Obese children have poor vitamin D genes (CYP27A1, CYP2R1, CYP27B1) – March 2024

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Decreased vitamin D bio-availability with altered DNA methylation of its metabolism genes in association with the metabolic disorders among the school-aged children with degree I, II and III obesity

The Journal of Nutritional Biochemistry, https://doi.org/10.1016/j.jnutbio.2024.109627
Xueyi Jiang 1 †, Lulu Xia 2 †, Tiantian Tang 1, Xiuqin Fan 1, Rui Wang 1, Meichen Wang 2, Wenli Yang 2, Jie Yan 2, Kemin Qi 1, Ping Li 1

Obesity is strongly associated with disturbances of vitamin D (VD) metabolites in the animal models. However, the related epidemiological evidence is still controversial, especially the different degrees of obesity. Hence, in this present representative cross-sectional study, 106 obesity school-age children aged 7-12 years were included and divided into different subgroups as degree I (the age- and sex-specific BMI≥95th percentile, n=45), II (BMI ≥120% percentile, n=34) and III (BMI ≥140% percentile, n=27) obesity groups across the ranges of body mass index (BMI).. While the age- and sex-matched subjects without obesity were as the control group. Notably, it was significantly different of body composition, anthropological and clinical characteristics among the above four subgroups with the dose-response relationships (P<0.05). Moreover, comparing with the control group, the serum VD concentrations were higher, VD metabolites like 25(OH)D, 25(OH)D3 and 1,25(OH)2D, and related hydroxylases as CYP27A1, CYP2R1 and CYP27B1 were lower in the degree I, II and III obesity subgroups (P<0.05), which were more disorder with the anthropological and clinical characteristics as the obesity was worsen in a BMI-independent manner (P<0.05). However, there was a significant increase of CYP27B1 in the degree III obesity group than those in the degree I and II obesity subgroups. Furthermore, the methylation patterns on the genome-wide (Methylation/Hydroxymethylation) and VD metabolism genes (CYP27A1, CYP2R1 and CYP27B1) were negatively correlated with the worse obesity and their related expressions (P<0.05). In summary, these results indicated that obesity could affect the homeostasis of VD metabolism related genes such as CYP27A1, CYP2R1, CYP27B1 and etc through abnormal DNA methylation, resulting in the disorders of VD related metabolites to decrease VD bio-availability with the BMI-independent manner. In turn, the lower levels of VD metabolites would affect the liver function to exacerbate the progression of obesity, as the Degree II and III obesity subgroups.

Introduction
Obesity in the children and adolescents aged 5 to 19 year represents a global health issue with an ever-increasing prevalence, as WHO has estimates that it has nearly tripled since 1975 to 2018 [1,2]. In China, the children obesity similarly increases rapidly [3]. As we all known, obesity is associated with a strong predisposition towards metabolic diseases and even cancer among the childhood and even adulthood [3,4]. Thus, early prevention of children obesity is of great significance for minimizing the adverse consequences through the entire life cycle. Notably, the main characteristics of obesity is the abnormal expansion of adipose depots, which can be driven by the imbalance of hypertrophy and hyperplasia [5]. Generally, the hyperplasia of adipocytes plays an important role on the early stages of obesity to be considered healthy and adaptive by maintaining proper vascularization, increasing many metabolism modulatory adipokines and so on [5,6]. However, with the further progresses of obesity, the hypertrophy of adipocytes with massively expanded expansion posits the most important outcomes, which is associated with the limitations of oxygen diffusion and angiogenesis to elevate the lipolysis, increase the secretion of inflammatory cytokines and reduce the adipokines to persistently contribute the occurrence of metabolic diseases [7,8]. Here, in this situation, it is becoming more crucial to explore the different grades of childhood obesity as the degree I, II and III obesity clearly for their clinical interventions using more detailed researches.

