TNFRSF1A Research Articles

Whole exome sequencing in patients with recurrent pericarditis

Abstract Background/Introduction About 10% of patients with recurrences may have a family history of pericarditis suggesting a genetic predisposition, that has been confirmed in selected cases with idiopathic recurrent pericarditis with a focused analysis of genes involved in autoinflammatory diseases (TNFRSF1A, MVK, NLRP3 genes, and variants at the Interleukin 1 Gene Locus). Purpose Aim of the present paper is to describe the frequency of detected variants of causative genes by Whole Exome Sequencing (WES) in patients with idiopathic recurrent pericarditis. Methods This is an observational, cohort study including all consecutive adult patients (>18 years) with > 2 episodes of recurrent pericarditis. All patients performed WES. We analyzed the frequency of genetic variants in: 1) known or predicted to be at least likely pathogenic (LP) variants already related to recurrent pericarditis, 2) predicted at least of unknown significance in genes not yet associated with recurrent pericarditis. Moreover, we highlighted incidental findings. Results We studied 108 patients with recurrent pericarditis (median age of 32 years, IQR 18.5; 67.6% females, all Caucasian, idiopathic etiology in 71.1%). The median number of recurrences was 5 (IQR 2). Eleven gene variants related to recurrent pericarditis were detected by WES analysis in 11/108 patients (10.2%): 9 variants were pathogenic/likely pathogenic (P/LP) according the ACMG guidelines and involved NLRP3 and TNFRSF1A genes, 2 VUS associated with familial Mediterranean fever (gene MEFV) (figure 1). We also recorded 3 P/LP variants and 2 VUS related to the inflammatory response (IFIH1, NFKBIA, JAK1, NOD2 and ALPK1 genes); 7 P/LP variants and 3 VUS related to heart’s electrical system (SCN5A, CALM2, TRPM4, KCND2, KCNQ1 genes); 12 P/LP variants and 10 VUS related to heart structural genes (figure 2). Conclusion(s) A substantial proportion of patients with multiple recurrences of pericarditis carries LP or P variants suggesting the importance of genetic predisposition evaluation in this population.

Genetic variants in the TNF pathway impact TNFi response in a mixed population with rheumatoid arthritis

Rheumatoid arthritis (RA) is a multifactorial autoimmune inflammatory disease that mainly affects the joints, on reducing functional capacity and impacting quality of life. Cytokines such as tumor necrosis factor (TNF) and interleukin 6 (IL-6) are crucial in the pathogenesis and treatment of this disease. Some patients using TNF inhibitors (TNFi) do not respond or lose their response to these medications. Clinical, sociodemographic, and genetic data were used to evaluate the associations of single nucleotide polymorphisms (SNP) in TNF, TNFRSF1A, and TNFRSF1B genes with the diagnosis of RA, standardized score results, laboratory tests, and response to TNFi. In one subsample, TNF and IL-6 serum levels cytokines were performed. A total of 654 subjects (360 healthy controls and 294 diagnosed with RA) were included in the analysis. Higher levels of TNF have been found in individuals diagnosed with RA. IL-6 levels were higher in individuals who did not respond to TNFi treatment, while responders had levels comparable to those without the disease. No associations were found between the SNPs studied and the diagnosis of RA; however, rs767455-C seems to play a role in the response to golimumab treatment, being related to better therapeutic response and lower mean serum leukocyte levels. In addition, rs1061622-G was associated with poorer functional capacity and rs1800629-A was associated with higher leukocyte values and serum transaminase levels. The rs1061622-G and rs767455-C may play a role in the response to TNFi treatment, especially for patients using golimumab, although they do not seem to be associated with the diagnosis of RA. Polymosphisms in the TNF pathway may impact baseline levels of immune cells and markers of renal and hepatic function in RA patients. Our results highlight the importance of evaluating the impact of these polymorphisms on TNFi response and safety, particularly in larger-scale studies.

A rare missense p.C125Y mutation in the TNFRSF1A gene identified in a Chinese family with tumor necrosis factor receptor-associated periodic fever syndrome.

