Rab4A-directed endosome traffic shapes pro-inflammatory mitochondrial metabolism in T cells via mitophagy, CD98 expression, and kynurenine-sensitive mTOR activation.

Activation of the mechanistic target of rapamycin (mTOR) is a key metabolic checkpoint of pro-inflammatory T-cell development that contributes to the pathogenesis of autoimmune diseases, such as systemic lupus erythematosus (SLE), however, the underlying mechanisms remain poorly understood. Here, we identify a functional role for Rab4A-directed endosome traffic in CD98 receptor recycling, mTOR activation, and accumulation of mitochondria that connect metabolic pathways with immune cell lineage development and lupus pathogenesis. Based on integrated analyses of gene expression, receptor traffic, and stable isotope tracing of metabolic pathways, constitutively active Rab4AQ72L exerts cell type-specific control over metabolic networks, dominantly impacting CD98-dependent kynurenine production, mTOR activation, mitochondrial electron transport and flux through the tricarboxylic acid cycle and thus expands CD4+ and CD3+CD4-CD8- double-negative T cells over CD8+ T cells, enhancing B cell activation, plasma cell development, antinuclear and antiphospholipid autoantibody production, and glomerulonephritis in lupus-prone mice. Rab4A deletion in T cells and pharmacological mTOR blockade restrain CD98 expression, mitochondrial metabolism and lineage skewing and attenuate glomerulonephritis. This study identifies Rab4A-directed endosome traffic as a multilevel regulator of T cell lineage specification during lupus pathogenesis.

Regulatory Effect of let-7f Transfection in Non-Small Cell Lung Cancer on its Candidate Target Genes.

Background: Let-7f has essential impacts on biological processes; however, its biological and molecular functions in lung cancer pathogenesis have yet been remained unclear. We aimed to investigate the expression level of let-7f and its candidate target genes both in lung cancer tissues and A549 cell line. Methods: Bioinformatics databases were first used to select candidate target genes of let-7f. Then the relative gene and protein expressions of let-7f and its target genes, including HMGA2, ARID3B, SMARCAD1, and FZD3, were measured in lung tissues of NSCLC patients and A549 cell line using qRT-PCR and Western blotting. The electroporation method was used to transfect A549 cells with let-7f mimic and microRNA inhibitor. The impact of let-7f transfection on the viability of A549 cells was assessed using MTT assay. The expression data of studied genes were analyzed statistically. Results: Results indicated significant downregulated expression level of let-7f-5p (p = 0.0013) and upregulated level of the HMGA2 and FZD3 in NSCLC cases (p < 0.05). In A549 cells, after transfection with let-7f mimic, the expression of both mRNA and protein levels of HMGA2, ARID3B, SMARCAD1, and FZD3 decreased. Also, the overexpression of let-7f significantly inhibited the A549 cell proliferation and viability (p = 0.017). Conclusion: Our findings exhibited the high value of let-7f and HMGA2 as biomarkers for NSCLC. The let-7f, as a major tumor suppressor regulatory factor via direct targeting genes (e.g. HMGA2), inhibits lung cancer cell viability and proliferation and could serve as a marker for the early diagnostic of NSCLC.

Evaluation of hsa-let-7d-5p, hsa-let-7g-5p and hsa-miR-15b-5p plasma levels in patients with Alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD) is a progressive neurodegenerative disorder MicroRNAs (miRNAs) may be promising diagnostic biomarkers for AD. Previous evidence shows that miR-15b-5p, hsa-let7g-5p and hsa-let7d-5p might confer potential blood biomarkers for timely diagnosis of AD. Therefore, in this replication study, we aimed to investigate the serum transcript level of these miRNAs to assess for their potential as diagnostic or prognostic biomarker in AD patients. METHODS: Blood samples were obtained from 50 AD patients and 50 age- and sex-matched healthy individuals. Then, total RNA was extracted from serum samples, cDNA was synthesized, and the expression level of miRNAs was measured by the real-time PCR method. RESULTS: The expression level of hsa-let7d-5p (fold change = 2.14, P = 0.007) and hsa-let7g-5p (fold change = 1.94; P = 0.013) was significantly increased in the AD patients compared to control individuals. However, the difference in the transcription of miR-15b-5p between the two groups was not statistically significant (fold change = 1.08; P = 0.76). The AROC for transcript levels of hsa-let-7d-5p was 0.68 (P = 0.014; 95% CI, 0.39-0.88) and it was 0.64 for hsa-let-7g-5p (P = 0.028; 95% CI, 0.27-0.89). The cut-off value for let-7d-5p had 0.82 sensitivity and 0.34 specificity. Moreover, the cut-off value for hsa-let-7g-5p indicated a 0.79 sensitivity and 0.28 specificity. CONCLUSION: Our findings suggest the potential of measuring the transcript levels of hsa-let7d-5p and hsa-let7g-5p miRNAs as a diagnostic biomarker for AD.

Prognostic and Diagnostic Values of miR-506 and SPON 1 in Colorectal Cancer with Clinicopathological Considerations.