As previously described, the causation of childhood obesity are complex and multifaceted, presenting the clinicians with myriad challenges. Moreover, a key contributor of micro-nutrients is an significant element to prevent the later chronic diseases of children obesity by arising the energy metabolism, adipogenesis process, inflammation status and etc in the adipocytes. Hence, it is necessary to study the relationships between the micro-nutrients intake and obesity [9,10], in which vitamin D (VD) deficiency has been reported commonly among the Chinese children and adolescents [11,12]. Recently, many researches have confirmed that both VD insufficient intake and its decreased bio-availability have been associated with features of metabolic syndrome using the animal experiments by affecting the insulin resistance, endocannabinoid tone, glucose and lipid metabolism, gut dysbiosis, chronic inflammation, differentiation of adipocytes and etc [13], [14], [15], However, it is still unknown about the explanations for the increasing risks of VD deficiency in the development of obesity. Of note, more than 50% patients of metabolic diseases have low circulating levels of total and free 25-hydroxyvitamin D [25(OH)D], including the 25(OH)D2 and 25(OH)D3, and 1,25 dihydroxyvitamin D [1,25(OH)2D][16]. However, different cohorts have put forward the inconsistent effectiveness on the cutoff points for VD deficiency on the progress of obesity, incompatible VD metabolites (25(OH)D, 1,25(OH)2D) due to no consensus of cause-effect relationships, different ages and different grades of obesity and so on [17], [18], [19]. Meanwhile, there is few researches on the dose-responses and causal relationships between the concentrations of VD metabolites and different grades of obesity. To go further in determining the impacts of VD status on the progression of obesity and its related metabolic disorders, we implemented a longitudinal study of school-aged children aged 7-12 years with degree I, II and III obesity to explore the VD bio-availability (total VD, 25(OH)D and 1,25(OH)2D) and its correlations with the clinical characteristics, which could provide the appropriate basis for personalizing the VD interventions among the school-aged children with different grades of obesity.

Notably, there are two hydroxylation steps on catalyzing the VD into their biologically active metabolites (25(OH)D and 1,25(OH)2D) [17], [18], [19]. On the first step, the endogenous or dietary VD is enzymatically hydroxylated to 25(OH)D, which is catalyzed by CYP2R1 (vitamin D 25-hydroxylase) and CYP27A1 (Cholesterol 27α hydroxylase) as the key enzymes of 25 hydroxylation. On the second step, the VDBP-bound 25(OH)D is then transported to the kidneys and various other organs to be metabolized into 1,25(OH)2D by CYP27B1 (25-hydroxyvitamin D-1-α hydroxylase). Then 1,25(OH)2D is attached to the vitamin D receptor (VDR), as a transcription factor in the steroid hormone receptor super-family in the cytoplasm, which is transported into the nucleus and acts as the ligand-dependent transcription factor to regulate the target genes to exert their corresponding biological functions [18].
Therefore, the disorders of these VD metabolic pathway genes (CYP27A1, CYP2R1, CYP27B1, VDR and VDBP) in the two hydroxylation steps could be related with the decreases of 25(OH)D and 1,25(OH)2D during the occurrence of many diseases such as Rheumatoid Arthritis, tuberculosis, type 2 diabetes, cancer and so on [20], [21], [22], [23].
Moreover, it suggests that DNA methylation is one of the most common epigenetic mechanisms for the changes in gene expression. High methylation levels in the promoter region of VD metabolic pathway genes may cause gene silencing to affect vitamin D response variation [20], [21], [22], [23]. However, the relationships between the altered DNA methylation on these VD metabolic pathway genes and progress of obesity still need to be further verified. Therefore, the aim of this study was to test the hypothesis that obesity could disrupt the VD homeostasis via modulating the related gene expressions by DNA methylation to conversely aggravate the metabolic disorders among the school-aged children under the degree I, II and III obesity.

