Exploring Interactions Between Retinoid and Vitamin D Signaling in Salivary Gland Homeostasis and Cancer

探索类视黄醇和维生素 D 信号在唾液腺稳态和癌症中的相互作用

• 批准号: 10388701
• 负责人: Kara A DeSantis
• 金额: $ 2.31万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10388701
• 关键词: Address; Adipocytes; Adult; Affect; Agonist; All-Trans-Retinol; Apoptosis; Archives; Area; Basal Cell; Breast; Cancer Model; Cell Differentiation process; Cell Fate Control; Cell Lineage; Cell Proliferation; Cell division; Cells; Cryoultramicrotomy; Cytokeratin; Data; Development; Ductal Epithelial Cell; Epithelial Cells; Genes; Homeostasis; Human; Immunohistochemistry; Incidence; Individual; Knock-out; Laboratories; Leukemic Cell; Ligands; Link; Malignant Neoplasms; Mammary Duct; Mammary Neoplasms; Monocytic leukemia; Mus; Myeloid Cells; Natural regeneration; Neurons; Nuclear; Nuclear Receptors; Oligodendroglia; Ontology; Organ; Osteoblasts; Oxides; Pathology; Pathway interactions; Patients; Pharmacologic Substance; Pharmacology; Physiological; Population; Prevention; Prognosis; Proliferation Marker; Property; Protein Isoforms; RNA; Receptor Signaling; Regulation; Reporter; Research; Retinoic Acid Receptor; Retinoids; Role; Salivary; Salivary Gland Neoplasms; Salivary Gland Tissue; Salivary Glands; Sampling; Signal Pathway; Signal Transduction; Skeletal Muscle; Squamous Cell; System; Testing; Tissue Microarray; Tissues; Tretinoin; Vitamin A; Vitamin D; Vitamin D3 Receptor; Vitamins; Work; acute myeloid leukemia cell; aldehyde dehydrogenases; cancer stem cell; cell motility; dimethylbenzanthracene; druggable target; granulocyte; human disease; human population study; irradiation; malignant breast neoplasm; mammary; mammary epithelium; neoplastic cell; novel therapeutic intervention; receptor; receptor expression; refractory cancer; retinoic acid receptor alpha; salivary cell; small molecule; stem cells; synergism; targeted delivery; three dimensional cell culture; transcriptome sequencing; tumor; tumorigenesis; tumorigenic

Abstract: Basal cell derived salivary and breast cancers are correlated with poor patient prognosis, increased cancer reoccurrence and lack of definitive therapies 1. In both branching organs, cytokeratin 5-positive (K5+) basal cells contribute to ductal cells during normal development 2–4 and recent research has highlighted similarities in the K5+ basal cell origin of mammary and salivary squamous cells 5,6. In mammary tumors of basal origin, irradiation- resistant cancer stem cells (CSCs) demonstrate K5 positivity 1. Both Vitamin D and Vitamin A signaling pathways have been implicated in prevention of tumorigenesis 7,8. In the developing 9 salivary gland, we and others 10 have recently demonstrated that retinoic acid receptor (RAR) signaling contributes to expansion of K5+ basal cells, a phenomena conserved in the adult salivary gland 11. In both mammary 12 and salivary glands 13, CSCs display an upregulated level of ALDH, an aldehyde dehydrogenase that oxidizes Vitamin A to retinol, the precursor to retinoic acid and signaling through RARs. The relationship between RAR signaling and K5+ basal cells during tumorigenesis has yet to be explored. Work in the Welsh laboratory has shown that deletion of the Vitamin D receptor (VDR) in mice leads to increased cell proliferation and tumorigenesis in the breast 7 where VDR activity is linked to K5+ cell fate transition 4. In human population studies, low levels of Vitamin D are associated with increased cancer incidence and progression 14–16. In the submandibular salivary gland (SMG), progenitor cell populations are highly positive for VDR 17 and in humans, VDR localizes to epithelial cells 18; yet, Vitamin D signaling has not been specifically explored in the salivary gland. We will test the hypothesis that co- activating VDR and RAR signaling will reduce K5+ basal cell expansion and tumorigenesis in the SMG. In this proposal we will address the following questions: 1) What is the role of VDR signaling in K5+ basal cell expansion and differentiation in the SMG? 2) Does VDR signaling synergize with RARα signaling to negatively regulate K5+ basal cell expansion in the SMG? 3) Will loss of VDR signaling synergize with inhibition of RARα signaling to enhance K5+ tumorigenesis in the SMG? 4) Will activation of VDR and RARα signaling in K5- TdTomato-positive tumor cells using directed delivery of vitamin derivatives and small molecules decrease tumorigenic cell properties? To determine if RARα and VDR signaling synergize in negatively regulating the K5+ population in the SMG, in Aim 1 we will inhibit RARα signaling in a VDR knockout K5 reporter mouse and trace K5+ basal cell fate. In Aim 2, we will determine if VDR and RARα pathways synergize to regulate K5+ cell expansion in basal salivary tumors. This aim will be achieved with targeted pharmacological inhibition of RARα signaling in a VDR knockout K5 reporter mouse induced for tumorigenesis with DMBA. Additionally, we will determine if activation of VDR and RARα signaling in K5-TdTomato-positive tumor cells using small molecules and vitamin derivatives in 3D culture will affect cell division and motility.

