Mechanisms of melanoma migration and survival in 3D microenvironments

 描述（由申请人提供）：更深入地了解介导黑色素瘤转移的机制对于治疗晚期疾病是必要的。因为所有的黑色素瘤细胞在转移扩散过程中必须在非天然组织微环境中移动并存活，所以在这些微环境中迁移和存活的能力是至关重要的 转移的可能性我们和其他人已经观察到包裹在模拟软组织的三维细胞外基质中的黑色素瘤细胞表现出自发和连续的非凋亡膜起泡。这些观察结果使我们想知道膜起泡是否与通过软组织存活和迁移的能力有关。我们建议研究的机制，可能使黑色素瘤的迁移和生存在转移性传播的功能膜起泡。因此，本项目的科学目标是确定水泡与黑色素瘤转移性播散的关系。在这个项目完成时，我们将确定与表现出较低效率的III期样本相比，在表现出有效转移的III期黑色素瘤样本中是否存在丰富的起泡。 转移，并且我们将确定转移效率的这些差异是否由ERM家族蛋白ezrin的支架功能介导。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基质中迁移的膜泡，并且我们将确定膜泡形成是否通过调节ezrin在膜和胞质隔室之间的空间分配来促进存活信号传导。本建议的培训目标是培训Erik Welf博士成为一名独立研究者所需的技能，专门从事定量方法，以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尖端分子生物学，成像和体内方法的培训来补充Welf博士以前在计算建模和分析方面的经验，这些方法是研究现实微环境中转移性传播过程所必需的。拟议的项目利用教师导师Gaudenz Danuser和Sean莫里森的专业知识，他们分别是活细胞成像和体内方法领域的专家。分析方法和实验方法之间的动态相互作用是Welf博士作为独立研究人员应用的方法的基础，这种培训经验将完成促进这种方法的技能。虽然很少有现有的实验室可以在与黑色素瘤相关的分析和实验方法方面拥有专业知识，但拟议的导师团队将使Welf博士能够发展在这奋进取得成功所需的技能。

 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.