EMMANUEL DIAS NETO
Projetos de Pesquisa
Unidades Organizacionais
LIM/27 - Laboratório de Neurociências, Hospital das Clínicas, Faculdade de Medicina
9 resultados
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Agora exibindo 1 - 9 de 9
- Towards a transcriptome-based theranostic platform for unfavorable breast cancer phenotypes(2016) DOBROFF, Andrey S.; D'ANGELO, Sara; ECKHARDT, Bedrich L.; FERRARA, Fortunato; STAQUICINI, Daniela I.; CARDO-VILA, Marina; STAQUICINI, Fernanda I.; NUNES, Diana N.; KIM, Kisu; DRIESSEN, Wouter H. P.; HAJITOU, Amin; LOMO, Lesley C.; BARRY, Marc; KRISHNAMURTHY, Savitri; SAHIN, Aysegul; WOODWARD, Wendy A.; PROSSNITZ, Eric R.; ANDERSON, Robin L.; DIAS-NETO, Emmanuel; BROWN-GLABERMAN, Ursa A.; ROYCE, Melanie E.; UENO, Naoto T.; CRISTOFANILLI, Massimo; HORTOBAGYI, Gabriel N.; MARCHIO, Serena; GELOVANI, Juri G.; SIDMAN, Richard L.; ARAP, Wadih; PASQUALINI, RenataInflammatory breast carcinoma (IBC) is one of the most lethal forms of human breast cancer, and effective treatment for IBC is an unmet clinical need in contemporary oncology. Tumor-targeted theranostic approaches are emerging in precision medicine, but only a few specific biomarkers are available. Here we report up-regulation of the 78-kDa glucose-regulated protein (GRP78) in two independent discovery and validation sets of specimens derived from IBC patients, suggesting translational promise for clinical applications. We show that a GRP78-binding motif displayed on either bacteriophage or adeno-associated virus/phage (AAVP) particles or loop-grafted onto a human antibody fragment specifically targets orthotopic IBC and other aggressive breast cancer models in vivo. To evaluate the theranostic value, we used GRP78-targeting AAVP particles to deliver the human Herpes simplex virus thymidine kinase type-1 (HSVtk) transgene, obtaining simultaneous in vivo diagnosis through PET imaging and tumor treatment by selective activation of the prodrug ganciclovir at tumor sites. Translation of this AAVP system is expected simultaneously to image, monitor, and treat the IBC phenotype and possibly other aggressive (e.g., invasive and/or metastatic) subtypes of breast cancer, based on the inducible cell-surface expression of the stress-response chaperone GRP78, and possibily other cell-surface receptors in human tumors.
- Synchronous down-modulation of miR-17 family members is an early causative event in the retinal angiogenic switch(2015) NUNES, Diana N.; DIAS-NETO, Emmanuel; CARDO-VILA, Marina; EDWARDS, Julianna K.; DOBROFF, Andrey S.; GIORDANO, Ricardo J.; MANDELIN, Jami; BRENTANI, Helena P.; HASSELGREN, Catrin; YAO, Virginia J.; MARCHIO, Serena; PEREIRA, Carlos A. B.; PASSETTI, Fabio; CALIN, George A.; SIDMAN, Richard L.; ARAP, Wadih; PASQUALINI, RenataSix members of the microRNA-17 (miR-17) family were mapped to three different chromosomes, although they share the same seed sequence and are predicted to target common genes, among which are those encoding hypoxia-inducible factor-1 alpha (HIF1A) and VEGFA. Here, we evaluated the in vivo expression profile of the miR-17 family in the murine retinopathy of prematurity (ROP) model, whereby Vegfa expression is highly enhanced at the early stage of retinal neovascularization, and we found simultaneous reduction of all miR-17 family members at this stage. Using gene reporter assays, we observed binding of these miRs to specific sites in the 3' UTRs of Hif1a and Vegfa. Furthermore, overexpression of these miRs decreased HIF1A and VEGFA expression in vitro. Our data indicate that this miR-17 family elicits a regulatory synergistic down-regulation of Hif1a and Vegfa expression in this biological model. We propose the existence of a coordinated regulatory network, in which diverse miRs are synchronously regulated to target the Hif1a transcription factor, which in turn, potentiates and reinforces the regulatory effects of the miRs on Vegfa to trigger and sustain a significant physiological response.
