LIZANDRE KEREN RAMOS DA SILVEIRA
Projetos de Pesquisa
Unidades Organizacionais
Instituto de Ortopedia e Traumatologia, Hospital das Clínicas, Faculdade de Medicina
LIM/17 - Laboratório de Investigação em Reumatologia, Hospital das Clínicas, Faculdade de Medicina
LIM/17 - Laboratório de Investigação em Reumatologia, Hospital das Clínicas, Faculdade de Medicina
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- Effects of different fluid management on lung and kidney during pressure-controlled and pressure-support ventilation in experimental acute lung injury(2022) CARVALHO, Eduardo Butturini de; FONSECA, Ana Carolina Fernandes; MAGALHAES, Raquel Ferreira; PINTO, Eliete Ferreira; SAMARY, Cynthia dos Santos; ANTUNES, Mariana Alves; BALDAVIRA, Camila Machado; SILVEIRA, Lizandre Keren Ramos da; TEODORO, Walcy Rosolia; ABREU, Marcelo Gama de; CAPELOZZI, Vera Luiza; FELIX, Nathane Santanna; PELOSI, Paolo; ROCCO, Patricia Rieken Macedo; SILVA, Pedro LemeOptimal fluid management is critical during mechanical ventilation to mitigate lung damage. Under normovolemia and protective ventilation, pulmonary tensile stress during pressure-support ventilation (PSV) results in comparable lung protection to compressive stress during pressure-controlled ventilation (PCV) in experimental acute lung injury (ALI). It is not yet known whether tensile stress can lead to comparable protection to compressive stress in ALI under a liberal fluid strategy (LF). A conservative fluid strategy (CF) was compared with LF during PSV and PCV on lungs and kidneys in an established model of ALI. Twenty-eight male Wistar rats received endotoxin intratracheally. After 24 h, they were treated with CF (minimum volume of Ringer's lactate to maintain normovolemia and mean arterial pressure >= 70 mmHg) or LF (similar to 4 times higher than CF) combined with PSV or PCV (VT = 6 ml/kg, PEEP = 3 cmH(2)O) for 1 h. Nonventilated animals (n = 4) were used for molecular biology analyses. CF-PSV compared with LF-PSV: (1) decreased the diffuse alveolar damage score (10 [7.8-12] vs. 25 [23-31.5], p = 0.006), mainly due to edema in axial and alveolar parenchyma; (2) increased birefringence for occludin and claudin-4 in lung tissue and expression of zonula-occludens-1 and metalloproteinase-9 in lung. LF compared with CF reduced neutrophil gelatinase-associated lipocalin and interleukin-6 expression in the kidneys in PSV and PCV. In conclusion, CF compared with LF combined with PSV yielded less lung epithelial cell damage in the current model of ALI. However, LF compared with CF resulted in less kidney injury markers, regardless of the ventilatory strategy.
