TSGA10, as a cancer/testis gene: Review article

Cancer/Testis antigens (CTAs) as a group of tumor antigens are the novel subjects for developing cancer vaccine and immunotherapy approaches. They aberrantly express in tumors with highest normal expression in testis, and limited or no expression in normal tissues. There are important similarities between the processes of germ-cell and cancer cell development Spermatogenesis begins at puberty when expression of novel cell-surface antigens occurs when the immune system has been refined the ability to distinguish self from non-self. Whereas macrophage and lymphocytes are commonly found within interstitial spaces of the testis, these antigen-presenting cells are rarely seen within the seminiferous tubules. These observations have led to the concept of the immune privileged site for testis. Localized normal expression of the CT genes in testis that makes them immunogenic for immune system, in one side, and their abnormal expression in different kinds of cancer cells, in the other side, has make them as promising target for developing cancer vaccines and new cancer therapeutics approaches. In malignancies, gene regulation is disrupted which results aberrant expression of CT antigen in a proportion of tumors of various types. For some CTAs, data support their fundamental role in tumorigenesis. Several authors believe it is not clear whether they have an essential role in tumorigenesis or they are by-products of chromatin variations in cancer. There is a growing list of CTAs within them advanced clinical trials are running by using some of them in cancers like lung cancer, malignant melanoma and neuroblastoma. In this review we discuss the gene TSGA10 as an example of CT genes. TSGA10 expresses in its highest levels in elongating spermatids and localized in the fibrous sheath of mature sperm. This gene is proposed as a serological biomarker in cutaneous lymphoma. Its abnormal expression has been reported in different cancers such as acute lymphoblastic leukemia, breast, brain, gastrointestinal and a range of other cancers either in mRNA or protein levels. It has an important role in angiogenesis in cancer tumors because of its effects in the gene hypoxia-inducible factor (HIF1). Absence or lack of TSGA10 expression has been reported in ascosporic infertile men

Effects of Indigenous Spore-Forming Probiotic as Feed Supplement on Performance and Safety in Broilers

ΔΕΝ ΔΙΑΤΙΘΕΤΑΙ ΠΕΡΙΛΗΨΗProbiotics colonize the intestine of animals and birds and provide useful effects on their performance and immune status. This study describes a high throughput screening and characterization of spore-forming bacteria from Iranian poultry farms with the aim to identify potential probiotic native Bacillus spp. and determine its effects on growth performance, hemato-biochemical parameters, immunity, intestinal microflora, morphology and MUC2 gene expression of broiler chickens. A total of 300 one-day-old female Ross 308 broilers (42.6 ± 0.6 g) were used in a 6-wk study. Broilers were randomly allotted to 1 of 3 dietary treatments consisting of 4 replicate cages with 25 broilers each: 1- Control (Corn-soy-based diet: C), 2- C + 200 g/ton of the GalliPro® (Bacillus subtilis DSM 17299, 4×109 CFU/g, as positive control group: PC), 3- C + 200 g/ton of the native probiotic (B. tequilensis K03, 4×109 CFU/g: NP) identified in this study. During the experiment parameters were measured weekly. The results revealed that birds of the NP and PC groups exhibited improved feed conversion ratio (FCR) and increased body weight (BW), carcass and breast meat yield compared with the birds of the C group (P<0.05). Also, lymphocytes level, antibody titers against Newcastle diseases virus (NDV) and infectious bronchitis virus (IBV) of vaccinated birds were increased, while serum triglycerides, total cholesterol levels and abdominal fat of birds fed NP and PC were decreased compared to birds of the C group (P<0.05). The villus height, the relative expression of MUC2 gene and Bacillus spp. populations were increased, while E. coli was significantly decreased in the ileum content of treated groups (P<0.05). These results indicate that the identified native B.tequilensis K03 strain can improve immunity and broiler performance by modifying intestinal microflora and morphology. Studied native probiotic Bacillus tequilensis K03 has useful effects on health status and it can be used as poultry feed supplement

