Imunoterapia com células CAR-T como nova perspectiva de tratamento da leucemia linfoblástica aguda recidivada/refratária / Immunotherapy with CAR-T cells as a new treatment perspective for relapsed/refractory acute lymphoblastic leukemia

Introdução: O tratamento da leucemia linfoblástica aguda (LLA) atualmente baseia-se em quimioterapia e/ou transplante de células tronco hematopoiéticas; entretanto, uma nova terapia vem se tornando promissora: a imunoterapia com células T modificadas geneticamente que expressam um receptor de antígeno quimérico (CAR-T) visando antígenos específicos presente em blastos de LLA, gerando resultados promissores em crianças e adultos com doença recidivada e refratária (r/r). Objetivo: Discorrer sobre a LLA e descrever a imunoterapia com CAR-T, como inovação terapêutica no tratamento da LLA de linhagem B. Método: Foi realizada uma revisão bibliográfica por meio de publicações indexadas nas bases de dados Scielo e Pubmed, utilizando os descritores: leucemia linfoblástica aguda de células B; células CAR-T; receptores de antígeno quimérico, recidivados/refratários; imunoterapia. Resultados: As altas taxas de remissão completa (42% até 100%) e parcial (28,5%) da LLA (r/r) tratadas com CAR-T, possibilitam um aumento considerável da sobrevida geral comparado a outros tratamentos convencionais. Efeitos desfavoráveis, tais como síndrome da liberação de citocinas (CRS) (0 até 90%) e neurotoxicidade (NT) (0 até 29%) podem ser vistos, sendo manejáveis, não prejudicando o desfecho do tratamento. Conclusão: A LLA é uma doença grave, de difícil tratamento e prognóstico reservado. A imunoterapia vêm se mostrando promissora à essa enfermidade, principalmente em casos de doença r/r se mostrado uma ferramenta poderosa que permite o foco específico de células malignas por meio de engenharia de células T

Introduction: The treatment of acute lymphoblastic leukemia (ALL) is currently based on chemotherapy and/or hematopoietic stem cell transplantation; however, a new therapy is becoming promising: immunotherapy with genetically modified T cells that express a chimeric antigen receptor (CAR-T) targeting specific antigens present on ALL blasts, reaching promising results in children and adults with relapsed and refractory disease (r/r). Objective: To discuss ALL and describe immunotherapy with CAR-T as a therapeutic innovation in the treatment of B-lineage ALL. Method: A literature review was carried out through publications indexed in the Scielo and Pubmed databases, using the following descriptors: B-cell acute lymphoblastic leukemia; CAR-T cells; chimeric antigen receptors, relapsed/refractory; immunotherapy. Results: The high rates of complete (42% to 100%) and partial remission (28.5%) of ALL (r/r) treated with CAR-T allows a considerable increase in overall survival compared to other conventional treatments. Unfavorable effects such as cytokine release syndrome (CRS) (0 to 90%) and neurotoxicity (NT) (0 to 29%) can be seen, being manageable, not impairing the treatment outcome. Conclusion: ALL is a serious disease, with a difficult treatment and poor prognosis. Immunotherapy has shown benefits for this disease, especially in cases of r/r ALL, showing itself to be a powerful tool that allows the specific focus of malignant cells through T cell engineering.

Implant-induced inflammatory angiogenesis is up-regulated in obese mice.