conferenceObject Usefulness of type V collagen and alpha 2 beta 1-integrin in the cytological diagnosis of pleural liquid biopsy(2019) ANTONANGELO, L.; TEODORO, W. Rosolia; FARIA, C. Silverio; QUEIROZ, Z. A.; SILVEIRA, L. Ramos da; VELOSA, A. P.; CAPELOZZI, V.- Photobiomodulation therapy increases collagen II after tendon experimental injury(2021) AKAMATSU, Flayia Emi; TEODORO, Walcy Rosolia; ITEZEROTE, Ana Maria; SILVEIRA, Lizandre Keren Ramos da; SALEH, Samir; MARTINEZ, Carlos Augusto Real; RIBEIRO, Marcelo Lima; PEREIRA, Jose Aires; HOJAIJ, Flavio; ANDRADE, Mauro; JACOMO, Alfredo LuizA tendon is a mechanosensitive tissue that transmits muscle-derived forces to bones. Photobiomodulation (PBM), also known as low-level laser therapy (LLLT), has been used in therapeutic approaches in tendon lesions, but uncertainties regarding its mechanisms of action have prevented its widespread use. We investigated the response of PBM therapy in experimental lesions of the Achilles tendon in rats. Thirty adult male Wistar rats weighing 250 to 300 g were surgically submitted to bilateral partial transverse section of the Achilles tendon. The right tendon was treated with PBM, whereas the left tendon served as a control. On the third postoperative day, the rats were divided into three experimental groups consisting of ten rats each, which were treated with PBM (Konf, Aculas HB 750), 780 nm and 80 mW for 20 seconds, three times/week for 7, 14 and 28 days. The rats were sacrificed at the end of the therapeutic time period. The Sca-1 was examined by immunohistochemistry and histomorphometry, and COLA1, COLA2 and COLA3 gene expression was examined by qRT-PCR. COLA2 gene expression was higher in PBM treated tendons than in the control group. The histomorphometric analysis coincided with increased number of mesenchymal cells, characterized by Sca-1 expression in the lesion region (p<0.001). PBM effectively interferes in tendon tissue repair after injury by stimulating mesenchymal cell proliferation and the synthesis of collagen type II, which is suggested to provide structural support to the interstitial tissues during the healing process of the Achilles tendon. Further studies are needed to confirm the role of PBM in tendon healing.
conferenceObject In situ and humoral sensitivity to native type V collagen in early synovial fibrosis of a rat model of arthritis(2019) SILVEIRA, L. K. Ramos da; VELOSA, A. P. Pereira; CATANOZI, S.; SANTOS FILHO, A.; FERNEZLIAN, S.; EHER, E.; MIRANDA, J. Tomaz de; CAPELOZZI, V. L.; TEODORO, W. Rosolia- Decreased Bone Type I Collagen in the Early Stages of Chronic Obstructive Pulmonary Disease (COPD)(2020) JUNQUEIRA, Jader Joel Machado; LOURENCO, Juliana Dias; SILVA, Kaique Rodrigues da; CERVILHA, Daniela Aparecida de Brito; SILVEIRA, Lizandre Keren Ramos da; CORREIA, Aristides Tadeu; SILVA, Larissa Emidio de Franca; TEODORO, Walcy Rosolia; TIBERIO, Iolanda de Fatima Lopes Calvo; BARBOSA, Alexandre Povoa; LOPES, Fernanda Degobbi Tenorio Quirino dos SantosSmoking is the main risk factor for the development of chronic obstructive pulmonary disease (COPD) and is known to have deleterious effects on bone metabolism. However, the effects on bone collagen matrix during the development of COPD are unclear. The aim of this study was to evaluate the temporal effect of cigarette smoke exposure on bone type I collagen during COPD development in a cigarette smoke-induced model. C57BL/6 mice were allocated to three groups: control (C), animals exposed to filtered air for 1, 3 and 6 months; cigarette smoke (S), animals exposed to cigarette smoke for 1, 3 and 6 months; provisional smoking (PS), animals exposed to cigarette smoke for 3 months, followed by another 3 months of filtered air exposure. Evaluation of the respiratory mechanics and alveolar enlargement were performed. Femoral and tibial extraction was also performed to evaluate the type I collagen by immunofluorescence andCOL1A1gene expression. Exposure to cigarette smoke led to an alveolar enlargement and progressive reduction in lung tissue resistance and elastance, progressive reduction of type I collagen and reduction inCOL1A1gene expression. Although we did not observe any improvement in the functional and histological parameters in the provisional smoking group, we detected an increase inCOL1A1gene expression. A worsening in bone collagen matrix is part of the initial physiopathological events during COPD development and the smoking cessation induced an evident recovery ofCOL1A1expression, possibly to attempt at tissue repair.
