The long non-coding RNA - microRNA (lncRNA–miRNA) regulatory axis is a key modulator of immune and inflammatory pathways, and growing evidence supports its contribution to therapeutic response variability. In our previous study, we identified the lncRNA MEG3 as potentially involved in the response to biological drugs, specifically TNFα- and IL17A-inhibitors, in psoriatic arthritis (PsA). To further characterize its role, we performed a bioinformatic analysis to identify MEG3-targeted miRNAs, followed by an exploratory expression profiling analysis using qRT-PCR in a cohort of 54 PsA patients at baseline (T0) and after 12 months of therapy (T12), compared with 15 healthy controls (CTRLs). We identified four candidate miRNA targets: hsa-miR-21-5p, hsa-miR-19b-3p, hsa-miR-19a-3p, and hsa-miR-17-5p. Among these, hsa-miR-17-5p was significantly upregulated in PsA patients at T0 versus CTRLs. Most importantly, we found a significant decrease in hsa-miR-17-5p and hsa-miR-19b-3p levels in Responder patients at 12-month follow-up. We then explored whether MEG3 genetic variability contributes to miRNA modulation. First, we observed that the MEG3 rs941576 genetic variant appears to influence the expression levels of hsa-miR-19a-3p and hsa-miR-19b-3p. Next, in silico analyses indicated that this variant lies within an immune-active enhancer, altering the binding affinity for the transcription factors HIF1A/ARNT2, and suggested a link between MEG3 enhancer activity and hypoxia-responsive regulation of these miRNAs. Lastly, pathway enrichment analysis highlighted that both hsa-miR-17-5p and hsa-miR-19b-3p converge on the TGF-β signalling pathway. Our findings suggest the involvement of a specific MEG3-miRNA network in modulating the response to biological therapies in PsA.

AIMS:Genome-wide association studies (GWAS) have identified the IGF2BP2 (Insulin-like Growth Factor 2 mRNA binding protein 2) gene as a susceptibility locus for Type 2 diabetes mellitus (T2D). This study aimed to evaluate the IGF2BP2 mRNA levels in the blood of a cohort of T2D patients and to quantify the expression levels of non-coding RNAs (ncRNAs) that are predicted to interact with it. METHODS:We extracted RNA from peripheral blood mononuclear cells of 50 T2D patients and 30 healthy controls (CTRL) and quantified, by qPCR, the IGF2BP2 expression levels. Using bioinformatics tools, we predicted its main ncRNAs target and quantified it. RESULTS:The expression study showed a significantly higher IGF2BP2 level in T2D subjects than in CTRL. In silico analysis identified hsa-let7b-5p as a potential microRNA regulator of IGF2BP2, with reduced expression levels observed in T2D patients. Additionally, three lncRNAs (SNHG5, HOTAIR, and MEG3) were predicted as potential targets of IGF2BP2. Their expression levels were significantly elevated in T2D patients. In vitro assays demonstrated that inhibiting hsa-let7b-5p in HeLa cells resulted in increased expression of IGF2BP2 and the three lncRNAs. CONCLUSION:These findings highlight a potential regulatory network involving IGF2BP2, hsa-let7b-5p, and lncRNAs. This network may contribute to dysregulation of insulin/IGF signaling and glucose metabolism pathways, providing insights into T2D pathogenesis.

The aim of this study was to investigate possible associations between the KCNJ11 rs2285676 and T2D susceptibility and/or clinical manifestations. The KCNJ11 rs2285676 genotypic/allelic distribution and the case/control association analysis were evaluated in one hundred and eighty-one T2D patients with type 2 diabetes (T2D), and one hundred and ninety-nine healthy controls. Our analyses showed a significant association between the presence of diabetic retinopathy (DR) and the variant allele of the SNV, both at the genotypic and at the allelic level. Our results indicate a possible involvement of the KCNJ11 gene in DR development and suggests that variants of this gene could help in the prediction of DR susceptibility.