Section snippets
Selection of the subjects and subgroups
This representative cross-sectional study, including 106 obesity children who were adjusted by the body mass index (BMI) and 45 sex- and age-matched subjects without obesity as the control group, was carried out at Beijing Children's Hospital from July 2018 to April 2021 under the strict inclusion and exclusion criteria. Specifically, the criteria for participation were as follows: The inclusion criteria consisted that the subjects were 7-12 year healthy school-age children without obvious

Basic characteristics of the subjects in different degrees of obese children and controls
The basic characteristics of 151 participants (106 obese children and 45 healthy children) by the category of BMI were shown in Table 1. Overall, the number of no-obese children was 45 as the control group. While the 45 (42.45%), 34 (32.08%) and 27 (25.47%) obese children were respectively categorized as the degree I, II and III obesity subgroups. Comparing with the control group (weight: 55.02±2.34kg, BMI: 22.99±0.76kg/m2, waist hip ratio: 0.85±0.012, WAZ: 3.49±0.51 and BMI Z: 0.89±0.024), the

Discussion
Children obesity and its related disease complications reduce life quality and expectancy and increase health-care costs [27]. Many studies also have suggested that obesity always increases the risk of diabetes, liver injury cardiometabolic complications and even cancer with respect to the obesity-related metabolic dysfunctions [27]. Thus, the identification of individual metabolites and discrete pathways able to distinguish the different degrees of obesity with more refined studies help to

Funding
This study was supported by the National Natural Science Foundation of China (No. 82173524 to P.L.), Beijing Municipal Natural Science Foundation (No. 7174302 to P.L.) and Reform and Development Funds, Profession Quota Budget from Beijing Municipal Health Commission (bjsekyjs-yf to K.Q.).

Some of the 42 References

• H Jebeile et.al. Obesity in children and adolescents: epidemiology, causes, assessment, and management -Lancet Diabetes Endocrinol (2022)
• M de Onis et.al. Global prevalence and trends of overweight and obesity among preschool children - Am J Clin Nutr (2010)
• Y Dong et.al. Economic development and the nutritional status of Chinese school-aged children and adolescents from 1995 to 2014: an analysis of five successive national surveys - Lancet Diabetes Endocrinol (2019)
• C Iacobini et.al. Metabolically healthy versus metabolically unhealthy obesity - Metabolism (2019)
• S Kumar et.al. Review of Childhood Obesity: From Epidemiology, Etiology, and Comorbidities to Clinical Assessment and Treatment - Mayo Clin Proc (2017)
• HM Hussein et.al. Vitamin D mitigates diabetes-associated metabolic and cognitive dysfunction by modulating gut microbiota and colonic cannabinoid receptor 1 - Eur J Pharm Sci (2022)
• L Bonnet et.al. Diet induced obesity modifies vitamin D metabolism and adipose tissue storage in mice - J Steroid Biochem Mol Biol (2019)
• VI Fiamenghi et.al. Vitamin D deficiency in children and adolescents with obesity: a meta-analysis - J Pediatr (Rio J) (2021)
• A Bassatne et.al. Vitamin D supplementation in obesity and during weight loss: A review of randomized controlled trials - Metabolism (2019)
• M Wang et.al. Vitamin D and the promoter methylation of its metabolic pathway genes in association with the risk and prognosis of tuberculosis - Clin Epigenetics (2018)
• A Elagizi et.al. An Overview and Update on Obesity and the Obesity Paradox in Cardiovascular Diseases - Prog Cardiovasc Dis (2018)
• KF Faridi et.al. Serum vitamin D and change in lipid levels over 5y: The Atherosclerosis Risk in Communities study - Nutrition (2017)
• J Wortsman et.al. Decreased bioavailability of vitamin D in obesity - Am J Clin Nutr (2000)
• AG Need et.al. Effects of skin thickness, age, body fat, and sunlight on serum 25-hydroxyvitamin D - Am J Clin Nutr (1993)
• KJ Dick et.al. DNA methylation and body-mass index: a genome-wide analysis - Lancet (2014)
• Y Zhou et.al. DNA methylation levels of CYP2R1 and CYP24A1 predict vitamin D response variation - J Steroid Biochem Mol Biol (2014)
• DS Freedman et.al. Relationship of childhood obesity to coronary heart disease risk factors in adulthood: the Bogalusa Heart Study - Pediatrics (2001)
• AL Ghaben et.al. Adipogenesis and metabolic health - Nat Rev Mol Cell Biol (2019)
• J Laurencikiene et.al. Regulation of lipolysis in small and large fat cells of the same subject - J Clin Endocrinol Metab (2011)
• V Cifarelli et.al. Decreased adipose tissue oxygenation associates with insulin resistance in individuals with obesity - J Clin Invest (2020)
• JD Smith et.al. Prevention and Management of Childhood Obesity and Its Psychological and Health Comorbidities - Annu Rev Clin Psychol (2020)

