Identification of potential urinary protein biomarkers in colorectal cancer: a pilot study using a proteomic approach

Colorectal cancer (CRC) is among the most diagnosed malignancies worldwide, and it is also the second leading cause of cancer-related deaths. Despite recent progress in screening programs, noninvasive accurate biomarkers are still needed in the CRC field. In this study, we evaluated and compared the urinary proteomic profiles of patients with colorectal adenocarcinoma and patients without cancer, aiming to identify potential biomarker proteins. Urine samples were collected from 9 patients with CRC and 9 patients with normal colonoscopy results. Mass spectrometry (label-free LC—MS/MS) was used to characterize the proteomic profile of the groups. Ten proteins that were differentially regulated were identified between patients in the experimental group and in the control group, with statistical significance with a p value ≤ 0.05. The only protein that presented upregulation in the CRC group was beta-2-microglobulin (B2M). Subsequent studies are needed to evaluate patients through different analysis approaches to independently verify and validate these biomarker candidates in a larger cohort sample. (AU)

Acute Cellular Rejection in Heart Transplant Patients: Insights of Global Longitudinal Strain, Myocardial Work, and an Exclusive Group of Chagas Disease.

Background: Echocardiographic markers associated with asymptomatic acute cellular rejection (ACR) in patients with orthotopic heart transplant (HT) are still under investigation. The aim of our study was to determine clinical and myocardial strain imaging (MSI) variables evaluated by echocardiography associated with ACR in the first year of HT. A separate analysis was performed to compare variables during the first 6 months of HT, when ACR has a prevalence in 60% of patients. Another analysis evaluated an exclusive population with Chagas disease as the cause of HT. Methods: We prospectively studied 67 patients with less than 1 year of HT, 36 patients without ACR (41% men, age 49 ± 12 years, 52% Chagas disease as the cause of heart failure), and 31 patients with ACR (59% men, age 55 ± 8 years, 74% Chagas disease as the cause of heart failure). Conventional echocardiographic measurements and MSI by global longitudinal strain (GLS) from the left ventricle (LV) and right ventricle free wall (RV-FWLS) and myocardial work (MW) from the left ventricle were obtained by experienced echocardiologists. Clinical variables, such as the presence of diabetes, hypertension, and immunosuppressant drugs, were compared between groups. Results: HT patients with ACR were older and used more cyclosporine for immunosuppression. The positive ACR group had an increased relative wall thickness and LV mass index and similar LVGLS and RV-FWLS compared to the negative ACR group. Nevertheless, MW analysis observed increased global work efficiency (GWE) in positive ACR. Multivariate analysis identified older age, cyclosporine use, LV mass index, and GWE as independent predictors for detecting rejection. A separate analysis was performed for patients with less than 6 months of HT. Similar MSI was observed in both groups, with a trend for increased GWE in patients with ACR and significantly increased LV mass index in the ACR group. An exclusive group of Chagas patients as the primary cause of HT was analyzed, and similar MSI results for LVGLS, RV-FWLS, and MW were observed for both ACR and the no rejection groups. Additionally, the survival rates at 2 years were similar between the Chagas disease groups. Conclusion: LVGLS and RV-FWLS were similar between patients with or without ACR in the first year after HT. Conversely, GWE, a derivative of LVGLS, and LV mass index were increased in positive ACR and could be markers for rejection. Increased LV mass index was also found in a subgroup analysis of patients less than 6 months after HT; however, MSI was similar regardless of ACR. For chagasic patients, rejection in the first year did not increase mortality at the 2-year follow-up, and MSI parameters were similar between patients with or without ACR. In a multivariate analysis to predict ACR, the independent parameters in this study were older age, cyclosporine use, LV mass index, and GWE.

The impact of mass spectrometry application to screen new proteomics biomarkers in Ophthalmology.

INTRODUCTION: In the search for molecular markers that aid in the early diagnosis and treatment of various human diseases, many studies have focused on changes in genes, their transcripts and protein products. Recent advances in proteomic methodologies, such as mass spectrometry (MS), generate new opportunities to obtain relevant information on normal and abnormal processes that occur in many important cell pathways. The human eye is a highly specialized and compartmentalized organ, and the interpretation of molecular biomarkers helps to evaluate its cellular structure, providing a broader molecular understanding that corroborates in the pathophysiology of ophthalmological diseases, with marked improvements in their diagnosis, prognosis and treatment. This review summarizes the most important protein biomarkers in Ophthalmology screened by MS tools. CONCLUSION: The use of translational medicine techniques (as MS), integrating basic and clinical research, still transforms scientific findings, from laboratory researches to clinical applications, from the bedside into the community.

Novel Possible Protein Targets in Neovascular Age-Related Macular Degeneration: A Pilot Study Experiment.