Tumor necrosis factor receptor-associated periodic syndrome (TRAPS) is a rare autosomal dominant disorder with a low incidence in Asia. The most frequent clinical manifestations include fever, rash, myalgia, joint pain and abdominal pain. Misdiagnosis rates are high because of the clinical and genetic variability of the disease. The pathogenesis of TRAPS is complex and yet to be fully defined. Early genetic diagnosis is the key to precise treatment. In this study, a Chinese family with suspected TRAPS were analyzed by genome-wide SNP genotyping, linkage analysis and targeted sequencing for identification of mutations in causative genes. To study the pathogenicity of the identified gene mutation, we performed a conservation analysis of the mutation site and protein structure analysis. Flow cytometry was used to detect TNFRSF1A shedding and quantitative real-time PCR were used to assess the activation of unfolded protein response (UPR) in the mutation carriers and healthy individuals. A typical TRAPS family history, with a pattern of autosomal dominant inheritance, led to the identification of a rare mutation in the TNFRSF1A gene (c.G374A [p.Cys125Tyr]) with unknown significance. The patient responded well to corticosteroids, and long-term therapy with colchicine effectively reduced the inflammatory attacks. No amyloid complications occurred during the 6-year follow-up. In silico protein analysis showed that the mutation site is highly conversed and the mutation prevents the formation of intrachain disulfide bonds in the protein. Despite a normal shedding of the TNFRSF1A protein from stimulated monocytes in the TRAPS patients with p.C125Y mutation, the expression of CHOP and the splicing of XBP1 was significantly higher than healthy controls, suggesting the presence of an activation UPR. This is the first report of a Chinese family with the rare p.C125Y mutation in TNFRSF1A. The p.C125Y mutation does not result in aberrant receptor shedding, but instead is associated with an activated UPR in these TRAPS patients, which may provide new insights into the pathogenesis of this rare mutation in TRAPS.

Association of polymorphisms in the TNFA, TNFRSF1A and TNFRSF1B genes with lepromatous leprosy in Western Mexican patients.

Studies in different populations have shown that single-nucleotide polymorphisms (SNPs) of tumor necrosis factor-alpha (TNFα) and TNF receptors 1 and 2 (TNFR1 and TNFR2) may be involved in the pathogenesis of lepromatous leprosy (LL). To further explore the results in a Mexican population, we compared the frequencies of the polymorphisms in - 308 G>A TNFA (rs1800629), - 383 A>C TNFRS1A (rs2234649), and + 196 T >G TNFSR1B (rs1061622) genes in LL patients (n = 133) and healthy subjects (n = 198). The genotyping was performed with the polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) technique. Statistical analysis was performed using the χ2 test, within the 95% confidence interval. Odds ratios (OR) were calculated and Hardy-Weinberg equilibrium was verified for all control subjects and patients. We found an association between the TNFSR1 -383 A>C genotype and the risk of lepromatous leprosy when leprosy patients were compared to controls (OR = 1.71, CI: 1.08-2.69, p = 0.02). Furthermore, it was also associated with the risk of LL in a dominant model (AC + CC vs AA, OR: 1.65, 95% CI: 1.05-2.057, p = 0.02). Similar genotype and allele frequencies for the SNPs TNFA - 308 G>A and TNFSR2 + 196 T>G were observed between leprosy patients and healthy subjects. The TNFSR1 -383 A>C could be a potential marker for the identification of high-risk populations. However, additional studies, using larger samples of different ethnic populations, are required.

Knockouts of TNFRSF1A and TNFRSF1B Genes in K562 Cell Line Lead to Diverse Long-Lasting Responses to TNF-α.

This research delves into the intricate landscape of tumor necrosis factor-alpha (TNF-α) signaling, a multi-functional cytokine known for its diverse cellular effects. Specifically, we investigate the roles of two TNF receptors, TNFR1 and TNFR2, in mediating TNF-α-induced transcriptional responses. Using human K562 cell lines with TNFR1 and TNFR2 knockouts, we explore changes in gene expression patterns following TNF-α stimulation. Our findings reveal distinct transcriptional profiles in TNFR1 and TNFR2 knockout cells, shedding light on the unique contributions of these receptors to TNF-α signaling. Notably, several key pathways associated with inflammation, apoptosis, and cell proliferation exhibit altered regulation in the absence of TNFR1 or TNFR2. This study provides valuable insights into the intricate mechanisms governing TNF-α signaling and its diverse cellular effects, with potential implications for targeted therapeutic strategies.