- Modeling extracellular matrix through histo-molecular gradient in NSCLC for clinical decisions(2022) BALDAVIRA, Camila Machado; PRIETO, Tabatha Gutierrez; MACHADO-RUGOLO, Juliana; MIRANDA, Jurandir Tomaz de; OLIVEIRA, Lizandre Keren Ramos de; VELOSA, Ana Paula Pereira; TEODORO, Walcy Rosolia; AB'SABER, Alexandre; TAKAGAKI, Teresa; CAPELOZZI, Vera LuizaLung cancer still represents a global health problem, being the main type of tumor responsible for cancer deaths. In this context, the tumor microenvironment, and the extracellular matrix (ECM) pose as extremely relevant. Thus, this study aimed to explore the prognostic value of epithelial-to-mesenchymal transition (EMT), Wnt signaling, and ECM proteins expression in patients with non-small-cell lung carcinoma (NSCLC) with clinical stages I-IIIA. For that, we used 120 tissue sections from patients and evaluated the immunohistochemical, immunofluorescence, and transmission electron microscopy (TEM) to each of these markers. We also used in silico analysis to validate our data. We found a strong expression of E-cadherin and beta-catenin, which reflects the differential ECM invasion process. Therefore, we also noticed a strong expression of chondroitin sulfate (CS) and collagens III and V. This suggests that, after EMT, the basal membrane (BM) enhanced the motility of invasive cells. EMT proteins were directly associated with WNT5A, and collagens III and V, which suggests that the WNT pathway drives them. On the other hand, heparan sulfate (HS) was associated with WNT3A and SPARC, while WNT1 was associated with CS. Interestingly, the association between WNT1 and Col IV suggested negative feedback of WNT1 along the BM. In our cohort, WNT3A, WNT5A, heparan sulfate and SPARC played an important role in the Cox regression model, influencing the overall survival (OS) of patients, be it directly or indirectly, with the SPARC expression stratifying the OS into two groups: 97 months for high expression; and 65 for low expression. In conclusion, the present study identified a set of proteins that may play a significant role in predicting the prognosis of NSCLC patients with clinical stages I-IIIA.
conferenceObject Imbalance between types I and VI collagen promotes skin fragility in human and experimental diabetes(2019) MIRANDA, J. T. De; TOLEDO, V. P.; QUEIROZ, Z. A.; SILVEIRA, L. K. Ramos da; SANTOS FILHO, A. dos; VELOSA, A. P. Pereira; CATANOZI, S.; CAPELOZZI, V. L.; TEODORO, W. Rosolia- In Situ Overexpression of Matricellular Mechanical Proteins Demands Functional Immune Signature and Mitigates Non-Small Cell Lung Cancer Progression(2021) YAEGASHI, Lygia Bertalha; BALDAVIRA, Camila Machado; PRIETO, Tabatha Gutierrez; MACHADO-RUGOLO, Juliana; VELOSA, Ana Paula Pereira; SILVEIRA, Lizandre Keren Ramos da; ASSATO, Aline; AB'SABER, Alexandre Muxfeldt; FALZONI, Roberto; TAKAGAKI, Teresa; SILVA, Pedro Leme; TEODORO, Walcy Rosolia; CAPELOZZI, Vera LuizaNon-small cell lung carcinoma (NSCLC) is a complex cancer biome composed of malignant cells embedded in a sophisticated tumor microenvironment (TME) combined with different initiating cell types, including immune cells and cancer-associated fibroblasts (CAFs), and extracellular matrix (ECM) proteins. However, little is known about these tumors' immune-matricellular relationship as functional and mechanical barriers. This study investigated 120 patients with NSCLC to describe the immune-matricellular phenotypes of their TME and their relationship with malignant cells. Immunohistochemistry (IHC) was performed to characterize immune checkpoints (PD-L1, LAG-3, CTLA-4+, VISTA 1), T cells (CD3+), cytotoxic T cells (CD8(+), Granzyme B), macrophages (CD68+), regulatory T cells (FOXP3+, CD4+), natural killer cells (CD57+), and B lymphocytes (CD20+), whereas CAFs and collagen types I, III, and V were characterized by immunofluorescence (IF). We observed two distinct functional immune-cellular barriers-the first of which showed proximity between malignant cells and cytotoxic T cells, and the second of which showed distant proximity between non-cohesive nests of malignant cells and regulatory T cells. We also identified three tumor-associated matricellular barriers: the first, with a localized increase in CAFs and a low deposition of Col V, the second with increased CAFs, Col III and Col I fibers, and the third with a high amount of Col fibers and CAFs bundled and aligned perpendicularly to the tumor border. The Cox regression analysis was designed in two steps. First, we investigated the relationship between the immune-matricellular components and tumor pathological stage (I, II, and IIIA), and better survival rates were seen in patients whose tumors expressed collagen type III > 24.89 fibers/mm(2). Then, we included patients who had progressed to pathological stage IV and found an association between poor survival and tumor VISTA 1 expression > 52.86 cells/mm(2) and CD3+ <= 278.5 cells/mm(2). We thus concluded that differential patterns in the distribution of immune-matricellular phenotypes in the TME of NSCLC patients could be used in translational studies to predict new treatment strategies and improve patient outcome. These data raise the possibility that proteins with mechanical barrier function in NSCLC may be used by cancer cells to protect them from immune cell infiltration and immune-mediated destruction, which can otherwise be targeted effectively with immunotherapy or collagen therapy.