PURPOSE: One of the most common cancers in the world is colorectal cancer, which has increased significantly in recent decades. In the carcinogenicity process in the colon, there are genes involved in various cellular processes, such as cell cycle, apoptosis, and cell migration. According to studies carried out, both miR-506 and SPON 1 genes are involved in the process which initiates and promotes cancer. In this study, alterations in the expression of target genes from the viewpoint of carcinogenicity were studied and evaluated. METHODS: Fifty tumor tissues and normal marginal tissue were collected from patients who were undergoing colorectal cancer surgery. After the extraction of RNA, the real-time PCR method was performed to evaluate the gene expression. Also, alterations of gene expression in response to defined amounts of chemotherapeutic drugs (IC50) were evaluated. P < 0.05 was considered statistically significant. RESULTS: The relative expression level of miR-506 in tumor tissues was significantly decreased in comparison with healthy marginal tissues (P = 0.044). On the other hand, the SPON1 gene expression level was decreased too in tumor tissues in comparison with healthy marginal tissues (P = 0.019). There was also a significant relationship between the expression of target genes and clinicopathological involvement. However, there was no significant alteration in these genes along with the chemotherapeutic drug. CONCLUSION: These findings suggest that the relative expression of miR-506 and SPON 1 gene can be considered as a diagnostic or predictive biomarker for colorectal cancer. However, further studies on protein levels should be conducted.

Aberrant Expression of miR-103, miR-184, miR-378, miR-497 and miR-506 in Tumor Tissue from Patients with Oral Squamous Cell Carcinoma Regulates the Clinical Picture of the Patients.

BACKGROUND: This study aimed to evaluate the expression patterns of miR-103, miR-184, miR-378, miR497 and in squamous cell carcinoma (SCC) of the tongue and to be compared with normal peripheral tissues. METHODS: Tumor and marginal tissues were obtained from 50 patients with OSCC. After RNA extraction, expression level of miR-103, miR-184, miR-378, miR497, and miR506 was estimated using SYBR green master mix and real-time quantitative PCR. RESULTS: It was observed that, there was no detectable difference in expression level of miR-103 between tumoral and marginal tissues. However, expression level of miR-184, and miR-378 showed significant increase in tumor tissue samples compared to marginal tissue samples. MiR-497 and miR-506 demonstrated considerable decrease in tumoral cells in comparison with peripheral tissues. Moreover, the expression level of miRNAs was associated with clinicopathological features of the patients. CONCLUSIONS: Our data indicated that miR-184, miR-378, miR-497, and miR-506 can be used as a potential diagnostic and prognostic biomarker in OSCC. Nevertheless, more studies are needed to confirm this claim.
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Dendritic cell therapy in cancer treatment; the state-of-the-art.

Dendritic cells (DCs) are considered as professional antigen presenting cells (APCs), containing a variety of subsets, that can be resident in organs or migrating among the lymphoid and non-lymphoid organs. In a normal steady condition, DCs concomitantly process and present antigens on major histocompatibility complex (MHC) class I and II. However, they are further activated after exposing to antigens. Recently, several approaches have been exerted to improve antigen presentation potency, to elicit powerful immune responses against tumor cells. In DC-based cancer immunotherapy, DC is obtained from patient and modulated ex vivo in order to entice the immune system toward tumor elimination. Several approaches have been on the evaluation for long-term anti-tumor immune responses by DCs. On the other side, combination of DC vaccines with other cancer therapies, like chemotherapy and monoclonal antibodies could confer efficient cancer therapeutics. In this review article, we first go through the immunobiology of DC, and development of DC vaccines. Then, we concentrate on the DC immunotherapy by highlighting combinational approaches to enhance the efficacy of cancer treatment strategies.

The lipoprotein lipase S447X and cholesteryl ester transfer protein rs5882 polymorphisms and their relationship with lipid profile in human serum of obese individuals.

BACKGROUND: Obesity is often associated with an alter lipid profile, e.g., raised serum triglycerides (TG) and low high-density lipoprotein (HDL) cholesterol levels, both important risk factor for cardiovascular-diseases. The aim of current study was to explore the association of a polymorphism of the lipoprotein lipase (LPL) rs328 and cholesteryl-ester-transfer-protein (CETP) rs5882 genes in relation to lipid profile in subjects with/without obesity. SUBJECTS/METHODS: Genotyping was carried out in 271 individuals, (151 obese subjects and 120 non-obese). Univariate/multivariate analyses were conducted to evaluate the association of these genetic-polymorphisms with obesity and lipid components. RESULTS: Obese subjects had a significantly (P<0.05) higher level of triglyceride (TG), blood pressure, waist-circumference and fasting-blood-glucose, and lower level of HDL-C. LPL and CETP polymorphisms were not associated with obesity in our population. However, the LPL rs328-GG-GC genotype was significantly related to a higher concentration of TG, compared to the CC wild-type; and a higher HDL-C level in the obesity-group with respect to the control group. Moreover, obese-subjects carrying the G allele of CETP had a significantly lower level of HDL-C (P<0.05) compared to those with C allele. CONCLUSION: We demonstrate a significant association of LPL and CETP polymorphisms with serum triglycerides and HDL-cholesterol.