Age-related macular degeneration (AMD) is among the world's leading causes of blindness. In its neovascular form (nAMD), around 25% of patients present further anatomical and visual deterioration due to persistence of neovascular activity, despite gold-standard treatment protocols using intravitreal anti-VEGF medications. Thus, to comprehend, the molecular pathways that drive choroidal neoangiogenesis, associated with the vascular endothelial growth factor (VEGF), are important steps to elucidate the mechanistic events underneath the disease development. This is a pilot study, a prospective, translational experiment, in a real-life context aiming to evaluate the protein profiles of the aqueous humor of 15 patients divided into three groups: group 1, composed of patients with nAMD, who demonstrated a good response to anti-VEGF intravitreal injections during follow-up (good responsive); group 2, composed of patients with anti-VEGF-resistant nAMD, who demonstrated choroidal neovascularization activity during follow-up (poor/non-responsive); and group 3, composed of control patients without systemic diseases or signs of retinopathy. For proteomic characterization of the groups, mass spectrometry (label-free LC-MS/MS) was used. A total of 2,336 proteins were identified, of which 185 were distinctly regulated and allowed the differentiation of the clinical conditions analyzed. Among those, 39 proteins, including some novel ones, were analyzed as potential disease effectors through their pathophysiological implications in lipid metabolism, oxidative stress, complement system, inflammatory pathways, and angiogenesis. So, this study suggests the participation of other promising biomarkers in neovascular AMD, in addition to the known VEGF.

Biomarkers in Colorectal Cancer: The Role of Translational Proteomics Research.

Colorectal cancer is one of the most common cancers in the world, and it is one of the leading causes of cancer-related death. Despite recent progress in the development of screening programs and in the management of patients with colorectal cancer, there are still many gaps to fill, ranging from the prevention and early diagnosis to the determination of prognosis factors and treatment of metastatic disease, to establish a personalized approach. The genetic profile approach has been increasingly used in the decision-making process, especially in the choice of targeted therapies and in the prediction of drug response, but there are still few validated biomarkers of colorectal cancer for clinical practice. The discovery of non-invasive, sensitive, and specific biomarkers is an urgent need, and translational proteomics play a key role in this process, as they enable better comprehension of colorectal carcinogenesis, identification of potential markers, and subsequent validation. This review provides an overview of recent advances in the search for colorectal cancer biomarkers through proteomics studies according to biomarker function and clinical application.

Absence of the Caspases 1/11 Modulates Liver Global Lipid Profile and Gut Microbiota in High-Fat-Diet-Induced Obese Mice.

Obesity is a chronic disease with rising worldwide prevalence and largely associated with several other comorbidities, such as cancer, non-alcoholic fatty liver disease (NAFLD), and metabolic syndrome. Hepatic steatosis, a hallmark of NAFLD, is strongly correlated with obesity and has been correlated with changes in the gut microbiota, which can promote its development through the production of short-chain fatty acids (SCFAs) that regulate insulin resistance, bile acid, choline metabolism, and inflammation. Recent studies have suggested a controversial role for the inflammasome/caspase-1 in the development of obesity and non-alcoholic steatohepatitis (NASH). Here, we evaluated the role of inflammasome NLRP3 and caspases 1/11 in the establishment of obesity and hepatic steatosis in diet-induced obese mice, correlating them with the global lipid profile of the liver and gut microbiota diversity. After feeding wild-type, caspases 1/11, and NLRP3 knockout mice with a standard fat diet (SFD) or a high-fat diet (HFD), we found that the caspases 1/11 knockout mice, but not NLRP3 knockout mice, were more susceptible to HFD-induced obesity, and developed enhanced hepatic steatosis even under SFD conditions. Lipidomics analysis of the liver, assessed by MALDI-MS analysis, revealed that the HFD triggered a significant change in global lipid profile in the liver of WT mice compared to those fed an SFD, and this profile was modified by the lack of caspases 1/11 and NLRP3. The absence of caspases 1/11 was also correlated with an increased presence of triacylglycerol in the liver. Gut microbial diversity analysis, using 16S rRNA gene sequencing, showed that there was also an increase of Proteobacteria and a higher Firmicutes/Bacteroidetes ratio in the gut of caspases 1/11 knockout mice fed an HFD. Overall, mice without caspases 1/11 harbored gut bacterial phyla involved with weight gain, obesity, and hepatic steatosis. Taken together, our data suggest an important role for caspases 1/11 in the lipid composition of the liver and in the modulation of the gut microbial community composition. Our results further suggest that HFD-induced obesity and the absence of caspases 1/11 may regulate both lipid metabolism and gut microbial diversity, and therefore may be associated with NAFLD and obesity.