SERGIO PAULO BYDLOWSKI
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Clínica Médica, Faculdade de Medicina - Docente
LIM/19 - Laboratório de Histocompatibilidade e Imunidade Celular, Hospital das Clínicas, Faculdade de Medicina - Líder
LIM/19 - Laboratório de Histocompatibilidade e Imunidade Celular, Hospital das Clínicas, Faculdade de Medicina - Líder
8 resultados
Resultados de Busca
Agora exibindo 1 - 8 de 8
- LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk(2020) ARALDI, Rodrigo Pinheiro; MELO, Thatiana Correa de; LEVY, Debora; SOUZA, Dener Madeiro de; MAURICIO, Beatriz; COLOZZA-GAMA, Gabriel Avelar; BYDLOWSKI, Sergio Paulo; PENG, Hongzhuang; III, Frank J. Rauscher; CERUTTI, Janete MariaAlthough papillary thyroid carcinoma (PTC) has a good prognosis, 20-90% of patients show metastasis to regional lymph nodes and 10-15% of patients show metastasis to distant sites. Metastatic disease represents the main clinical challenge that impacts survival rate. We previously showed that LIMD2 was a novel metastasis-associated gene. In this study, to interrogate the role of LIMD2 in cancer invasion and metastasis, we used CRISPR-mediated knockout (KO) of LIMD2 in PTC cells (BCPAP and TPC1). Western blot and high-content screening (HCS) analysis confirmed functional KO of LIMD2. LIMD2 KO reduced in vitro invasion and migration. Ultrastructural analyses showed that cell polarity and mitochondria function and morphology were restored in LIMD2 KO cells. To unveil the signals supervising these phenotypic changes, we employed phospho-protein array. Several members of the MAPK superfamily showed robust reduction in phosphorylation. A Venn diagram displayed the overlap of kinases with reduced phosphorylation in both cell lines and showed that they were able to initiate or sustain the epithelial-mesenchymal transition (EMT) and DNA damage checkpoint. Flow cytometry and HCS validation analyses further corroborated the phospho-protein array data. Collectively, our findings show that LIMD2 enhances phosphorylation of kinases associated with EMT and invasion. Through cooperation with different kinases, it contributes to the increased genomic instability that ultimately promotes PTC progression.
- Beneficial effects of IL-4 and IL-6 on rat neonatal target cardiac cells(2020) ZOGBI, Camila; OLIVEIRA, Nathalia C.; LEVY, Debora; BYDLOWSKI, Sergio P.; BASSANEZE, Vinicius; NERI, Elida A.; KRIEGER, Jose E.The nature of the early post-natal immune response in rodents appears to influence cardiac regeneration even though the underlying molecules remain poorly understood. Consistent with this idea, we show now significant changes in the expression of immune and cell movement gene pathways in heart samples from 1- and 7-day-old rats with ventricle resection. We then tested whether conditioned media from adult M2 anti-inflammatory macrophages target neonatal cardiac cells to a pro-regenerative like phenotype compared to the M1 pro-inflammatory macrophages. We found that M2 compared to M1 macrophage-conditioned media upregulates neonatal cardiomyocyte proliferation, suppresses myofibroblast-induced differentiation and stimulates endothelial cell tube formation. Using a cytokine array, we selected four candidate cytokine molecules uniquely expressed in M2 macrophage-conditioned media and showed that two of them (IL-4 and IL-6) induce endothelial cell proliferation whilst IL-4 promotes proliferation in neonatal cardiomyocytes and prevents myofibroblast-induced collagen type I secretion. Altogether, we provided evidence that adult M2 macrophage-conditioned media displays a paracrine beneficial pro-regenerative response in target cardiac cells and that IL-4 and IL-6 recapitulate, at least in part, these pleiotropic effects. Further characterization of macrophage immune phenotypes and their secreted molecules may give rise to novel therapeutic approaches for post-natal cardiac repair.