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VitaminDWiki – Genetics category contains

343 articles in the Genetics category

see also

• Vitamin D Receptor has 530 items
• Vitamin D Binding Protein = GC has 178 items
• CYP27B1 has 63 items
• CYP24A1 in title of 39+ items
• CYP2R1 25+ items
• Calcidiol has 48 items
• Calcitriol has 62 items
• Topical Vitamin D
• Nanoemulsion Vitamin D may be a substantially better form
• 1289 genes changed with higher doses of Vitamin D - RCT Dec 2019
• CYP3A4 (7 as of Dec 2022)
• Getting Vitamin D into your body
• 

Vitamin D blood test misses a lot

• Vitamin D from coming from tissues (vs blood) was speculated to be 50% in 2014, and by 2017 was speculated to be 90%
• Note: Good blood test results (> 40 ng) does not mean that a good amount of Vitamin D actually gets to cells
• A Vitamin D test in cells rather than blood was feasible (2017 personal communication)   Commercially available 2019
  • However, test results would vary in each tissue due to multiple genes
• Good clues that Vitamin D is being restricted from getting to the cells
  1) A vitamin D-related health problem runs in the family
      especially if it is one of 51+ diseases related to Vitamin D Receptor
  2) Slightly increasing Vitamin D shows benefits (even if conventional Vitamin D test shows an increase)
  3) DNA and VDR tests - 100 to 200 dollars $100 to $250
  4) PTH bottoms out ( shows that parathyroid cells are getting Vitamin d)
     Genes are good, have enough Magnesium, etc.
  5) Back Pain
     probably want at least 2 clues before taking adding vitamin D, Omega-3, Magnesium, Resveratrol, etc
  • The founder of VitaminDWiki took action with clues #3&5

VitaminDWiki – Some diseases reduce vitamin D getting to blood or cells

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VitaminDWiki – Cancers might alter CYP24A1 gene

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VitaminDWiki – CYP27B1 category contains

The CYP27B1 gene activates Vitamin D in the Kidney,    Skin,    Lungs,    Brain,   Eyes   Breasts   etc.
Poor CYP27B1 is assocated with COVID, Miscarriage,   Lupus,   Alz, Parkinson, MSA,   Rickets

CYtochrome P450 family 27 subfamily B member 1    = 25-Hydroxyvitamin D3 1-alpha-hydroxylase

• 63 items in CYP27B1 category
• 343 articles in the Genetics
• 530 articles in Vitamin D Receptor
• 178 articles in Vitamin D Binding Protein
• CYP27B1 and other genes are less activated in seniors
• CYP27B causes many health problems – March 2020
• Every Parkinson’s brain had a poor CYP27B1 gene

What can be done if have a poor CYP27B1

• Larger doses of Vitamin D
• More Bio-available: Gut-friendly form, Topical form, taken with fatty meal, taken with evening meal
• Additional sources: UV
• Increase Vitamin D metabolism: additional Magnesium, Omega-3
  • All cytochrome P450 enzymes require Mg++ as a cofactor
• Increase the amount of Vitamin D in the blood that gets to cells: increase activation of VDR
  • See: Reasons for low response to vitamin D

Vitamin D blood test misses CYP27B1 and other genes

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VitaminDWiki – Nanoemulsion improves CYP27A1 gene activation