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/摘要（项目2） 肿瘤细胞的固有应力，包括癌基因激活以及外在应力，例如低 氧/营养物的可用性，引起内质网中的扰动（ER）。此外，在肿瘤上 转化的细胞面对通过增强的生物合成途径和恢复新陈代谢的增强伯嫩的增加 满足快速扩散施加的需求。适应确保压力和重新建立 通过激活协调的信号转导计划，可实现细胞体内稳态的稳态。 综合应力反应（ISR）。在上一个资助的时期，我们证明了利率提高 致癌MYC引起的蛋白质合成，激活ISR的PERK/GCN2→EIF2手臂，从而激活 支持MYC诱导的细胞转化。在初步未发表的研究中，我们积累了强大的 支持ISR效应子和eIF2ATF4的目标的基本作用的证据，在转型和 肿瘤发生，特别是在激活MYC的肿瘤中。但是，ATF4如何引起差异响应 在依赖MYC的转型背景下，各种压力是一个关键问题，仍然存在 未得到答复。我们将检验以下假设：ISR激活ATF4在MYC-中起关键作用 通过促进代谢和转化适应来诱导的转化和肿瘤进展 通过关注三个特定目标，与MYC协调。在AIM 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协同调节。 具体而言，我们将描述ATF4和 确定GLUT1和EIF4E在调节性糖酵解，翻译和生存过程中的功能需求 MYC依赖于体外和体内的转化。在AIM 2下，我们将描述共同的机制 通过淋巴瘤中的CHIP-seq分析对ATF4和MYC之间的转录靶标进行调节 （CRC）和前列腺（PCA）癌细胞表达MYC的诱导形式，并分析协调调节 基因。然后，我们将确定已确定的共同调节基因的敲除/敲除的影响 新确定的靶标在MYC依赖性增殖，凋亡和肿瘤生长中。最后，在AIM 3下， 我们将确定AT​​F4在PCA和CRC中依赖MYC依赖性转化和肿瘤发生中的作用 使用条件敲除ATF4模型与PTENFL/FL：MYCTG和鼠标CRC模型交叉的肿瘤 （原位，合成和赞助）以及3D CRC器官。我们还将与项目1合作 分析BMAL1和时钟基因的ATF4依赖性下调的调节及其在 翻译和淋巴作用。最后，通过项目3，我们将分析ATF4消融对 I型干扰素途径和肿瘤浸润细胞毒性T淋巴细胞的活力/效应子功能。 这些目标的完成将更好地理解ATF4在MYC依赖性中的关键作用 促肿瘤的过程，可能会发现这些恶性肿瘤治疗干预的新靶标。