The SASP factor IL-6 sustains cell-autonomous senescent cells via a cGAS-STING-NFκB intracrine senescent noncanonical pathway.

Senescent cells produce a Senescence-Associated Secretory Phenotype (SASP) that involves factors with diverse and sometimes contradictory activities. One key SASP factor, interleukin-6 (IL-6), has the potential to amplify cellular senescence in the SASP-producing cells in an autocrine action, while simultaneously inducing proliferation in the neighboring cells. The underlying mechanisms for the contrasting actions remain unclear. We found that the senescence action does not involve IL-6 secretion nor the interaction with the receptor expressed in the membrane but is amplified through an intracrine mechanism. IL-6 sustains intracrine senescence interacting with the intracellular IL-6 receptor located in anterograde traffic specialized structures, with cytosolic DNA, cGAS-STING, and NFκB activation. This pathway triggered by intracellular IL-6 significantly contributes to cell-autonomous induction of senescence and impacts in tumor growth control. Inactivation of IL-6 in somatotrophic senescent cells transforms them into strongly tumorigenic in NOD/SCID mice, while re-expression of IL-6 restores senescence control of tumor growth. The intracrine senescent IL-6 pathway is further evidenced in three human cellular models of therapy-induced senescence. The compartmentalization of the intracellular signaling, in contrast to the paracrine tumorigenic action, provides a pathway for IL-6 to sustain cell-autonomous senescent cells, driving the SASP, and opens new avenues for clinical consideration to senescence-based therapies.

Expression of RSUME is Associated With Poor Prognosis in Clear Cell Renal Carcinoma: Involvement of ROS Related Metabolism.

INTRODUCTION: RSUME (RWD domain-containing protein SUMO Enhancer), RWD domain containing 3 (RWDD3) gene product, is upregulated by hypoxia and expressed in organs prone to develop von Hippel-Lindau (VHL) syndrome tumors. MATERIALS AND METHODS: We evaluated RSUME prognostic value in clear cell renal cell carcinoma (ccRCC) based mainly on the dataset (KIRC) from patients in The Cancer Genome Atlas (TCGA). Wilcoxon signed-rank test and one-way analysis of variance (ANOVA) followed by Tukey's test were used to evaluate relationships between clinicopathological features and RSUME expression and univariate and multivariate Cox regression analysis methods were used to evaluate prognostic factors. The biological function of RSUME was assessed by gene set enrichment analysis (GSEA). For validation, total amount of ROS was detected in ccRCC cell lines using dichlorofluorescin diacetate. RESULTS: RSUME is highly expressed in tumor tissues compared with normal tissues (P = .006, P = .039, P = .002, P = .036, P < .001) and associates with tumor T (P = .018) and tumor M (P = .036) advanced stages and higher extent cysts (P = .005). RSUME expression appears to be an independent risk factor for overall survival (OS) (P = .002) and disease-specific survival (DSS) (P = .026) in ccRCC patients. GSEA showed enrichment of relevant glycerophospholipid- and ROS-related pathways in RSUME high-expression phenotype. ROS diminished levels in RSUME-silenced ccRCC cell lines validated RSUME relevance in ROS-related pathways. CONCLUSION: RSUME high expression may predict poor prognosis in ccRCC and impact through its action on metabolism and ROS related pathways.

Impact of RSUME Actions on Biomolecular Modifications in Physio-Pathological Processes.

The small RWD domain-containing protein called RSUME or RWDD3 was cloned from pituitary tumor cells with increasing tumorigenic and angiogenic proficiency. RSUME expression is induced under hypoxia or heat shock and is upregulated, at several pathophysiological stages, in tissues like pituitary, kidney, heart, pancreas, or adrenal gland. To date, several factors with essential roles in endocrine-related cancer appear to be modulated by RWDD3. RSUME regulates, through its post-translational (PTM) modification, pituitary tumor transforming gene (PTTG) protein stability in pituitary tumors. Interestingly, in these tumors, another PTM, the regulation of EGFR levels by USP8, plays a pathogenic role. Furthermore, RSUME suppresses ubiquitin conjugation to hypoxia-inducible factor (HIF) by blocking VHL E3-ubiquitin ligase activity, contributing to the development of von Hippel-Lindau disease. RSUME enhances protein SUMOylation of specific targets involved in inflammation such as IkB and the glucocorticoid receptor. For many of its actions, RSUME associates with regulatory proteins of ubiquitin and SUMO cascades, such as the E2-SUMO conjugase Ubc9 or the E3 ubiquitin ligase VHL. New evidence about RSUME involvement in inflammatory and hypoxic conditions, such as cardiac tissue response to ischemia and neuropathic pain, and its role in several developmental processes, is discussed as well. Given the modulation of PTMs by RSUME in neuroendocrine tumors, we focus on its interactors and its mode of action. Insights into functional implications and molecular mechanisms of RSUME action on biomolecular modifications of key factors of pituitary adenomas and renal cell carcinoma provide renewed information about new targets to treat these pathologies.

