Mechanisms of FIP200 regulation of breast cancer through its autophagy and non-autophagy functions

FIP200通过自噬和非自噬功能调控乳腺癌的机制

基本信息

批准号：
9927485
负责人：
JUN-LIN GUAN
金额：
$ 36.71万
依托单位：
UNIVERSITY OF CINCINNATI
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-15 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9927485
关键词：
Ablation Acute Alleles Animal Model Apoptosis Autophagocytosis BRCA1 gene Binding Biological Process Breast Cancer Cell Breast Cancer Model Breast Cancer therapy Breast Epithelial Cells Breast cancer metastasis Cancer Relapse Cell physiology Cells Characteristics Complement Complex DNA Sequence Alteration Data Development Disease Embryo Epidermal Growth Factor Receptor Family Goals Growth Health Homeostasis Human Impairment Incidence Knock-in Knock-in Mouse Knowledge Laboratories Maintenance Malignant Neoplasms Mammary Neoplasms Mediating Modeling Molecular Mutant Strains Mice Mutate Mutation Neoplasm Metastasis PTK2 gene Patients Play Population Property Proteins Recurrent disease Regulation Relapse Role Signal Transduction Starvation Stat3 Signaling Pathway System TNF gene Transforming Growth Factor Beta 2 Transforming Growth Factor beta Tumorigenicity Woman aldehyde dehydrogenases antitumor effect base breast cancer progression conditional mutant genetic analysis genetic approach immune function in vivo insight malignant breast neoplasm mouse genetics mouse model mutant neoplastic cell novel novel therapeutics response stem-like cell targeted treatment tool transforming growth factor beta3 tumor tumor growth tumor progression tumorigenic

项目摘要

Breast cancer is the most common malignancy among US women and remains a major health threat with high incidence and lethality. Despite the remarkable progress made recently in deciphering the genetic mutations associated with breast cancer, our understanding of the mechanistic basis for metastasis and relapse of breast cancer is still very limited. The long-term goal of the proposed studies is to understand the molecular and cellular mechanisms involved in these properties of breast cancer cells that are ultimately responsible for patient lethality. FIP200 (FAK-family Interacting Protein of 200 kDa) was initially identified in our laboratory and subsequently shown to be a component of the ULK1/Atg13/FIP200 complex essential for the induction of autophagy. We have shown recently that FIP200 ablation inhibited the development and progression of breast cancer, providing the first evidence of a pro-tumorigenic role for autophagy in animal models with intact immune functions. We also developed an inducible system to delete FIP200 after tumor development in vivo and demonstrated that acute disruption of autophagy by FIP200 deletion in established tumors blocked their growth. In preliminary studies, we identified two distinct populations of breast cancer stem-like cells (BCSCs) and showed that FIP200 ablation impaired the maintenance and tumorigenicity of both BCSCs. Our preliminary data implicated TGF-β/Smad and EGFR/Stat3 signaling pathways in mediating FIP200 regulation of these two BCSCs respectively. In another set of preliminary studies, we identified residues 582-585 in FIP200 for binding to Atg13 and generated a new FIP200 knock-in mutant mouse with these residues mutated to Ala (designated as KI allele, encoding FIP200-4A lacking binding to Atg13). Analysis of KI/KI mice and MEFs derived from KI/KI embryos showed both autophagy and non-autophagy functions of FIP200 in regulating different biological processes. These studies further revealed a new non-autophagy function of FIP200 to protect normal and transformed MEFs from TNFα-induced apoptosis. Thus this unique mutant KI allele as well as additional mouse models prepared in preliminary studies provides us with powerful tools for the genetic analysis of mechanisms of FIP200 regulation of breast cancer through its autophagy and non-autophagy functions in vivo. Based on these strong preliminary data and using our unique novel mouse models, we propose to 1) determine the mechanisms of FIP200 regulation of CD29hiCD61+ and ALDH+ BCSCs in PyMT and BRCA1-deficient mouse models of breast cancer; 2) examine autophagy and non-autophagy functions of FIP200 in the regulation of BCSCs and breast cancer development and progression; and 3) explore the strategies of targeting FIP200 autophagy and non-autophagy functions in BCSCs for breast cancer therapy. Together, these studies will provide significant insights into the molecular and cellular mechanisms of breast cancer metastasis and relapse that may contribute to novel therapies for this devastating disease.

乳腺癌是美国女性最常见的恶性肿瘤，并且仍然是一个主要的健康威胁尽管最近在破译遗传基因方面取得了显着进展。与乳腺癌相关的突变，我们对转移机制基础的理解和乳腺癌复发的可能性仍然非常有限。拟议研究的长期目标是了解乳腺癌的复发情况。涉及乳腺癌细胞这些特性的分子和细胞机制最终是 FIP200（200 kDa 的 FAK 家族相互作用蛋白）最初是在我们的研究中发现的。实验室，随后被证明是 ULK1/Atg13/FIP200 复合体的组成部分，对于我们最近表明 FIP200 消融抑制了自噬的发展和发生。乳腺癌的进展，为动物自噬的促肿瘤作用提供了第一个证据我们还开发了一种在肿瘤后删除 FIP200 的模型。体内发育并证明 FIP200 缺失对自噬的急性破坏已建立在初步研究中，我们发现了两个不同的乳腺癌群体。干细胞样细胞 (BCSC) 并表明 FIP200 消融损害了两者的维持和致瘤性我们的初步数据表明 TGF-β/Smad 和 EGFR/Stat3 信号通路参与介导。在另一组初步研究中，我们确定了 FIP200 对这两种 BCSC 的分别调节。 FIP200 中的残基 582-585 与 Atg13 结合，并生成新的 FIP200 敲入突变小鼠这些残基突变为 Ala（称为 KI 等位基因，编码缺乏与 Atg13 结合的 FIP200-4A）。 KI/KI 小鼠和源自 KI/KI 胚胎的 MEF 均表现出自噬和非自噬功能这些研究进一步揭示了FIP200在调节不同生物过程中的作用。 FIP200 的功能是保护正常和转化的 MEF 免受 TNFα 诱导的细胞凋亡。突变KI等位基因以及初步研究中制备的其他小鼠模型为我们提供了强大的 FIP200 通过自噬和调控乳腺癌机制的遗传分析工具基于这些强有力的初步数据并使用我们独特的新型小鼠体内非自噬功能。模型中，我们建议 1) 确定 FIP200 对 CD29hiCD61+ 和 ALDH+ BCSC 的调节机制在 PyMT 和 BRCA1 缺陷型乳腺癌小鼠模型中 2) 检查自噬和非自噬； FIP200 在调节 BCSC 和乳腺癌发生和进展中的功能；3) 探索靶向BCSCs中FIP200自噬和非自噬功能治疗乳腺癌的策略总之，这些研究将为分子和细胞机制提供重要的见解。乳腺癌转移和复发可能有助于针对这种破坏性疾病的新疗法。