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Since the COVID-19 pandemic started, mesenchymal stromal cells (MSC) appeared as a therapeutic option to reduce the over-activated inflammatory response and promote recovery of lung damage. Most clinical studies use intravenous injection for MSC delivery, raising several concerns of thrombogenic risk due to MSC procoagulant activity (PCA) linked to the expression of tissue factor (TF/CD142). This is the first study that demonstrated procoagulant activity of TF+ human immature dental pulp stromal cells (hIDPSC, NestaCell® product) with the percentage of TF+ cells varied from 0.2% to 63.9% in plasma of healthy donors and COVID-19 heparin-treated patients. Thrombogenic risk of TF+ hIDPSCs was evaluated by rotational thromboelastometry (in vitro) and in critically ill COVID-19 patients (clinical trial). We showed that the thromboelastography is not enough to predict the risk of TF+ MSC therapies. Using TF-negative HUVEC cells, we demonstrated that TF is not a unique factor responsible for the cell's procoagulant activity. However, heparin treatment minimizes MSC procoagulant (in vitro). We also showed that the intravenous infusion of hIDPSCs with prophylactic enoxaparin administration in moderate to critically ill COVID-19 patients did not change the values of D-dimer, neither in the PT and PTT times. Our COVID-19 clinical study measured and selected the therapeutic cells with low TF (less than 25% of TF+ hIDPSCs). Our data indicate that the concomitant administration of enoxaparin and low TF-loaded is safe even for critically ill COVID-19 patients.

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Cancer is one of the most important health problems and the second leading cause of death worldwide. Despite the advances in oncology, cancer heterogeneity remains challenging to therapeutics. This is because the exosome-mediated crosstalk between cancer and non-cancer cells within the tumor microenvironment (TME) contributes to the acquisition of all hallmarks of cancer and leads to the formation of cancer stem cells (CSCs), which exhibit resistance to a range of anticancer drugs. Thus, this review aims to summarize the role of TME-derived exosomes in cancer biology and explore the clinical potential of mesenchymal stem-cell-derived exosomes as a cancer treatment, discussing future prospects of cell-free therapy for cancer treatment and challenges to be overcome.

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Medullary thyroid carcinoma (MTC) is a malignant tumor originating from thyroid C-cells that can occur either in sporadic (70-80%) or hereditary (20-30%) form. In this study we aimed to identify recurrent copy number alterations (CNA) that might be related to the pathogenesis or progression of MTC. We used Affymetrix SNP array 6.0 on MTC and paired-blood samples to identify CNA using PennCNV and Genotyping Console software. The algorithms identified recurrent copy number gains in chromosomes 15q, 10q, 14q and 22q in MTC, whereas 4q cumulated losses. Coding genes were identified within CNA regions. The quantitative PCR analysis performed in an independent series of MTCs (n = 51) confirmed focal recurrent copy number gains encompassing the DLK1 (14q32.2) and AIFM3 (22q11.21) genes. Immunohistochemistry confirmed AIFM3 and DLK1 expression in MTC cases, while no expression was found in normal thyroid tissues and few MTC samples were found with normal copy numbers. The functional relevance of CNA was also assessed by in silico analysis. CNA status correlated with protein expression (DLK1, p = 0.01), tumor size (DLK1, p = 0.04) and AJCC staging (AIFM3p = 0.01 and DLK1p = 0.05). These data provide a novel insight into MTC biology, and suggest a common CNA landscape, regardless of if it is sporadic or hereditary MTC.

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Cancer is one of the most important health problems and the second leading cause of death worldwide. Despite the advances in oncology, cancer heterogeneity remains challenging to therapeutics. This is because the exosome-mediated crosstalk between cancer and non-cancer cells within the tumor microenvironment (TME) contributes to the acquisition of all hallmarks of cancer and leads to the formation of cancer stem cells (CSCs), which exhibit resistance to a range of anticancer drugs. Thus, this review aims to summarize the role of TME-derived exosomes in cancer biology and explore the clinical potential of mesenchymal stem-cell-derived exosomes as a cancer treatment, discussing future prospects of cell-free therapy for cancer treatment and challenges to be overcome.

RESUMO

The recent reclassification of a follicular variant of papillary thyroid carcinoma (FVPTC), subset as noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP), aims to avoid overtreatment of patients with an indolent lesion. The diagnosis of NIFTP has recently been revisited using more rigid criteria. This study presents histological and molecular findings and a long clinical follow-up of 94 FVPTC, 40 cases of follicular adenoma (FTA) and 22 cases of follicular carcinoma (FTC) that were classified before the advent of the NIFTP reclassification. All slides were reviewed using these rigid criteria and analysis of numerous sections of paraffin blocks and reclassified as 7 NIFTPs, 2 EFVPTCs, 29 infiltrative FVPTC (IFVPTCs), 57 invasive EFVPTC (I-EFVPTCs), 39 FTAs and 22 FTCs. Remarkably, EFVPTC and NIFTP patients were all free of disease at the end of follow-up and showed no BRAF mutation. Only one NIFTP sample harbored mutations, an NRAS Q61R. PAX8/PPARG fusion was found in I-EFVPTCs and FTC. Although additional studies are needed to identify a specific molecular profile to aid in the diagnosis of lesions with borderline morphological characteristics, we confirmed that the BRAF V600E mutation is an important tool to exclude the diagnosis of NIFTP. We also show that rigorous histopathological criteria should be strongly followed to avoid missing lesions in which more aggressive behavior is present, mainly via the analysis of capsule or vascular invasion and the presence of papillary structures.

