Project 2- The ISR effector ATF4 in metabolic reprogramming and survival during Myc-induced tumorigenesis

项目 2 - ISR 效应子 ATF4 在 Myc 诱导的肿瘤发生过程中代谢重编程和存活中的作用

• 批准号: 10247662
• 负责人: Constantinos Koumenis
• 金额: $ 28.8万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-18 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10247662
• 关键词: 3-Dimensional; ARNTL gene; Ablation; Acute; Allografting; Amino Acids; Apoptosis; Attenuated; Autophagocytosis; B-Cell Lymphomas; Binding; Cells; ChIP-seq; Collaborations; Colorectal; Colorectal Cancer; Colorectal Neoplasms; Cytotoxic T-Lymphocytes; Development; Down-Regulation; Endoplasmic Reticulum; Enzymes; FRAP1 gene; Face; Funding; Genes; Genetic; Genetic Engineering; Genetic Transcription; Glucose Transporter; Glycolysis; Glycolysis Inhibition; Hematopoietic stem cells; Homeostasis; Immunologic Surveillance; In Vitro; Interferon Type I; Investigation; Knock-out; Large Intestine Carcinoma; Lymphoma; Lymphoma cell; Lymphomagenesis; Malignant - descriptor; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Metabolic; Metabolism; Modeling; Molecular; Mus; Nutrient; Oncogene Activation; Oncogenic; Organoids; Oxygen; Pathway interactions; Patients; Pharmacology; Phosphorylation; Play; Process; Proliferating; Prostate carcinoma; Prostatic Neoplasms; Protein Biosynthesis; Proteins; Regulation; Role; SLC2A1 gene; Signal Transduction; Stress; Testing; Text; Therapeutic Intervention; Transcriptional Activation; Transcriptional Regulation; Transfer RNA; Transgenic Model; Translational Regulation; Translations; Transplantation; Up-Regulation; Work; Xenograft procedure; arm; biological adaptation to stress; c-myc Genes; cell growth; cell transformation; colon carcinogenesis; conditional knockout; endoplasmic reticulum stress; experimental study; glucose uptake; glycosylation; in vivo; inhibitor/antagonist; knock-down; neoplastic cell; new therapeutic target; novel; programs; promoter; prostate cancer cell; response; sensor; transplant model; tumor; tumor growth; tumor microenvironment; tumor progression; tumorigenesis; tumorigenic

ABSTRACT/SUMMARY (Project 2) Tumor cell intrinsic stress including oncogene activation, as well as extrinsic stresses, such as low oxygen/nutrient availability, elicit perturbations in the endoplasmic reticulum (ER). Moreover, oncogenically transformed cells face increased burden placed by augmented biosynthetic pathways and rewired metabolism to meet the demands imposed by rapid proliferation. Adaptation to the ensuing stress and re-establishment of cellular homeostasis is achieved via activation of a coordinated signal transduction program termed the Integrated Stress Response (ISR). During the previous funded period, we demonstrated that increased rates of protein synthesis elicited by oncogenic MYC, activate the PERK/GCN2→eIF2 arm of the ISR, thereby supporting MYC-induced cell transformation. In preliminary unpublished studies, we have accumulated strong evidence supporting an essential role for the ISR effector and target of eIF2 ATF4, in transformation and tumorigenesis, particularly in tumors with activated MYC. However, how ATF4 elicits differential responses to various stresses in the context of MYC-dependent transformation is a critical question that remains unanswered. We will test the hypothesis that activation of ATF4 by the ISR plays a critical role in MYC- induced transformation and tumor progression by promoting metabolic and translational adaptation in coordination with MYC by focusing on three specific Aims. In Aim 1 we will identify critical nodes in cellular metabolism and translational regulation which are coordinately regulated by both ATF4 and c-MYC. Specifically, we will delineate the mechanism of Glut-1 and eIF4E transcriptional activation by ATF4 and determine functional requirements of GLUT1 and eIF4E in regulating glycolysis, translation and survival during MYC-dependent transformation in vitro and in vivo. Under Aim 2, we will delineate the mechanism of co- regulation of transcriptional targets between ATF4 and MYC by ChIP-seq analysis in lymphoma, colorectal (CRC) and prostate (PCa) cancer cells expressing inducible forms of MYC and analyze coordinately regulated genes. We will then determine the effects of knockout/knockdown of the identified co-regulated genes and newly identified targets in MYC-dependent proliferation, apoptosis and tumor growth. Finally, under Aim 3, we will determine the role of ATF4 in MYC-dependent transformation and tumorigenesis in PCa and CRC tumors using a conditional knockout ATF4 model crossed with PTENfl/fl:MycTg, and mouse CRC models (orthotopic, syngeneic and spontaneous) as well as 3D CRC organoids. We will also work with Project 1 to analyze the regulation of ATF4-dependent downregulation of BMAL1 and Clock genes and their role in translation and lymphomagenesis. Finally, with Project 3, we will analyze the effects of ATF4 ablation on type I interferon pathway and viability/effector functions of tumor-infiltrating cytotoxic T lymphocytes. Completion of these aims will provide a better understanding of the critical role of ATF4 in MYC-dependent pro-tumorigenic processes and may uncover new targets for therapeutic intervention in these malignancies.

