BLRD Research Career Scientist Award Application

我的实验室正在进行的研究集中在三个广泛的领域：1）了解分子 驱动黑色素瘤肿瘤发展和进展的机制，2）定义皮肤的作用 微环境对黑色素瘤发生的影响，以及3）鉴定诊断为黑色素瘤的患者的预后生物标志物。 早期黑素瘤由VA BLR & D Merit Review Award资助的研究项目#1的目标， 是为了了解EPAC的作用，介导替代cAMP信号传导的蛋白质，在促进生长， 原发性黑色素瘤和代谢适应机制，消除EPAC依赖 在肿瘤进展过程中。我们正在探索在原发性黑色素瘤中抑制EPAC信号传导， 转移性黑色素瘤中的EPAC依赖性分别作为预防和治疗 黑色素瘤在退伍军人以及一般人群。在项目#2中，我们使用 人类皮肤芯片该项目的基本原理是，虽然转基因小鼠模型和人类 虽然体外黑色素瘤细胞系模型是有用的，但它们不能完全模拟在黑色素瘤细胞系中发生的复杂相互作用， 在完整的人类皮肤微环境中的黑色素瘤发生。DoD Peer Reviewed Medical的目标 研究计划资助的研究是了解表皮角质形成细胞和真皮成纤维细胞的作用 在黑色素瘤的发生，制定战略，黑色素瘤预防现役军人和退伍军人， 增加患黑色素瘤的风险。这些研究项目深入关注细胞和分子方面， 黑色素瘤还涉及使用固定的和石蜡包埋的人黑素瘤的回顾性分析的转化研究。 原发性黑素瘤肿瘤组织。我未来研究的主要重点是将我们的发现转化为前瞻性的研究。 对被诊断患有早期皮肤黑色素瘤的退伍军人进行调查。 最近，与Gunasekaran博士和Jose Ayuso博士合作，他们是生物医学工程师， 生物传感器和微流体技术，分别使我们能够开发一种敏感的生物传感器， 建议用于针对退伍军人中循环黑色素瘤细胞检测的前瞻性研究 被诊断为早期黑素瘤在原理验证研究中，我们展示了选择性和灵敏度检测 患者血液中的细胞。我们表明，这种免疫传感器很容易适应，在阵列格式， 同时检测多种生物标志物，并且可以并入微流体装置中并多路复用 基于CTC的细胞表面标志物来鉴定和捕获CTC的亚群， 特征化项目#3的目标是识别和表征肿瘤细胞中的循环肿瘤细胞（CTC）。 外周血作为转移性黑色素瘤和残留疾病的风险指标。拟议的具体目标 a）基于表面标志物异质性的黑素瘤细胞的检测和捕获，B） 使用多重微流体免疫传感器阵列检测和表征CTC异质性，和c） CTC的表型和分子表征。这些研究将得到VA CSR & D Merit Review的支持 获奖申请已被选中。 在研究职业科学家奖期间，我计划整合我们对分子生物学的理解， 黑色素瘤进展机制的前瞻性研究，往往需要长期监测， 患者我计划利用微流控平台开发人类皮肤芯片来研究这种关系 环境因素与退伍军人患黑色素瘤风险之间的关系。更重要的是，研究生涯 科学家奖的支持将使我们能够收集关键的额外数据，以支持我的下一轮优异奖 通过超越循环黑色素瘤细胞的检测来审查应用，以利用液体活检， 监测退伍军人转移性黑色素瘤的风险。

Ongoing research in my laboratory is focused on three broad areas: 1) understanding the molecular mechanisms that drive melanoma tumor development and progression, 2) defining the role of skin microenvironment on melanomagenesis, and 3) identifying prognostic biomarkers for patients diagnosed with early-stage melanoma. The goal of the research project #1, which is funded by VA BLR&D Merit Review Award, is to understand the role of EPACs, proteins that mediate the alternative cAMP signaling, in promoting the growth of primary melanoma and the mechanism involved in metabolic adaptation that abolishes EPAC dependency during tumor progression. We are exploring inhibition of EPAC signaling in primary melanoma and restoration of EPAC dependency in metastatic melanoma as strategies, respectively, for prevention and treatment of melanoma in Veterans as well as general population. In project #2, we are modeling melanomagenesis using human skin-on-a-chip. The rationale for this project is that while genetically modified mouse models and human melanoma cell lines models in vitro are useful, they do not fully mimic the complex interactions that occur during melanomagenesis in the intact human skin microenvironment. The goal of the DoD Peer Reviewed Medical Research Program-funded research is to understand the role of epidermal keratinocytes and dermal fibroblasts in melanomagenesis to devise strategies for melanoma prevention in active service members and Veterans with increased risk of melanoma. These research projects with in-depth focus on cell and molecular aspects of melanoma also involve translational research using retrospective analysis of fixed and paraffin embedded human primary melanoma tumor tissues. The major focus of my future research is to translate our findings in prospective investigations in Veterans diagnosed with early-stage cutaneous melanoma. Recently, a collaboration with Drs. Gunasekaran and Jose Ayuso, biomedical engineers with expertise in biosensors and microfluidic technology, respectively, allowed us to develop a sensitive biosensor that we propose to employ for prospective studies targeted to detection of circulating melanoma cells in Veteran diagnosed with early-stage melanoma. In proof-of-principle studies, we showed selective and sensitive detection of cells in patient blood. We show that this immunosensor is readily adaptable, in an arrayed format, for simultaneous detection of multiple biomarkers and can be incorporated into a microfluidic device and multiplexed to identify and capture subsets of CTC based on their cell surface markers for phenotypic and molecular characterization. The goal of project #3 is to identify and characterize circulating tumor cells (CTC) in the peripheral blood as indicators of risk of metastatic melanoma and residual disease. The proposed specific aims of this project are a) detection and capture of melanoma cells based on surface marker heterogeneity, b) detection and characterization of CTC heterogeneity using multiplexed microfluidic immuno-sensor array and c) phenotypic and molecular characterization of CTC. These studies will be supported by VA CSR&D Merit Review Award application selected for funding. During the Research Career Scientist Award period, I plan to integrate our understanding of molecular mechanisms in melanoma progression with prospective studies that often require long-term monitoring of the patients. I plan to leverage the microfluidic platform to develop human skin-on-chip to investigate the relationship between environmental factors and risk of melanoma in Veterans. More importantly, the Research Career Scientist Award support will allow us to collect the critical additional data to support my next round of Merit Review application by going beyond detection of circulating melanoma cells to leverage the liquid biopsy for monitoring risk of metastatic melanoma in Veterans.