Defining the role of Taiman, the Drosophila homolog of AIB1, as a super-competitor in developing epithelia

定义 Taiman（AIB1 的果蝇同源物）作为上皮细胞发育中超级竞争者的作用

• 批准号: 10626071
• 负责人: Colby Kristina Schweibenz
• 金额: $ 4.77万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10626071
• 关键词: Affinity Chromatography; Alleles; Apoptosis; Binding; Biochemical; Biological Assay; Breast; Cancer Biology; Cell Survival; Cells; Cessation of life; Classification; Complex; Data; Development; Diagnosis; Disease; Dose; Drosophila genus; Drosophila melanogaster; Ecdysone; Elements; Embryo; Ensure; Environment; Epiblast; Epithelial Cells; Epithelium; Estrogens; Genes; Genetic Transcription; Goals; Growth; Homologous Gene; Hormones; Human; Immunofluorescence Immunologic; Interruption; Invaded; Knowledge; Learning; Ligands; Link; Localized Malignant Neoplasm; MYC gene; Malignant Neoplasms; Measures; Mediating; Messenger RNA; Methods; Modeling; Modification; Molecular; NCOA3 gene; Neoplasm Metastasis; Nuclear Hormone Receptors; Oncogene Activation; Oncogenes; Outcome; Pathway interactions; Population; Process; Production; Progesterone; Proliferating; Proteins; Proteome; Publishing; RNA Interference; Receptor Signaling; Research; Ribosomes; Role; Scientist; Signal Transduction; System; Testing; Therapeutic; Tissues; Transcription Coactivator; Translating; Tumor Suppressor Proteins; United States; Wing; Woman; Work; breast cancer progression; cancer cell; cancer therapy; cell type; cofactor; cost; design; dosage; ecdysone receptor; effective therapy; experimental study; fitness; genetic variant; hormonal signals; insight; mRNA Translation; malignant breast neoplasm; model organism; novel; novel therapeutics; overexpression; pro-apoptotic protein; receptor; statistics; steroid hormone; steroid hormone receptor; tool

PROJECT SUMMARY Invasive breast cancer is a deadly disease, killing ~42,000 women each year and costing the United States over $16.5 billion annually. The complicated disease mechanisms that drive breast cancer are still insufficiently understood to confidently design therapeutics. One mechanism emerging as a potential driver for local invasion in breast and other cancers is ‘cell competition.’ This phenomenon occurs when two different cell populations with different ‘fitness’ levels are juxtaposed in the same tissue. High fitness cells (called ‘winners’) grow more rapidly and kill off slower growing neighbors (called ‘losers’) by apoptosis. Cancer cells acquire ‘winner’ status by activating oncogenes. Oncogenes that confer ‘winner’ status are called ‘super-competitors.’ Our lab uses the model organism, Drosophila melanogaster, to study conserved growth and proliferation pathways that are altered in human cancer. In previous work we showed that Drosophila Taiman (Tai, AIB1 in humans), a co-activator of the Ecdysone steroid hormone receptor (EcR), is a candidate super-competitor and imparts ‘winner’ status to cells via production of the secreted, pro-apoptotic protein Spätzle (Spz), a Toll receptor ligand. We have also shown that Tai binds the Yorkie (Yki) coactivator protein, the main target of the Hippo tumor suppressor pathway, and that Yki:Tai collaboratively drive expression of pro-growth genes. However, we do not fully understand how Tai drives neighbor killing, or whether it requires interactions with EcR and/or Yki. Moreover, classifying Tai as a ‘super-competitor’ requires evidence that lowering the Tai dose confers ‘loser’ status relative to wildtype cells. In the following three Aims, I will test my hypothesis that Tai acts as a super-competitor through either its interaction with Yki (Hippo pathway) or EcR (ecdysone pathway), test the relative fitness of Tai-overexpressing (Taihigh), wildtype (Taiwt), and Tai hypomorphic (Tailow) cells, and carry out experiments to identify cell competition factors regulated by Tai. In Aim 1, I will assess the effects of elevated or reduced Tai on cell survival in homotypic vs. heterotypic environments. To link to our published data, I will also investigate the requirement of the Spz/Toll pathway in these contexts. In Aim 2, I will use candidate-based approaches to test the requirements for Yki/Hippo and EcR signaling in Tai-driven killing of neighbor cells. Finally in Aim 3, I will use the unbiased, discovery-based method Translating Ribosome Affinity Purification with sequencing (TRAP-Seq) to identify the Tai-induced translated proteome in wing cells and candidate ‘competition’ factors within it. These aims will define the molecular mechanisms underlying Tai-regulated cell competition in epithelial tissue. This work could also reveal a link between cell competition and steroid hormone signaling, which could be a novel element of cancer biology. The pathways that will be uncovered by this work will have long-term impacts, as knowledge of cell competition mechanisms could be applied to mammalian models to learn more about cancer emergence and progression. Specifically, knowledge gained from this research could lead to greater understanding of the role of the human homolog AIB1 in breast cancer and lead to the development of potential new therapeutics.