摘要/总结（项目 2） 肿瘤细胞内在应激包括癌基因激活，以及外在应激，例如低 氧气/营养物质的可用性，引起内质网（ER）的扰动。 转化细胞面临着生物合成途径增强和新陈代谢重新连接带来的更大负担 以满足快速扩散所带来的需求，以适应随之而来的压力和重建。 细胞稳态的实现是通过激活协调信号转导程序来实现的，该程序称为 综合压力反应（ISR） 在上一个资助期间，我们证明了比率的增加。 致癌 MYC 引起的蛋白质合成，激活 ISR 的 PERK/GCN2→eIF2 臂，从而 在未发表的初步研究中，我们已经积累了强大的支持 MYC 诱导的细胞转化的能力。 有证据支持 ISR 效应子和 eIF2 ATF4 靶标在转化和 肿瘤发生，特别是在 MYC 激活的肿瘤中，ATF4 如何引发不同的反应。 MYC 依赖性转化背景下的各种压力仍然是一个关键问题 我们将检验 ISR 激活 ATF4 在 MYC- 中发挥关键作用的假设。 通过促进代谢和翻译适应诱导转化和肿瘤进展 与 MYC 协调，重点关注三个具体目标 在目标 1 中，我们将确定关键节点。 细胞代谢和翻译调节由 ATF4 和 c-MYC 协调调节。 具体来说，我们将描述 ATF4 和 eIF4E 转录激活 Glut-1 和 eIF4E 的机制 确定 GLUT1 和 eIF4E 在调节糖酵解、翻译和存活过程中的功能需求 在目标 2 下，我们将描述 MYC 依赖性体外和体内转化的机制。 通过 ChIP-seq 分析淋巴瘤、结直肠癌中 ATF4 和 MYC 之间转录靶标的调控 (CRC) 和前列腺 (PCa) 癌细胞表达可诱导形式的 MYC 并分析协调调节 然后，我们将确定已识别的共同调控基因的敲除/敲低的影响和 新确定的 MYC 依赖性增殖、细胞凋亡和肿瘤生长靶标最后，在目标 3 下， 我们将确定 ATF4 在 PCa 和 CRC 中 MYC 依赖性转化和肿瘤发生中的作用 使用与 PTENfl/fl:MycTg 杂交的条件性敲除 ATF4 模型和小鼠 CRC 模型进行的肿瘤 （原位、同基因和自发）以及 3D CRC 类器官我们还将与项目 1 合作。 分析 ATF4 依赖性 BMAL1 和 Clock 基因下调的调节及其在 最后，在项目 3 中，我们将分析 ATF4 消融对淋巴瘤发生的影响。 I 型干扰素途径和肿瘤浸润细胞毒性 T 淋巴细胞的活力/效应器功能。 完成这些目标将有助于更好地理解 ATF4 在 MYC 依赖性中的关键作用 促肿瘤发生过程，并可能发现这些恶性肿瘤治疗干预的新靶点。