Polymorphisms of plasminogen activator inhibitor-1, angiotensin converting enzyme and coagulation factor XIII genes in patients with recurrent spontaneous abortion

We investigated polymorphisms of plasminogen activator inhibitor-1 (PAI-1), angiotensin converting enzyme (ACE ) and coagulation factor XIII (FXIII) genes and their association with recurrent spontaneous abortion (RSA) in Iranian patients and normal healthy controls. Ten (18.5%) patients were homozygote (4G/4G) for PAI-1 polymorphism, in contrast with two (2%) controls (p = 0.001). Patients with homozygote 4G mutation were significantly more prone to RSA in contrast to others (odds ratio: 11.0, 95% CI: 2.3-52.4). Nineteen (30.2%) patients and 25 (26.6%) controls were homozygote (DD) for ACE polymorphism. We observed only two patients and one control with homozygosity (34leu) for FXIII polymorphism. 4G/4G polymorphism for PAI-1 gene could be a thrombophilic mutation leading to abortion in Iranian population

Differential gene-expression of metallothionein 1M and 1G in response to zinc in sertoli TM4 cells

Background: Zinc (Zn) as an important trace element is essential for testicular development and spermatogenesis. Molecular mechanism of Zn action in the reproductive system may be related to metal binding low-molecular weight proteins, metallothioneins (MT). Our objective was to determine the effect of Zn on two important isoforms of MT, MT1M and MT1G genes expression on testicular sertoli cells. Methods: Cultured sertoli TM4 cells were exposed to different concentrations of Zn at different time points. Cellular uptake of Zn was tested using flame atomic absorption spectrometry. The cellular viability and gene expression were assessed by MTT and real-time PCR methods, respectively. Results: The treated cells resulted in higher Zn concentration and cellular viability. The expression of MT1M and MT1G genes in the treated cells were greater than those of the untreated cells (P<0.05). In the high dosage treated group (100 and 500 μM), Zn concentration and expression of MT1M and MT1G genes increased three h after treatment; MT1G gene expression increased more at sixth h. At 18th h of treatment, the expression of both genes especially MT1G, increased dramatically while Zn concentration decreased. Conclusion: Since the increase of MT1G mRNA was coincident with cellular Zn level, it seems that MT1G has a more prominent role than MT1M in the homeostasis of Zn. In addition, Zn at dosage of 50 μM (pharmacologic concentration) may protect cells by increasing the expression of MT genes at longer periods

Upregulation of circulating cancer stem cell marker, DCLK1 but not Lgr5, in chemoradiotherapy-treated colorectal cancer patients

Cancer stem cell (CSC) markers have attracted considerable attention in tumor diagnostic, prognostic, and therapeutic implications. Detection of cancer stem cells in circulating blood using cancer stem cell markers has received remarkable attention recently. In this study, we aimed to investigate the messenger RNA (mRNA) expression level of Lgr5 and DCLK1 as most proposed colorectal CSC markers in blood circulation also determine the subsequent association to patients� clinical and pathological findings. Peripheral blood mononuclear cells (PBMCs) of 58 patients with colorectal cancer at stage I�IV with 33 out of 58 patients undergoing preoperative chemoradiotherapy (CRT), as well as 58 healthy controls have been isolated and the extracted RNAs were analyzed using real-time PCR. The mRNA expression pattern of CSC markers of patients and controls was compared using ��Ct method. The expression level of Lgr5 was significantly higher in colorectal cancer (CRC) patients comparing to healthy group (4.8-fold change, p < 0.001). Also there was a significant increase in expression level of Lgr5 in patients at stages III and IV comparing to stages I and II (p = 0.031) and higher grades (p = 0.039) of CRC. The expression of DCLK1 was also elevated in patients significantly (2.7-fold change, p < 0.001) and the related expression was increased by increasing disease stage (p = 0.025). Combination of DCLK1 and Lgr5 markers was analyzed by logistic regression and proved to be a slightly better marker compared to each marker alone. Interestingly the DCLK1 expression level was significantly higher in patients undergoing preoperative CRT (p = 0.041); however, no association to neoadjuvant CRT was observed for Lgr5. Considering the over-expression of DCLK1 and Lgr5 in circulating blood of CRC patients comparing to controls, our results might emphasize on the presence of CSCs in blood of these patients which might be attributed to their clinical and pathological characteristics and may lead to apply in future clinical implications. Moreover, the higher expression level of DCLK1 in patients undergoing CRT can propose it as a more relevant candidate among CSC markers comparing to Lgr5 for CRC patients. © 2015, International Society of Oncology and BioMarkers (ISOBM)