Clinical analysis of 10 cases of multi-center tumor necrosis factor receptor-associated periodic syndrome

Objective: To summarize the clinical characteristics of tumour necrosis factor receptor-associated periodic syndrome (TRAPS) in children. Methods: The clinical manifestations, laboratory tests, genetic testing and follow-up of 10 children with TRAPS from May 2011 to May 2021 in 6 hospitals in China were retrospectively analyzed. Results: Among the 10 patients with TRAPS, including 8 boys and 2 girls. The age of onset was 2 (1, 5) years, the age of diagnosis was (8±4) years, and the time from onset to diagnosis was 3 (1, 7) years. A total of 7 types of TNFRSF1A gene variants were detected, including 5 paternal variations, 1 maternal variation and 4 de novo variations. Six children had a family history of related diseases. Clinical manifestations included recurrent fever in 10 cases, rash in 4 cases, abdominal pain in 6 cases, joint involvement in 6 cases, periorbital edema in 1 case, and myalgia in 4 cases. Two patients had hematological system involvement. The erythrocyte sedimentation rate and C-reactive protein were significantly increased in 10 cases. All patients were negative for autoantibodies. In the course of treatment, 5 cases were treated with glucocorticoids, 7 cases with immunosuppressants, and 7 cases with biological agents. Conclusions: TRAPS is clinically characterized by recurrent fever accompanied by joint, gastrointestinal, skin, and muscle involvement. Inflammatory markers are elevated, and autoantibodies are mostly negative. Treatment mainly involves glucocorticoids, immunosuppressants, and biological agents.

The Role of TNF-α, TNFRSF1A, and CD40 Gene Polymorphisms in Multiple Sclerosis in the Tomsk Region

The Role of TNF-α, TNFRSF1A, and CD40 Gene Polymorphisms in Multiple Sclerosis in the Tomsk Region

Marine Natural Compound (Neviotin A) Displays Anticancer Efficacy by Triggering Transcriptomic Alterations and Cell Death in MCF-7 Cells.

We investigated the anticancer mechanism of a chloroform extract of marine sponge (Haliclona fascigera) (sample C) in human breast adenocarcinoma (MCF-7) cells. Viability analysis using MTT and neutral red uptake (NRU) assays showed that sample C exposure decreased the proliferation of cells. Flow cytometric data exhibited reactive oxygen species (ROS), nitric oxide (NO), dysfunction of mitochondrial potential, and apoptosis in sample C-treated MCF-7 cells. A qPCR array of sample C-treated MCF-7 cells showed crosstalk between different pathways of apoptosis, especially BIRC5, BCL2L2, and TNFRSF1A genes. Immunofluorescence analysis affirmed the localization of p53, bax, bcl2, MAPKPK2, PARP-1, and caspase-3 proteins in exposed cells. Bioassay-guided fractionation of sample C revealed Neviotin A as the most active compound triggering maximum cell death in MCF-7, indicating its pharmacological potency for the development of a drug for the treatment of human breast cancer.

Apoptosis and inflammatory genes variants in primary non-response to anti-TNF therapy in Crohn's disease patients.

Anti-TNF therapy has indeed revolutionized the treatment of Crohn's disease, leading to higher rates of response and remission in patients. However, a significant proportion of 20-40% of patients do not respond to the initial therapy, others experience a secondary loss of response with ongoing treatment. Adverse drug reactions also occur in some patients. The effectiveness of anti-TNF treatment may be influenced by genetic variability, including FCGR3A, ADAM17, TNFRSF1A, TNFRSF1B, FAS, FASL, IL1B, CASP9 , and MIF genes. In this article, we provide an overview of the current knowledge and findings in the pharmacogenetics of anti-TNF drugs in CD focusing on the aspect of apoptosis and inflammatory genes variants in primary non-response. Pharmacogenetic investigations have been conducted to identify genetic markers that can predict response to anti-TNF therapy. However, large multi-center validation studies and multi-loci algorithms development are required to effectively prognose the treatment effect. The identification of predictive markers of response to anti-TNF therapy can help clinicians make informed decisions about treatment options and minimize adverse drug reactions in patients.

A family case of a rare autoinflammatory disease associated with mutations in the NLRP3 and TNFRSF1A genes in the practice of a rheumatologist

The article presents a clinical case of a rare autoinflammatory disease – a family case of Muckle – Wells syndrome. The diversity of clinical manifestations and the impossibility of confirming the diagnosis without a genetic study by DNA sequencing determine the complexity of and delay in the diagnosis. The development of severe complications and, as a consequence, a fatal outcome necessitates early diagnosis. The described clinical case demonstrates the importance of DNA sequencing for the timely diagnosis of the disease, the features of the disease course, and the familial nature of the disease. The diagnosis of Mackle – Wells syndrome in young family members before the development of severe complications will allow to start adequate and timely treatment and prevent the development of amyloidosis.