The damaging effects of obesity extend to multiple pre-existing tissue/organs. However, the influence of this condition on key components (inflammation and angiogenesis) of fibrovascular connective proliferating tissue, essential in repair processes, has been neglected. Our objective in this study was to investigate whether obesity would influence inflammatory-angiogenesis induced by synthetic matrix of polyether-polyurethane implanted subcutaneously in high-fat-fed obese C57/BL6 mice. Fourteen days after implantation, the inflammatory and angiogenic components of the newly formed tissue intra-implant were evaluated. The pro-inflammatory enzyme activities, myeloperoxidase (MPO) and N-acetyl-ß-D-glucosaminidase (NAG), the levels of TNF-α, CXCL1/KC and CCL2 and NF-κB transcription factor were examined. Angiogenesis was determined by morphometric analysis of implant blood vessels, intra-implant levels of hemoglobin content, VEGF levels, and western blot for VEGFR2. All inflammatory and angiogenic markers were increased in the implants of obese mice compared with their non-obese counterparts. Similarly, activation of the NF-κB pathway and phosphorylation of VEGFR2 were higher in implants of obese mice (1.60 ± 0.28 Np65/Cp65; 0.96 ± 0.08 p-VEGFR2/VEGFR2-T) compared with implants of non-obese animals (1.40 ± 0.14; 0.49 ± 0.08). These observations suggest that obesity exerts critical role in sponge-induced inflammatory-angiogenesis, possibly by activating fibrovascular components in the inflamed microenvironment. Thus, this pathological condition causes damage not only to pre-existing tissues/organs but also to newly formed proliferating fibrovascular tissue. This is relevant to the development of therapeutic approaches to improve healing processes in patients with obesity.

Cytokine Production Is Differentially Modulated in Malignant and Non-malignant Tissues in ST2-Receptor Deficient Mice.

IL-33/ST2 axis has been shown to exert both pro- and anti- effects in wound healing and tumor development. To further understand the role of this cytokine complex, we characterized comparatively the inflammatory component of a malignant tissue and non-malignant tissue in mice lacking ST2 receptor (ST2-KO). KO mice and their wild-type (WT) counterparts were either implanted subcutaneously with polyether-polyurethane sponge discs to induce non-malignant fibrovascular tissue growth or inoculated with 4T1 cells to induce mammary tumor. Loss of ST2 receptor in mice resulted in enhanced mammary tumor and fibrovascular tissue relative to the WT animals. The inflammatory parameters (MPO and NAG activities, levels of the cytokines CXCL1/KC, CCL2, TNF-α, TGF-ß1, and mast cell number) were differentially modulated in both tissues. In tumors, these parameters were, overall, lower compared with those in tumors of WT mice. In KO implants, CXCL1/KC and TNF-α levels increased; MPO, NAG, and CCL2 levels decreased relative to the WT implants. In addition, deletion of ST2 receptor inhibited mast cell recruitment but had no effect on TGF-ß1 levels in implants. Our study has shown antitumorigenic effect of ST2 in mammary tumor and this may be mediated by downregulation of pro-inflammatory cytokines (CXCL1/KC, CCL2, TNF-α, and TGF-ß1). Conversely, in the fibrovascular tissue, lack of ST2 receptor resulted in differential modulation of cytokine production. Differential signaling mechanisms may be activated by IL-33/ST2 axis to modulate cytokine production in malignant and non-malignant proliferative processes.

Upregulation of Foreign Body Response in Obese Mice.

OBJECTIVE: Obesity is a highly prevalent multifactorial metabolic condition in which the need for functional bioengineered substitutes (e.g., scaffolds for tissue engineering) is likely to occur. However, the adverse foreign body response (FBR) that invariably takes place adjacent to implant devices impairing their function is poorly characterized in this condition. This study investigated the influence of obesity on the host response to a synthetic matrix implanted subcutaneously in high-fat-fed obese mice. METHODS: Histological analysis of 14-day-old implants was performed to identify collagen deposition, capsule thickness, fibroblast-like cells, foreign body giant cells, and mast cells. In addition, transforming growth factor ß1 (TGF-ß1) levels in the implants and serum were determined. RESULTS: All fibrogenic markers (and TGF-ß1 levels) increased in the implants of obese mice compared with their nonobese counterparts. Particularly relevant was the fibrous capsule thickness in implants of obese mice (234.2 ± 22.1 µm vs. 109.2 ± 13.4 µm in implants of nonobese animals). CONCLUSIONS: The study results showing that obesity upregulates the main features of the FBR induced by subcutaneous implants in mice may be relevant in understanding biomaterial integration and performance in this condition. This is crucial to the development of strategies to maintain the integrity and function of implantable devices.

Kinetics of pancreatic tissue proliferation in a polymeric platform in mice.

