Autophagy and Epithelial Cell Fate During Anoikis and 3D Morphogenesis

失巢凋亡和 3D 形态发生过程中的自噬和上皮细胞命运

基本信息

批准号：
8071994
负责人：
Jayanta Debnath
金额：
$ 27.44万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8071994
关键词：
5&apos -AMP-activated protein kinase ATP Synthesis Pathway Address Adhesions Affect Anoikis Antithymoglobulin Apoptosis Autophagocytosis Autophagosome Biochemical Pathway Biological Assay Carcinoma Cell Death Cell Proliferation Cell Survival Cells Cessation of life DNA Damage Data Development Digestion Dominant-Negative Mutation Eating Epithelial Epithelial Cells Eukaryotic Cell Eukaryotic Initiation Factor-2 Extracellular Matrix Fibroblasts Genomic Instability Goals Growth Factor Human Integrin-mediated Cell Adhesion Pathway Lead Learning Link MAP Kinase Gene Maintenance Malignant Neoplasms Mammary gland Mediating Mitochondria Modeling Morphogenesis Neoplasm Metastasis Normal Cell Nutrient Oncogene Activation Oncogenes Oncogenic Pathway interactions Phenotype Phosphorylation Process Protein Biosynthesis Proteolysis RNA Interference Role Self-control as a personality trait Signal Transduction Staging Starvation Stress Structure System TSC2 gene Testing Therapeutic Time Tumor Suppressor Proteins Withdrawal Work base cancer cell cancer initiation cell transformation deprivation follow-up inhibition of autophagy inhibitor/antagonist innovation insight interest killings mTOR protein neoplastic cell novel oxidative damage prevent research study response segregation treatment strategy tumor tumor progression tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Autophagy is a tightly regulated lysosomal degradation process in which a cell digests its own contents under various forms of duress. In eukaryotic cells, autophagy primarily functions as a protective response during nutrient depletion or stress. Overall, much remains to be learned about how this fundamental process affects cancer initiation, progression and response to therapy. In this proposal, we will test the hypothesis that autophagy influences epithelial cancer development through two mechanisms-1) enhancing the ability of epithelial cells to survive ECM detachment-induced stress and apoptosis (anoikis) and 2) limiting proliferation. This hypothesis is based on initial studies using a unique 3D epithelial culture system to examine how oncogenic insults affect the formation of the normally hollow lumen in glandular structures. In this model, lumen formation involves the selective death of central cells lacking direct contact with extracellular matrix (ECM). Recently, we have discovered high levels of autophagy during lumen formation, indicating that this critical process also contributes to epithelial cell fate. In subsequent work, we are the first to establish that autophagy is strongly induced during ECM deprivation (anoikis). Based on this preliminary data, in Aim 1, we will evaluate whether autophagy promotes cell survival via mitigating the stresses of ECM detachment during anoikis and 3D lumen formation. In addition, we have obtained preliminary evidence that the autophagy regulatory molecule Beclin/ATG6 (a known tumor suppressor) inhibits cell proliferation in 3D culture. In Aim 2. we will interrogate whether other autophagy regulators (called ATGs) inhibit proliferation during growth factor withdrawal and 3D morphogenesis in order to definitively establish that autophagy can restrain epithelial proliferation and to identify new autophagy regulators that are able to inhibit neoplastic cell expansion. In Aim 3. we will delineate how modulating autophagy influences survival and proliferation in oncogene-expressing cells in order to determine how autophagy can be exploited to therapeutically target tumor cells harboring perturbations in known cancer pathways. Overall, the proposed experiments will provide unique information on the precise roles of autophagy during the development and progression of human carcinomas. Ultimately, these studies may lead to new treatment strategies to prevent the uncontrolled expansion of cancer cells via manipulating autophagy.

描述（由申请人提供）：自噬是一个严格调节的溶酶体降解过程，其中细胞在各种形式的胁迫下消化了自己的内容。在真核细胞中，自噬主要是养分耗竭或应激期间的保护性反应。总体而言，关于这一基本过程如何影响癌症的开始，进展和对治疗的反应，还有很多待了解。在该提案中，我们将检验以下假设：自噬通过两种机制影响上皮癌的发展1）增强上皮细胞在ECM脱离引起的应激和凋亡（AnoikiS）和2）限制增殖的能力。该假设基于使用独特的3D上皮培养系统的初步研究，以检查致癌性损伤如何影响腺体结构中正常空心管腔的形成。在此模型中，管腔形成涉及中枢细胞的选择性死亡，缺乏与细胞外基质（ECM）直接接触的中心细胞。最近，我们在管腔形成过程中发现了高水平的自噬，表明这种关键过程也有助于上皮细胞的命运。在随后的工作中，我们是第一个确定在ECM剥夺期间强烈诱导自噬（Anoikis）的人。基于此初步数据，在AIM 1中，我们将通过减轻Anoiki和3D管腔形成过程中ECM脱离的应力来评估自噬是否促进细胞存活。此外，我们获得了初步证据，表明自噬调节分子beclin/atg6（已知的肿瘤抑制剂）抑制了3D培养的细胞增殖。在AIM 2中。我们将询问其他自噬调节剂（称为ATG）是否会抑制生长因子戒断和3D形态发生过程中是否抑制增殖，以确定确定自噬可以抑制上皮增生并识别能够抑制神经肿瘤细胞扩张的新自噬调节剂。在AIM 3中。我们将描述自噬如何在表达癌基因的细胞中影响生存和增殖，以确定如何利用自噬来靶向具有障碍的肿瘤细胞，这些肿瘤细胞具有已知癌症途径的扰动。总体而言，拟议的实验将提供有关自噬在人癌发展和进展过程中自噬的确切作用的独特信息。最终，这些研究可能会导致新的治疗策略，以防止通过操纵自噬来防止癌细胞的不受控制。