Aims: To assess whether, besides "traditional" risk factors, overall oxidative stress, oxidized lipoproteins, and glycemic variability are associated with early macro-vascular damage in type 1 diabetes (T1D). Methods: In 267 children/adolescents with T1D (130 girls, age 9.1-23.0 years) we evaluated: derivatives of reactive oxygen metabolites [d-ROMs], serum total antioxidant capacity [TAC] and oxidized LDL-cholesterol [oxLDL]; markers of early vascular damage (Lipoprotein-associated phospholipase A2 [Lp-PLA2], z-score of carotid intima-media thickness [z-cIMT] and carotid-femoral pulse wave velocity [z-PWV]); CGM metrics of four weeks preceding the visit, central systolic/diastolic blood pressures (cSBP/cDBP), and HbA1c, z-score of BP (zSBP/z-DBP) and circulating lipids longitudinally collected since T1D onset.. Three general linear models were built with z-cIMT, z-PWV adjusted for current cDBP, and Lp-PLA2 as independent variables. Results: The z-cIMT was associated with male gender (B = 0.491, .92 = 0.029, p = 0.005), cSBP (B = 0.023, .92 = 0.026, p = 0.008) and oxLDL (B = 0.022, .92 = 0.022, p = 0.014). The z-PWV was associated with diabetes duration (B = 0.054, .92 = 0.024, p = 0.016), daily insulin dose (B = 0.52, .92 = 0.018, p = 0.045), longitudinal zSBP (B = 0.18, .92 = 0.018, p = 0.045) and dROMs (B = 0.003, .92 = 0.037, p = 0.004). Lp-PLA2 was associated with age (B = 0.221, .92 = 0.079, p = 3*10-6), oxLDL (B = 0.081, .92 = 0.050, p = 2*10-4), longitudinal LDLcholesterol (B = 0.031, .92 = 0.043, p = 0.001) and male gender (B = -1.62, .92 = 0.10, p = 1.3*107). Conclusions: Oxidative stress, male gender, insulin dose, diabetes duration and longitudinal lipids and blood pressure, contributed to the variance of early vascular damage in young patients with T1D.

Introduction: Type 1 diabetes (T1D) is associated with an increased risk of cardiovascular disease. Insulin resistance is an important cardiovascular risk factor (CVRF), also in subjects with T1D, but the influence of the genetic predisposition of insulin resistance on cardiovascular risk is still unknown in T1D. We aimed to determine whether a genetic score composed of six variants, previously associated with insulin resistance and type 2 diabetes (T2D) risk, associates with insulin sensitivity and known CVRFs in children and adolescents with T1D. Materials and Methods: 330 children and adolescents (174 males; mean age 15.7 ± 3.5 years) with T1D were genotyped for the following genetic variants: rs1801278 (IRS1), rs1044498 (ENPP1), rs2295490 (TRIB3), rs1801282 (PPARG), rs780094 (GCKR), and rs35767 (IGF1). An additive genetic risk score (GRS) and cardiovascular risk score (CVRS) were calculated. Anthropometric, glycemic control, insulin sensitivity, blood pressure, and biochemical parameters were assessed. Multivariate regression between evaluated phenotypes and GRS was performed. Results: We found a significant association between the GRS and estimated insulin sensitivity (β = −0.027 [−0.040 to −0.013], R2 = 0.86, p≤ 0.001), diastolic blood pressure (β = 0.68 [0.08–1.27], R2 = 0.20, p = 0.026), triglycerides (β = 4.26 [1.74–6.77], R2 = 0.13, p = 0.001), waist to height ratio (β = 0.003 [0.001–0.006], R2 = 0.75, p = 0.010), non-HDL-cholesterol (β = 3.63 [1.39–5.87], R2 = 0.12, p = 0.002), and CVRS (β = 0.063 [0.008–0.118], R2 = 0.19, p = 0.025), independent of age, sex, BMI, pubertal stage, diabetes duration, glycated hemoglobin, type of treatment, and total insulin requirement. The addition of the GRS to established clinical risk factors significantly improved the discriminatory capability of the regression model for predicting subjects with more CVRFs (C-statistic 0.89 [95% CI: 0.84–0.95] versus 0.83 (0.73–0.93); p = 0.037). Conclusions: Insulin resistance and T2D risk-associated genetic variants influence insulin sensitivity and known CVRFs in children and adolescents with T1D.