Genotype-phenotype correlation and description of two novel mutations in Iranian patients with glycogen storage disease 1b (GSD1b)

Background: Glycogen storage disease (GSD) is a rare inborn error of the synthesis or degradation of glycogen metabolism. GSD1, the most common type of GSD, is categorized into GSD1a and GSD1b which caused by the deficiency of glucose-6-phosphatase (G6PC) and glucose-6-phosphate transporter (SLC37A4), respectively. The high rates of consanguineous marriages in Iran provide a desirable context to facilitate finding the homozygous pathogenic mutations. This study designates to evaluate the clinical and genetic characteristics of patients with GSD1b to assess the possible genotype-phenotype correlation. Results: Autozygosity mapping was performed on nineteen GSD suspected families to suggest the causative loci. The mapping was done using two panels of short tandem repeat (STR) markers linked to the corresponding genes. The patients with autozygous haplotype block for the markers flanking the genes were selected for direct sequencing. Six patients showed autozygosity in the candidate markers for SLC37A4. Three causative variants were detected. The recurrent mutation of c.10421043delCT (p.Leu348Valfs*53) and a novel missense mutation of c.365G > A (p.G122E) in the homozygous state were identified in the SLC37A4. In silico analysis was performed to predict the pathogenicity of the variants. A novel whole SLC37A4 gene deletion using long-range PCR and sequencing was confirmed as well. Severe and moderate neutropenia was observed in patients with frameshift and missense variants, respectively. The sibling with the whole gene deletion has shown both severe neutropenia and leukopenia. Conclusions: The results showed that the hematological findings may have an appropriate correlation with the genotype findings. However, for a definite genotype-phenotype correlation, specifically for the clinical and biochemical phenotype, further studies with larger sample sizes are needed. © 2020 The Author(s)

Whole-Exome Sequencing Uncovers Novel Causative Variants and Additional Findings in Three Patients Affected by Glycogen Storage Disease Type VI and Fanconi�Bickel Syndrome

Glycogen storage diseases (GSDs) are the heterogeneous group of disorders caused by mutations in at least 30 different genes. Different types of GSDs, especially liver GSDs, take overlapping symptoms and can be clinically indistinguishable. This survey evaluated the use of whole-exome sequencing (WES) for the genetic analysis of the liver GSD-suspected patients in three unrelated families. An in-house filtering pipeline was used to assess rare pathogenic variants in GSD-associated genes, autosomal recessive/mendelian disorder genes (carrier status for genetic counseling subjects), and the ACMG�s list of 59 actionable genes. For the interpretation of the causative variants and the incidental/secondary findings, ACMG guidelines were applied. Additionally, we have explored PharmGKB class IA/IB pharmacogenetic variants. The segregation analysis was performed using Sanger sequencing for the novel causative variants. Bioinformatics analysis of the exome data in three individuals revealed three novel homozygous causative variants in the GSD-associated genes. The first variant, c.298307delATGATCAACC in PYGL gene has related to HERS disease (GSD VI). Both variants of c.1043dupT and c.613-1G > C in SLC2A2 gene have been associated with Fanconi-Bickel syndrome (GSDXI). Eight pathogenic/likely pathogenic medical actionable findings in Mendelian disease genes and 10 pharmacogenetic variants with underlying drug response phenotypes have been identified. No known/expected pathogenic variants were detected in the ACMG�s list of 59 actionable genes. The logical filtering steps can help in finding other medical actionable secondary/incidental findings as well as effectively identifying the causative variants in heterogeneous conditions such as GSDs. Three novel variants related to GSD genes recognized in liver GSD-suspected patients with early infantile and childhood-age onset. © Copyright © 2021 Eghbali, Fatemi, Salehpour, Abiri, Saei, Talebi, Olyaei, Yassaee and Modarressi