The Role of <i>TNF</i>-α, <i>TNFRSF1A</i>, and <i>CD40</i> Genes Polymorfisms in Multiple Sclerosis in Tomsk Region

We studied the role of polymorphisms rs1800629 of the TNF-α gene; rs4149584 of the TNFRSF1A gene; rs6074022, rs1883832, rs1535045, rs11086996 of the CD40 gene in the onset, clinical course and response to treatment in multiple sclerosis (MS) in a group of 152 patients, living in Tomsk region. 707 volunteers without autoimmune diseases and pathology of the nervous system were included in control group. The allele C of the rs6074022 polymorphism of CD40 gene was associated with the risk of MS and contributed to the high rate of disease progression. The T allele of the rs6074022 polymorphism of CD40 gene showed a significant association with the average rate of disease progression, and the GA genotype of rs1800629 polymorphism of TNF-α gene was associated with a higher frequency of MS exacerbations. Other polymorphisms did not demonstrate an association with both the risk of disease, the clinical features and response to treatment.

TNFα Causes a Shift in Gene Expression of TNFRSF1A and TNFRSF1B Isoforms.

Alternative splicing is a part of mRNA processing that expands the diversity of proteins encoded by a single gene. Studying the full range of proteins-products of translation of alternatively spliced mRNA is extremely important for understanding the interactions between receptor proteins and ligands since different receptor protein isoforms can provide variation in the activation of signaling pathways. In this study, we investigated the expression of isoforms of TNFR1 and TNFR2 receptors before and after exposure to TNFα in two cell lines that had previously demonstrated diverse effects on cell proliferation under TNFα incubation using RT-qPCR. We found that after incubation with TNFα: (1) expression of isoform 3 of the TNFRSF1A gene was increased in both cell lines; (2) the cell line with increased proliferation, K562, had decreased expression of isoforms 1 and 4 of the TNFRSF1A gene and expression of isoform 2 of TNFRSF1B gene was absent at all; (3) the cell line with decreased proliferation-MCF-7 had significantly increased expression of isoform 2 of TNFRSF1B gene. Thus, we can conclude that TNFα exposure to the K562 and MCF-7 cell lines leads to changes in the expression of TNFα receptor isoforms, which, in turn, can appear via diverse proliferative effects.

ETV7 reduces inflammatory responses in breast cancer cells by repressing the TNFR1/NF-κB axis

The transcription factor ETV7 is an oncoprotein that is up-regulated in all breast cancer (BC) types. We have recently demonstrated that ETV7 promoted breast cancer progression by increasing cancer cell proliferation and stemness and was also involved in the development of chemo- and radio-resistance. However, the roles of ETV7 in breast cancer inflammation have yet to be studied. Gene ontology analysis previously performed on BC cells stably over-expressing ETV7 demonstrated that ETV7 was involved in the suppression of innate immune and inflammatory responses. To better decipher the involvement of ETV7 in these signaling pathways, in this study, we identified TNFRSF1A, encoding for the main receptor of TNF-α, TNFR1, as one of the genes down-regulated by ETV7. We demonstrated that ETV7 directly binds to the intron I of this gene, and we showed that the ETV7-mediated down-regulation of TNFRSF1A reduced the activation of NF-κB signaling. Furthermore, in this study, we unveiled a potential crosstalk between ETV7 and STAT3, another master regulator of inflammation. While it is known that STAT3 directly up-regulates the expression of TNFRSF1A, here we demonstrated that ETV7 reduces the ability of STAT3 to bind to the TNFRSF1A gene via a competitive mechanism, recruiting repressive chromatin remodelers, which results in the repression of its transcription. The inverse correlation between ETV7 and TNFRSF1A was confirmed also in different cohorts of BC patients. These results suggest that ETV7 can reduce the inflammatory responses in breast cancer through the down-regulation of TNFRSF1A.