Mass Spectrometry-Based Metabolic Fingerprinting Contributes to Unveil the Role of RSUME in Renal Cell Carcinoma Cell Metabolism.

Clear cell renal cell carcinoma (ccRCC) is a heterogeneous disease with 50-80% patients exhibiting mutations in the von Hippel-Lindau (VHL) gene. RSUME (RWD domain (termed after three major RWD-containing proteins: RING finger-containing proteins, WD-repeat-containing proteins, and yeast DEAD (DEXD)-like helicases)-containing protein small ubiquitin-related modifier (SUMO) enhancer) acts as a negative regulator of VHL function in normoxia. A discovery-based metabolomics approach was developed by means of ultraperformance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (MS) for fingerprinting the endometabolome of a human ccRCC cell line 786-O and three other transformed cell systems (n = 102) with different expressions of RSUME and VHL. Cross-validated orthogonal projection to latent structures discriminant analysis models were built on positive, negative, and a combination of positive- and negative-ion mode MS data sets. Discriminant feature panels selected by an iterative multivariate classification allowed differentiating cells with different expressions of RSUME and VHL. Fifteen identified discriminant metabolites with level 1, including glutathione, butyrylcarnitine, and acetylcarnitine, contributed to understand the role of RSUME in ccRCC. Altered pathways associated with the RSUME expression were validated by biological and bioinformatics analyses. Combined results showed that in the absence of VHL, RSUME is involved in the downregulation of the antioxidant defense system, whereas in the presence of VHL, it acts in rerouting energy-related pathways, negatively modulating the lipid utilization, and positively modulating the fatty acid synthesis, which may promote deposition in droplets.

Modifications in the cellular proteome and their clinical application.

Post-translational modifications (PTMs) are covalent modifications in proteins during or after their synthesis. Among them, the best known are phosphorylation, methylation, acetylation, and also cleavage or binding of small peptides (ubiquitination, SUMOylation and NEDDylation). Often the protein is modified in multiple sites and these modifications are coordinated generating a PTMs crosstalk. Altered patterns of PTMs have been related to several pathologies. Currently, advances in mass spectrometry have made it possible to study multiple PTMs simultaneously. Oncology is one of the disciplines that incorporated these technologies for the need to better characterize tumors. In cancer, several alterations related to the ubiquitinlike PTMs have been described, such as SUMOylation. In particular, the interaction between different PTMs with SUMOylation has been studied in the context of the von Hippel Lindau (VHL) multitumoral syndrome, generating new putative biomarkers for the evolution of these tumors. RSUME or RWDD3, an enhancer of SUMOylation that acts on VHL and HIF proteins, shows a correlation with malignant parameters in this type of tumors, such as angiogenesis. Regulators of PTMs are becoming relevant as biomarkers in cancer.

Las modificaciones postraduccionales (PTMs por sus siglas en inglés) son modificaciones covalentes en las proteínas durante o posteriormente a su síntesis. Las más conocidas son fosforilación, metilación y acetilación, también clivajes o unión de pequeños péptidos (ubiquitinación, SUMOilación y NEDDilación). Frecuentemente la proteína es modificada en múltiples sitios y estas modificaciones se coordinan generando una interacción de PTMs. Patrones alterados de PTMs han sido relacionados con varias enfermedades. En la actualidad los avances en la espectrometría de masas han hecho posible estudiar en simultáneo múltiples PTMs. La oncología es una de las disciplinas que ha incorporado estas tecnologías por su necesidad de caracterizar a los tumores. En cáncer se han descripto varias alteraciones relacionadas a las PTMs del tipo ubiquitina como la SUMOilación. En particular la interacción entre distintas PTMs con la SUMOilación ha sido estudiada en el contexto de la enfermedad multitumoral de von Hippel Lindau, generando posibles nuevos biomarcadores para la evolución de estos tumores. RSUME o RWDD3, un enhancer de SUMOilación que actúa sobre las proteínas VHL y HIF, ha mostrado una correlación con parámetros malignos en este tipo de tumores, como la angiogénesis. Los reguladores de las PTMs están cobrando relevancia como biomarcadores en el cáncer.

Low Doses of CPS49 and Flavopiridol Combination as Potential Treatment for Advanced Prostate Cancer.

