Somatic Reversion of a Novel IL2RG Mutation Resulting in Atypical X-Linked Combined Immunodeficiency

Mutations of the IL2RG gene, which encodes for the interleukin-2 receptor common gamma chain (γC, CD132), can lead to X-linked severe combined immunodeficiency (X-SCID) associated with a T−B+NK− phenotype as a result of dysfunctional γC-JAK3-STAT5 signaling. Lately, hypomorphic mutations of the IL2RG gene have been described causing atypical SCID with a milder phenotype. Here, we report three brothers with low-normal lymphocyte counts and susceptibility to recurrent respiratory infections and cutaneous warts. The clinical presentation combined with dysgammaglobulinemia suspected an inherited immunity disorder, which has been proven by Next Generation Sequencing as a novel c.458T > C; p.Ile153Thr IL2RG missense-mutation. Subsequent functional characterization revealed impaired T-cell proliferation, low TREC levels and a skewed TCR Vβ repertoire in all three patients. Interestingly, investigation of various subpopulations showed normal expression of CD132 but with partially impaired STAT5 phosphorylation compared to healthy controls. Additionally, we performed precise genetic analysis of subpopulations revealing spontaneous somatic reversion, predominately in lymphoid derived CD3+, CD4+ and CD8+ T cells. Our data demonstrate that the atypical SCID phenotype noticed in these three brothers is due to the combination of hypomorphic IL-2RG function and somatic reversion.

Initial Evaluation of Uroplakins UPIIIa and UPII in Selected Benign Urological Diseases

Background: Uroplakins (UPs) are glycoproteins that play a specific role in the structure and function of the urothelium. Disorders which affect the normal expression of UPs are associated with the pathogenesis of infections and neoplasms of the urinary tract, primary vesicoureteral reflux, hydronephrosis and renal dysfunction. The appearance of uroplakins in the urine and/or plasma may be of potential importance in the detection of urinary tract dysfunction. The aim of the present study was to investigate uroplakin IIIa (UPIIIa) and uroplakin II (UPII) expression in patients with selected urological diseases. Methods: Plasma and urine from patients with benign prostatic hyperplasia (BPH), urethral stricture (US), urinary tract infection (UTI) and urolithiasis were compared to healthy people without urological disorders. UPs concentrations were measured by the immunoenzymatic method. Results: In patients with BPH and UTI, concentrations of UPIIIa in urine and plasma, as well as UPII in urine, were statistically significantly higher than in the control groups. In the US group, only the plasma UPIIIa concentration differed significantly from the control. Conclusion: The conducted research shows that benign urological diseases may affect the state of the urothelium, as manifested by increased concentrations of both UPs in patients’ urine and plasma, especially in BPH and UTI.

Mining sponge phenomena in RNA expression data

In the last few years, the interactions among competing endogenous RNAs (ceRNAs) have been recognized as a key post-transcriptional regulatory mechanism in cell differentiation, tissue development, and disease. Notably, such sponge phenomena substracting active microRNAs from their silencing targets have been recognized as having a potential oncosuppressive, or oncogenic, role in several cancer types. Hence, the ability to predict sponges from the analysis of large expression data sets (e.g. from international cancer projects) has become an important data mining task in bioinformatics. We present a technique designed to mine sponge phenomena whose presence or absence may discriminate between healthy and unhealthy populations of samples in tumoral or normal expression data sets, thus providing lists of candidates potentially relevant in the pathology. With this aim, we search for pairs of elements acting as ceRNA for a given miRNA, namely, we aim at discovering miRNA-RNA pairs involved in phenomena which are clearly present in one population and almost absent in the other one. The results on tumoral expression data, concerning five different cancer types, confirmed the effectiveness of the approach in mining interesting knowledge. Indeed, 32 out of 33 miRNAs and 22 out of 25 protein-coding genes identified as top scoring in our analysis are corroborated by having been similarly associated with cancer processes in independent studies. In fact, the subset of miRNAs selected by the sponge analysis results in a significant enrichment of annotation for the KEGG32 pathway “microRNAs in cancer” when tested with the commonly used bioinformatic resource DAVID. Moreover, often the cancer datasets where our sponge analysis identified a miRNA as top scoring match the one reported already in the pertaining literature.