conferenceObject Oral Collagen Type V Supplementation Inhibits Cartilage Degeneration in Experimental Arthritis(2019) SILVEIRA, Lizandre Keren; RODRIGUES, Jose Eduardo; ATAYDE, Silvana; CATANOZI, Sergio; SANTOS FILHO, Antonio dos; CAPELOZZI, Vera Luiza; FULLER, Ricardo; VELOSA, Ana Paula; TEODORO, Walcy- Immunization with SARS-CoV-2 Nucleocapsid protein triggers a pulmonary immune response in rats(2022) SILVA, E. K. V. B.; BOMFIM, C. G.; BARBOSA, A. P.; NODA, P.; NORONHA, I. L.; FERNANDES, B. H. V.; MACHADO, R. R. G.; DURIGON, E. L.; CATANOZI, S.; RODRIGUES, L. G.; PIERONI, F.; LIMA, S. G.; TEODORO, W. R.; QUEIROZ, Z. A. J.; SILVEIRA, L. K. R.; CHARLIE-SILVA, I.; CAPELOZZI, V. L.; GUZZO, C. R.; FANELLI, C.The SARS-CoV-2 pandemic have been affecting millions of people worldwide, since the beginning of 2020. COVID-19 can cause a wide range of clinical symptoms, which varies from asymptomatic presentation to severe respiratory insufficiency, exacerbation of immune response, disseminated microthrombosis and multiple organ failure, which may lead to dead. Due to the rapid spread of SARS-CoV-2, the development of vaccines to minimize COVID-19 severity in the world population is imperious. One of the employed techniques to produce vaccines against emerging viruses is the synthesis of recombinant proteins, which can be used as immunizing agents. Based on the exposed, the aim of the present study was to verify the systemic and immunological effects of IM administration of recombinant Nucleocapsid protein (NP), derived from SARS-CoV-2 and produced by this research group, in 2 different strains of rats (Rattus norvegicus); Wistar and Lewis. For this purpose, experimental animals received 4 injections of NP, once a week, and were submitted to biochemical and histological analysis. Our results showed that NP inoculations were safe for the animals, which presented no clinical symptoms of worrying side effects, nor laboratorial alterations in the main biochemical and histological parameters, suggesting the absence of toxicity induced by NP. Moreover, NP injections successfully triggered the production of specific anti-SARS-CoV-2 IgG antibodies by both Wistar and Lewis rats, showing the sensitization to have been well sufficient for the immunization of these strains of rats. Additionally, we observed the local lung activation of the Bronchus-Associated Lymphoid Tissue (BALT) of rats in the NP groups, suggesting that NP elicits specific lung immune response. Although pre-clinical and clinical studies are still required, our data support the recombinant NP produced by this research group as a potential immunizing agent for massive vaccination, and may represent advantages upon other recombinant proteins, since it seems to induce specific pulmonary protection. © 2022 Silva et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.