PiRNA and miRNA Can Suppress the Expression of Multiple Sclerosis Candidate Genes

Multiple sclerosis (MS) is a common inflammatory demyelinating disease with a high mortality rate. MS is caused by many candidate genes whose specific involvement has yet to be established. The aim of our study was to identify endogenous miRNAs and piRNAs involved in the regulation of MS candidate gene expression using bioinformatic methods. A program was used to quantify the interaction of miRNA and piRNA nucleotides with mRNA of the target genes. We used 7310 miRNAs from three databases and 40,000 piRNAs. The mRNAs of the candidate genes revealed miRNA binding sites (BSs), which were located separately or formed clusters of BSs with overlapping nucleotide sequences. The miRNAs from the studied databases were generally bound to mRNAs in different combinations, but miRNAs from only one database were bound to the mRNAs of some genes. For the first time, a direct interaction between the complete sequence of piRNA nucleotides and the nucleotides of their mRNA BSs of target genes was shown. One to several clusters of BSs of miRNA and piRNA were identified in the mRNA of ADAM17, AHI1, CD226, EOMES, EVI5, IL12B, IL2RA, KIF21B, MGAT5, MLANA, SOX8, TNFRSF1A, and ZBTB46 MS candidate genes. These piRNAs form the expression regulation system of the MS candidate genes to coordinate the synthesis of their proteins. Based on these findings, associations of miRNAs, piRNAs, and candidate genes for MS diagnosis are recommended.

OS08.4.A Analysis of melanoma brain metastasis immune microenvironment through multiplex gene expression profiling

Abstract Background Novel immunotherapies based on targeting of specific immune checkpoints enabled a significant improvement of melanoma outcome, but melanoma brain metastases (BM) remain an unmet oncological need with an overall 2-year survival rate lower than 10%. Tumour immune microenvironment has been demonstrated to play a key role in BM establishment and development, but data regarding the specific milieu of melanoma BM is limited. Material and Methods Gene expression profiles of 55 samples of primary melanoma and BM were evaluated using the nCounter PanCancer IO 360 Panel (NanoString Technologies) targeting 770 mRNA involved in tumor immune microenvironment modulation. The case series consisted of 10 primary melanomas and their 10 matched BM, 25 unmatched BM, and 10 locally advanced control melanomas without evidence of BM after >5 year follow up. Results Among BM samples, most patients (25/45) were males and median age at BM diagnosis was 61,2 years with a median time to BM development of 2,1 years. Median OS from BM diagnosis was 1,3 years. Several genes resulted significantly downregulated in BM compared to primary melanomas, including SERPINB5 (p<0.001), ARG1 (p=0.0067), S100A8 (p<0.001), S100A9 (p<0.001), S100A12 (p=0.0037), IL1RN (p=0.0012), CCL21 (p=0.0012), CCL22 (p=0.0012) and CCL13 (p=0.037) and SELE (p=0.026); conversely, C7 was upregulated (p<0.001). Downregulated signatures in BM involved those associated with multiple immune cell populations, including neutrophils, dendritic cells, mast cells and Treg, as well as inflammatory chemokines, the CTLA4 immune checkpoint and ARG1 enzyme function; conversely, MAGEs-related signature was upregulated. Comparison between primary melanomas which developed BM and those which did not showed a significant overexpression of RRM2 (p=0.0247) and TNFRSF1A (p=0.032) genes in the latter group and an upregulation of the PD-1 pathway. Analysis according to tumour mutational status showed an upregulation of signatures associated with inflammatory chemokines, dendritic and myeloid cells and neutrophils. No differences were observed according to time to BM development and survival from BM diagnosis. Conclusion Our findings show that melanoma BM harbor distinct immunosuppressive mechanisms compared to primary tumors: this data elicits the importance of investigating the heterogeneity of BM microenvironment. Genes and pathways selectively overexpressed or downregulated in melanoma BM should be validated to be possibly considered as novel therapeutic targets.

КЛИНИКО-ГЕНЕТИЧЕСКАЯ ХАРАКТЕРИСТИКА БОЛЬНЫХ С TRAPS (ПЕРИОДИЧЕСКИЙ СИНДРОМ, АССОЦИИРОВАННЫЙ С РЕЦЕПТОРОМ ФАКТОРА НЕКРОЗА ОПУХОЛИ) ПО ДАННЫМ ФЕДЕРАЛЬНОГО РЕВМАТОЛОГИЧЕСКОГО ЦЕНТРА

