Lung cancer is the leading cause of cancer-related morbidity and mortality worldwide. The initial treatment of lung cancer depends on the definition of the tumor type and its staging. The most common ...treatment is chemotherapy, and the first-line treatment is a combination of carboplatin and paclitaxel. Although this treatment has good efficacy, there is a high prevalence of adverse events, particularly hematological reactions. Studies on new biomarkers related to these adverse events, such as circulating microRNAs (miRNAs/miRs), are important for optimizing the quality of life of patients. miRNAs have high stability in several biological fluids and they have specific expressions in different tissues or pathologies. Thus, the present study aimed to assess the relationship between circulating miRNAs and adverse hematologic reactions caused by treatment with carboplatin + paclitaxel in patients with lung cancer. Blood was collected from patients before and 15 days after chemotherapy for hematological adverse reaction analysis, microarray and quantitative (q)PCR validation. Adverse reactions were classified according to the Common Terminology Criteria for Adverse Events v4.0. Microarray analysis was performed using plasma from six patients without anemia and six patients with anemia, and nine miRNAs were differentially expressed. miR-1273g-3p, miR-3613-5p and miR-455-3p, identified using microarray, were assessed using qPCR in 20 patients without anemia and 26 patients with anemia. Bioinformatic analyses of miR-455-3p were performed using miRWalk, the Database for Annotation, Visualization and Integrated Discovery and GeneMania software. Microarray analysis of patients with and without anemia revealed nine significant differentially- expressed plasma miRNAs among these patients. Of these, miR-1273g-3p, miR-3613-5p and miR-455-3p were chosen for further assessment. Only miR-455-3p demonstrated a significant reduction in expression (P=0.04) between the groups before chemotherapy with carboplatin + paclitaxel. Bioinformatics analysis of miR-455-3p revealed a relationship between this miRNA and the hematopoietic pathway, particularly with respect to the RUNX family transcription factor 1 (RUNX1) and TAL bHLH transcription factor 1, erythroid differentiation factor (TAL1) genes. The most prevalent adverse reactions in patients with lung cancer treated with carboplatin + paclitaxel were hematological, particularly anemia. This adverse reaction, caused by dysfunction of the hematopoietic system, may be explained by a possible association between the important genes in this system, RUNX1 and TAL1, and hsa-miR-455-3p. Key words: carboplatin, paclitaxel, hematological adverse reactions, microRNAs, lung cancer
Gefitinib is a selective inhibitor of the epidermal growth factor receptor that is used to treat advanced and metastatic non-small cell lung cancer (NSCLC). Dermatological adverse reactions are most ...commonly associated with gefitinib treatment. The cause of adverse reactions in individuals is multifactorial. Pharmacogenetics is an effective tool to detect such adverse reactions. This case report describes a female patient with NSCLC who was administered gefitinib at a dose of 250 mg/day. However, due to severe adverse dermatological reactions, the treatment was interrupted for 15 d and antibiotic therapy was administered to manage the skin rashes, maculopapular rashes, and hyperpigmentation. Treatment adherence was adequate, and no drug interactions were detected. A pharmacogenetic analysis revealed homozygosity in the ATP-binding cassette (ABC)-B1 rs1128503 (c.1236A>G), heterozygosity in ABCG2 rs2231142 (c.421G>T) and rs2622604 (c.-20+614T>C), and a non-functional variant of the cytochrome P450 family 3, subfamily A, member 5 (CYP3A5). The relationship between altered genetic variants and the presence of adverse reactions induced by gefitinib is still controversial. Overall, this case report highlights the importance of continuing to study pharmacogenetics as predictors of adverse drug reactions.
