Mechanisms of melanoma migration and survival in 3D microenvironments

 描述(由申请人提供)：对黑色素瘤转移的机制有更深入的了解对于晚期疾病的治疗是必要的。由于所有黑色素瘤细胞在转移扩散过程中都必须通过非自然组织微环境并在其中存活，因此在这些微环境中迁移和存活的能力至关重要。 有转移潜能。我们和其他人观察到，被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的和持续的非凋亡性膜气泡。这些观察结果让我们问，膜起泡是否与软组织的存活和迁移能力有关。我们建议研究黑色素瘤在转移扩散过程中迁移和存活的机制，作为细胞膜起泡的功能。因此，该项目的科学目标是确定起泡与黑色素瘤转移扩散的关系。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期黑色素瘤样本中的起泡是否丰富。 我们将确定这些转移效率的差异是否由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，我们将确定膜泡是否通过调节Ezrin在膜和细胞质之间的空间分配来促进生存信号。这项提议的培训目标是培训埃里克·韦尔夫博士所需的技能，使其成为一名独立的研究人员，专门从事分析黑色素瘤转移性扩散机制的定量方法。拟议的培训活动将通过提供研究现实微环境中转移扩散过程所需的尖端分子生物学、成像和体内方法的培训，补充韦尔夫博士以前在计算建模和分析方面的经验。拟议的项目利用了教师导师高登兹·丹尼瑟和肖恩·莫里森的专业知识，他们分别是活细胞成像和活体方法领域的专家。分析方法和实验方法之间的动态相互作用是韦尔夫博士将作为独立研究员应用的方法的基础，而这一培训经验将完成促进这一方法的技能。虽然现有的实验室几乎没有能够同时具备与黑色素瘤相关的分析和实验方法的专业知识，但拟议中的导师团队将使韦尔夫博士能够培养在这项工作中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.