TET1-Mediated Tumor Inhibition and Dox Resistance in Colon Cancer by Blocking WNT / β-Catenin Pathway

BackgroundAs DNA demethylation protein, Ten-eleven translocation 1 (TET1) has been widely reported that is related to tumorigenesis and tumor metastasis. This study is to investigate the role and regulation mechanism of TET1 in colon cancer.Methods The TET1 and Catenin beta-1 (CTNNB1) expression level in colon cancer samples and cancer cell lines HCT116/SW480 were observed to discover the relationship between these two genes. Knockdown and overexpression of TET1 through shRNA and CRISPR technology were used to elucidate the effect of TET1 on WNT/β-catenin pathway. The 5-hmC/5-mC level were explored by bisulfate sequencing (BSP) and Chromatin immunoprecipitation (ChIP) to further explain the regulation mechanism. Combined with the reverse assay and transwell invasion assay, the cell migration and invasion ability were tested. Finally, the role of TET1 on DOX resistance was analyzed.Results TET1 downregulated in colon cancer and showed an opposite expression trend with WNT pathway associated gene CTNNB1. TET1 bound to CTNNB1 promotor and catalyzed demethylation to activate transcription of CTNNB1, inhibiting WNT/β‐catenin signaling pathways. Colon cancer cells proliferation was promoted by TET1 downregulation, which was further verified as shTET1 could upregulate the tumor invasion. The DOX addition could rescue the cell migration, compared with normal expression of TET1. Meanwhile, TET1 down-regulation was related to DOX resistances.Conclusion TET1 played as a DNA hydroxymethylation activates inhibitors of the WNT/β-catenin signaling pathway in colon tumor and TET1 down-regulation contributed to DOX-resistance, which might provide reference to targeting therapy in clinical practice.

Transcriptome Profiling of Peripheral Blood Mononuclear Cells Reveals COVID-19 Patients Are Not Recovered to Normal After Discharge for 5 Months

BackgroundHighly pathogenic coronavirus disease-2019 (COVID-19) initiated by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection has swiftly expanded throughout the world, and the fatality rate is still expanding due to the second wave in 2020 winters. This ongoing epidemic threatens public health with its new strain that emerged in some countries and might cause devastating deaths. Therefore, the host transcriptomic profile from patients during recovery is important for understanding this disease. MethodsWe performed transcriptome profiling of the RNAs isolated from the peripheral blood mononuclear cells (PBMCs) of recovered COVID-19 patients at hospital discharge of three months and five months respectively.ResultsOur results exposed diverse inflammatory genes and cytokine profiles to infection in recovered patients, and emphasize the highly expressed genes in COVID-19 patients like CCL4, CCL3, CXCL9, CXCL16, IL10, CSF2, VEGFA showed a decreasing trend in recovered patients. Furthermore, the integrated analysis predicted that JUN, CTSL, DDIT4, RRAS, BIRC5, CTSZ, CCNB2, CDK1, OAS1/2, IFIT3, RSAD2, and TP53I3 genes may be valuable for the recovery of COVID-19 patients. ConclusionsOur analysis confirms the presence of some inflammatory genes in recovered patients, suggesting COVID-19 patients did not return to their normal expression even after 5-months of discharge. Identification of transcriptome profiling of recovered patients provides useful information regarding its pathogenesis and might help for the development of better treatment for COVID-19.

Therapeutic Potential of SphK1 Inhibitors Based on Abnormal Expression of SphK1 in Inflammatory Immune Related-Diseases

Sphingosine kinase 1(SphK1) a key enzyme that catalyzes the conversion of sphingosine (Sph) to sphingosine 1-phosphate (S1P), so as to maintain the dynamic balance of sphingolipid-rheostat in cells and participate in cell growth and death, proliferation and migration, vasoconstriction and remodeling, inflammation and metabolism. The normal expression of SphK1 maintains the balance of physiological and pathological states, which is reflected in the regulation of inflammatory factor secretion, immune response in traditional immune cells and non-traditional immune cells, and complex signal transduction. However, abnormal SphK1 expression and activity are found in various inflammatory and immune related-diseases, such as hypertension, atherosclerosis, Alzheimer’s disease, inflammatory bowel disease and rheumatoid arthritis. In view of the therapeutic potential of regulating SphK1 and its signal, the current research is aimed at SphK1 inhibitors, such as SphK1 selective inhibitors and dual SphK1/2 inhibitor, and other compounds with inhibitory potency. This review explores the regulatory role of over-expressed SphK1 in inflammatory and immune related-diseases, and investigate the latest progress of SphK1 inhibitors and the improvement of disease or pathological state.