As SARS-CoV-2 continues to produce new variants, the demand for diagnostics and a better understanding of COVID-19 remain key topics in healthcare. Skin manifestations have been widely reported in ...cases of COVID-19, but the mechanisms and markers of these symptoms are poorly described. In this cross-sectional study, 101 patients (64 COVID-19 positive patients and 37 controls) were enrolled between April and June 2020, during the first wave of COVID-19, in São Paulo, Brazil. Enrolled patients had skin imprints sampled non-invasively using silica plates; plasma samples were also collected. Samples were used for untargeted lipidomics/metabolomics through high-resolution mass spectrometry. We identified 558 molecular ions, with lipids comprising most of them. We found 245 plasma ions that were significant for COVID-19 diagnosis, compared to 61 from the skin imprints. Plasma samples outperformed skin imprints in distinguishing patients with COVID-19 from controls, with F1-scores of 91.9% and 84.3%, respectively. Skin imprints were excellent for assessing disease severity, exhibiting an F1-score of 93.5% when discriminating between patient hospitalization and home care statuses. Specifically, oleamide and linoleamide were the most discriminative biomarkers for identifying hospitalized patients through skin imprinting, and palmitic amides and N-acylethanolamine 18:0 were also identified as significant biomarkers. These observations underscore the importance of primary fatty acid amides and N-acylethanolamines in immunomodulatory processes and metabolic disorders. These findings confirm the potential utility of skin imprinting as a valuable non-invasive sampling method for COVID-19 screening; a method that may also be applied in the evaluation of other medical conditions.
Key messages
Skin imprints complement plasma in disease metabolomics.
The annotated markers have a role in immunomodulation and metabolic diseases.
Skin imprints outperformed plasma samples at assessing disease severity.
Skin imprints have potential as non-invasive sampling strategy for COVID-19.
Coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Recent research has demonstrated how epigenetic mechanisms regulate the host–virus ...interactions in COVID-19. It has also shown that microRNAs (miRNAs) are one of the three fundamental mechanisms of the epigenetic regulation of gene expression and play an important role in viral infections. A pilot study published by our research group identified, through next-generation sequencing (NGS), that miR-4433b-5p, miR-320b, and miR-16–2-3p are differentially expressed between patients with COVID-19 and controls. Thus, the objectives of this study were to validate the expression of these miRNAs using quantitative real-time polymerase chain reaction (qRT-PCR) and to perform in silico analyses. Patients with COVID-19 (n = 90) and healthy volunteers (n = 40) were recruited. MiRNAs were extracted from plasma samples and validated using qRT-PCR. In addition, in silico analyses were performed using mirPath v.3 software. MiR-320b was the only miRNA upregulated in the case group com-pared to the control group. The in silico analyses indicated the role of miR-320b in the regulation of the KITLG gene and consequently in the inflammatory process. This study confirmed that miR-320b can distinguish patients with COVID-19 from control participants; however, further research is needed to determine whether this miRNA can be used as a target or a biomarker.
Dendritic cell (DC) vaccines have demonstrated good efficacy in preventing relapse and in increasing survival of patients affected by a variety of both solid and hematological tumors. Most protocols ...used to generate these cells involve the automated separation of peripheral blood monocytes from patients. This approach requires specialized equipment, which elevates the cost of this type of therapy, potentially limiting the widespread access to patients. Method: In this study, we compare the yield and quality of dendritic cells generated from monocytes and isolated by an automated method or by manual methods using gradient centrifugation.
The results demonstrate the equivalence of the 3 methods in relation to the yield and final quality of the product, however with considerable differences between the costs of these procedures. In addition, this study also demonstrates the feasibility of the antigenic pulse with autologous tumor cell lysates, constituting a source of antigens, not only easily obtained and manipulated, but also specific to the patient's tumor.
These findings may have important implications for emerging centers interested in using this medical approach and potentially increase the access of a greater number of patients to this therapeutic option.
COVID-19 is still placing a heavy health and financial burden worldwide. Impairment in patient screening and risk management plays a fundamental role on how governments and authorities are directing ...resources, planning reopening, as well as sanitary countermeasures, especially in regions where poverty is a major component in the equation. An efficient diagnostic method must be highly accurate, while having a cost-effective profile. We combined a machine learning-based algorithm with mass spectrometry to create an expeditious platform that discriminate COVID-19 in plasma samples within minutes, while also providing tools for risk assessment, to assist healthcare professionals in patient management and decision-making. A cross-sectional study enrolled 815 patients (442 COVID-19, 350 controls and 23 COVID-19 suspicious) from three Brazilian epicenters from April to July 2020. We were able to elect and identify 19 molecules related to the disease’s pathophysiology and several discriminating features to patient’s health-related outcomes. The method applied for COVID-19 diagnosis showed specificity >96% and sensitivity >83%, and specificity >80% and sensitivity >85% during risk assessment, both from blinded data. Our method introduced a new approach for COVID-19 screening, providing the indirect detection of infection through metabolites and contextualizing the findings with the disease’s pathophysiology. The pairwise analysis of biomarkers brought robustness to the model developed using machine learning algorithms, transforming this screening approach in a tool with great potential for real-world application.