The tumor necrosis factor receptor-associated periodic syndrome (TRAPS) is a rare autosomal dominant disease associated with a mutation in the TNFRSF1A gene. It belongs to the group of monogenic autoinflammatory diseases (mAIDs) characterized by the repeated and prolonged episodes of fever, skin rashes, musculoskeletal, ophthalmic symptoms and an increase in the level of acute phase markers. The main targeted therapy is inhibitors of interleukin-1 (IL-1). The purpose of the study was to present the clinical and genetic characteristics of patients with TRAPS according to the data from the V.A. Nasonova Research Institute of Rheumatology (Moscow, Russia). The study included 28 patients from 2013 till 2021 with a clinical diagnosis of TRAPS from a selection of 162 (both pediatric and adult) patients with mAIDs: 17 females/11 males aged 2.5 to 65 years old (Me 9.0 y/o [7.9; 14.5]); 7 adults/21 children. All of them have underwent molecular genetic analysis for mutations in the TNFRSF1A gene. Results: the TRAPS was diagnosed in 28 (17.3%) among 162 patients with mAIDs. The age of onset ranged from 1 month till 28 years old (Me 4.0 y/o [1.0; 7.1]). Most patients fell ill prior to the age of 10 years old (89.3%), of which 8 (28.6%) before 1 year old, 7 (25%) from 1 to 5 years old, 1 (3.6%) from 5 to 10 years old, 10 (35.7%) from 10 to 18 years old and 2 (7.1%) from 21 to 28 years old. The duration of the disease and the delay in diagnosis at the time of inclusion ranged from 3 months till 59 years old. The diagnosis was made within the first year of illness in 4 (14.3%) of observed cases; within 1 to 10 years in 18 (64.3%); in 8 (28.6%) there was a delay in diagnosis of over 10 years; and in 4 (14.3%) the delay was more than 20 years of illness. The duration of attacks varied from 10 to 30 days, the intervals between the attacks ranged from 2 weeks till 6 months. The clinical manifestations were as follows: fever in 96.4% of cases, skin rashes in 71.4%, musculoskeletal symptoms in 18 (64.3%) cases, oligoarthritis in 9 (32.1%), polyarthritis in 2 (7.1% ), gastrointestinal manifestations in 18 (64.3%), lymphadenopathy of various groups in 10 (35.7%), ophthalmological manifestations in 8 (28.6%). The less often were the different nervous system manifestations, which accounted for 6% to 21% of cases, pharyngitis in 4 (14.3%) and stomatitis in 2 (7.1%). An increase in the sedimentation rate of erythrocytes was noted in 23 (82.1%), C-reactive protein in 20 (71.4%), leukocytosis in 11 (39.3%). The diagnosis in all patients was confirmed genetically. 10 patients had a low-penetrance mutation of р.Arg121Gln (р.Arg92Gln according to the old nomenclature). 5 familial cases were identified, the total number of sick family members was 12, from 2 to 4 in each family; 4 cases of amyloidosis (the all four were adults). Treatment: glucocorticoids in 19 (67.9%) cases, Cyclosporine A in 1, Colchicine in 5. 15 patients (53.6%) received genetically engineered biological preparations, the IL-1 (Canakinumab) was prescribed in 10 (35.7%) patients with the duration of admission from 1 to 9 years, Etanercept in 1, Adalimumab in 1 and Tocilizumab in 3. Conclusion: due to the similarity of clinical and laboratory symptoms with manifestations of rheumatic diseases, patients with TRAPS can be encountered in the rheumatologists’ practice. Most patients in the studied cohort had an early onset of the disease (prior to 10 years old), and the diagnosis delay of 10+ years was noted in 1/3 of patients. All patients had a specialized inflammatory phenotype coupled with genetic mutations. About half had required the appointment of biological drugs, IL-1 mainly. A complete response to the therapy in most patients and a good tolerability of treatment were noted, which allows the authors to consider this as the basis in the treatment and management of patients with TRAPS.

Heterozygous E148Q Mutation in an Indian Family with a Milder Form of Familial Mediterranean Fever

