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      A preliminary transcriptome analysis suggests a transitory effect of vitamin D on mitochondrial function in obese young Finnish subjects

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          Abstract

          Objective

          The effect of vitamin D at the transcriptome level is poorly understood, and furthermore, it is unclear if it differs between obese and normal-weight subjects. The objective of the study was to explore the transcriptome effects of vitamin D supplementation.

          Design and methods

          We analysed peripheral blood gene expression using GlobinLock oligonucleotides followed by RNA sequencing in individuals participating in a 12-week randomised double-blinded placebo-controlled vitamin D intervention study. The study involved 18 obese and 18 normal-weight subjects (of which 20 males) with mean (± s.d.) age 20.4 (±2.5) years and BMIs 36 (±10) and 23 (±4) kg/m 2, respectively. The supplemental daily vitamin D dose was 50 µg (2000 IU). Data were available at baseline, 6- and 12-week time points and comparisons were performed between the vitamin D and placebo groups separately in obese and normal-weight subjects.

          Results

          Significant transcriptomic changes were observed at 6 weeks, and only in the obese subjects: 1724 genes were significantly upregulated and 186 genes were downregulated in the vitamin D group compared with placebo. Further analyses showed several enriched gene categories connected to mitochondrial function and metabolism, and the most significantly enriched pathway was related to oxidative phosphorylation (adjusted P value 3.08 × 10 −14). Taken together, our data suggest an effect of vitamin D supplementation on mitochondrial function in obese subjects.

          Conclusions

          Vitamin D supplementation affects gene expression in obese, but not in normal-weight subjects. The altered genes are enriched in pathways related to mitochondrial function. The present study increases the understanding of the effects of vitamin D at the transcriptome level.

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          Most cited references24

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          Physiology of leptin: energy homeostasis, neuroendocrine function and metabolism.

          Leptin is secreted by adipose tissue and regulates energy homeostasis, neuroendocrine function, metabolism, immune function and other systems through its effects on the central nervous system and peripheral tissues. Leptin administration has been shown to restore metabolic and neuroendocrine abnormalities in individuals with leptin-deficient states, including hypothalamic amenorrhea and lipoatrophy. In contrast, obese individuals are resistant to leptin. Recombinant leptin is beneficial in patients with congenital leptin deficiency or generalized lipodystrophy. However, further research on molecular mediators of leptin resistance is needed for the development of targeted leptin sensitizing therapies for obesity and related metabolic diseases. Copyright © 2015 Elsevier Inc. All rights reserved.
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            Highly multiplexed and strand-specific single-cell RNA 5' end sequencing.

            Single-cell analysis of gene expression is increasingly important for the analysis of complex tissues, including cancer, developing organs and adult stem cell niches. Here we present a detailed protocol for quantitative gene expression analysis in single cells, by the sequencing of mRNA 5' ends. In all, 96 cells are lysed, and their mRNA is converted to cDNA. By using a template-switching mechanism, a bar code and an upstream primer-binding sequence are introduced simultaneously with reverse transcription. All cDNA is pooled and then prepared for 5' end sequencing, including fragmentation, adapter ligation and PCR amplification. The chief advantage of this approach is the great reduction in cost and time, afforded by the early bar-coding strategy. Compared with previous methods, it is more suitable for large-scale quantitative analysis, as well as for the characterization of transcription start sites, but it is unsuitable for the detection of alternatively spliced transcripts. Sample preparation takes 3 d, and two sets of 96 cells can be prepared in parallel. Finally, the sequencing and data analysis can take an additional 4 d altogether.
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              Relationships between fat and bone.

              I. D. Reid (2008)
              Body weight impacts both bone turnover and bone density, making it, therefore, an important risk factor for vertebral and hip fractures and ranking it alongside age in importance. The effect of body weight is probably contributed to by both fat mass and lean mass, though in postmenopausal women, fat mass has been more consistently demonstrated to be important. A number of mechanisms for the fat-bone relationship exist and include the effect of soft tissue mass on skeletal loading, the association of fat mass with the secretion of bone active hormones from the pancreatic beta cell (including insulin, amylin, and preptin), and the secretion of bone active hormones (e.g., estrogens and leptin) from the adipocyte. These factors alone probably do not fully explain the observed clinical associations, and study of the actions on bone of novel hormones related to nutrition is an important area of further research. An understanding of this aspect of bone biology may open the way for new treatments of osteoporosis. More immediately, the role of weight maintenance in the prevention of osteoporosis is an important public health message that needs to be more widely appreciated.
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                Author and article information

                Journal
                Endocr Connect
                Endocr Connect
                EC
                Endocrine Connections
                Bioscientifica Ltd (Bristol )
                2049-3614
                May 2019
                09 April 2019
                : 8
                : 5
                : 559-570
                Affiliations
                [1 ]Folkhälsan Institute of Genetics , University of Helsinki, Helsinki, Finland
                [2 ]Molecular Neurology Research Program , University of Helsinki, Helsinki, Finland
                [3 ]Department of Biosciences and Nutrition , Karolinska Institutet, Huddinge, Sweden
                [4 ]Children’s Hospital , University of Helsinki and Helsinki University Hospital, Helsinki, Finland
                [5 ]Competence Centre on Health Technologies , Tartu, Estonia
                [6 ]School of Basic and Medical Biosciences , King’s College London, Guy’s Hospital, London, United Kingdom
                [7 ]Department of Molecular Medicine and Surgery and Center for Molecular Medicine , Karolinska Institutet, Stockholm, Sweden
                [8 ]Department of Clinical Genetics , Karolinska University Hospital, Stockholm, Sweden
                [9 ]Department of Food and Environmental Sciences , University of Helsinki, Helsinki, Finland
                Author notes
                Correspondence should be addressed to H Viljakainen: heli.viljakainen@ 123456helsinki.fi

                (E Einarsdottir is now at Department of Gene Technology, Science for Life Laboratory, KTH-Royal Institute of Technology, Solna, Sweden)

                Article
                EC-18-0537
                10.1530/EC-18-0537
                6499919
                30965285
                ea6540ca-4226-469a-ad0d-5aa563441ea5
                © 2019 The authors

                This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

                History
                : 04 April 2019
                : 09 April 2019
                Categories
                Research

                vitamin d,gene expression,obesity,transcriptome,mitochondrial function,intervention

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