项目摘要 侵入性乳腺癌是一种致命的疾病，每年造成约42,000名女性，使美国造成 每年165亿美元。驱动乳腺癌的复杂疾病机制仍然不足 理解自信设计理论。一种机制作为当地入侵的潜在驱动力 在乳腺癌和其他癌症中是“细胞竞争”。当两个不同的细胞种群时，就会发生这种现象 具有不同的“适应性”水平在同一组织中并置。高健身细胞（称为“获奖者”）成长更多 迅速地杀死了凋亡的缓慢成长的邻居（称为“失败者”）。癌细胞获得“获胜者”状态 通过激活癌基因。会议“获胜者”身份的致癌基因称为“超级竞争者”。我们的实验室使用 模型有机体，果蝇Melanogaster，以研究改变的保守生长和增殖途径 在人类癌症中。在以前的工作中，我们表明果蝇taiman（人类的AIB1，AIB1） ecdysone类固醇马酮受体（ECR）是候选超级竞争者，并将“获胜者”身份授予 通过生产分泌的促凋亡蛋白Spätzle（SPZ）的细胞，Toll受体配体。我们也有 表明TAI结合了约克（YKI）共激活因子蛋白，这是河马肿瘤抑制途径的主要目标， YKI：TAI合作推动了促增长基因的表达。但是，我们不完全了解 太极驾驶邻居杀人，还是需要与ECR和/或YKI进行互动。此外，将泰归类为 “超级竞争者”需要证据表明，相对于野生型细胞，降低了TAI剂量贡献“失败者”状态。 在以下三个目标中，我将测试我的假设，即太极通过它是超级竞争者 与YKI（河马途径）或ECR（ECDYSONE途径）的相互作用，测试Tai-Over表达的相对适应性 （Taihigh），Wildtype（Taiwt）和Tai hydolodlic（Tailow）细胞，并进行实验以鉴定细胞竞争 由太极监管的因素。在AIM 1中，我将评估TAI升高或降低TAI对同型细胞存活的影响 与异型环境。要链接到我们已发布的数据，我还将调查SPZ/TOLL的要求 在这些情况下的途径。在AIM 2中，我将使用基于候选的方法来测试YKI/HIPPO的要求 在邻居细胞中杀死泰驱动的ECR信号传导。最后，在AIM 3中，我将使用公正的，基于发现的基于发现的 用测序（TRAP-SEQ）翻译核糖体亲和力纯化以识别Tai诱导的 在其中翻译了机翼细胞中的蛋白质组和其中的候选“竞争”因素。这些目标将定义 TAI调节细胞竞争的分子机制在上皮组织中。这项工作也可以揭示 细胞竞争与类固醇骑马信号传导之间的联系，这可能是癌症生物学的新元素。 随着细胞竞争的了解，这项工作将会带来的途径将产生长期影响 机制可以应用于哺乳动物模型，以了解有关癌症出现和进展的更多信息。 具体而言，从这项研究中获得的知识可能会进一步了解人类的作用 同源物AIB1在乳腺癌中，导致潜在的新疗法的发展。