Light-sheet micro-aspiration microscopy for the isolation and analysis of rare tumor cells from intact clinical specimens

光片微抽吸显微镜用于从完整的临床标本中分离和分析稀有肿瘤细胞

• 批准号: 10451940
• 负责人: ADAM K GLASER
• 金额: $ 24.83万
• 依托单位: ALLEN INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10451940
• 关键词: Light; sheet; micro; aspiration; microscopy

PROJECT SUMMARY This grant application responds to Program Announcement Number PA-18-398 and outlines a training and research program which will prepare me for a long-term research career focused on the development of optical imaging methods to better understand, diagnose, and treat human prostate cancer. The proposal outlines a mentored-training program which places me under the direct guidance of a team of NIH funded researchers from the University of Washington (UW) and the neighboring Fred Hutchinson Cancer Research Center (FHCRC). My training plan involves three complimentary areas of focus: (1) receiving further training in tissue clearing and labeling, (2) obtaining educational and hands-on training in single-cell isolation and analysis, and (3) developing a knowledge-base in molecular assays and prostate cancer to facilitate meaningful future collaborations with experts in these fields. To achieve these goals, I will use the funding to dedicate time for attending graduate courses, workshops, training seminars, grand rounds lectures, and most importantly regular meetings with my entire career award team, as outlined in my career development plan. The research project component of the proposal is driven by the fact that rare cell populations are known to play an outsized role in many of the most challenging problems in cancer, and downstream profiling (i.e., genomics, proteomics, and transcriptomics) of these individual cells can therefore allow for more-effective therapies targeted against these often-hidden lynchpins of the disease. Despite this, single-cell isolation (the process by which individual cells are targeted and collected for further study) is still technically challenging with current methods, particularly for rare tumor cells. This is particularly problematic, as these rare tumor cells often dictate metastasis, treatment resistance, and recurrence, all of which ultimately result in patient mortality. While this challenge exists for nearly all forms of cancer, this proposal will focus specifically on metastatic tumor cells which have invaded the lympho-vascular space in prostate cancer, as the isolation and analysis of these rare cells can enable the development of new diagnostic and treatment strategies. To achieve this, I propose to build upon my post- doctoral research, and with the necessary training above, develop a novel light-sheet micro-aspiration microscope which will enable the isolation of these rare tumor cells from intact clinical specimens for the first time. The project is comprised of three specific aims: (1) development of a multi-resolution open-top light-sheet microscope system for identifying rare tumor cells; (2) combined demonstration of light-sheet microscopy with micro-aspiration for isolating rare tumor cells; and (3) proof-of-concept clinical validation of the system for isolating rare invading tumor cells from human prostate tissues with lympho-vascular invasion. The prostate cancer research environment at UW and the FHCRC is an ideal place for me to train and pursue this research project, and with the guidance of my mentorship team, I will successfully complete this career development award and pursue my long-term career goal of becoming an independent investigator. !

项目摘要 本赠款申请响应计划公告号PA-18-398，并概述了培训和 研究计划，这将为我准备一个长期的研究生涯，重点是光学的发展 成像方法，以更好地了解，诊断和治疗人类前列腺癌。该提案概述了 我参加了一个指导培训项目，该项目将我置于一个由NIH资助的研究人员团队的直接指导下 来自华盛顿大学和邻近的弗雷德哈钦森癌症研究中心 （FHCRC）。我的培训计划包括三个重点领域：（1）接受组织方面的进一步培训 清除和标记，（2）获得单细胞分离和分析方面的教育和实践培训，以及 (3)开发分子检测和前列腺癌的知识库，以促进有意义的未来 与这些领域的专家合作。为了实现这些目标，我将利用资金投入时间， 参加研究生课程，研讨会，培训研讨会，大轮讲座，最重要的是定期 与我的整个职业奖励团队会面，正如我的职业发展计划中所概述的那样。该研究项目 该提案的一个组成部分是由这样一个事实驱动的，即已知稀有细胞群在 癌症中的许多最具挑战性的问题，以及下游分析（即，基因组学，蛋白质组学， 因此，这些单个细胞的转录组学）可以允许针对这些细胞的更有效的疗法。 往往隐藏的疾病的关键。尽管如此，单细胞分离（单个细胞分离的过程） 目标和收集用于进一步研究）对于当前方法仍然具有技术挑战性，特别是对于 罕见的肿瘤细胞这是特别成问题的，因为这些罕见的肿瘤细胞通常决定转移、治疗和预后。 耐药和复发，所有这些最终导致患者死亡。虽然这一挑战存在于 几乎所有形式的癌症，这项建议将特别集中在转移性肿瘤细胞已侵入 前列腺癌的淋巴血管间隙，因为这些罕见细胞的分离和分析可以使 开发新的诊断和治疗策略。为了实现这一目标，我建议在我的职位上- 博士研究，并与上述必要的培训，开发一种新型的光片微抽吸 显微镜，这将使这些罕见的肿瘤细胞分离从完整的临床标本，为第一次 时间该项目包括三个具体目标：（1）开发多分辨率开顶光片 显微镜系统，用于识别罕见的肿瘤细胞;（2）结合光片显微镜的演示， 用于分离罕见肿瘤细胞的微抽吸;以及（3）用于以下目的的系统的概念验证临床验证： 从具有淋巴-血管侵袭的人前列腺组织中分离罕见的侵袭性肿瘤细胞。前列腺 华盛顿大学和FHCRC的癌症研究环境是我培训和从事这项研究的理想场所 项目，并与我的导师团队的指导下，我将成功地完成这一职业发展 奖励和追求我的长期职业目标，成为一名独立调查员。 !