Dear Editor, Familial Mediterranean fever (FMF, OMIM no. 249100) is an inherited autoinflammatory disorder resulting from MEFV gene mutations. A 15-year-old girl from Kerala, South India, born of a nonconsanguineous union presented with recurrent fever, diffuse abdominal pain, and left knee joint pain for 1 year. She had fever for 3 days every week associated with left knee arthritis. Some of the episodes were associated with diffuse abdominal pain, oral ulcers, and an erythematous facial rash. She also had right hip and ankle pain with swelling on a few occasions. On examination, she had swollen and warm left knee joint. Her 12-year-old sibling also had similar problems (recurrent fever, diffuse abdominal pain, and joint pain) for 10 months. Both the parents were asymptomatic. Blood tests showed neutrophilic leukocytosis, high erythrocyte sedimentation rate, and C-reactive protein. Ultrasonography of the abdomen showed mild ascites. Immunological testing showed normal C3 complement and the absence of rheumatoid factor, antinuclear, anti-double-stranded DNA, and anti-neutrophilic cytoplasmic antibodies. The periodicity of symptoms and positive family history made us consider inherited autoinflammatory disorder. Targeted mutational analysis showed the MEFV variant E148Q in the heterozygous state in both siblings. No pathogenic variants were identified in the MEFV exon, TNFRSF1A, and MVK genes [Table 1]. The patient and her younger sister were managed with colchicine (1.0 mg daily), and on follow-up up to 1 year, the patients were asymptomatic. Genetic counseling was given to the patient family.Table 1: The results of targeted mutational analysis of the MEFV, TNFRSF1A, and MVK genes in the two siblingsFMF is common among certain ethnic groups of the Eastern Mediterranean region. More than 300 different mutations of the MEFV gene related to FMF have been identified, although only a few are pathogenic.[1] Previous studies testing the pathogenicity of E148Q alteration showed conflicting results. In a study of 646 FMF patients by Tirosh et al., 1% had a heterozygous E148Q variant with a milder clinical phenotype compared with patients having M694V homozygosity.[2] A Turkish FMF cohort of 153 patients had 33 (21.6%) patients with E148Q variant and 34 compound heterozygous patients (with E148Q and exon 10 mutations).[3] E148Q allele frequency among FMF patients was found to be 25% in Jewish, Arab, and Druze patients living in Israel, 20% in Turkish, 34% in Japanese, 17% in Azeri Turk patients living in Iran, and 2% in Armenian patients.[3] Several studies noted that the E148Q mutation was associated with a milder FMF phenotype. However, there was no significant difference in the frequency of clinical findings among the Turkish cohort.[3] Two patients from a South Indian family with recurrent fever, diffuse abdominal pain, and left knee joint pain for 1 year were found to have the MEFV variant E148Q in the heterozygous state. Both of them responded well to colchicine. The diagnosis of a milder phenotype of FMF was thus confirmed. Even though autoinflammatory disorders were reported from the Indian subcontinent,[4,5] we do not have data regarding the E148Q allele frequency and other mutations causing FMF. We report FMF in two members of a family with E148Q mutation which was not previously reported from the Indian subcontinent. This case reminds the readers to consider hereditary periodic fever syndromes when evaluating recurrent fever and the importance of E148Q mutation in the Indian population. Ethics approval All procedures performed in studies involving human participants were by the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Declaration of patient consent The authors certify that informed consent was taken from the father of both patients and assent was obtained from the children for participating in the study. The informed consent from the father and assent from the children were also obtained for using the clinical information to be reported in the journal. The patients and parents understand that their names and initials will not be published and due efforts will be made to conceal their identity, but anonymity cannot be guaranteed. Financial support and sponsorship Nil. Conflicts of interest There are no conflicts of interest.

Variant rs4149584 (R92Q) of the TNFRSF1A gene in patients with familial multiple sclerosis

IntroductionGenomic studies have identified numerous genetic variants associated with susceptibility to multiple sclerosis (MS); however, each one explains only a small percentage of the risk of developing the disease. These variants are located in genes involved in specific pathways, which supports the hypothesis that the risk of developing MS may be linked to alterations in these pathways, rather than in specific genes. We analyzed the role of the TNFRSF1A gene, which encodes one of the TNF-α receptors involved in a signaling pathway previously linked to autoimmune disease. MethodsWe included 138 individuals from 23 families including at least 2 members with MS, and analyzed the presence of exonic variants of TNFRSF1A through whole-exome sequencing. We also conducted a functional study to analyze the pathogenic mechanism of variant rs4149584 (-g.6442643C > G, NM_001065.4:c.362 G > A, R92Q) by plasmid transfection into human oligodendroglioma (HOG) cells, which behave like oligodendrocyte lineage cells; protein labeling was used to locate the protein within cells. We also analyzed the ability of transfected HOG cells to proliferate and differentiate into oligodendrocytes. ResultsVariant rs4149584 was found in 2 patients with MS (3.85%), one patient with another autoimmune disease (7.6%), and in 5 unaffected individuals (7.46%). The 2 patients with MS and variant rs4149584 were homozygous carriers and belonged to the same family, whereas the remaining individuals presented the variant in heterozygosis. The study of HOG cells transfected with the mutation showed that the protein does not reach the cell membrane, but rather accumulates in the cytoplasm, particularly in the endoplasmic reticulum and near the nucleus; this suggests that, in the cells presenting the mutation, TNFRSF1 does not act as a transmembrane protein, which may alter its signaling pathway. The study of cell proliferation and differentiation found that transfected cells continue to be able to differentiate into oligodendrocytes and are probably still capable of producing myelin, although they present a lower rate of proliferation than wild-type cells. ConclusionsVariant rs4149584 is associated with risk of developing MS. We analyzed its functional role in oligodendrocyte lineage cells and found an association with MS in homozygous carriers. However, the associated molecular alterations do not influence the differentiation into oligodendrocytes; we were therefore unable to confirm whether this variant alone is pathogenic in MS, at least in heterozygosis.