- AUTOLOGOUS CHONDROCYTE IMPLANTATION IN BRAZIL(2020) GIGLIO, Pedro Nogueira; LIZIER, Nelson Foresto; LEVY, Debora; SOBRADO, Marcel Faraco; GOBBI, Riccardo Gomes; PECORA, Jose Ricardo; BYDLOWSKI, Sergio Paulo; DEMANGE, Marco KawamuraObjective: To describe the first series of cases of autologous chondrocyte implantation (ACI) in collagen membrane performed in Brazil. Methods: ACI was performed in 12 knees of 11 patients, aged 32.1 +/- 10.9 years, with 5.3 +/- 2.6 cm(2) full-thickness knee cartilage lesions, with a six-month minimum follow-up. Two surgical procedures were performed: arthroscopic cartilage biopsy for isolation and expansion of chondrocytes, which were seeded onto collagen membrane and implanted in the lesion site; the characterization of cultured cells and implant was performed using immunofluorescence for type II collagen (COL2) for cell viability and electron microscopy of the implant. Clinical safety, KOOS and IKDC scores and magnetic resonance imaging were evaluated. We used repeated-measures ANOVA and post-hoc comparisons at alpha = 5%. Results: COL2 was identified in the cellular cytoplasm, cell viability was higher than 95% and adequate distribution and cell adhesion were found in the membrane. The median follow-up was 10.9 months (7 to 19). We had two cases of arthrofibrosis, one of graft hypertrophy and one of superficial infection as complications, but none compromising clinical improvement. KOOS and IKDC ranged from 71.2 +/- 11.44 and 50.72 +/- 14.10, in preoperative period, to 85.0 +/- 4.4 and 70.5 +/- 8.0, at 6 months (p = 0.007 and 0.005). MRI showed regenerated tissue compatible with hyaline cartilage. Conclusion: ACI in collagen membrane was feasible and safe in a short-term follow- up, presenting regenerated formation visualized by magnetic resonance imaging and improved clinical function.
- Uptake of silver, gold, and hybrids silver-iron, gold-iron and silver-gold aminolevulinic acid nanoparticles by MCF-7 breast cancer cells(2020) GONCALVES, Karina de Oliveira; VIEIRA, Daniel Perez; LEVY, Debora; BYDLOWSKI, Sergio Paulo; COURROL, Lilia CoronatoBackground: Nanoparticles show promise for theranostic applications in cancer. The metal-based nanoparticles can be used both as photosensitizers and delivery vehicles. In bimetallic particles based on gold or silver and iron, a combination of the plasmonic features of the gold or silver components with the magnetic properties of the iron makes these hybrid nanomaterials suitable for both imaging and therapeutic applications. Herein, we discuss toxicity and cell internalization of metallic (silver and gold) and bimetallic (silver-iron, gold-iron, and silver-gold) aminolevulinic acid (ALA) nanoparticles. ALA can control the production of an intracellular photosensitizer, protoporphyrin IX (PpIX), commonly used in photodynamic therapy. Methods: Nanoparticles were synthesized by photoreduction method and characterized by UV/Vis spectra, Zeta potential, FTIR, XRD, and transmission electron microscopy. The amount of singlet oxygen generation by a yellow LED, and ultrasound was studied for gold, gold-iron, and silver-gold nanoparticles. Cytotoxicity assays of MCF-7 in the presence of nanoparticles were performed, and PpIX fluorescence was quantified by high content screening (HCS). Results: Red fluorescence observed after 24 h of nanoparticles incubation on MCF-7 cells, indicated that the ALA in surface of nanoparticles was efficiently converted to PpIX. The best results for singlet oxygen generation with LED or ultrasound irradiation were obtained with ALA:AgAuNPs. Conclusions: The studied nanoparticles present the potential to deliver aminolevulinic acid to breast cancer cells efficiently, generate singlet oxygen, and convert ALA into PpIX inside the cells allowing photodiagnosis and therapies such as photodynamic and sonodynamic therapies.