Background/Objectives: Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease strongly influenced by genetic factors. Genome-wide association studies (GWASs) have identified numerous non-coding susceptibility loci, but their functional roles remain poorly understood. The single-nucleotide variant (SNV) rs2836882, located in an enhancer near the ETS2 proto-oncogene, has been implicated in immune regulation, though its contribution to SLE is unclear. Methods: We analyzed rs2836882 in 246 Italian patients with SLE and 216 matched controls using TaqMan genotyping. A weighted genetic risk score (wGRS) combining rs2836882 with other known SLE variants was calculated. ETS2 mRNA expression was quantified by RT-qPCR in PBMCs from 60 individuals, and in silico analyses assessed the variant's functional context. Results: The rs2836882 risk allele was significantly associated with SLE (OR = 1.54, p = 0.02). Patients showed a markedly higher wGRS than controls (p < 0.00001), confirming an additive genetic burden. In silico data indicated that rs2836882 lies within an active enhancer region (H3K4me1/H3K27ac+) containing PU.1 binding motifs and functions as an expression quantitative trait locus (eQTL) for ETS2. Expression analysis demonstrated that carriers of the risk allele exhibited significantly increased ETS2 expression compared to non-carriers (p = 0.002) in both groups. Conclusions: In conclusion, rs2836882 is a functional regulatory variant that enhances ETS2 transcription and contributes to increased SLE susceptibility. These findings establish a mechanistic link between a non-coding GWAS locus and disease risk, emphasizing the role of regulatory variants in autoimmune pathogenesis and supporting the integration of functional non-coding variants into genetic risk models for improved patient stratification.

Aims We aimed to analyse Sirtuin 1 (SIRT1) and Vitamin D receptor (VDR) expression levels in the peripheral blood of patients with type 2 diabetes (T2D), characterized for the presence of diabetic neuropathy (DN), and to evaluate possible genetic factors that could influence the VDR expression levels. Methods Fifty-one participants with T2D, who underwent neurological assessment for DN were recruited. We quantified the mRNA levels of SIRT1 and VDR in peripheral blood mononuclear cells. Moreover, we analysed the methylation status and the rs2228570 genetic variant of VDR promoter. Results Patients with DN ( n = 32) showed lower expression of SIRT1 (p corr =0.018) and VDR (p corr =0.009), compared to those without DN. Furthermore, we observed a positive correlation between the mRNA levels of SIRT1 and VDR ( p = 0.01). The expression levels of these genes negatively correlated with the score based on cardiovascular reflex tests (CARTs score). Moreover, the variant allele of rs2228570 in the VDR gene was associated with higher expression of this gene compared to the wild-type allele ( p = 0.003). Conclusion In patients with DN, both SIRT1 and VDR expression levels are reduced and interrelated. Low VDR expression levels could negatively affect SIRT1 transcription, thus influencing all the most pathogenetic pathways of DN regulated by this protein.

Inflammation and cellular senescence are two interconnected biological processes critical for several chronic diseases. Pro-inflammatory factors promote the accumulation of reactive oxygen species, which synergistically accelerate the deterioration process. This creates a feedback loop that exacerbates telomere attrition and mitochondrial dysfunction, hallmarks of cellular senescence. Conventional therapies have been shown to influence oxidative stress and consequently cellular senescence, while the impact of biologic and targeted synthetic disease-modifying antirheumatic drugs (b/tsDMARDs) is poorly investigated. Our aim was to explore biomarkers of cellular senescence in a population of psoriatic arthritis (PsA) patients treated with different b/tsDMARDs; we evaluated telomere length (TL), mtDNA copy numbers and oxidative damage at the beginning of therapy and after 12 months. We enrolled 50 PsA patients starting a b/tsDMARDs treatment and monitored the response to treatment for 12 months, based on the Disease Activity Index for PsA score, to identify subgroups of patients: responders and non-responders. We collected a blood sample for each patient at the beginning of therapy and after 12 months. In addition, we collected blood samples of 34 age- and sex-matched controls (CTRLs). We evaluated the cellular senescence biomarkers by qPCR. PsA patients at T0 showed a lower TL and fewer mtDNA copy numbers with respect to CTRLs (P < 0.001). Moreover, oxidative telomeric and mitochondrial damage seems to correlate positively with BMI (P < 0.05). We performed a combined ROC curve analysis to evaluate their ability to discriminate the two groups, showing an AUC of 0.828 with 59.2