Background
Coronavirus disease 2019 (COVID-19) is caused by a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). It is known that host microRNAs (miRNAs) can be ...modulated to favor viral infection or to protect the host. Herein, we report preliminary results of a study aiming at identifying differentially expressed plasmatic miRNAs in Brazilian patients with COVID-19.
Methods and results
miRNAs were extracted from the plasma of eight patients with COVID-19 (four patients with mild COVID-19 and four patients with severe/critical COVID-19) and four healthy controls. Patients and controls were matched for sex and age. miRNA expression levels were detected using high-throughput sequencing. Differential miRNA expression and enrichment analyses were further evaluated. A total of 18 miRNAs were differentially expressed between patients with COVID-19 and controls. miR-4433b-5p, miR-6780b-3p, miR-6883-3p, miR-320b, miR-7111-3p, miR-4755-3p, miR-320c, and miR-6511a-3p were the most important miRNAs significantly involved in the PI3K/AKT, Wnt/β-catenin, and STAT3 signaling pathways. Moreover, 42 miRNAs were differentially expressed between severe/critical and mild patients with COVID-19. miR-451a, miR-101-3p, miR-185-5p, miR-30d-5p, miR-25-3p, miR-342-3p, miR-30e-5p, miR-150-5p, miR-15b-5p, and miR-29c-3p were the most important miRNAs significantly involved in the Wnt/β-catenin, NF-κβ, and STAT3 signaling pathways.
Conclusions
If validated by quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in a larger number of participants, the miRNAs identified in this study might be used as possible biomarkers for the diagnosis and severity of COVID-19.
Lung cancer remains a major global cause of mortality. While 18F-FDG PET/CT is widely utilized for detection, staging, and monitoring, detecting increased glucose metabolism in tumor cells, it lacks ...theranostic potential. The prostate-specific membrane antigen (PSMA) tracer, initially linked to prostate cancer, is also recognized as a marker of neoangiogenesis, accumulating in various neoplasms and showing promising theranostic capabilities.
This study aims to compare the uptake patterns of 18F-FDG and 18F-PSMA in primary and metastatic lesions of non-small cell lung cancer.
Four male patients diagnosed with non-small cell lung cancer (including two adenocarcinomas, one squamous cell carcinoma, and one unspecified type), aged between 58 and 71 years, underwent PET/CT imaging. 18F-FDG PET/CT scans were performed 60 minutes after intravenous administration of 0.1 mCi/kg of 18F-FDG, while 18F-PSMA PET/CT scans were obtained 90 minutes after intravenous injection of 0.1 mCi/kg of 18F-PSMA. Imaging data were analyzed by two nuclear medicine physicians and one radiologist. The maximum standardized uptake value (SUVmax) of each lesion was measured for both radiotracers in the primary tumor, lymph nodes, and metastatic sites identified through visual analysis, considering values obtained above the cardiac blood pool measured in the left atrium.
A total of 100 lesions were detected, with 82 identified using 18F-FDG and 92 using 18F-PSMA. Eight lesions were exclusively detected by 18F-FDG, while 14 were only identified by 18F-PSMA. The median SUVmax of lesions in 18F-FDG and 18F-PSMA images was 5.3 (1.7 – 25.0) and 3.7 (0.7 - 12.4), respectively. Brain lesions were more readily identified on 18F-PSMA images, whereas liver lesions were more notable on 18F-FDG images due to the intense physiological uptake of 18F-FDG and 18F-PSMA in the brain and liver, respectively.