The ubiquitin-proteasome system (UPS) plays an important role in maintaining cellular homeostasis by degrading a multitude of key regulatory proteins. FBXW11, also known as b-TrCP2, belongs to the F-box family, which targets the proteins to be degraded by UPS. Transcription factors or proteins associated with cell cycle can be modulated by FBXW11, which may stimulate or inhibit cellular proliferation. Although FBXW11 has been investigated in embryogenesis and cancer, its expression has not been evaluated in osteogenic cells. With the aim to explore FBXW11gene expression modulation in the osteogenic lineage we performed molecular investigations in mesenchymal stem cells (MSCs) and osteogenic cells in normal and pathological conditions. In vitro experiments as well as ex vivo investigations have been performed. In particular, we explored the FBXW11 expression in normal osteogenic cells as well as in cells of cleidocranial dysplasia (CCD) patients or osteosarcoma cells. Our data showed that FBXW11 expression is modulated during osteogenesis and overexpressed in circulating MSCs and in osteogenically stimulated cells of CCD patients. In addition, FBXW11 is post-transcriptionally regulated in osteosarcoma cells leading to increased levels of beta-catenin. In conclusion, our findings show the modulation of FBXW11 in osteogenic lineage and its dysregulation in impaired osteogenic cells.

Cleidocranial dysplasia (CCD), a dominantly inherited skeletal disease, is characterized by a variable phenotype ranging from dental alterations to severe skeletal defects. Either de novo or inherited mutations in the RUNX2 gene have been identified in most CCD patients. Transcription factor RUNX2, the osteogenic master gene, plays a central role in the commitment of mesenchymal stem cells to osteoblast lineage. With the aim to analyse the effects of RUNX2 mutations in CCD patients, we investigated RUNX2 gene expression and the osteogenic potential of two CCD patients' cells. In addition, with the aim to better understand how RUNX2 mutations interfere with osteogenic differentiation, we performed string analyses to identify proteins interacting with RUNX2 and analysed p53 expression levels. Our findings demonstrated for the first time that, in addition to the alteration of downstream gene expression, RUNX2 mutations impair p53 expression affecting osteogenic maturation. In conclusion, the present work provides new insights into the role of RUNX2 mutations in CCD patients and suggests that an in-depth analysis of the RUNX2-associated gene network may contribute to better understand the complex molecular and phenotypic alterations in mutant subjects.

Aims: We assessed whether oxidative stress (OS) is increased in children/adolescents with type 1 diabetes (T1D) compared to healthy peers. Moreover, we searched for OS predictors in the T1D population. Methods: We compared the concentration of serum derivatives of reactive oxygen metabolites (d-ROMs) in 412 children/adolescents with T1D (3.6-23.5 years old) to that of 138 healthy children/adolescents (1.2-19.2 years old) by ANOVA adjusted for age, gender, and BMI z-score (z-BMI). Applying a general linear model, in a subgroup of 331 patients using continuous glucose monitoring, we searched for predictors of d-ROMs among 3-day, 2-week, and 4-week metrics of glucose control and variability, such as mean blood glucose, percent time in range (70-180 mg/dl,TIR70-180), coefficient of variation, and others, as well as among conventional cardiovascular risk factors like current and average HbA1c, z-BMI, blood pressure percentiles, and lipid concentrations recorded retrospectively over the entire follow-up period. Results: D-ROMs levels were significantly higher in children/adolescents with T1D compared to controls [371.9 (64.2) versus 324.9 (46.3), p < 10(-16)]. Sex (B = 49.1, eta(2) = 0.14, p = 1.3 * 10(-9)), age < 12 years in boys (B = 79.4, eta(2) = 0.074, p = 10(-7)),3-day TIR70-180 (B =-0.87, eta(2) = 0.048, p = 6.5 * 10(-5)), and z-BMI (B = 7.4, eta(2) = 0.016, p = 0.022) predicted d-ROMs with an overall R-2 of 0.278. Conclusions: OS is increased in youth with T1D and only partially predicted by gender, age, glucose control, and anthropometry. Other potential determinants of OS in this population should be targeted in future studies. (C) 2021 Elsevier B.V. All rights reserved.