Due to some inconsistencies in the figures provided by the first author that have come to light, and after a thorough investigation we would like to retract our paper: "Low doses of CPS49 and flavopiridol combination as potential treatment for advanced prostate cancer. By: Zalazar F, De Luca P, Gardner K, Figg WD, Meiss R, Spallanzani RG, Vallecorsa P, Elguero B, Cotignola J, Vazquez E, De Siervi A. Curr. Pharm. Biotechnol., 2015, 16(6), 553-63. Submission of a manuscript to the respective journals implies that all authors have read and agreed to the content of the Copyright Letter or the Terms and Conditions. As such this article represents a severe abuse of the scientific publishing system. Bentham Science Publishers takes a very strong view on this matter and apologizes to the readers of the journal for any inconvenience this may cause.

von Hippel-Lindau mutants in renal cell carcinoma are regulated by increased expression of RSUME.

Renal cell carcinoma (RCC) is the major cause of death among patients with von Hippel-Lindau (VHL) disease. Resistance to therapies targeting tumor angiogenesis opens the question about the underlying mechanisms. Previously we have described that RWDD3 or RSUME (RWD domain-containing protein SUMO Enhancer) sumoylates and binds VHL protein and negatively regulates HIF degradation, leading to xenograft RCC tumor growth in mice. In this study, we performed a bioinformatics analysis in a ccRCC dataset showing an association of RSUME levels with VHL mutations and tumor progression, and we demonstrate the molecular mechanism by which RSUME regulates the pathologic angiogenic phenotype of VHL missense mutations. We report that VHL mutants fail to downregulate RSUME protein levels accounting for the increased RSUME expression found in RCC tumors. Furthermore, we prove that targeting RSUME in RCC cell line clones carrying missense VHL mutants results in decreased early tumor angiogenesis. The mechanism we describe is that RSUME sumoylates VHL mutants and beyond its sumoylation capacity, interacts with Type 2 VHL mutants, reduces HIF-2α-VHL mutants binding, and negatively regulates the assembly of the Type 2 VHL, Elongins and Cullins (ECV) complex. Altogether these results show RSUME involvement in VHL mutants deregulation that leads to the angiogenic phenotype of RCC tumors.

Atrofia progressiva de retina associada a mutação no gene PRCD - método de detecção genética em cães / Atrofia progesiva de retina asociada con la mutación en el gen PRCD-metodo de detección genética en los caninos / Progressive retinal atrophy caused by a PRCD gene mutation-genetic-based detection method in dogs

Nos cães, a atrofia progressiva de retina é uma doença causada por uma mutação pontual no gene PRCD, que leva à cegueira no seu estágio final. Na Argentina, o diagnóstico é realizado atualmente por meio de exame clínico e de eletrorretinografia. Essa enfermidade apresenta herança recessiva, com portadores assintomáticos capazes de transmitir a mutação para os seus filhotes, o que evidencia a necessidade de um diagnóstico por meio de teste genético, que permita a detecção precoce de portadores e doentes. No presente trabalho, descreve-se uma técnica para a detecção genética da APR-PRCD, por meio de método de sequenciamento, que determina a sucessão de nucleotídeos na região de PRCD onde se localiza a mutação. Foram utilizadas amostras de suabe bucal de dezoito cães (quinze poodles e três cockers) com atrofia de retina diagnosticada por exame clínico. Como controle foram utilizadas amostras de dez animais das mesmas raças, que não possuíam alterações oftalmológicas.(AU)

Canine progressive retinal atrophy is a disease caused by a mutation in the PRCD gene, which progresses toward end-stage blindness. In Argentina, the current method of diagnosis is by clinical examination and electroretinography. The fact that this condition is a recessive trait, with unaffected carriers that can transmit the mutation to their offspring, highlights the importance of a genetic screening procedure to detect carriers and affected individuals. This paper shows a PRA-PRCD genetic test based on sequencing the nucleotides around the PRCD mutation, allowing the most accurate diagnosis. Oral swab samples from eighteen dogs (fifteen poodles and three cockers) clinically diagnosed with retinal atrophy were studied. The control group consisted of ten animals of the same breeds without ophthalmic diseases.(AU)

La atrofia progresiva de retina en los perros es una enfermedad causada por una mutación puntual en el gen PRCD que conduce en su estadio final a la ceguera. En Argentina el diagnóstico se realiza actualmente a través el examen clínico y la electroretinografía. Presenta herencia recesiva, con portadores asintomáticos capaces de trasmitir la mutación a sus crías, esto torna necesario el desarrollo de un test genético para realizar la detección temprana de portadores y afectados. En el siguiente trabajo se describe una técnica para la detección genética de APR-PRCD a través del método de secuenciación, que determina la sucesión de nucleótidos en la región de PRCD que contiene la mutación. Se emplearon muestras de hisopado bucal provenientes de dieciocho caninos (quince caniches y tres cockers) con atrofia de retina diagnosticada clínicamente y como control se estudiaron muestras de diez animales de las mismas razas sin enfermedad oftalmológica.(AU)