Both 18F-FDG-PET/CT and 18F-PSMA-PET/CT have the ability to identify most lesions of non-small cell lung cancer. Although 18F-PSMA images detected a greater number of lesions, the uptake intensity is generally higher with 18F-FDG. These findings suggest that the two radiopharmaceuticals may have complementary roles in lung cancer by independently detecting lesions with higher glycolytic activity or greater neoangiogenesis, or both simultaneously, which could contribute to a more personalized approach in managing these patients. The results also suggest a possible theranostic approach in selected patients with high uptake of PSMA. Further investigations involving a larger patient cohort are essential to validate these findings.
Orientadores: Lair Zambon, Fernando Ferreira Costa
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas
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Previous issue date: 2002
Resumo: Considerada uma doença rara no início do século XX, o carcinoma de pulmão hoje é a neoplasia visceral mais comum e a principal causa de morte por câncer. O estudo dos eventos moleculares envolvidos no câncer de pulmão é importante para o conhecimento do processo de carcinogênese, e para a determinação do espectro mutacional dos genes relacionados ao carcinoma brônquico. No futuro poderemos empregar as técnicas de biologia molecular no diagnóstico precoce e na correlação entre variáveis clínicas, tumorais e genéticas para definir os fatores de prognóstico e a abordagem terapêutica. Realizamos a pesquisa de mutações nos genes p53 e K-ras em pacientes com neoplasia de pulmão. Em relação ao gene p53, pesquisamos mutações nos exons 5 a 10 em 38 pacientes com carcinoma de pulmão não pequenas células, e nos exons 5 a 9 em nove pacientes com carcinoma de pequenas células. A freqüência de mutações foi, respectivamente, de 21 e 33%. A pesquisa de mutações no gene K-ras foi realizada em 8 pacientes com adenocarcinoma e em quatro com carcinoma de grandes células. Não encontramos mutações, provavelmente, devido ao tamanho da amostra. Nos pacientes com carcinoma não pequenas células a presença de mutação no gene p53 não interferiu na sobrevida (p=0,53), no estadiamento (p=0,67) e no tempo de sintomatologia (p=0,15). Comparamos as mutações encontradas no gene p53 com o banco de dados mantido pela Organização Mundial de Saúde (International Agency for Research on Cancer). Considerando que o banco de dados da IARC foi atualizado em março/2002 (16285 entradas), provavelmente somos o primeiro grupo a descrever a mutação no codon 237 (ATGàAAG) em carcinoma brônquico, independente do tipo histológico; da mutação no codon 337 (CGCàCAC) em carcinoma de grandes células e da mutação silenciosa no codon 295 (CCT>CCC) (IARC, 2002)
Abstract: Considered a rare illness in the beginning of 20th century, the lung cancer today is the more common visceral neoplasia and the main cause of death by cancer. The study of the molecular events involved in lung cancer is important for the knowledge of the process of carcinogenesis and for the determination of the mutational spectrum of the genes related to lung cancer. In the future we will be able to use the techniques of molecular biology in the early diagnosis, and the correlation between clinical, tumoral and genetic variables will be useful for us to define the factors of prognostic and therapeutic approach. We carried out the study of mutations in the genes p53 and K-ras in patients with lung cancer. In relation to the p53 gene, we have looked for mutations in exons 5 to 10 in 38 patients with non-small cell lung cancer, and in exons 5 to 9 in nine patients with small cell lung cancer. The frequency of mutations was, respectively, 21 and 33%. The analysis of mutations in the K-ras gene was carried out in 8 patients with adenocarcinoma and four with large cell carcinoma. We didn¿t find mutations, probably, due to the size of the sample. In patients with non-small cell lung cancer, the presence of mutation in the p53 gene did not correlate with survival (p=0,53), staging (p=0,67) and time of symptoms (p=0,15). We compared the mutations found in the p53 gene with the database kept by the World Health Organization (International Agency for Research on Cancer). Taking into account that the IARC data bank was updated in March/2002 (16,285 entries), we are probably the first group to describe the transversion on codon 237 (ATGàAAG) in bronchial cancer, independent of the histological type and the mutation on codon 337 in large cell cancer (IARC, 2002)
Mestrado
Clinica Medica
Mestre em Clinica Medica
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