Protrusion plasticity during in vivo tumor cell migration

体内肿瘤细胞迁移过程中的突出可塑性

• 批准号: 9534544
• 负责人: Jose Javier Bravo-Cordero
• 金额: $ 19.22万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9534544
• 关键词: Actins; Address; Advisory Committees; Affect; Anatomy; Animal Model; Area; Behavior; Biology; Biophotonics; Biosensor; Blood Vessels; Breast Cancer Cell; Breast Cancer Model; Breast cancer metastasis; Cell Communication; Cell Polarity; Cells; Cellular Morphology; Cellular biology; Chemotaxis; Collaborations; Collagen Fiber; Complex; Cues; Custom; Cytoskeleton; Data; Development; Diagnosis; Dimensions; Disease; Early treatment; Environment; Event; Exposure to; Extracellular Matrix; Faculty; Fluorescence Resonance Energy Transfer; Goals; Guanosine Triphosphate Phosphohydrolases; Homeostasis; Image; Imaging Techniques; In Vitro; Institution; Knowledge; Laboratories; Lead; Link; Macrophage Colony-Stimulating Factor Receptor; Malignant Neoplasms; Mediating; Membrane; Mentors; Microscope; Microscopy; Modality; Molds; Molecular; Monitor; Movement; Neoplasm Metastasis; Outcome; Output; Pathway interactions; Pattern; Peer Review; Phenotype; Physiological; Play; Postdoctoral Fellow; Primary Neoplasm; Process; Protein Family; Proteins; Pseudopodia; Publications; Receptor Signaling; Regulation; Research; Resolution; Resources; Role; Shapes; Signal Pathway; Signal Transduction; Signaling Protein; Spain; Stromal Cells; System; Techniques; Tertiary Protein Structure; Training; Translating; Tumor Cell Invasion; Tumor Cell Migration; Universities; Work; Xenograft Model; beta Actin; cancer cell; career development; cell behavior; cell motility; design; drug development; experimental study; extracellular; human disease; imaging system; improved; in vitro Model; in vivo; in vivo imaging; innovation; instructor; interest; intravital imaging; macrophage; member; migration; multidisciplinary; multiphoton imaging; neoplastic cell; novel; post-doctoral training; public health relevance; recruit; spatiotemporal; targeted treatment; tumor; tumor microenvironment; tumor progression

 DESCRIPTION (provided by applicant): Dr. Bravo-Cordero obtained his degree in biology at Autonoma University of Madrid, Spain. After completing his doctorate work, in which he used high-resolution confocal imaging techniques to study cancer cell biology, he continued his training in the area of state-of-the-art imaging in the laboratories of Dr. John Condeelis and Dr. Louis Hodgson. During postdoctoral training, Dr. Bravo-Cordero focused on understanding how RhoGTPases are spatiotemporally regulated during breast cancer cell migration and invasion. He w a s trained in techniques such as FRET microscopy as well as FRET biosensors imaging to address these questions. His work to date has resulted in 23 peer-review publications. Recent work has shown that metastasis of tumor cells is affected by the extracellular microenvironment in which the cells are located. In order to understand the mechanisms of tumor cell metastasis and the activation of the intracellular signals, high-resolution microscopy techniques like multiphoton imaging is an ideal modality to observe tumor cells inside their physiological environment. In vitro models are limited in their complexity, thus using animal models that recapitulate the disease, will be a more effective way to address questions that could not be addressed with in vitro systems. Dr. Bravo Cordero has been trained in techniques such as multiphoton in vivo imaging and FRET microscopy in order to understand cell signaling in vivo. This training makes it possible to lead a laboratory that integrates animal models, multiphoton imaging and FRET- biosensor imaging in vivo to understand mechanisms of tumor cell metastasis. Environment: Advisory committee of the PI included Dr. John Condeelis, Co-Chair of Anatomy Department and Biophotonic Center at Einstein. His lab and the Center create a multidisciplinary environment focused on answering mechanisms of human diseases, such as cancer, through use of microscopy. The Center is well known for its shared imaging resources and Innovation Laboratory, in which new microscopes are custom-built to accommodate specific needs of different projects. Other members of the advisory committee are: Dr. Louis Hodgson, he is an expert in FRET biosensor imaging and FRET biosensor design and Dr. Richard Stanley, he is an expert on macrophages biology and CSF-1 receptor signaling, he also studies F-Bar domain proteins in the context of chemotaxis. Dr. Richard Stanley is also a renowned mentor. Einstein is an institution that values collaboration and insists on career development of postdoctoral fellows, instructors and junior faculty. Research: Motility and invasion are crucial steps for multiple processes from development and homeostasis to metastasis. In order for cells to move, they must form membrane extensions to propel themselves through the extracellular matrix. Thus, understanding the molecular pathways that drive spatiotemporal control of protrusion formation is a fundamental question to be answered. The tumor microenvironment is composed of collagen fibers, stromal cells and blood vessels that, in combination, will influence the motility behavior of tumor cells. RhoGTPases are master regulators of cytoskeleton dynamics being tightly regulated by multiple proteins. A family of proteins containing Bar-domains acts at the interface between membrane plasticity and RhoGTPases signaling, and these proteins have emerged as important regulators of GTPases and membrane shape. The final migratory output of a tumor cell will be dictated by the extracellular matrix conditions and that will be translated through a complex signaling system that include BAR proteins and RhoGTPases to induce cytoskeleton rearrangements. Signaling pathways through RhoGTPases have been widely studied in vitro, but the mechanism that regulates GTPase activation in vivo is still unknown. To address the link between tumor microenvironment, motility behavior and RhoGTPases signaling is necessary to combine multiphoton intravital imaging with FRET-biosensors imaging. Two different types of protrusion have been shown to mediate tumor invasion, lamellipodia and invadopodia. To date, it is not clear the contribution of each of them to motility in vivo and tumor intravasation. My preliminary results have shown that tumor cells expressing β-actin-TagRFP-T as a marker for pseudopodia protrusions show that cell extending membrane protrusion in order to move have enriched in action. Aim 1 will explore how signaling mediated by the GTPases RhoA and RhoC determine the formation of invadopodia and pseudopodia protrusions depending on the extracellular matrix context. By using FRET biosensors in vivo, the activation pattern of these GTPases will be analyzed in these different protrusions. In preliminary experiments the Bar protein srGAP1, which regulates RhoGTPases, is recruited to pseudopodia and invadopodia protrusions of tumor cells. Aim 2 will explore the role of srGAP1 in establishing lamellipodia and invadopodia protrusions through RhoGTPases regulation. Aim 3 will explore the role of srGAP1 in tumor cell dissemination and metastasis in vivo. Results of this study will lead to a better understanding of the interplay among microenvironment components, GTPase signaling and cytoskeleton rearrangements during tumor progression and the results will be used to improve diagnosis and treatment of early metastasis.

