Rdh10 and retinoic acid effects on differentiation

Rdh10 和视黄酸对分化的影响

基本信息

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
项目类别：
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

项目摘要

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) 催化全反式视黄酸 (atRA) 的第一步和限速步骤通过生物合成视黄醇生成视黄醛，视黄醛脱氢酶将其转化为 atRA。大多数Rdh10 研究重点是视觉周期或胚胎发育。在产后方面所做的工作有限 Rdh10 函数。然而，atRA 控制间充质干细胞 (MSC) 分化为脂肪细胞与分化为脂肪细胞的命运。成人中的成骨细胞调节肥胖和骨骼健康。 atRA 指导 MSC 的作用机制分化和 atRA 生物合成的调节仍不清楚。文献中包含相互矛盾的内容报告了 atRA 和其他分化刺激调节的主要靶基因。大多数 atRA 研究脂肪细胞与成骨细胞之间的平衡调节已经在细胞系中完成，并且重点关注单个基因通常在分化后期控制一个干细胞的命运。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，因为细胞系确实不能忠实地模拟原代细胞，彼此不同，甚至具有相同标称名称的细胞系往往由于长期文化的影响而有所不同。该提案通过以下方式解决 atRA 指导 MSC 命运的功能：提议研究 atRA 不足的体内模型，该模型将伴随产生初级小鼠胚胎成纤维细胞产生全面的机制见解来确定脂肪细胞与脂肪细胞成骨细胞的命运。体内模型依赖于具有表型的杂合子消融 Rdh10 (Rdh10+/-) 生命早期骨髓中的肥胖和脂肪细胞增殖增加，并且钙化减少成骨细胞。 Rdh10+/- 模型（纯合子是胚胎致死的）提供一致的减毒维生素 A 在体内通过可重复还原的 atRA 发挥作用。这是一种创新模式，将提供新的深入了解 atRA 调节脂肪细胞与成骨细胞分化的功能，以及新的见解 Rdh10 功能和 atRA 生物发生。了解 Rdh10 和 atRA 在肥胖和肥胖中的作用脂肪生成有望直接应用于人类健康。这一期望得到了二十六条的支持与体重指数和体重增加相关的基因间特征与人类 Rdh10 接近。目标1将确定减少 Rdh10 表达和 atRA 浓度的表型和代谢后果体内（Rdh10+/- 小鼠），重点关注脂肪细胞和成骨细胞分化。该目标将检验假设 Rdh10 是控制 atRA 稳态以调节脂肪细胞中类视黄醇功能的主要酶，成骨细胞分化。目标 2 将确定 Rdh10+/- 小鼠表型背后的机制尊重细胞命运的决定。该目标将使用两组 MEF：1) 来自 Rdh10+/- 小鼠； 2）永垂不朽完全敲除 Rdh10 的 MEF 来测试 Rdh10 生成的 atRA 指导成熟的假设 MSC很早就通过调节基因表达来决定细胞命运，并且会识别出反应基因。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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
影响因子：
0
作者：
J. L. Napoli
通讯作者：
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;
通讯作者：

Retinoic Acid: The Autacoid for All Seasons.