摘要： 基底细胞来源的唾液腺癌和乳腺癌与患者预后差、癌症增加相关。 复发和缺乏明确的治疗1。在两个分支器官中，细胞角蛋白5阳性（K5+）基底细胞 在正常发育过程中对导管细胞有贡献2-4，最近的研究强调了 K5+乳腺和唾液鳞状细胞的基底细胞起源5，6。在基底部乳腺肿瘤中，照射- 耐药癌症干细胞（CSC）显示K5阳性1。维生素D和维生素A信号通路 与预防肿瘤发生有关7，8。在发育中的9个唾液腺中，我们和其他10个有 最近证实，视黄酸受体（RAR）信号有助于K5+基底细胞的扩增， 在成人唾液腺中保守的现象11.在乳腺12和唾液腺13两者中，CSC显示出与乳腺12和唾液腺13相似的免疫反应。 ALDH水平上调，ALDH是一种将维生素A氧化为视黄醇的醛脱氢酶， 视黄酸和通过RAR的信号传导。RAR信号通路与K5+基底细胞凋亡的关系 肿瘤发生还有待探索。威尔士实验室的工作表明， 小鼠中的VDR受体（VDR）导致乳腺中细胞增殖和肿瘤发生增加7，其中VDR活性 与K5+细胞命运转换4有关。在人群研究中，低水平的维生素D与 增加癌症发病率和进展14-16.在下颌下腺（SMG），祖细胞 人群对VDR 17高度阳性，并且在人类中，VDR定位于上皮细胞18;然而，维生素D 在唾液腺中还没有专门研究信号传导。我们将测试假设，共同- 激活VDR和RAR信号传导将减少SMG中的K5+基底细胞扩增和肿瘤发生。 在本研究中，我们将解决以下问题：1）VDR信号在K5+基底细胞中的作用 在SMG的扩张和分化？2)VDR信号是否与RARα信号协同作用， 调节SMG中K5+基底细胞扩增？3)VDR信号的丢失是否会与RARα的抑制协同作用 在SMG中增强K5+肿瘤发生的信号传导？4)在K5-HT细胞中，VDR和RARα信号转导的激活是否会影响细胞的增殖？ 使用维生素衍生物和小分子的定向递送的TdTomato阳性肿瘤细胞减少 致瘤细胞的特性为了确定RARα和VDR信号传导是否协同负调节K5+， 在目标1中，我们将在VDR敲除K5报告基因小鼠中抑制RARα信号传导，并跟踪 K5+基底细胞命运。在目的2中，我们将确定VDR和RARα通路是否协同调节K5+细胞， 基底部唾液腺肿瘤的扩张。这一目标将通过靶向药理学抑制RARα来实现 在用DMBA诱导肿瘤发生的VDR敲除K5报告小鼠中的信号传导。此外，我们将 确定是否使用小分子在K5-TdTomato阳性肿瘤细胞中激活VDR和RARα信号传导 而维生素衍生物在3D培养中会影响细胞分裂和运动。