- PRUNE2 and PCA3 expression in paired non-malignant and tumor specimens from radical prostatectomy patients with gleason score 7 prostate cancer.(2017) LAUER, Richard C.; BARRY, Marc; WU, Jin; LEE, Ji-Hyun; MCCANCE, Dennis J.; DU, Ruofei; RAO, Arpit; DOBROFF, Andrey S.; DIAS-NETO, Emmanuel; CHEN, Isan; PASQUALINI, Renata; ARAP, Wadih
- PCA3 upregulation in prostate cancer: Analysis in a cohort of 497 subjects from TCGA.(2017) DIAS-NETO, Emmanuel; NUNES, Diana N.; THOMAS, Andrew M.; SMITH, Tracey L.; RAO, Arpit; LAUER, Richard C.; CHEN, Isan; ARAP, Wadih; PASQUALINI, Renata
- PRUNE2 is a human prostate cancer suppressor regulated by the intronic long noncoding RNA PCA3(2015) SALAMEH, Ahmad; LEE, Alessandro K.; CARDO-VILA, Marina; NUNES, Diana N.; EFSTATHIOU, Eleni; STAQUICINI, Fernanda I.; DOBROFF, Andrey S.; MARCHIO, Serena; NAVONE, Nora M.; HOSOYA, Hitomi; LAUER, Richard C.; WEN, Sijin; SALMERON, Carolina C.; Anh Hoang; NEWSHAM, Irene; LIMA, Leandro A.; CARRARO, Dirce M.; OLIVIERO, Salvatore; KOLONIN, Mikhail G.; SIDMAN, Richard L.; Kim-Anh Do; TRONCOSO, Patricia; LOGOTHETIS, Christopher J.; BRENTANI, Ricardo R.; CALIN, George A.; CAVENEE, Webster K.; DIAS-NETO, Emmanuel; PASQUALINI, Renata; ARAP, WadihProstate cancer antigen 3 (PCA3) is the most specific prostate cancer biomarker but its function remains unknown. Here we identify PRUNE2, a target protein-coding gene variant, which harbors the PCA3 locus, thereby classifying PCA3 as an antisense intronic long noncoding (lnc) RNA. We show that PCA3 controls PRUNE2 levels via a unique regulatory mechanism involving formation of a PRUNE2/PCA3 double-stranded RNA that undergoes adenosine deaminase acting on RNA (ADAR)-dependent adenosine-to-inosine RNA editing. PRUNE2 expression or silencing in prostate cancer cells decreased and increased cell proliferation, respectively. Moreover, PRUNE2 and PCA3 elicited opposite effects on tumor growth in immunodeficient tumor-bearing mice. Coregulation and RNA editing of PRUNE2 and PCA3 were confirmed in human prostate cancer specimens, supporting the medical relevance of our findings. These results establish PCA3 as a dominant-negative oncogene and PRUNE2 as an unrecognized tumor suppressor gene in human prostate cancer, and their regulatory axis represents a unique molecular target for diagnostic and therapeutic intervention.