Interstitial lung disease (ILD) is a common extra-articular manifestation of rheumatoid arthritis (RA). The inflammatory response in lung disease is characterized by severe oxidative stress, which enhances cellular senescence. Telomeric shortening and mitochondria dysregulation represent two hallmarks of cellular senescence. The maintenance of telomere length (TL) and mitochondrial DNA (mtDNA) copy number is preserved by many proteins, such as TERF1 and TFAM, respectively. Our aim was to evaluate the TL, the mtDNA copy number and the expression of two regulator gene factors in RA patients with (RA-ILD) and without lung involvement (RA-NILD). Eighty-five RA patients and 21 healthy subjects were enrolled. Relative TL, mtDNA copy number, and expression analysis of TERF1 and TFAM genes were measured using qPCR assay. All RA patients present a statistically significant telomere shortening; in particular, RA-ILD patients show shorter TL compared to both controls and RA-NILD. Patients with Usual Interstitial Pneumonia pattern show a more evident shortening of TL. Lastly, both RA-ILD and RA-NILD patients present a significant decrease in mtDNA copy number compared to controls. The analysis of regulatory genes showed an increase in TERF1 expression in RA patients compared to controls, also after stratification in the two subgroups, and a decrease in TFAM expression in RA patients compared to controls. These results show that the alteration of TL and mtDNA copy number in RA patients is more evident in the presence of ILD. The hypothesis is that, in these patients, oxidative stress could accelerate the shortening of telomeres and the decrease of mtDNA copy number.

We aimed to identify an expression profile of lncRNAs potentially related to treatment response in Psoriatic arthritis (PsA) patients, to be used as potential genomic biomarkers predictors of drug treatment effectiveness. In addition, we evaluated a possible association between lncRNAs genetic variants and the response to therapy using the clinical parameter of Disease Activity Index. For the expression study, we collected 48 treated PsA patients, monitoring the treatment response for 12 months. We initially used PCR Array and, then, we validated the results with qRT-PCR. We also retrospectively genotyped 163 treated PsA patients. Firstly, we observed a significant difference in the expression level between Responder and non-Responder patients, of 4 lncRNAs in the group of PsA patients treated with TNFi and of 3 lncRNAs in the group of patients treated with IL17i. Then, we confirmed a significant decrease of MEG3 expression in non-Responder patients compared to Responders, also considering separately the single groups of patients treated with TNFi and IL17i. In addition, our results seem to highlight a potential dose-dependent effect of rs941576 (MEG3) variant allele on Disease Activity Index. Our study suggests a possible role of the lncRNA MEG3 in the treatment response to biological drugs.

Background & aims: This study investigated a possible correlation between three circulating miRNAs, previously observed to be associated to diabetic polyneuropathy, and the obesity condition. Methods & results: The expression levels of miR-128a, miR-155 and miR499a were evaluated in 49 participants with Type 2 diabetes, divided into different groups based on the presence or absence of obesity and central obesity. The analyses revealed a significant decrease of miR-155 and miR-499a expression levels in obese subjects. In particular, the reduction appears to be even more significant in Type 2 diabetes subjects with central obesity. Conclusion: The results suggest that these miRNAs could be involved in obesity-driven pathogenetic mechanisms.

Primary Sjögren's syndrome (pSS) is a chronic, systemic autoimmune disease characterized by a wide spectrum of glandular and extra-glandular features. Genetic and epigenetic factors play an important role in the disease susceptibility and phenotype. There are a multitude of genes that have been identified as implicated in the pathogenesis of pSS, both in HLA and extra-HLA regions with a strong contribution given by genes in interferon signalling pathways. Among the HLA alleles, the most consistent associations have been found with DR2 and DR3 alleles at the DRB1 locus. Moreover, several gene variants outside the MHC locus are in genes involved in NF-κB signalling, B- and T-cell function and methylation processes possibly responsible for lymphomagenesis. There is still a lack of knowledge on precise genetic patterns and prediction models of diseases, and data on pharmacogenetics is scarce. A comprehensive summary of the common genetic factors and an extensive analysis of novel epigenetic aspects is provided, together with a view on the relationships between novel therapeutic agents for pSS and genetic targets in signalling pathways, aiming at improving tailored treatment strategies in the view of a more personalized medicine.