 描述(申请人提供)：Bravo-Cordero博士在西班牙马德里的Autonoma大学获得生物学学位。在完成博士学位后，他使用高分辨率共聚焦成像技术研究癌细胞生物学，并在约翰·康迪利斯博士和路易斯·霍奇森博士的实验室继续进行最先进的成像领域的培训。在博士后培训期间，Bravo-Cordero博士专注于了解RhoGTP酶如何在乳腺癌细胞迁移和侵袭过程中进行时空调控。他是一位S，受过FRET显微镜和FRET生物传感器成像等技术的培训，以解决这些问题。到目前为止，他的工作已经出版了23本同行评议出版物。最近的工作表明，肿瘤细胞的转移受到细胞所处的细胞外微环境的影响。为了了解肿瘤细胞转移的机制和细胞内信号的激活，多光子成像等高分辨率显微技术是观察肿瘤细胞内部生理环境的理想手段。体外模型的复杂性有限，因此使用概括疾病的动物模型将是解决体外系统无法解决的问题的更有效的方法。布拉沃·科德罗博士曾接受过体内多光子成像和FRET显微镜等技术的培训，以了解体内的细胞信号。这种训练使其有可能领导一个整合了动物模型、多光子成像和体内FRET生物传感器成像的实验室，以了解肿瘤细胞转移的机制。环境：国际和平研究所顾问委员会成员包括约翰·康迪利斯博士，他是爱因斯坦解剖学和生物光子学中心的联合主席。他的实验室和该中心创建了一个多学科环境，专注于通过使用显微镜回答人类疾病(如癌症)的机制。该中心以共享成像资源和创新实验室而闻名，在该实验室中，新的显微镜是定制的，以适应不同项目的特定需求。咨询委员会的其他成员包括：Louis Hodgson博士，他是FRET生物传感器成像和FRET生物传感器设计方面的专家，Richard Stanley博士，他是巨噬细胞生物学和CSF-1受体信号方面的专家，他还在趋化作用的背景下研究F-Bar结构域蛋白质。理查德·斯坦利博士也是一位著名的导师。爱因斯坦是一所重视合作的机构，坚持博士后研究员、讲师和初级教员的职业发展。研究：运动和侵袭是从发育、动态平衡到转移的多个过程的关键步骤。为了让细胞移动，它们必须形成膜延伸，以推动自己穿过细胞外基质。因此，了解驱动突起形成的时空控制的分子途径是一个需要回答的基本问题。肿瘤微环境由胶原纤维、间质细胞和血管组成，它们结合在一起会影响肿瘤细胞的运动行为。RhoGTP酶是细胞骨架动力学的主要调节者，受到多种蛋白质的严格调控。一个含有Bar结构域的蛋白质家族在膜可塑性和RhoGTP酶信号之间起着重要的作用，这些蛋白质已经成为GTP酶和膜形状的重要调节因子。肿瘤细胞的最终迁移输出将由细胞外基质条件决定，并将通过包括bar蛋白和RhoGTP酶在内的复杂信号系统进行翻译，以诱导细胞骨架重排。RhoGTP酶的信号通路在体外已经得到了广泛的研究，但体内调节GTP酶激活的机制仍不清楚。为了解决肿瘤微环境、运动行为和RhoGTP酶信号之间的联系，需要将多光子活体成像与FRET生物传感器成像相结合。有两种不同类型的突起被证明可以介导肿瘤的侵袭，片状脂膜和内翻足。到目前为止，还不清楚它们在体内运动和肿瘤血管内转移中的作用。我的初步结果显示，肿瘤细胞表达β-肌动蛋白-TagRFP-T作为伪足突起的标志，表明细胞为了移动而延长膜突起的作用有所增强。目的1将探索由GTP酶RhoA和RhoC介导的信号如何根据细胞外基质环境决定内陷和伪足突起的形成。通过在体内使用FRET生物传感器，将分析这些GTP酶在这些不同突起中的激活模式。在初步实验中，调节RhoGTP酶的Bar蛋白srGAP1被招募到肿瘤细胞的伪足和伪足突起中。目的2探讨srGAP1通过调节RhoGTP酶在片状脂体和内侧足突起形成中的作用。目的3探讨srGAP1在肿瘤细胞体内扩散和转移中的作用。这项研究的结果将有助于更好地了解肿瘤进展过程中微环境成分、GTP酶信号转导和细胞骨架重排之间的相互作用，并将其用于改善早期转移的诊断和治疗。