Rdh10 and retinoic acid effects on differentiation

Rdh10 和视黄酸对分化的影响

• 批准号: 10217113
• 负责人: JOSEPH L NAPOLI
• 金额: $ 37.61万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-18 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10217113
• 关键词: 11 cis Retinal; Ablation; Address; Adipocytes; Adult; All-Trans-Retinol; Anabolism; Animals; Astrocytes; Attenuated; BMP2 gene; Biogenesis; Body mass index; Bone Marrow; Cell Line; Cell Nucleus; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Conflict (Psychology); Dendritic Cells; Diet; Embryo; Embryonic Development; Enzymes; Equilibrium; Event; FOXO1A gene; Fat-Restricted Diet; Female; Fibroblasts; Gene Abnormality; Gene Expression; Genes; Health; Heterozygote; High Fat Diet; Homeostasis; Homozygote; Hormones; Human; Insulin; Knock-out; Life; Literature; Maps; Mesenchymal; Mesenchymal Stem Cells; Messenger RNA; Metabolic; Modeling; Mus; Nuclear; Obesity; Osteoblasts; Osteoporosis; Outcome; Pathology; Pharmacologic Actions; Phenotype; Physiological; Reaction; Reaction Time; Regulation; Reporting; Research; Retina; Retinal dehydrogenase; Retinoids; Retinol dehydrogenase; Role; Skin; Stimulus; Testing; Testis; Time; Tretinoin; Vitamin A; Weight Gain; Work; adipocyte differentiation; age related; aged; bone; bone health; calcification; energy balance; established cell line; expectation; health application; in vivo; in vivo Model; innovation; insight; lipid biosynthesis; lipid metabolism; male; osteoblast differentiation; postnatal; programs; stem cell differentiation; stem cell fate; stem cells; trait; visual cycle

Retinol dehydrogenase 10 (Rdh10) catalyzes the first and rate-limiting step in all-trans-retinoic acid (atRA) biosynthesis from retinol by generating retinal, which retinal dehydrogenases convert into atRA. Most Rdh10 research has focused on the visual cycle or embryonic development. Limited work has been done on postnatal Rdh10 function. Yet, atRA controls the fate of mesenchymal stem cell (MSC) differentiation into adipocytes vs osteoblasts in the adult to regulate adiposity and bone health. Mechanisms of atRA action in directing MSC differentiation remain unclear as does regulation of atRA biosynthesis. The literature contains conflicting reports as to the primary target genes regulated by atRA and other differentiation stimuli. Most studies of atRA regulation of the balance between adipocytes vs osteoblasts has been done in cell lines, and has focused on single genes controlling in one stem cell fate, often later during differentiation. The use of cell lines (e.g. 3T3- L1, F442A, C2C12, MC3T3-E1) most likely has contributed to the conflicting conclusions, because cell lines do not faithfully model primary cells, differ from each other, and even cell lines with the same nominal designation often differ as a result of long-term culture. This proposal addresses atRA function in directing MSC fate by proposing study of an in vivo model of atRA insufficiency, which will be accompanied by generating primary mouse embryonic fibroblasts to produce comprehensive mechanistic insight into determining adipocyte vs. osteoblast cell fates. The in vivo model relies on heterozygote ablated Rdh10 (Rdh10+/-) that has a phenotype of increased adiposity and adipocyte proliferation in bone marrow early in life, with decreased calcification of osteoblasts. The Rdh10+/- model (the homozygote is embryonic lethal) provides for consistently attenuated vitamin A function in vivo through reproducibly reduced atRA. This is an innovative model that will provide new insight into atRA function concerning regulating differentiation of adipocytes vs osteoblasts, and new insight into Rdh10 function and atRA biogenesis. Understanding the roles of Rdh10 and atRA in obesity and adipogenesis promises direct human health applications. This expectation is supported by the twenty-six intergenic traits related to body mass index and weight gain that map close to human Rdh10. Aim 1 will determine the phenotype and metabolic consequences of reducing Rdh10 expression and atRA concentrations in vivo (Rdh10+/- mice), focusing on adipocyte and osteoblast differentiation. This aim will test the hypothesis that Rdh10 is a major enzyme that controls atRA homeostasis to regulate retinoid function in adipocyte and osteoblast differentiation. Aim 2 will determine mechanism(s) that underlie the phenotype of Rdh10+/- mice with respect to cell fate determination. This aim will use two sets of MEF: 1) from Rdh10+/- mice; 2) immortalized MEF with a total knockout of Rdh10 to test the hypothesis that atRA generated by Rdh10 directs maturation of MSC by regulating gene expression very early in cell fate determination, and will identify the responsive genes.