Background: Telomeres are specific regions of repetitive nucleotide sequences that protect chromosome ends and preserve genetic information. In each cell division, telomeres shorten, leading finally to apoptosis and cell cycle arrest when reaching a critical point. The telomere-associated protein (telomeric repeat-binding factor 1 [TERF1]) is essential for the maintenance of telomeres, acting as an inhibitor of telomerase, and its levels correlate with telomere length (TL).Shortened telomeres have been demonstrated in Rheumatoid Arthritis (RA). Furthermore, they are a recognized risk factor for idiopathic pulmonary fibrosis. Few data are present in the literature regarding TL in RA-associated interstitial lung disease (RA-ILD), while no data are present regarding TERF1 in RA and RA-ILD. Objectives: We aimed to evaluate the TL and TERF1 expression levels in RA-ILD. Methods: TL and TERF1 expression levels were evaluated in patients with RA-ILD compared to age-matched RA patients without lung involvement (RA-non-ILD) and healthy controls. Genomic DNA and total RNA were isolated from peripheral blood mononuclear cells. Relative TL was measured using real-time quantitative polymerase chain reaction (qPCR) assay, which quantifies a ratio of telomeric repeat copy signal and a reference single-copy gene (human beta globin) signal. Expression analysis of TERF1 was performed by qPCR assay. T-test was used to compare mean TL and TERF1 expression data among the different phenotypic groups. RA patients were divided according to whether their TL fell within or above the first quartile of the cohort. A multivariate logistic regression analysis was used to correct the p-value for sex, age and disease duration. Results: Eighty-nine RA patients were included (mean age 63.6 ± 13.8 years, median disease duration 9 [IQR 8.4-11.6] years): 42 RA-ILD and 47 RA-non-ILD. 21 age- and sex-matched controls were collected. RA-ILD patients were older and with minor disease duration than RA-non-ILD patients. They exhibited higher disease activity, CRP levels, positivity for RF and ACPA, and were more treated with bDMARDs than RA-non-ILD patients (Table 1). TL in all patients was significantly shorter compared to controls (p = 0.0016). RA-ILD patients presented significantly shorter TL compared to controls (p = 0.00001) and compared to RA-non-ILD (p = 0.0006) (Figure 1A). In the multivariate regression analysis, TL was reduced in RA-ILD compared with RA-non-ILD when adjusted for sex, age and disease duration (p<0.001). After patients stratification according to their TL, it is observed that the prevalence of ILD was significantly higher in patients with short vs normal-length telomeres (82.6% vs 34.8% p=0.00008). In RA-ILD, TL correlated negatively with disease duration (p= 0.007 r= -0.408). TERF1 expression levels were reduced in RA compared with controls (p= 2.17E-17). Both RA-ILD patients and RA-non-ILD patients exhibited reduced TERF1 expression levels than controls (p= 3.37E-10 and p= 2.78E-10, respectively. Figure 1B). TERF1 levels correlated positively with TL (p= 0.004 r= 0.328). Conclusion: Telomere shortening is a feature of immunosenescence and it has been linked to more severe articular disease. Shorter TL and reduced TERF1 expression levels characterize RA. In particular, RA-ILD patients displayed higher disease activity, negative prognostic factors, received more biologic treatments, and exhibited shorten TL than RA-non-ILD patients suggesting that TL might represent a hallmark of a more aggressive disease with lung involvement.Table 1. Acknowledgements: NIL. Disclosure of Interests: None declared.Figure 1

BACKGROUND:The PCSK3 gene encodes for the protease enzyme Furin, which promotes proteolytic maturation of important regulators of the immune response, and also enhances the secretion of interferon-γ (IFN). Several studies have suggested its possible involvement in the pathogenesis of chronic inflammatory diseases.METHODS:We investigated the PCSK3 gene expression level in peripheral blood mononuclear cells isolated from Sjögren's Syndrome (SS) patients and healthy controls and we evaluated a possible correlation with IFN-γ gene expression. Moreover, we also explored the variability of two PCSK3 genetic polymorphisms (rs4932178 and rs4702) to evaluate a possible association between these polymorphisms and the expression levels of this gene.RESULTS:We observed, by RT-qPCR, that the PCSK3 expression level was significantly higher in SS patients compared to the controls (p = 0.028), and we confirmed a positive correlation between PCSK3 and IFN-γ expression levels (p < 0.001). Moreover, we reported that the variant homozygous genotype of rs4932178 SNP is associated with a higher expression of the PCSK3 gene (p = 0.038) and with the SS susceptibility (p = 0.016).CONCLUSIONS:Our data suggest that Furin could play a role in SS development, also promoting IFN-γ secretion.

