Project 1- Micro-RNA-dependent signaling by the UPR

摘要 快速增殖的癌细胞必须在微环境中茁壮成长，其中代谢营养物， 当肿瘤体积扩大超过既定的体积时，葡萄糖、氧气和生长因子变得有限。 组织的血管分布。在正常细胞中，营养可用性的限制触发生长停滞和/或凋亡 从而防止细胞在这种条件下扩张。我们的科学前提是， PERK的活性反映了蛋白质合成的PERK依赖性差异控制的综合影响 和基因表达。在上一个资助周期中，我们将miR-211和miR-216 b鉴定为PERK-ATF 4。 调节细胞存活的微RNA。我们确定了三个至关重要的miR-211靶点。 第一个是促凋亡转录因子Chop。第二个靶标是Bmal 1-Clock异二聚体 作为昼夜节律基因表达的主要驱动因子， 振荡Bmal 1和Clock都是miR-211的直接靶点，在细胞凋亡过程中被miR-211抑制。 普遍定期审议。通过分析PERK-miR-211对Bmal 1和生物钟的调节，我们发现， 1)昼夜节律基因表达被UPR破坏，2）Bmal 1缺失有助于PERK依赖性的 mRNA翻译的调控。我们提出了一个假设，其中UPR诱导的micro-RNA（miR- 211，miR-217）通过沉默关键靶点如Bmal 1来调节细胞命运，从而有助于PERK-1的表达。 依赖的翻译调节和细胞存活。为了解决这个问题，我们提出了三个具体的建议。 目标。目标1，将定义PERK-miR 211-Bmal 1调节的关键靶点及其对细胞的贡献 生存目的2将确定miR-217在拮抗miR-211对其转录调控中的作用， 目标的目的3将明确miR-211依赖性调节Bmal 1对发育和分化的作用。 B细胞淋巴瘤的进展这个建议和项目2之间有明显的交叉点 其重点是UPR如何拮抗myc诱导的细胞凋亡和ATF 4依赖的调节， 肿瘤细胞代谢通过与项目2的合作，我们将评估miR-211和Bmal 1的作用。 作为肿瘤细胞代谢和存活的介质。通过与项目3的合作，我们将 确定miR-211、miR-217和Bmal 1如何调节IFNAR 1信号传导和细胞质T细胞中的调节。 淋巴细胞本文所提出的工作结果将为设计 新的抗癌治疗剂。

Abstract Rapidly proliferating cancer cells must thrive in a microenvironment wherein metabolic nutrients such as glucose, oxygen and growth factors become limiting as tumor volume expands beyond the established vascularity of the tissue. In normal cells, limits in nutrient availability trigger growth arrest and/or apoptosis thereby preventing cellular expansion under such conditions. Our scientific premise is that the pro-survival activities of PERK reflect the combined impact of PERK-dependent differential control of protein synthesis and gene expression. In the previous funding cycle, we identified miR-211 and miR-216b as a PERK-ATF4 regulated micro-RNAs that regulate cell survival. We identified three critically important miR-211 targets. The first is the pro-apoptotic transcription factor, Chop. The second target is the Bmal1-Clock heterodimeric transcription factor that functions as the primary driver of circadian gene expression and thus circadian oscillation. Both Bmal1 and Clock are direct targets of miR-211 and are repressed by miR-211 during a UPR. Through analysis of PERK-miR-211 regulation of Bmal1 and the circadian clock, we discovered that 1) circadian gene expression is disrupted by the UPR, 2) Bmal1 loss contributes to PERK-dependent regulation of mRNA translation. We propose a hypothesis wherein the UPR inducible micro-RNAs (miR- 211, miR-217) regulate cell fate by silencing key targets such as Bmal1 which thereby contributes to PERK- dependent translational regulation and cell survival. To address this hypothesis, we propose three specific aims. Aim 1, will defined the critical targets of PERK-miR211-Bmal1 regulation and their contribution to cell survival. Aim 2 will define the role of miR-217 in antagonizing miR-211 regulation of its transcriptional targets. Aim 3 will define the role of miR-211-dependent regulation of Bmal1 to the development and progression of B-cell lymphoma. There are obvious points of cross-talk between this proposal and Project 2 which focuses on how the UPR antagonizes myc induced apoptosis and ATF4-dependent regulation of tumor cell metabolism. Through collaboration with Project 2, we will assess the role of miR-211 and Bmal1 as a mediator of tumor cell metabolism and survival. Through collaboration with Project 3, we will determine how miR-211, miR-217 and Bmal1 regulate IFNAR1 signaling and regulation in cytotic T- lymphocytes. The findings steming from work proposed herein will provide a foundation for the design of novel anti-cancer therpeutics.