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: 9321473
• 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/9321473
• 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; 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; Recruitment Activity; 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; 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（由申请人提供）：Bravo-Cordero 博士在西班牙马德里自治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来研究癌细胞生物学，之后，他在 John Condeelis 博士和 Louis Hodgson 博士的实验室中继续接受最先进成像领域的培训。在博士后培训期间，Bravo-Cordero 博士专注于了解 RhoGTPase 在乳腺癌细胞迁移和侵袭过程中如何进行时空调控。他接受了 FRET 显微镜和 FRET 生物传感器成像等技术培训，以解决这些问题。迄今为止，他的工作已发表 23 篇同行评审出版物。 最近的工作表明，肿瘤细胞的转移受到细胞所在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镜技术是观察生理环境内肿瘤细胞的理想方式。体外模型的复杂性有限，因此使用重现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方法。 Bravo Cordero 博士接受过多光子体内成像和 FRET 显微镜等技术的培训，以便了解体内细胞信号传导。这项培训使领导一个将动物模型、多光子成像和 FRET 生物传感器体内成像相结合的实验室来了解肿瘤细胞转移的机制成为可能。环境：PI 的咨询委员会包括爱因斯坦解剖学系和生物光子中心联合主席 John Condeelis 博士。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来解答癌症等人类疾病的机制。该中心以其共享的成像资源和创新实验室而闻名，其中新的显微镜是定制的，以满足不同项目的特定需求。顾问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 受体信号传导方面的专家，他还研究趋化性背景下的 F-Bar 结构域蛋白。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协作并坚持博士后、导师和初级教师职业发展的机构。研究：运动和侵袭是从发育、稳态到转移的多个过程的关键步骤。为了使细胞移动，它们必须形成膜延伸以推动自身穿过细胞外基质。因此，了解驱动突起形成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、基质细胞和血管组成，它们共同影响肿瘤细胞的运动行为。 RhoGTPases 是细胞骨架动力学的主要调节因子，受到多种蛋白质的严格调节。含有 Bar 结构域的蛋白质家族在膜可塑性和 RhoGTPases 信号传导之间的界面发挥作用，这些蛋白质已成为 GTPase 和膜形状的重要调节剂。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。通过 RhoGTPase 的信号通路已在体外得到广泛研究，但调节体内 GTPase 激活的机制仍不清楚。为了解决肿瘤微环境、运动行为和 RhoGTPases 信号传导之间的联系，有必要将多光子活体成像与 FRET 生物传感器成像相结合。两种不同类型的突出已被证明可介导肿瘤侵袭，即板状伪足和侵袭伪足。迄今为止，尚不清楚它们各自对体内运动和肿瘤内渗的贡献。我的初步结果表明，表达β-肌动蛋白-TagRFP-T作为伪足突起标志物的肿瘤细胞表明，为了移动而延伸的膜突起的作用已经丰富。目标 1 将探讨 GTP 酶 RhoA 和 RhoC 介导的信号传导如何根据细胞外基质环境决定侵袭伪足和伪足突起的形成。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 被募集到肿瘤细胞的伪足和侵袭伪足突起上。目标 2 将探讨 srGAP1 通过 RhoGTPase 调节在建立片状伪足和侵袭伪足突起中的作用。目标3将探讨srGAP1在肿瘤细胞体内传播和转移中的作用。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、GTPase 信号传导和细胞骨架重排之间的相互作用，并且结果将用于改善早期转移的诊断和治疗。