The vitamin D receptor (VDR), binding to the active form of the vitamin, promotes the transcription of numerous genes involved in the proliferation of immune cells, cytokine production and lymphocyte activation. It is known that vitamin D deficiency can influence the risk of developing rheumatoid arthritis (RA) or modulate its disease activity. The aim of this study was to investigate a possible association between the rs11568820 (C > T) polymorphism in the promoter region of VDR gene and the response to therapy with anti-TNF drugs in patients with RA. A total of 178 consecutive Italian patients with RA treated with anti-TNF, naïve for biological therapy, were recruited. Disease activity data were evaluated using specific indices such as DAS28, CDAI and SDAI, measured at the start of therapy and subsequently at 22, 52, 104 and 240 weeks. A statistically significant association emerged between the rs11568820 variant allele of VDR gene and failure to remission assessed by CDAI and SDAI at 52 weeks, and by DAS28, CDAI and SDAI at 104 weeks of follow-up. Furthermore, the variant allele of this polymorphism was observed more frequently in patients who did not undergo sustained remission calculated by CDAI and SDAI. The variant T allele of rs11568820 in VDR gene is associated with a reduced remission rate with anti-TNFα drugs. These data suggest the role of VDR genetic variability in the response to therapy and in the achievement of remission.

Background: Psoriatic arthritis (PsA) is a chronic inflammatory disease, characterized by heterogeneous articular and periarticular manifestations. The achievement of remission or low disease activity is the goal of therapy. However, some patients experience primary failure and lack or loss of response to cs-, b- and ts-DMARDs. The treatment response could be affected by multiple factors, including epigenetic factors. Recently, some studies have suggested the possible involvement of lncRNAs in modulating treatment response [1]. In this context, the identification of genetic and epigenetic factors associated to treatment response could help to define new biomarkers for a more effective and personalized therapy. Objectives: The main aim of this study was to prospectively investigate lncRNAs potentially related to treatment response in a cohort of PsA patients treated with TNFi and IL17i, to identify potential predictors of drug treatment effectiveness. In addition, we analysed retrospectively a cohort of PsA patients treated with TNFi to look for possible association between lncRNAs genetic variants and the response after 6 and 12 months of TNFi. Methods: For the expression study, a cohort of 48 PsA patients starting a TNFi (n=28) or IL17i (n=20) drugs was recruited, monitoring their treatment response for 12 months in order to identify subgroup of patients Reponder e Non-Responder. For the genotyping study, we retrospectively analysed 163 PsA patients treated with TNFi. For each patient was estimated the Disease Activity in Psoriatic Arthritis (DAPsA) score at 6 and 12 months after the beginning of therapy. The expression level of lncRNA was analysed in a panel of 84 lncRNAs, after the extraction of total RNA from PBMCs. Then we validated the differentially expressed lncRNAs, resulted from the array experiments, by qRT-PCR using specific primer assay. Web-based data analysis tool (NPInter v4.0 and DIANALncBase v3) were used to confirm miRNA target genes of the validated lncRNAs. For the genotyping study, we extracted genomic DNA from PBMCs and we performed allelic discrimination assay by TaqMan assays. We evaluated a possible association between the selected SNPs and the response to therapy at 6 and 12 months from the beginning of the TNFi treatment, using the clinical parameter of DAPsA value. Results: We observed a significant difference in the expression level between Responder and non-Responder patients, of 4 lncRNAs in the group of PsA patients treated with TNFi and of 3 lncRNAs in the group of patients treated with IL17i. Then, we confirmed a significant decrease of MEG3 expression in non-Responder patients compared to Responder patients, both considering the whole cohort (P= 0.01) and stratifying patients by drugs (P= 0.05 and P= 0.03, respectively for TNFi and IL17i) (Figure 1). In addition, we observed an association between the variant allele of rs7158663 and a lower expression of MEG3 compared to the wild-type allele, although without statistical significance. We also observed that the variant allele of rs941576 (MEG3) was associated with a better response at T6 and T12, with a linear decrease of mean DAPsA value among wildtype, heterozygous and homozygous variant patients. Interestingly, we noticed that the variant allele of rs941576 SNV resulted associated with a better response regarding joints involvement. Indeed, the number of TJ and SJ decreases more in patients carrying the variant allele, both at T6 and T12 (P= 0.0006 and P= 0.032, respectively) (Table 1). Conclusion: Our study suggests a possible role, both in terms of genetic variability and expression, of the lncRNA MEG3 in the treatment response to TNFi and IL17i in PsA patients. REFERENCES: [1] De Benedittis G, Latini A, Ciccacci C, et al. Impact of TRAF3IP2, IL10 and HCP5 Genetic Polymorphisms in the Response to TNF-i Treatment in Patients with Psoriatic Arthritis. J Pers Med. 2022;12(7):1094. Acknowledgements: NIL. Disclosure of Interests: None declared.