TNFRSF1A Gene Polymorphism (−610 T > G, rs4149570) as a Predictor of Malnutrition and a Prognostic Factor in Patients Subjected to Intensity-Modulated Radiation Therapy Due to Head and Neck Cancer

Simple SummaryMalnutrition is frequently related to the increased level of proinflammatory cytokines. Tumor necrosis factor α is a proinflammatory cytokine that plays a pivotal role in the development of malnutrition and cachexia in cancer patients. This study aimed to assess the relationship between a functional polymorphism of the TNFRSF1A gene and the occurrence of nutritional disorders in patients subjected to radiotherapy due to head and neck cancer. Multivariable analysis revealed that the TT genotype of the TNFRSF1A gene (−610 T > G) was independently correlated with a higher risk of nutritional disorders. Determination of this polymorphism may be useful in the assessment of the risk of nutritional deficiencies in patients subjected to radiotherapy due to head and neck cancer.Background: Malnutrition is a nutritional disorder observed in 52% of patients with head and neck cancer (HNC). Malnutrition is frequently related to the increased level of proinflammatory cytokines. In turn, ongoing inflammation is associated with increased catabolism of skeletal muscle and lipolysis. Tumor necrosis factor α (TNF-α) is a proinflammatory cytokine that plays a pivotal role in the development of malnutrition and cachexia in cancer patients. The aim of the study was to assess the relationship between a functional single-nucleotide polymorphism (SNP) −610 T > G (rs4149570) of the TNFRSF1A gene and the occurrence of nutritional disorders in patients subjected to RT due to HNC. Methods: The study group consisted of 77 patients with HNC treated at the Oncology Department of the Medical University in Lublin. Genotyping of the TNFRSF1A gene was performed using capillary electrophoresis (Genetic Analyzer 3500). Results: Multivariable analysis revealed that the TT genotype of the TNFRSF1A gene (−610 T > G) was an independent predictor of severe malnutrition (odds ratio—OR = 5.05; p = 0.0350). Moreover, the TT genotype of this gene was independently related to a higher risk of critical weight loss (CWL) (OR = 24.85; p = 0.0009). Conclusions: SNP (−610 T > G) of the TNFRSF1A may be a useful marker in the assessment of the risk of nutritional deficiencies in HNC patients treated with intensity-modulated radiotherapy (IMRT).

Proteomic Signatures of Monocytes in Hereditary Recurrent Fevers.

Hereditary periodic recurrent fevers (HRF) are monogenic autoinflammatory associated to mutations of some genes, such as diseases caused by mutations of including MEFV, TNFRSF1A and MVK genes. Despite the identification of the causative genes, the intracellular implications related to each gene variant are still largely unknown. A large –scale proteomic analysis on monocytes of these patients is aimed to identify with an unbiased approach the mean proteins and molecular interaction networks involved in the pathogenesis of these conditions. Monocytes from HRF 15 patients (5 with MFV, 5 TNFRSF1A and 5with MVK gene mutation) and 15 healthy donors (HDs) were analyzed by liquid chromatography and tandem mass spectrometry before and after lipopolysaccharide (LPS) stimulation. Significant proteins were analyzed through a Cytoscape analysis using the ClueGo app to identify molecular interaction networks. Protein networks for each HRF were performed through a STRING database analysis integrated with a DISEAE database query. About 5000 proteins for each HRF were identified. LPS treatment maximizes differences between up-regulated proteins in monocytes of HRF patients and HDs, independently from the disease’s activity and ongoing treatments. Proteins significantly modulated in monocytes of the different HRF allowed creating a disease-specific proteomic signatures and interactive protein network. Proteomic analysis is able to dissect the different intracellular pathways involved in the inflammatory response of circulating monocytes in HRF patients. The present data may help to identify a “monocyte proteomic signature” for each condition and unravel new possible unexplored intracellular pathways possibly involved in their pathogenesis. These data will be also useful to identify possible differences and similarities between the different HRFs and some multifactorial recurrent fevers.