视黄醇脱氢酶10（RDH10）催化了全反式反激酸（ATRA）中的第一个和速率限制步骤 视网膜产生视网膜的生物合成，视网膜脱氢酶转化为ATRA。大多数RDH10 研究的重点是视觉周期或胚胎发育。有限的工作是在产后完成的 RDH10功能。然而，ATRA控制间充质干细胞（MSC）的命运分化为脂肪细胞与 成年人的成骨细胞调节肥胖和骨骼健康。 ATRA动作指导MSC的机制 分化尚不清楚，就像对ATRA生物合成的调节一样。文献包含矛盾 关于由ATRA和其他分化刺激调节的主要靶基因的报告。大多数ATRA研究 调节脂肪细胞与成骨细胞之间的平衡已在细胞系中进行，并集中在 在一个干细胞命运中控制的单个基因，通常在分化过程中经常。使用细胞系（例如3T3- L1，F442A，C2C12，MC3T3-E1）很可能导致了相互矛盾的结论，因为细胞系DO 不忠实地对主要单元格建模，彼此不同，甚至具有相同名称的细胞系 长期文化通常会有所不同。该建议涉及指导MSC命运的ATRA功能 提出研究ATRA功能不全的体内模型，该模型将伴随 小鼠胚胎成纤维细胞可为确定脂肪细胞VS产生全面的机械洞察力。 成骨细胞命运。体内模型依赖于具有表型的杂合子烧蚀的RDH10（RDH10 +/-） 生命早期骨髓中肥胖和脂肪细胞增殖的增加，钙化降低 成骨细胞。 RDH10 +/-型号（纯合子是胚胎致死）提供了一致减弱的 维生素A通过可重复降低的ATRA在体内功能。这是一个创新的模型，将提供新的 洞悉有关调节脂肪细胞与成骨细胞的分化的ATRA功能和新洞察力 进入RDH10功能和ATRA生物发生。了解RDH10和ATRA在肥胖和 脂肪形成有望直接人类健康应用。这种期望得到了二十六个 与体重指数和体重增加有关的基因间特征，该图与人RDH10接近。目标1意志 确定降低RDH10表达和ATRA浓度的表型和代谢后果 体内（RDH10 +/-小鼠），重点是脂肪细胞和成骨细胞分化。这个目标将检验假设 该RDH10是一种主要酶，控制ATRA稳态以调节脂肪细胞和 成骨细胞分化。 AIM 2将确定带有RDH10 +/-小鼠表型的机制 尊重细胞命运的确定。此目标将使用RDH10 +/-鼠标的两组MEF：1）； 2）永生 MEF总敲除RDH10的MEF测试了RDH10产生的ATRA的假设指导成熟 MSC通过在细胞命运确定的早期调节基因表达，并将鉴定响应基因。

项目成果

Energy status regulates levels of the RAR/RXR ligand 9-cis-retinoic acid in mammalian tissues: Glucose reduces its synthesis in β-cells.

• DOI: 10.1016/j.jbc.2023.105255
• 发表时间: 2023-10
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Yoo, Hong Sik;Moss, Kristin Obrochta;Cockrum, Michael A.;Woo, Wonsik;Napoli, Joseph L.
• 通讯作者: Napoli, Joseph L.

Cyp26a1 supports postnatal retinoic acid homeostasis and glucoregulatory control.

• DOI: 10.1016/j.jbc.2023.104669
• 发表时间: 2023-05
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Yoo, Hong Sik;Cockrum, Michael A.;Napoli, Joseph L.
• 通讯作者: Napoli, Joseph L.

Post-natal all-trans-retinoic acid biosynthesis.

• DOI: 10.1016/bs.mie.2020.02.003
• 发表时间: 2020
• 期刊: Methods in enzymology
• 作者: J. L. Napoli

Examination of Fluconazole-Induced Alopecia in an Animal Model and Human Cohort.

在动物模型和人类队列中检查氟康唑引起的脱发。

• DOI: 10.1128/aac.01384-18
• 发表时间: 2019
• 期刊: Antimicrobial agents and chemotherapy
• 影响因子: 4.9
• 作者: Thompson3rd,GeorgeR;Krois,CharlesR;Affolter,VerenaK;Everett,AngelaD;Varjonen,EKatarina;Sharon,VictoriaR;Singapuri,Anil;Dennis,Michael;McHardy,Ian;Yoo,HongSik;Fedor,DawnM;Wiederhold,NathanP;Aaron,PhyliciaA;Gelli,Angie

Retinoic Acid: The Autacoid for All Seasons.

• DOI: 10.3390/nu14214526
• 发表时间: 2022-10-27
• 期刊: Nutrients
• 影响因子: 5.9
• 作者: Napoli JL