BACKGROUND/OBJECTIVES: Pancreas regenerative capacity after injury is not always sufficient to comply with the body's requirement of digestive enzymes and hormones. We present an alternative system to induce pancreas parenchyma proliferation (exocrine and endocrine components), rather than regeneration or remodeling in normoglycemic mice. METHODS: Porous discs of polyether-polyurethane were surgically placed adjacent to the native pancreas and removed at days 15, 30 and 45 after implantation. No exogenous growth factors or extracellular matrix components were added to the platform. The synthetic matrix provided a platform that was filled with parenchymal and non-parenchymal pancreas tissue as detected by histological analysis. Immunohistochemistry analysis were performed to identify insulin positive cells in the newly formed tissue. In addition, angiogenic, inflammatory and metabolic parameters were carried out in those mice. RESULTS: At day 15, the pores of the platform were filled with inflammatory cells, spindled-shaped like fibroblasts, extracellular matrix components, blood vessels and clusters of pancreatic parenchyma (acini, ducts and islet-like structures). At days 30 and 45 the pancreas features remained well organized; its organization resembled that of a native pancreas. Interestingly, besides islet-like structures that showed positive cells to insulin, some ductal cells were also positive for insulin immunostaining. No significant differences in serum glucose and c-peptide concentrations during the experimental period were detected. CONCLUSIONS: The plain synthetic porous platform (without addition of exogenous molecules) placed adjacent to the native organ exhibits potential to restore and/or expand exocrine (acini, ducts) and endocrine (ß-cell mass) components in pancreatic injuries and in high metabolic demand.

Induction of liver proliferation using a polymeric platform in mice.

AIMS: Currently, animal models of liver regeneration are based on extensive lesions of the native organ and on cellular approaches using biomaterials to host growth factors and extracellular components to create artificial liver systems. We report a polymeric biological platform, minimally invasive, that induced sequential proliferation of liver parenchyma inside the scaffold in mice. MAIN METHODS: Porous discs of polyether-polyurethane were surgically placed under the left liver lobe and removed at days 4, 8, 12 and 25 after implantation. No exogenous growth factors or extracellular matrix components were added to the scaffold. Histological analysis of the implants was performed to identify hepatocytes, liver vascular structures and bile ducts in the newly formed tissue. In addition, systemic markers for hepatic function were determined. KEY FINDINGS: This biohybrid device provided a scaffold that was gradually filled with parenchymal and non-parenchymal liver tissue as detected by histological analysis. At day 4, the pores of the scaffold were filled with inflammatory cells and spindled-shaped like fibroblasts, and extracellular matrix components. At day 8, hepatocytes clusters, central lobular hepatic veins, portal space containing arteries, veins and biliary ducts were detected. By days 12 and 25 a liver-like structure filled 2/3 of the scaffold. Its organization resembled that of a mature liver. Serum concentration of ALT increased three-fold initially after implantation, returning gradually to control levels. SIGNIFICANCE: The plain synthetic scaffold (without addition of exogenous molecules) placed under the intact left liver lobe exhibits the potential to investigate physiological mechanisms that regulate liver parenchyma proliferation.

Synthetic matrix of polyether-polyurethane as a biological platform for pancreatic regeneration.

AIMS: Several alternative cellular approaches using biomaterials to host insulin-producing cells derived from stem cells have been developed to overcome the limitations of type 1 diabetes treatment (exogenous insulin injection). However, none seem to fulfill all requirements needed to induce pancreatic cells successful colonization of the scaffolds. Here, we report a polymeric platform adherent to the native mice pancreas filled with human adipose stem cells (hASCs) that was able to induce growth of pancreatic parenchyma. MAIN METHODS: Synthetic polyether-polyurethane discs were placed adjacent to pancreas of normoglycemic and streptozotocin-induced diabetic mice. At day 4 post implantation, 1×106 hASCs were injected intra-implant in groups of normoglycemic and diabetic mice. Immunohistochemistry analysis of the implants was performed to identify insulin positive cells in the newly formed tissue. In addition, metabolic, inflammatory and angiogenic parameters were carried out in those mice. KEY FINDINGS: This study provides evidence of the ability of a biohybrid device to induce the growth of differentiated pancreas parenchyma in both normoglycemic and streptozotocin-induced diabetic mice as detected by histological analysis. Glucose metabolism and body weight of hyperglycemic mice bearing hASCs implants improved. SIGNIFICANCE: The synthetic porous scaffold bearing hASC cells placed adjacent to the native animal pancreas exhibits the potential to be exploited in future cell-based type 1 diabetes therapies.