In vitro effects of glutathione on Transforming Growth Factor beta and epidermal Growth Factor Receptor genes expression in the protoscoleces and strobilated worms of Echinococcus granulosus

Cystic Echinococcosis (CE) caused by the small taeniid cestode Echinococcus granulosus, is a globally distributed zoonosis. Administration of some chemicals or natural compounds could lead to significant effects on the expression of some developmentally important genes including Transforming Growth Factor beta (TGF-β) and Epidermal Growth Factor Receptor (EGFR) in other parasitic organisms. The main purpose of this study was to describe the effect of glutathione (GSH) on the expression of TGF-β and EGFR genes in different developmental stages of E. granulosus. Protoscoleces of hydatid cysts collected from naturally infected sheep liver were cultured in diphasic CMRL1066 medium. Glutathione Mono-ethyl Ester (GME) at 250 μg/ml concentration was applied on the invaginated protoscoleces (PSCi), evaginated protoscoleces (PSCe) and strobilated worms (SW3) in vitro. TGF-β and EGFR genes expression were evaluated by using Real Time qPCR analysis compared to the controls. In response to GME treatment TGF-β expression was affected, however no significant effect was observed in EGFR expression. The results indicate a significant difference of TGF-β expression in the intact protoscoleces and the strobilated worms comparing to the controls. In intact invaginated protoscoleces TGF-β expression was significantly increased (p < 0.01) while in the strobilated worms a significant decrease was observed comparing to no-treatment controls (p < 0.001). None of the three developmental stages of E. granulosus demonstrated significant changes in EGFR expression. The results indicated that administration of GSH modified TGF-β expression in the protoscoleces and strobilar stages of E. granulosus. To improve our understanding of the physiology and biochemistry of the parasite more in depth in vitro and in vivo studies on the morphological and molecular effects of glutathione on the parasite is recommended. Further investigation on the gene profiles in other stages including microcysts and germinal layer cells is also suggested. This paves the way for the effective treatment and control of cystic echinococcosis. © 2020 Elsevier Inc

Expression of the testis-specific gene, TSGA10, in Iranian patients with acute lymphoblastic leukemia (ALL)

Testis-specific gene antigen (TSGA10) is expressed in fetus, testis and frequently in human solid cancers and acute leukemias, making it a candidate for immunotherapy and for detection of minimal residual disease (MRD). This gene is considered as a member of cancer-testis (CT) genes. We previously demonstrated TSGA10 expression during spermatogenesis. There is also evidence for potential TSGA10 involvement in cell proliferation. TSGA10 expression has been observed in a wide spectrum of cancers but not in hematopoietic neoplasm. Here we demonstrated expression of TSGA10 by semi-quantitative RT-PCR in 44 (84.6) out of 52 bone marrow samples and all peripheral blood samples from patients with acute lymphoblastic leukemia (ALL). Twenty-seven (52) cases had high level of gene expression and 16 (30.7) cases had a lower expression level of the gene in the patients bone marrow. Presence of TSGA10 expression in ALL may open a window to functional study of mitotic checkpoint proteins in leukemia. RT-PCR of TSGA10 may help in detection of residual clonal cells leading to early diagnosis and better prognostic qualification of the disease. © 2005 Elsevier Ltd. All rights reserved