Impact of prediabetes with a high risk of diabetes stratified by glycated hemoglobin level on the severity of coronavirus disease 2019 during admission.

Objective: Prediabetes with a glycated hemoglobin (HbA1c) level of 5.7 - 6.4% is associated with a poor prognosis of coronavirus disease 2019 (COVID-19), but whether the degree of glycemic control is associated with COVID-19 severity is unknown. The aim of this study was to evaluate the association between the degree of glycemic control and COVID-19 severity in patients with prediabetes. Materials and methods: We reviewed 254 patients with COVID-19 admitted to our hospital between April 2020 and September 2021. Based on their HbA1c level, patients were classified into low (HbA1c level < 5.7%), moderate (HbA1c level, 5.7 - 5.9%), and high risk of diabetes (HbA1c level, 6.0 - 6.4%). The association between risk of diabetes and the worst COVID-19 symptom in terms of severity during admission was evaluated using binary logistic regression analysis. Results: Seventy-one and 88 patients had moderate and high risks of diabetes, respectively. Sixty-three and seven patients presented severe (requiring non-invasive oxygen therapy) or critical (intensive care unit admission or artificial respiratory management) COVID-19. The multivariate logistic regression analysis showed that a high risk of diabetes was correlated with severe COVID-19 (P = 0.01) after adjusting for baseline characteristics, whereas a moderate risk of diabetes was not (P = 0.17). Conclusion: Prediabetes with a high risk of diabetes is associated with the worst COVID-19 symptom in terms of severity during admission. Our findings could aid in more efficient allocation of healthcare resources to a narrower population of prediabetic patients. Supplementary Information: The online version contains supplementary material available at 10.1007/s13340-023-00643-z.

In Vivo Tissue-Specific DNA Demethylation in Mouse Liver Through a Hydrodynamic Tail Vein Injection.

A new technique called the dCas9-SunTag and scFv-TET1CD epigenome editing system has recently been developed to edit the DNA methylation status of specific genes. The transfection of an all-in-one vector containing this system into cells is feasible and induces the DNA demethylation of specific genes; however, due to the large size of the vector, difficulties are associated with its introduction into mice. We herein used a hydrodynamic tail vein injection (HTVi) to introduce the all-in-one vector into mice for in vivo epigenome editing. HTVi needs to be considered for inducing the targeted DNA demethylation of particular genes in the mouse liver.

Ascorbic acid during the suckling period is required for proper DNA demethylation in the liver.

Ascorbic acid (AA, vitamin C) serves as a cofactor for ten-eleven translocation (TET) enzymes and induces DNA demethylation in vitro. However, its role in DNA demethylation in vivo remains unclear. We previously reported that DNA demethylation in the mouse liver was enhanced during the suckling period. Therefore, we hypothesized that DNA demethylation is enhanced in an AA-dependent manner during the suckling period. To examine our hypothesis, we employed wild-type (WT) mice, which synthesize AA, and senescence marker protein-30/gluconolactonase (SMP30/GNL) knockout (KO) mice, which cannot synthesize AA, and analyzed the DNA methylation status in the livers of offspring in both the suckling period and adulthood. SMP30/GNL KO offspring showed DNA hypermethylation in the liver possibly due to low plasma and hepatic AA levels during the suckling period despite the administration of rescue-dose AA to dams. Furthermore, DNA hypermethylation of the fibroblast growth factor 21 gene (Fgf21), a PPARα target gene, persisted into adulthood. In contrast, a high-dose AA administration to SMP30/GNL KO dams during the lactation period restored DNA demethylation in the livers of offspring. Even though a slight increase was observed in plasma AA levels with the administration of rescue-dose AA to WT dams during the gestation and lactation periods, DNA demethylation in the livers of offspring was minimally enhanced. The present results demonstrate that AA intake during the suckling period is required for proper DNA demethylation in the liver.

Targeted DNA demethylation of the Fgf21 promoter by CRISPR/dCas9-mediated epigenome editing.