- Evaluation of the Dot-ELISA as a diagnostic test for human strongyloidiasis based on the detection of IgA in saliva(2020) BOSQUI, Larissa Rodrigues; CORRAL, Marcelo Andreetta; LEVY, Debora; BYDLOWSKI, Sergio Paulo; GRYSCHEK, Ronaldo Cesar Borges; CUSTODIO, Luiz Antonio; PAVANELLI, Wander Rogerio; CONCHON-COSTA, Ivete; COSTA-CRUZ, Julia Maria; PAULA, Fabiana Martins de; COSTA, Idessania NazarethThis study aimed to evaluate the use of saliva samples in the Dot-ELISA test for immunodiagnosis of human strongyloidiasis. The Dot-ELISA presented similar results to the ELISA test, with 70% and 60% sensitivity and 85% and 90% specificity, respectively, for IgA in the saliva. The Dot-ELISA with alternative saliva samples may be a suitable tool for diagnosing human strongyloidiasis, especially in populations with high levels of exposure to helminth.
- von Willebrand factor and factor VIII in a healthy Brazilian population. Association with ABO blood groups(2020) VALADARES, Daniela F.; SOARES, Rosangela R. P.; GIACOMO, Giovanna Di; ROCHA, Tania; REICHERT, Cadiele O.; BYDLOWSKI, Sergio P.
- Cell Cycle Changes, DNA Ploidy, and PTTG1 Gene Expression in HTLV-1 Patients(2020) LEVY, Debora; FERREIRA, Mari Cleia M. R.; REICHERT, Cadiele O.; ALMEIDA, Lis Vilela de; BROCARDO, Graciela; LAGE, Luis Alberto P. C.; CULLER, Hebert F.; NUKUI, Youko; BYDLOWSKI, Sergio P.; PEREIRA, JulianaHuman T-cell lymphotropic virus type-1 (HTLV-1) is a pathogenic retrovirus that is associated with adult T-cell leukemia/lymphoma (ATL). Genetic instability is the hallmark of ATL. Cell cycle progression is needed for virus particle reproduction. HTLV-1 encoded Tax protein ultimately disrupts the mitotic spindle checkpoint, leading to incorrect chromosome segregation, resulting in aneuploidy. Cell cycle abnormalities have been described in T cells transfected with HTLV-1 virusin vitro, but not in HTLV-1 asymptomatic carriers. PTTG1 and HTLV-1 viral protein Tax exhibit a cooperative transforming activity. Overexpressed PTTG1 results in chromosome instability and aneuploidy, which has been suggested as a mechanism underlying PTTG1 transforming activity. Here we aimed to investigate cell cycle, DNA ploidy and PTTG1 mRNA expression in CD4(+)and CD8(+)T cells in healthy subjects (HS), HTLV-1 asymptomatic carriers and ATL patients. We have identified that HTLV-1 asymptomatic carriers have shown DNA aneuploidy and cell cycle arrest at cell cycle phase G(0)/G(1)in CD4(+)T cells. CD8(+)T cells of HTLV-1 asymptomatic carriers also demonstrated DNA aneuploidy but without alteration in cell cycle. In ATL, CD4(+)and CD8(+)T cells present a higher number of cells in cell cycle S-phase and PTTG1 overexpression. These studies provide insight into malignant transformation of HTLV-1 asymptomatic carriers to ATL patients.
- Ferroptosis Mechanisms Involved in Neurodegenerative Diseases(2020) REICHERT, Cadiele Oliana; FREITAS, Fabio Alessandro de; SAMPAIO-SILVA, Juliana; ROKITA-ROSA, Leonardo; BARROS, Priscila de Lima; LEVY, Debora; BYDLOWSKI, Sergio PauloFerroptosis is a type of cell death that was described less than a decade ago. It is caused by the excess of free intracellular iron that leads to lipid (hydro) peroxidation. Iron is essential as a redox metal in several physiological functions. The brain is one of the organs known to be affected by iron homeostatic balance disruption. Since the 1960s, increased concentration of iron in the central nervous system has been associated with oxidative stress, oxidation of proteins and lipids, and cell death. Here, we review the main mechanisms involved in the process of ferroptosis such as lipid peroxidation, glutathione peroxidase 4 enzyme activity, and iron metabolism. Moreover, the association of ferroptosis with the pathophysiology of some neurodegenerative diseases, namely Alzheimer's, Parkinson's, and Huntington's diseases, has also been addressed.