BLRD Research Career Scientist Award Application

我的实验室正在进行的研究主要集中在三个方面：1)了解分子 驱动黑色素瘤发生和进展的机制，2)确定皮肤的作用 微环境对黑色素瘤发生的影响，以及3)确定诊断为黑色素瘤患者的预后生物标志物 早期黑色素瘤。由退伍军人事务部BLR&D功绩审查奖资助的研究项目#1的目标是， 是为了了解EPAC在促进生长中的作用，EPAC是介导另一种cAMP信号的蛋白质 原发性黑色素瘤的发病机制及其代谢适应机制 在肿瘤进展过程中。我们正在探索抑制原发性黑色素瘤的EPAC信号和修复 EPAC依赖于转移性黑色素瘤分别作为预防和治疗的策略 退伍军人和普通人群中的黑色素瘤。在项目2中，我们使用以下方法模拟黑色素瘤的发生 人类芯片上的皮肤。这个项目的基本原理是，虽然转基因小鼠模型和人类 黑色素瘤细胞系体外模型是有用的，它们不能完全模拟发生在 在完整的人类皮肤微环境中黑色素瘤的发生。美国国防部同行评审医学的目标 研究计划资助的研究是为了了解表皮角质形成细胞和真皮成纤维细胞的作用 在黑色素瘤发生方面为现役军人和退伍军人制定黑色素瘤预防策略 患黑色素瘤的风险增加。这些研究项目深入关注细胞和分子方面的 黑色素瘤还涉及对固定的和石蜡包埋的人类进行回顾性分析的翻译研究 原发黑色素瘤肿瘤组织。我未来研究的主要重点是将我们的发现翻译成前瞻性的 退伍军人早期皮肤黑色素瘤的调查。 最近，与Gunasekaran博士和Jose Ayuso博士合作的生物医学工程师在 生物传感器和微流体技术分别使我们能够开发出一种灵敏的生物传感器，我们可以 建议用于前瞻性研究，以检测退伍军人循环中的黑色素瘤细胞 被诊断为早期黑色素瘤。在原则证明研究中，我们显示了选择性和灵敏的检测 病人血液中的细胞。我们表明，这种免疫传感器以阵列的形式很容易地适应于 同时检测多个生物标志物，并可集成到微流控设备中并进行多路复用 根据细胞表型和分子标记鉴定和捕获CTC亚群 人物刻画。项目#3的目标是识别和表征循环中的肿瘤细胞(CTC) 外周血可作为转移性黑色素瘤和残留病风险的指标。拟议的具体目标 该项目包括a)基于表面标记异质性检测和捕获黑色素瘤细胞，b) 利用多路微流控免疫传感器阵列和c)检测和表征CTC的异质性 CTC的表型和分子特征。这些研究将得到退伍军人事务部CSR&D功绩审查的支持 获奖申请被选为资助对象。 在研究生涯科学家奖期间，我计划整合我们对分子的理解 黑色素瘤进展机制的前瞻性研究，通常需要长期监测 病人。我计划利用微流控平台开发人类皮肤芯片来研究两者之间的关系 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，以支持我的下一轮功绩 通过超越对循环黑色素瘤细胞的检测来审查应用程序，以利用液体活检来 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.