Recently, we reported PPARα-dependent DNA demethylation of the Fgf21 promoter in the postnatal mouse liver, where reduced DNA methylation is associated with enhanced gene expression after PPARα activation. However, there is no direct evidence for the effect of site-specific DNA methylation on gene expression. We employed the dCas9-SunTag and single-chain variable fragment (scFv)-TET1 catalytic domain (TET1CD) system to induce targeted DNA methylation of the Fgf21 promoter both in vitro and in vivo. We succeeded in targeted DNA demethylation of the Fgf 21 promoter both in Hepa1-6 cells and PPARα-deficient mice, with increased gene expression response to PPARα synthetic ligand administration and fasting, respectively. This study provides direct evidence that the DNA methylation status of a particular gene may determine the magnitude of the gene expression response to activation cues.

Epigenetic modulation of Fgf21 in the perinatal mouse liver ameliorates diet-induced obesity in adulthood.

The nutritional environment to which animals are exposed in early life can lead to epigenetic changes in the genome that influence the risk of obesity in later life. Here, we demonstrate that the fibroblast growth factor-21 gene (Fgf21) is subject to peroxisome proliferator-activated receptor (PPAR) α-dependent DNA demethylation in the liver during the postnatal period. Reductions in Fgf21 methylation can be enhanced via pharmacologic activation of PPARα during the suckling period. We also reveal that the DNA methylation status of Fgf21, once established in early life, is relatively stable and persists into adulthood. Reduced DNA methylation is associated with enhanced induction of hepatic FGF21 expression after PPARα activation, which may partly explain the attenuation of diet-induced obesity in adulthood. We propose that Fgf21 methylation represents a form of epigenetic memory that persists into adulthood, and it may have a role in the developmental programming of obesity.

Mild Maternal Hypothyroxinemia During Pregnancy Induces Persistent DNA Hypermethylation in the Hippocampal Brain-Derived Neurotrophic Factor Gene in Mouse Offspring.

BACKGROUND: Thyroid hormones are essential for normal development of the central nervous system (CNS). Experimental rodents have shown that even a subtle thyroid hormone insufficiency in circulating maternal thyroid hormones during pregnancy may adversely affect neurodevelopment in offspring, resulting in irreversible cognitive deficits. This may be due to the persistent reduced expression of the hippocampal brain-derived neurotrophic factor gene Bdnf, which plays a crucial role in CNS development. However, the underlying molecular mechanisms remain unclear. METHODS: Thiamazole (MMI; 0.025% [w/v]) was administered to dams from two weeks prior to conception until delivery, which succeeded in inducing mild maternal hypothyroxinemia during pregnancy. Serum thyroid hormone and thyrotropin levels of the offspring derived from dams with mild maternal hypothyroxinemia (M offspring) and the control offspring (C offspring) were measured. At 70 days after birth, several behavior tests were performed on the offspring. Gene expression and DNA methylation status were also evaluated in the promoter region of Bdnf exon IV, which is largely responsible for neural activity-dependent Bdnf gene expression, in the hippocampus of the offspring at day 28 and day 70. RESULTS: No significant differences in serum thyroid hormone or thyrotropin levels were found between M and C offspring at day 28 and day 70. M offspring showed an impaired learning capacity in the behavior tests. Hippocampal steady-state Bdnf exon IV expression was significantly weaker in M offspring than it was in C offspring at day 28. At day 70, hippocampal Bdnf exon IV expression at the basal level was comparable between M and C offspring. However, it was significantly weaker in M offspring than in C offspring after the behavior tests. Persistent DNA hypermethylation was also found in the promoter region of Bdnf exon IV in the hippocampus of M offspring compared to that of C offspring, which may cause the attenuation of Bdnf exon IV expression in M offspring. CONCLUSIONS: Mild maternal hypothyroxinemia induces persistent DNA hypermethylation in Bdnf exon IV in offspring as epigenetic memory, which may result in long-term cognitive disorders.