DFNA20/26 and Other ACTG1-Associated Phenotypes: A Case Report and Review of the Literature

Since the early 2000s, an ever-increasing subset of missense pathogenic variants in the ACTG1 gene has been associated with an autosomal-dominant, progressive, typically post-lingual non-syndromic hearing loss (NSHL) condition designed as DFNA20/26. ACTG1 gene encodes gamma actin, the predominant actin protein in the cytoskeleton of auditory hair cells; its normal expression and function are essential for the stereocilia maintenance. Different gain-of-function pathogenic variants of ACTG1 have been associated with two major phenotypes: DFNA20/26 and Baraitser–Winter syndrome, a multiple congenital anomaly disorder. Here, we report a novel ACTG1 variant [c.625G>A (p. Val209Met)] in an adult patient with moderate-severe NSHL characterized by a downsloping audiogram. The patient, who had a clinical history of slowly progressive NSHL and tinnitus, was referred to our laboratory for the analysis of a large panel of NSHL-associated genes by next generation sequencing. An extensive review of previously reported ACTG1 variants and their associated phenotypes was also performed.

Dynein Heavy Chain 64C Differentially Regulates Cell Survival and Proliferation of Wingless-Producing Cells in Drosophila melanogaster

Dynein is a multi-subunit motor protein that moves toward the minus-end of microtubules, and plays important roles in fly development. We identified Dhc64Cm115, a new mutant allele of the fly Dynein heavy chain 64C (Dhc64C) gene whose heterozygotes survive against lethality induced by overexpression of Sol narae (Sona). Sona is a secreted metalloprotease that positively regulates Wingless (Wg) signaling, and promotes cell survival and proliferation. Knockdown of Dhc64C in fly wings induced extensive cell death accompanied by widespread and disorganized expression of Wg. The disrupted pattern of the Wg protein was due to cell death of the Wg-producing cells at the DV midline and overproliferation of the Wg-producing cells at the hinge in disorganized ways. Coexpression of Dhc64C RNAi and p35 resulted in no cell death and normal pattern of Wg, demonstrating that cell death is responsible for all phenotypes induced by Dhc64C RNAi expression. The effect of Dhc64C on Wg-producing cells was unique among components of Dynein and other microtubule motors. We propose that Dhc64C differentially regulates survival of Wg-producing cells, which is essential for maintaining normal expression pattern of Wg for wing development.

Initial Evaluation of Uroplakins UPIIIa and UPII in Selected Benign Urological Diseases

BackgroundUroplakins (UPs) are glycoproteins that play a specific role in the structure and function of the urothelium. Disorders of normal expression of uroplakins are associated with the pathogenesis of infections and neoplasms of the urinary tract, primary vesicoureteral reflux, hydronephrosis and renal dysfunction. The appearance of uroplakins in the urine and/or plasma may be of potential importance in the detection of urinary tract dysfunction. The aim of the study was to investigate uroplakin IIIa (UPIIIa) and uroplakin II (UPII) expression in patients with selected urological diseases. Plasma and urine from patients with benign prostatic hyperplasia (BPH), urethral stricture (US), urinary tract infection (UTI) and urolithiasis, were compared to healthy people without urological disorders. MethodsA total 152 of human urine and plasma samples from normal and patients with selected benign urological diseases were analyzed. UPs concentration was measured by immunoenzymatic method. All calculations were done using the STATISTICA 13.3 (TIBCO software Inc.).ResultsIn patients with BPH and UTI, UPIIIa in urine and plasma also UPII in urine concentrations were statistically significantly higher than in the control groups. In the US group, only the plasma UPIIIa concentration differed significantly from the control. There were no significant differences between the concentrations of UPs compared to the controls in both the urine and plasma of patients with urolithiasis. ConclusionThe conducted research shows that benign urological diseases may affect the state of the urothelium, as manifested by an increased concentration of both UPs in patients’ urine and plasma, especially in BPH and UTI.

THE ANALYSIS OF a-CRYSTALLINE PROTEIN IN WHITE AND BRUNESCENT CATARACT

Objectives: The objectives of the study were to determine the difference of concentration and expression of α-crystalline protein in white and brunescent cataract lenses. Methods: The design of this study is cross-sectional comparative. The subject was cataract patients who underwent cataract surgery in Puskesmas Pariaman, West Sumatra, Indonesia. Lens examination was carried out at the Microbiology Laboratory of FK Unand from July 2019 to February 2020. The samples consisted of 36 subjects who met the inclusion criteria. ELISA examination was used to determine the concentration of α-crystalline protein and Western Blot examination was performed to see the expression of the α-crystalline protein in all subjects. Results: The difference in the concentration of α-crystalline protein in white cataract and brunescent cataract was not statistically significant, with p=0.129 (p>0.05). The result of Western blot examination was normal expression of α-crystalline protein in white cataract and under expression of α-crystalline protein in brunescent cataracts. Conclusion: The expression of α-crystalline protein appeared to be different between white and brunescent cataract lenses. In brunescent cataract, under expression of α-crystalline proteins was related to the decrease of chaperone activity. This change occurred allegedly because of photochemical reaction that happened inside the lens.