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We previously proposed that high expression of FAM129A can be used as a thyroid carcinoma biomarker in preoperative diagnostic exams of thyroid nodules. Here, we identify that FAM129A expression is increased under nutrient and growth factor depletion in a normal thyroid cell line (PCCL3), overlapping with increased expression of autophagy-related protein and inhibition of AKT/mTOR/p70S6K. Supplementation of insulin, TSH and serum to the medium was able to reduce the expression of both FAM129A and autophagy-related protein and reestablish the AKT/mTOR/p70S6K axis. To determine the direct role of FAM129A on autophagy, FAM129A was transfected into PCCL3 cells. Its overexpression induced autophagic vesicles formation, evidenced by transmission electron microscopy. Co-expression of FAM129A and mCherry-EGFP-LC3B in PCCL3 showed an increased yellow puncta formation, suggesting that FAM129Ainduces autophagy. To further confirm its role on autophagy, we knockdown FAM129A in two thyroid carcinoma cell lines (TPC1 and FTC-236). Unexpectedly, FAM129A silencing increased autophagic flux, suggesting that FAM129A inhibits autophagy in these models. We next co-transfected PCCL3 cells with FAM129A and RET/PTC1 and tested autophagy in this context. Co-expression of FAM129A and RET/PTC1 oncogene in PCCL3 cells, inhibited RET/PTC1-induced autophagy. Together, our data suggest that, in normal cells FAM129A induces autophagy in order to maintain cell homeostasis and provide substrates under starvation conditions. Instead, in cancer cells, decreased autophagy may help the cells to overcome cell death. FAM129A regulates autophagy in a cell- and/or context-dependent manner. Our data reinforce the concept that autophagy can be used as a strategy for cancer treatment.

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We have identified previously a panel of markers (C1orf24, ITM1 and PVALB) that can help to discriminate benign from malignant thyroid lesions. C1orf24 and ITM1 are specifically helpful for detecting a wide range of thyroid carcinomas, and PVALB is particularly valuable for detecting the benign Hürthle cell adenoma. Although these markers may ultimately help patient care, the current understanding of their biological functions remains largely unknown. In this article, we investigated whether PVALB is critical for the acquisition of Hürthle cell features and explored the molecular mechanism underlying the phenotypic changes. Through ectopic expression of PVALB in thyroid carcinoma cell lines (FTC-133 and WRO), we demonstrated that PVALB sequesters free cytoplasmic Ca(2+), which ultimately lowers calcium levels and precludes endoplasmic reticulum (ER) Ca(2+) refilling. These results were accompanied by induced expression of PERK, an ER stress marker. Additionally, forced expression of PVALB reduces Ca(2+) inflow in the mitochondria, which can in turn cause changes in mitochondria morphology, increase mitochondria number and alter subcellular localization. These findings share striking similarity to those observed in Hürthle cell tumors. Moreover, PVALB inhibits cell growth and induces cell death, most likely through the AKT/GSK-3ß. Finally, PVALB expression coincides with Ca(2+) deposits in HCA tissues. Our data support the hypothesis that the loss of PVALB plays a role in the pathogenesis of thyroid tumors.

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INTRODUÇÃO: Os esteroides anabólicos androgênicos são usados por pessoas que desejam aumentar sua massa muscular para obter um melhor desempenho nos esportes ou melhorar a aparência física. Os EAA são derivados sintéticos da testosterona, capazes de promover a hipertrofia das fibras musculares, aumentando a síntese proteica intracelular. A L-carnitina é um suplemento alimentar empregado para aumentar a produção energética por meio da oxidação de ácidos graxos. Embora haja trabalhos mostrando as propriedades fisiológicas dessas drogas, há poucos estudos sobre o potencial mutagênico das mesmas. OBJETIVOS: Este trabalho avaliou a clastogenicidade e genotoxicidade do decanoato de nandrolona, decanoato de testosterona e da L-carnitina, em diferentes tratamentos, através do teste do micronúcleo em eritrócitos policromáticos de ratos Wistar. MÉTODOS: Os animais foram submetidos a diferentes concentrações e associações de EAA. O controle positivo recebeu ciclofosfamida 50 mg/kg através de injeção intraperitoneal e o controle negativo, 1 ml de soro fisiológico por gavagem. Os ratos foram sacrificados após 36 horas da última aplicação, tendo seus fêmures removidos e a medula óssea extraída. O material foi homogeneizado e centrifugado. O botão de células foi pipetado e transferido para as lâminas, que foram coradas com Giemsa. Foram contados 1.000 eritrócitos policromáticos por animal, observando a frequência de micronúcleos. RESULTADOS: Foi realizado o teste de Kruskal-Wallis, com nível de significância de 5%, que demostrou que o decanoato de nandrolona - três doses de 0,2 mg/kg e 0,6 mg/kg, oito doses de 7,5 mg/kg, L-carnitina - sete doses de 0,4 ml/250g e 1,5 ml/250g, decanoato de testosterona - 28 doses de 0,075 mg/kg, decanoato de nandrolona - oito doses de 7,5 mg/kg associado a L-carnitina 1 ml e decanoato de nandrolona - oito doses de 7,5 mg/kg associado à decanoato de testosterona - oito doses de 7,5 mg/kg apresentaram potencial mutagênico. CONCLUSÃO: Os tratamentos revelaram-se clastogênicos, não sendo indicado como recurso ergogênico.

