Mechanisms of melanoma migration and survival in 3D microenvironments

3D 微环境中黑色素瘤迁移和存活的机制

• 批准号: 9883752
• 负责人: Erik S Welf
• 金额: $ 11.61万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9883752
• 关键词: 1-Phosphatidylinositol 3-Kinase; 3-Dimensional; Actins; Actomyosin; Affect; Binding; Binding Proteins; Biological; Bulla; CD44 gene; Cell Survival; Cell physiology; Cell surface; Clustered Regularly Interspaced Short Palindromic Repeats; Collagen Fiber; Complement; Computer Analysis; Computer Models; Data; Diglycerides; Encapsulated; Equilibrium; Exhibits; Extracellular Matrix; Family; Family member; Image; In Vitro; Laboratories; Measures; Mediating; Melanoma Cell; Membrane; Mentorship; Metastatic Melanoma; Metastatic to; Molecular; Molecular Conformation; Mus; Mutation; Neoplasm Metastasis; Neural Crest Cell; Pathway interactions; Patients; Phosphatidylinositol 4,5-Diphosphate; Phospholipids; Phosphorylation; Prevalence; Process; Property; Protein Family; Proteins; Research Personnel; Role; Sampling; Shapes; Signal Transduction; Structure; Structure of primordial sex cell; Surface; Testing; Threonine; Tissues; Training; Training Activity; Xenograft Model; Zebrafish; advanced disease; cell motility; experience; ezrin; faculty mentor; genome editing; in vivo; live cell imaging; melanoma; metastatic process; migration; moesin; mortality; neoplastic cell; non-Native; polymerization; pressure; public health relevance; radixin protein; receptor; scaffold; skills; soft tissue; spatiotemporal; therapy design; tumor progression

 DESCRIPTION (provided by applicant): A greater understanding of the mechanisms mediating melanoma metastasis is necessary for treatment of advanced disease. Because all melanoma cells must move through and survive in non-native tissue microenvironments during metastatic dissemination, the ability to migrate and survive in these microenvironments is critical for metastatic potential. We and others have observed that melanoma cells encapsulated in 3-dimensional extracellular matrices that mimic soft tissues exhibit spontaneous and continuous non-apoptotic membrane blebbing. These observations have led us to ask if membrane blebbing is associated with the ability to survive and migrate through soft tissues. We propose to study the mechanisms that may enable melanoma migration and survival during metastatic dissemination as a function of membrane blebbing. Thus, the scientific objective of this project is to determine how blebbing is related to metastatic dissemination in melanoma. At the completion of this project, we will have determined if blebbing is enriched in stage III melanoma samples that exhibit efficient metastasis compared to stage III samples that exhibit less efficient metastasis, and we will have determined if these differences in metastatic efficiency are mediated by the scaffolding function of the ERM family protein ezrin. We will have determined if the spatiotemporal organization of phospholipid signaling regulates ezrin phosphorylation to organize membrane blebs that facilitate cell migration in 3D matrices, and we will have determined if membrane blebbing facilitates survival signaling by regulating the spatial partitioning of ezrin between membrane and cytosolic compartments. The training objective of this proposal is to train Dr. Erik Welf in the skills required to become an independent investigato specializing in quantitative approaches to dissect the mechanisms underlying metastatic dissemination of melanoma. The proposed training activities will complement Dr. Welf's previous experience in computational modeling and analysis by providing training in the cutting edge molecular biological, imaging, and in vivo approaches necessary to study the processes of metastatic dissemination in realistic microenvironments. The proposed project leverages the expertise of faculty mentors Gaudenz Danuser and Sean Morrison, who are experts in the fields of live cell imaging and in vivo approaches, respectively. The dynamic interplay between analytical and experimental approaches is fundamental to the approach that Dr. Welf will apply as an independent researcher, and this training experience will complete the skills to facilitate this approach. Although few, if any, existing laboratories can boast expertise in both analytical and experimental approaches relevant to melanoma, the proposed mentorship team will enable Dr. Welf to develop the skills required to become successful in this endeavor.

 描述（由申请人提供）：对于晚期疾病的治疗，有必要更好地了解介导黑色素瘤转移的机制，因为所有黑色素瘤细胞在转移扩散期间必须穿过非天然组织微环境并在其中存活，因此具有迁移和存活的能力。这些微环境至关重要 我们和其他人观察到，封装在模仿软组织的三维细胞外基质中的黑色素瘤细胞表现出自发且持续的非凋亡膜起泡，这些观察结果使我们思考膜起泡是否与生存能力有关。我们建议研究转移扩散过程中黑色素瘤迁移和存活的机制，作为膜起泡的函数。因此，该项目的科学目标是确定起泡的方式。在该项目完成时，我们将确定与表现出较低效率的 III 期黑色素瘤样本相比，表现出有效转移的 III 期黑色素瘤样本中是否富含起泡。 我们将确定这些转移效率的差异是否是由 ERM 家族蛋白 ezrin 的支架功能介导的。我们将确定磷脂信号传导的时空组织是否调节 ezrin 磷酸化以组织促进细胞在 3D 中迁移的膜泡。矩阵，我们将确定膜起泡是否通过调节膜和胞质区室之间埃兹蛋白的空间分配来促进生存信号传导。该提案的目的是培训 Erik Welf 博士成为一名独立研究人员所需的技能，专门研究定量方法来剖析黑色素瘤转移性传播的机制。拟议的培训活动将通过提供培训来补充 Welf 博士之前在计算建模和分析方面的经验。该项目利用了研究现实微环境中转移扩散过程所需的尖端分子生物学、成像和体内方法，利用了导师 Gaudenz Danuser 和 Sean Morrison 的专业知识，他们是这方面的专家。分析方法和实验方法之间的动态相互作用是韦尔夫博士作为独立研究人员所采用的方法的基础，而这种培训经验将完善促进这种方法的技能。尽管现有实验室很少（如果有的话）能够拥有与黑色素瘤相关的分析和实验方法方面的专业知识，但拟议的导师团队将使韦尔夫博士能够培养在这一努力中取得成功所需的技能。

项目成果

An image-based assay to quantify changes in proliferation and viability upon drug treatment in 3D microenvironments.

一种基于图像的测定，用于量化 3D 微环境中药物治疗后增殖和活力的变化。

• 发表时间: 2019-05-28
• 影响因子: 3.8
• 作者: Murali, Vasanth S;Chang, Bo;Fiolka, Reto;Danuser, Gaudenz;Cobanoglu, Murat Can;Welf, Erik S
• 通讯作者: Welf, Erik S