Systemic lupus erythematosus (SLE) is a chronic multifactorial autoimmune disease. It is now widely demonstrated that oxidative stress (OS) plays an important role in the modulation of the pathogenesis of this disease. Mitochondrial DNA (mtDNA) is highly vulnerable to OS and it is known a decrease of mtDNA copy number in SLE patients. However, to date, it has not been investigated if this decrease is associated with a dysregulation of mitochondrial homeostasis genes. Our aim is to evaluate the amount of mtDNA copy number and the expression of the genes more involved in the mitochondrial homeostasis pathways, in peripheral blood mononuclear cells (PBMCs) of SLE patients and healthy controls. We analysed the amount of mtDNA in PBMCs of 72 SLE patients and 61 healthy controls by qPCR. Then, we investigated the expression variability of TFAM and SIRT1 (biogenesis), MFN1 and MFF (fusion/fission) and PRKN2 (mitophagy) genes in a subgroup of SLE patients and healthy controls. Interestingly, we have observed a highly significant decrease in mtDNA copies in SLE patients compared to healthy controls (P < 0.0001). In addition, we have shown that the expression levels of SIRT1, MFN1 and PRKN2 genes were significantly decreased in SLE patients with respect to healthy controls (P = 0.00001 for SIRT1, P = 0.0150 for MFN1 and P = 0.0009 for PRKN2). Lastly, we have reported a positive correlation between PRKN2 expression level and mtDNA copy number (P = 0.019, r = 0.475). In conclusion, our data confirm the impairment of mtDNA copy number in the disease and show for the first time a dysregulation of the mitochondrial homeostasis genes. These results could provide additional support to the important role of mitochondria in SLE development.

The identification of rare genetic variants associated to Systemic Lupus Erythematosus (SLE) could also help to understand the pathogenic mechanisms at the basis of the disease. In this study we have analyzed a cohort of 200 Italian SLE patients in order to explore the rare protein-coding variants in five genes (TNFAIP3, STAT4, IL10, TRAF3IP2, and HCP5) already investigated for commons variants found associated in our previous studies. Genomic DNA of 200 SLE patients was sequenced by whole exome sequencing. The identified variants were filtered by frequency and evaluated by in silico predictions. Allelic association analysis was performed with standard Fisher's exact test. Introducing a cutoff at MAF < 0.01, a total of 19 rare variants were identified. Seven of these variants were ultra-rare (MAF < 0.001) and six were absent in the GnomAD database. For TNFAIP3 gene, the variant c.A1939C was observed in 4 SLE patients and it is located in a region enriched in phosphorylation sites and affects the predict affinity of specific kinases. In TRAF3IP2 gene, we observed 5 different rare variants, including the novel variant c.G410A, located in the region that mediates interaction with TRAF6, and therefore a possible risk factor for SLE development. In STAT4 gene, we identified 6 different rare variants. Among these, three missense variants decrease the stability of this protein. Moreover, 3 novel rare variants were detected in 3 SLE patients. In particular, c.A767T variant was predicted as damaging by six prediction tools. Concluding, we have observed that even in genes whose common variability is associated with SLE susceptibility, it is possible to identify rare variants that could have a strong effect in the disease development and could therefore allow a better understanding of the functional domain involved.