INTRODUCTION: Anabolic androgenic steroids (AAS) are frequently used by people whose aim to increase muscle mass to obtain a better performance in sports or improve physical appearance. AAS are synthetic derivatives of testosterone, able to promote muscle fibers hypertrophy, increasing intracellular protein synthesis. L-carnitine is a food supplement used to increase energetic production by means of fat acids oxidation. Although there are several works about physiological properties of these drugs, there are few studies about their mutagenic potential. OBJECTIVES: This work evaluated the clastogenicity and genotoxicity of nandrolone decanoate, testosterone decanoate and L-carnitine, in different treatments through the micronucleus test in polychromatic erythrocytes of Wistar rats. METHODS: The animals were submitted to different concentrations and associations of AAS. The positive control received cyclophosphamide 50 mg/kg by intraperitoneal injection and negative control, one ml of saline solution by gavage. The rats were sacrificed after 36 hours of latest application, having the femurs removed and the bone marrow extracted. Material was homogenized and centrifuged. Button cell was pipetted and transferred to slides, which were stained by Giemsa. 1,000 polychromatic erythrocytes were counted per animal, noting the frequency of micronuclei. RESULTS: The Kruskal-Wallis test was performed, with a significance level of 5%, which demonstrated that nandrolone decanoate - three doses of 0,2 mg/kg and 0,6 mg/kg, eight doses of 7,5 mg/kg, L-carnitine - seven doses of 0,4 ml/250 g and 1,5 ml/250 g, testosterone decanoate - 28 doses of 0,075 mg/kg, nandrolone decanoate - eight doses of 7,5 mg/kg associated to L-carnitine and 1 mL and nandrolone decanoate - eight doses of 7,5 mg/kg associated to testosterone decanoate - eight doses of 7,5 mg/kg, showed mutagenic potential. CONCLUSION: The treatments proved to be clastogenic, not being indicated like ergogenic aid.

INTRODUCCIÓN: Los esteroides anabólicos androgénicos son usados por personas que quieren aumentar su masa muscular a fin de obtener un mejor desempeño en los deportes o para mejorar la apariencia física. Los EAA son derivados sintéticos de la testosterona, que pueden causar la hipertrofia de las fibras musculares, aumentando la síntesis proteica intracelular. La L-carnitina es un suplemento alimenticio utilizado para hacer crecer la producción energética por medio de la oxidación de ácidos grasos. Aunque hay trabajos que muestran las propiedades fisiológicas de esos medicamentos, hay pocos estudios sobre el potencial mutagénico de estos. OBJETIVOS: Este trabajo evaluó la clastogenicidad y la genotoxicidad del decanoato de nandrolona, decanoato de testosterona y de la L-carnitina, en diversos tratamientos, mediante la prueba del micronúcleo en eritrocitos policromáticos de ratones Wistar. MÉTODOS: Los animales fueron sometidos a diferentes concentraciones y asociaciones de EAA. El control positivo recibió ciclofosfamida 50 mg/kg a través de inyección intraperitoneal, y el control negativo 1 ml de suero fisiológico por gavaje. Los ratones fueron sacrificados después de 36 horas de la última aplicación, teniendo sus fémures removidos y la médula ósea extraída. El material fue homogeneizado y centrifugado. El botón de células fue tomado con pipetas y transferido para las láminas, a las cuales se les dio color con Giemsa. Fueron contados 1.000 eritrocitos policromáticos por animal, observándose la frecuencia de micronúcleos. RESULTADOS: Se realizó la prueba de Kruskal-Wallis, con nivel de significancia de 5%, la cual demostró que el decanoato de nandrolona - tres dosis de 0,2 mg/kg y 0,6 mg/kg, ocho dosis de 7,5 mg/kg, L-carnitina - siete dosis de 0,4 ml/250g y 1,5 ml/250g, decanoato de testosterona - 28 dosis de 0,075 mg/kg, decanoato de nandrolona - ocho dosis de 7,5 mg/kg asociado a L-carnitina 1 ml y decanoato de nandrolona - ocho dosis de 7,5 mg/kg vinculado al decanoato de testosterona - ocho dosis de 7,5 mg/kg, presentaron potencial mutagénico. CONCLUSIÓN: Los tratamientos se revelaron como siendo clastogénicos, no indicándose como recurso ergogénico.