Atrofia progressiva de retina associada a mutação no gene PRCD - método de detecção genética em cães / Atrofia progesiva de retina asociada con la mutación en el gen PRCD-metodo de detección genética en los caninos / Progressive retinal atrophy caused by a PRCD gene mutation-genetic-based detection method in dogs

Nos cães, a atrofia progressiva de retina é uma doença causada por uma mutação pontual no gene PRCD, que leva à cegueira no seu estágio final. Na Argentina, o diagnóstico é realizado atualmente por meio de exame clínico e de eletrorretinografia. Essa enfermidade apresenta herança recessiva, com portadores assintomáticos capazes de transmitir a mutação para os seus filhotes, o que evidencia a necessidade de um diagnóstico por meio de teste genético, que permita a detecção precoce de portadores e doentes. No presente trabalho, descreve-se uma técnica para a detecção genética da APR-PRCD, por meio de método de sequenciamento, que determina a sucessão de nucleotídeos na região de PRCD onde se localiza a mutação. Foram utilizadas amostras de suabe bucal de dezoito cães (quinze poodles e três cockers) com atrofia de retina diagnosticada por exame clínico. Como controle foram utilizadas amostras de dez animais das mesmas raças, que não possuíam alterações oftalmológicas.

Canine progressive retinal atrophy is a disease caused by a mutation in the PRCD gene, which progresses toward end-stage blindness. In Argentina, the current method of diagnosis is by clinical examination and electroretinography. The fact that this condition is a recessive trait, with unaffected carriers that can transmit the mutation to their offspring, highlights the importance of a genetic screening procedure to detect carriers and affected individuals. This paper shows a PRA-PRCD genetic test based on sequencing the nucleotides around the PRCD mutation, allowing the most accurate diagnosis. Oral swab samples from eighteen dogs (fifteen poodles and three cockers) clinically diagnosed with retinal atrophy were studied. The control group consisted of ten animals of the same breeds without ophthalmic diseases.

La atrofia progresiva de retina en los perros es una enfermedad causada por una mutación puntual en el gen PRCD que conduce en su estadio final a la ceguera. En Argentina el diagnóstico se realiza actualmente a través el examen clínico y la electroretinografía. Presenta herencia recesiva, con portadores asintomáticos capaces de trasmitir la mutación a sus crías, esto torna necesario el desarrollo de un test genético para realizar la detección temprana de portadores y afectados. En el siguiente trabajo se describe una técnica para la detección genética de APR-PRCD a través del método de secuenciación, que determina la sucesión de nucleótidos en la región de PRCD que contiene la mutación. Se emplearon muestras de hisopado bucal provenientes de dieciocho caninos (quince caniches y tres cockers) con atrofia de retina diagnosticada clínicamente y como control se estudiaron muestras de diez animales de las mismas razas sin enfermedad oftalmológica.

Critical role of endogenous heme oxygenase 1 as a tuner of the invasive potential of prostate cancer cells.

Prostate cancer (PCa) is the second leading cause of cancer-associated death in men. Inflammation has been recognized as a risk factor for this disease. Heme oxygenase 1 (HO-1), the inducible isoform of the rate-limiting enzyme in heme degradation, counteracts oxidative and inflammatory damage. Here, we investigated the regulated expression of HO-1 and its functional consequences in PCa. We studied the effect of genetic and pharmacologic disruption of HO-1 in the growth, invasion, and migration in androgen-sensitive (MDA PCa2b and LNCaP) and androgen-insensitive (PC3) PCa cell lines. Our results show that HO-1 levels are markedly decreased in PC3 compared with MDA PCa2b and LNCaP. Hemin treatment increased HO-1 at both protein and mRNA levels in all cell lines and decreased cell proliferation and invasion. Furthermore, overexpression of HO-1 in PC3 resulted in markedly reduced cell proliferation and migration. Accordingly, small interfering RNA-mediated silencing of HO-1 expression in MDA PCa2b cells resulted in increased proliferation and invasion. Using reverse transcription-quantitative PCR-generated gene array, a set of inflammatory and angiogenic genes were upregulated or downregulated in response to HO-1 overexpression identifying matrix metalloprotease 9 (MMP9) as a novel downstream target of HO-1. MMP9 production and activity was downregulated by HO-1 overexpression. Furthermore, PC3 cells stably transfected with HO-1 (PC3HO-1) and controls were injected into nu/nu mice for analysis of in vivo tumor xenograft phenotype. Tumor growth and MMP9 expression was significantly reduced in PC3HO-1 tumors compared with control xenografts. Taken together, these results implicate HO-1 in PCa cell migration and proliferation suggesting its potential role as a therapeutic target in clinical settings.