- Poly (A)(+) Transcriptome Assessment of ERBB2-Induced Alterations in Breast Cell Lines(2011) CARRARO, Dirce Maria; FERREIRA, Elisa Napolitano; MOLINA, Gustavo de Campos; PUGA, Renato David; ABRANTES, Eduardo Fernandes; TRAPE, Adriana Priscila; EKHARDT, Bedrich L.; NUNES, Diana Noronha; BRENTANI, Maria Mitzi; ARAP, Wadih; PASQUALINI, Renata; BRENTANI, Helena; DIAS-NETO, Emmanuel; BRENTANI, Ricardo RenzoWe report the first quantitative and qualitative analysis of the poly (A)(+) transcriptome of two human mammary cell lines, differentially expressing (human epidermal growth factor receptor) an oncogene over-expressed in approximately 25% of human breast tumors. Full-length cDNA populations from the two cell lines were digested enzymatically, individually tagged according to a customized method for library construction, and simultaneously sequenced by the use of the Titanium 454-Roche-platform. Comprehensive bioinformatics analysis followed by experimental validation confirmed novel genes, splicing variants, single nucleotide polymorphisms, and gene fusions indicated by RNA-seq data from both samples. Moreover, comparative analysis showed enrichment in alternative events, especially in the exon usage category, in ERBB2 over-expressing cells, data indicating regulation of alternative splicing mediated by the oncogene. Alterations in expression levels of genes, such as LOX, ATP5L, GALNT3, and MME revealed by large-scale sequencing were confirmed between cell lines as well as in tumor specimens with different ERBB2 backgrounds. This approach was shown to be suitable for structural, quantitative, and qualitative assessment of complex transcriptomes and revealed new events mediated by ERBB2 overexpression, in addition to potential molecular targets for breast cancer that are driven by this oncogene.
- A total transcriptome profiling method for plasma-derived extracellular vesicles: applications for liquid biopsies(2017) AMORIM, Maria G.; VALIERIS, Renan; DRUMMOND, Rodrigo D.; PIZZI, Melissa P.; FREITAS, Vanessa M.; SINIGAGLIA-COIMBRA, Rita; CALIN, George A.; PASQUALINI, Renata; ARAP, Wadih; SILVA, Israel T.; DIAS-NETO, Emmanuel; NUNES, Diana N.Extracellular vesicles (EVs) are key mediators of intercellular communication. Part of their biological effects can be attributed to the transfer of cargos of diverse types of RNAs, which are promising diagnostic and prognostic biomarkers. EVs found in human biofluids are a valuable source for the development of minimally invasive assays. However, the total transcriptional landscape of EVs is still largely unknown. Here we develop a new method for total transcriptome profiling of plasma-derived EVs by next generation sequencing (NGS) from limited quantities of patient-derived clinical samples, which enables the unbiased characterization of the complete RNA cargo, including both small- and long-RNAs, in a single library preparation step. This approach was applied to RNA extracted from EVs isolated by ultracentrifugation from the plasma of five healthy volunteers. Among the most abundant RNAs identified we found small RNAs such as tRNAs, miRNAs and miscellaneous RNAs, which have largely unknown functions. We also identified protein-coding and long noncoding transcripts, as well as circular RNA species that were also experimentally validated. This method enables, for the first time, the full spectrum of transcriptome data to be obtained from minute patient-derived samples, and will therefore potentially allow the identification of cell-to-cell communication mechanisms and biomarkers.
- Going viral? Linking the etiology of human prostate cancer to the PCA3 long noncoding RNA and oncogenic viruses(2017) TEIXEIRA, Andre A.; MARCHIO, Serena; DIAS-NETO, Emmanuel; NUNES, Diana N.; SILVA, Israel T. da; CHACKERIAN, Bryce; BARRY, Marc; LAUER, Richard C.; GIORDANO, Ricardo J.; SIDMAN, Richard L.; WHEELER, Cosette M.; CAVENEE, Webster K.; PASQUALINI, Renata; ARAP, Wadih
- Evaluation of PRUNE2 and PCA3 expression as metastasis predictors in Gleason 7 prostate cancer(2017) LAUER, Richard C.; BARRY, Marc; WU, Jin; LEE, Ji-Hyun; MCCANCE, Dennis J.; DU, Ruofei; RAO, Arpit; DOBROFF, Andrey S.; DIAS-NETO, Emmanuel; CHEN, Isan; PASQUALINI, Renata; ARAP, Wadih