Murine strain differences in inflammatory angiogenesis of internal wound in diabetes.

Genetic susceptibility is associated with inflammation, neovascularization, and diabetes phenotypes. However, to what extent this susceptibility influences inflammatory angiogenesis in internal injuries in diabetes has not been fully investigated. Using the subcutaneous implantation of a synthetic matrix as an internal wound model in Swiss, C57BL/6 and Balb/c mice, we have studied inflammation, angiogenesis, and cytokine production in the fibrovascular tissue induced by implants in diabetic animals. The hyperglycemic levels (mg/dl) after the diabetogenic treatment were 455.0±15 in Swiss, 393.0±22 in C57BL/6, and 190.0±10 in Balb/c mice. Angiogenesis in Swiss implants from non-diabetic animals were higher than those in the implants from the other strains. However, the angiogenic inducers VEGF and nitric oxide (NO) were higher in implants from non-diabetic Swiss and Balb/c mice. Strain-related differences were also observed in the angiogenic parameters in implants from diabetic mice. Hb content and number of vessels decreased more than 40% in Swiss implants. In contrast, Hb content did not alter in implants from Balb/c diabetic mice and the number of vessels decreased. VEGF levels increased in implants from Swiss and C57BL/6 diabetic mice, but decreased in Balb/c implants. The levels of pro-inflammatory markers intra-implant also varied among the strains in both conditions. In the hyperglycemic environment, almost all inflammatory markers increased in implants from diabetic Swiss mice. These findings demonstrate the major contribution of genetic background in the pattern of inflammatory angiogenesis components of internal injury, in both normoglycemic and hyperglycemic animals.

Envelhecimento Digno: inserção da Mulher na Universidade Aberta para Terceira Idade / Decent Aging: Women's insertion at the Open University for the Third Age

As Universidades Abertas para Terceira Idade (UATI) configuram-se como espaço de socialização e implementação de políticas de integração de questões do envelhecimento dentro dos preceitos estabelecidos pelo Plano de Ação Internacional de Madri, engajando-se em diálogo com a sociedade. Este estudo que trata da inserção da mulher na UATI verificou que as questões conceituais que despertaram maior interesse em uma abordagem sobre gênero, principalmente do estudo da questão do feminino na terceira idade, centraram-se na valorização da memória e da identidade.

The conceptual issues that aroused the greatest interest in the approach on gender, especially the study of the question of women in old age is presented in memory enhancement and identity. Universities Open for Third Age is configured as a space of socialization and implementation of policies to integrate aging issues within the rules established by the International Plan of Action Madrid engaging in dialogue with society.

Envelhecimento Digno: inserção da Mulher na Universidade Aberta para Terceira Idade / Decent Aging: Women's insertion at the Open University for the Third Age

As Universidades Abertas para Terceira Idade (UATI) configuram-se como espaço de socialização e implementação de políticas de integração de questões do envelhecimento dentro dos preceitos estabelecidos pelo Plano de Ação Internacional de Madri, engajando-se em diálogo com a sociedade. Este estudo que trata da inserção da mulher na UATI verificou que as questões conceituais que despertaram maior interesse em uma abordagem sobre gênero, principalmente do estudo da questão do feminino na terceira idade, centraram-se na valorização da memória e da identidade.(AU)

The conceptual issues that aroused the greatest interest in the approach on gender, especially the study of the question of women in old age is presented in memory enhancement and identity. Universities Open for Third Age is configured as a space of socialization and implementation of policies to integrate aging issues within the rules established by the International Plan of Action Madrid engaging in dialogue with society.(AU)