterization of exosome-enriched urinary extracellular vesicles from Dent’s disease type 1 Spanish patients

Background and objectives: Dent's disease type 1 (DD1) is a rare X-linked hereditary pathology caused by CLCN5 mutations that is characterized mainly by proximal tubule dysfunction, hypercalciuria, nephrolithiasis/nephrocalcinosis, progressive chronic kidney disease, and low-weight proteinuria, the molecular hallmark of the disease. Currently, there is no specific curative treatment, only symptomatic and does not prevent the progression of the disease. In this study we have isolated and characterized urinary extracellular vesicles (uEVs) enriched in exosomes that will allow us to identify biomarkers associated with DD1 progression and a better understanding of the pathophysiological bases of the disease. Materials and methods: Through a national call from the Spanish Society of Nephrology (SEN) and the Spanish Society of Pediatric Nephrology (AENP), urine samples were obtained from patients and controls from different Spanish hospitals, which were processed to obtain the uEVS. The data of these patients were provided by the respective nephrologists and/or extracted from the RENALTUBE registry. The uEVs were isolated by ultracentrifugation, morphologically characterized and their protein and microRNA content extracted. Results: 25 patients and 10 controls were recruited, from which the urine was processed to isolate the uEVs. Our results showed that the relative concentration of uEVs/mL is lower in patients compared to controls (0.26 × 106 uEVs/mL vs 1.19 × 106 uEVs/mL, p < 0.01). In addition, the uEVs of the patients were found to be significantly larger than those of the control subjects (mean diameter: 187.8 nm vs 143.6 nm, p < 0.01). Finally, our data demonstrated that RNA had been correctly extracted from both patient and control exosomes. Conclusions: In this work we describe the isolation and characterization of uEVs from patients with Dent 1 disease and healthy controls, that shall be useful for the subsequent study of differentially expressed cargo molecules in this pathology. Resumen: Antecedentes y objetivo: La enfermedad de Dent tipo 1 (DD1) es una patología hereditaria rara ligada al cromosoma X causado por mutaciones en el CLCN5 que se caracteriza principalmente por una disfunción del túbulo proximal, hipercalciuria, nefrolitiasis/nefrocalcinosis, enfermedad renal crónica progresiva y proteinuria de bajo peso molecular, rasgo distintivo de la enfermedad. Actualmente, no existe un tratamiento curativo específico, únicamente sintomático y no previene la progresión de la enfermedad. En este estudio hemos aislado y caracterizado las vesículas extracelulares urinarias (uEVs) enriquecidas en exosomas que nos permitirán identificar biomarcadores asociados a la progresión de DD1 y una mejor comprensión de las bases fisiopatológicas de la misma. Materiales y métodos: A través de una convocatoria nacional de la Sociedad Española de Nefrología (SEN) y la Sociedad Española de Nefrología Pediátrica (AENP), se obtuvieron orinas de pacientes y controles de distintos hospitales españoles, las cuales se procesaron para obtener los uEVS. Los datos de estos pacientes fueron proporcionados por los respectivos nefrólogos y/o extraídos del registro RENALTUBE. Los uEVs se aislaron mediante ultracentrifugación, fueron caracterizados morfológicamente y se extrajo su contenido de proteína y microRNA. Resultados: Se reclutaron 25 pacientes y 10 controles, de los cuales se procesaron las orinas para aislar los uEVs. Nuestros resultados mostraron que la concentración relativa de uEVs/mL es menor en pacientes comparado con controles (0,26 × 106 uEVs/mL vs 1,19 × 106 uEVs/mL, p < 0.01). Además, se vio que los uEVs de los pacientes eran significativamente más grandes que los de los sujetos control (diámetro medio: 187,8 nm vs 143,6 nm, p < 0.01). Finalmente, nuestros datos demostraron que se había extraído correctamente RNA tanto de los exosomas de pacientes como de controles. Conclusiones: En este trabajo describimos el aislamiento y caracterización de uEVs de pacientes de la enfermedad de Dent 1 y controles sanos, útiles para el posterior estudio de moléculas cargo diferencialmente expresadas en esta patología

Differences in the presentation and evolution of primary aldosteronism in elderly (≥65 years) and young patients (<65 years).

To compare the presentation and evolution of primary aldosteronism (PA) in the elderly (≥65 years) and young patients ( A retrospective multicenter study was performed in 20 Spanish hospitals of PA patients in follow-up between 2018 and 2021. Three hundred fifty-two patients with PA Older patients with PA have a worse cardiometabolic profile than young patients with PA that it is related to a longer duration of hypertension. However, the results of the AVS, and adrenalectomy are similar in both groups. Therefore, the management of elderly patients with PA should be based not only on age, but rather on the overall medical, physical, social, and mental characteristics of the patients

Comparison of pheochromocytoma-specific morbidity and mortality among adults with bilateral pheochromocytomas undergoing total adrenalectomy vs cortical-sparing adrenalectomy

Importance: Large studies investigating long-term outcomes of patients with bilateral pheochromocytomas treated with either total or cortical-sparing adrenalectomies are needed to inform clinical management. Objective: To determine the association of total vs cortical-sparing adrenalectomy with pheochromocytoma-specific mortality, the burden of primary adrenal insufficiency after bilateral adrenalectomy, and the risk of pheochromocytoma recurrence. Design, Setting, and Participants: This cohort study used data from a multicenter consortium-based registry for 625 patients treated for bilateral pheochromocytomas between 1950 and 2018. Data were analyzed from September 1, 2018, to June 1, 2019. Exposures: Total or cortical-sparing adrenalectomy. Main Outcomes and Measures: Primary adrenal insufficiency, recurrent pheochromocytoma, and mortality. Results: Of 625 patients (300 [48%] female) with a median (interquartile range [IQR]) age of 30 (22-40) years at diagnosis, 401 (64%) were diagnosed with synchronous bilateral pheochromocytomas and 224 (36%) were diagnosed with metachronous pheochromocytomas (median [IQR] interval to second adrenalectomy, 6 [1-13] years). In 505 of 526 tested patients (96%), germline mutations were detected in the genes RET (282 patients [54%]), VHL (184 patients [35%]), and other genes (39 patients [7%]). Of 849 adrenalectomies performed in 625 patients, 324 (52%) were planned as cortical sparing and were successful in 248 of 324 patients (76.5%). Primary adrenal insufficiency occurred in all patients treated with total adrenalectomy but only in 23.5% of patients treated with attempted cortical-sparing adrenalectomy. A third of patients with adrenal insufficiency developed complications, such as adrenal crisis or iatrogenic Cushing syndrome. Of 377 patients who became steroid dependent, 67 (18%) developed at least 1 adrenal crisis and 50 (13%) developed iatrogenic Cushing syndrome during median (IQR) follow-up of 8 (3-25) years. Two patients developed recurrent pheochromocytoma in the adrenal bed despite total adrenalectomy. In contrast, 33 patients (13%) treated with successful cortical-sparing adrenalectomy developed another pheochromocytoma within the remnant adrenal after a median (IQR) of 8 (4-13) years, all of which were successfully treated with another surgery. Cortical-sparing surgery was not associated with survival. Overall survival was associated with comorbidities unrelated to pheochromocytoma: of 63 patients who died, only 3 (5%) died of metastatic pheochromocytoma. Conclusions and Relevance: Patients undergoing cortical-sparing adrenalectomy did not demonstrate decreased survival, despite development of recurrent pheochromocytoma in 13%. Cortical-sparing adrenalectomy should be considered in all patients with hereditary pheochromocytoma.Published versio