Background and aims: Type 2 diabetes (T2D) hyperglycaemia alters basal autophagy. Since autophagy is an essential cellular process, our aim was to investigate the ATG5 (autophagy-related 5) gene expression level and genetic variants in a cohort of diabetic patients, characterized for the presence of microangiopathic complications. Methods and Results: the expression levels of ATG5 were evaluated in PBMCs from 48 T2D patients with an extensive evaluation for microangiopathic complications. Our analyses revealed a significant lower expression of ATG5 in T2D patients with retinopathy compared to those without retinopathy. We also highlighted a significant lower expression of ATG5 in T2D patients with early-cardiovascular autonomic neuropathy compared to those without it, after correction for sex, age, body mass index and levels of hemoglobin A1c. Conclusion: our results highlight that dysregulation in the autophagy process could be involved in the development of severe microangiopathic complications. (C) 2023 The Italian Diabetes Society, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.

The vitamin D receptor (VDR) regulates bone development and calcium homeostasis, suggesting a central role in musculoskeletal diseases such as osteoporosis (OP). Several studies have examined the contribution of VDR polymorphisms and epigenetic signatures in bone metabolism and OP risk, with sometimes inconclusive results. Our study aimed to explore the association between genetic variability, expression and the methylation pattern of VDR with the risk of OP in a cohort of Caucasian patients. Genomic DNA from 139 OP, 54 osteopenic (Ope) and 73 healthy (CTR) subjects were used for genotyping the rs731236 (TaqI), rs2228570 (FokI) and rs11568820 (Cdx2) polymorphisms of the VDR gene by an allelic discrimination assay. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of VDR expression levels and pyrosequencing analysis of a VDR promoter CpG island were carried out in a subcohort (25 OP and 25 CTR) of subjects. Data obtained showed a significantly higher OP risk for rs11568820 G/A and A/A genotypes (p = 0.05). qRT-PCR revealed lower VDR gene expression levels in the OP group compared to CTR subjects (p = 0.0009), also associated with both the rs11568820 A/A genotype (p = 0.03) and femoral fragility fractures (p = 0.05). No association was found between the methylation pattern of the region analyzed of the VDR promoter and its expression levels. Our results identify a significative association between Cdx2 rs11568820 polymorphism and OP risk. In addition, the VDR transcriptomic profile suggests a putative interconnection with OP progression, providing a useful tool to stratify OP phenotype and fragility fracture risk.

Objective: The purpose of this study was to determine the distribution of organ damage in a cohort of systemic lupus erythematosus (SLE) patients and to evaluate the roles of clinical and genetic factors in determining the development of chronic damage. Methods: Organ damage was assessed by the SLICC Damage Index (SDI). We analyzed a panel of 17 single-nucleotide polymorphism (SNPs) of genes already associated with SLE, and we performed a phenotype–genotype correlation analysis by evaluating specific domains of the SDI. Results: Among 175 Caucasian SLE patients, 105 (60%) exhibited damage (SDI ≥1), with a median value of 1.0 (IQR 3.0). The musculoskeletal (26.2%), neuropsychiatric (24.6%) and ocular domains (20.6%) were involved most frequently. The presence of damage was associated with higher age, longer disease duration, neuropsychiatric (NP) manifestations, anti-phospholipid syndrome and the positivity of anti-dsDNA. Concerning therapies, cyclophosphamide, mycophenolate mofetil and glucocorticoids were associated with the development of damage. The genotype–phenotype correlation analysis showed an association between renal damage, identified in 6.9% of patients, and rs2205960 of TNFSF4 (p = 0.001; OR 17.0). This SNP was significantly associated with end-stage renal disease (p = 0.018, OR 9.68) and estimated GFR < 50% (p = 0.025, OR 1.06). The rs1463335 of MIR1279 gene was associated with the development of NP damage (p = 0.029; OR 2.783). The multivariate logistic regression analysis confirmed the associations between TNFSF4 rs2205960 SNP and renal damage (p = 0.027, B = 2.47) and between NP damage and rs1463335 of MIR1279 gene (p = 0.014, B = 1.29). Conclusions: Our study could provide new insights into the role of genetic background in the development of renal and NP damage.