Development of Spectroscopic Biomarkers of Bone Quality in Metastatic Disease

描述(由申请人提供)：PI的长期职业目标是成为一名独立的研究人员，专门从事骨转移的定量癌症研究。主要的研究兴趣是开发用于癌症诊断或手术指导的新的光学方法，以及开发/评估骨转移的治疗方法。未来四年，PI的近期目标是获得必要的培训，获得癌症光子学方面的经验，建立独立研究，并与同事建立合作关系。这些目标将通过实现以下目标来实现：1)丰富她在癌症生物学方面的知识，2)扩大她在生物医学光学领域的专业，3)从事一项严肃的研究项目，从中她将获得生物医学研究经验并为未来的外部资金获得初步数据，5)通过范德比尔特大学的研究和教育资源建立合作关系并获得其他必要的职业发展培训。K25奖励将允许PI Protected有时间通过遵循详细的培训计划来实现这些目标，其中包括动手实验室生物技能培训、头两年的几个课程/研讨会，以及有指导的研究计划。 对于拟议的研究项目，PI寻求开发一种可与拉曼光纤探头结合使用的统计模型，对情况下转移的骨质量提供实时定量评估。这一目标将从三个方面实现。目的1)用拉曼光谱研究乳腺癌和前列腺癌转移骨的时间变化。这一目的将证明这项研究的总体假设，即转移性骨质量的变化可以通过拉曼光谱检测到。将分别在乳腺癌和前列腺癌的小鼠心脏内和胫骨内注射模型中研究乳癌和母细胞转移。对于完整的非肿瘤和有肿瘤的骨骼，将使用光纤探头界面拉曼光谱来评估骨基质的结构和成分特性。这些拉曼衍生的骨质量测量将与骨结构(通过X射线成像和Micro-CT分析)、肿瘤负荷(通过体内生物发光成像评估)、骨转换(通过血清生物标记物)和骨细胞活动(通过骨组织形态计量学)相关联。统计学评价将检验骨质量是否随着肿瘤的进展而改变，以及成骨细胞/破骨细胞活性和病理性骨转换与转移性骨质量变化的关系。目的2)建立一种能够实时、自动化地评价骨转移瘤质量的统计模型。将使用统计模式识别方法分析在目标1中收集的所有光谱数据，以提取诊断特征并将其归入相应的骨质量类别。目的3)将拉曼光谱生物标志物与骨转移瘤的微环境联系起来。这个目的是为了找出转移骨表面光谱特征(生物标记物)的生物学贡献因素。在使用常规共聚焦拉曼显微光谱学进行拉曼探测测量的标记位置将获得二维拉曼图谱，其空间分辨率在细胞水平。拉曼图上的多元因子分析可以提取样品中各组分(因子)的光谱和空间分布。每个组分的分子性质将通过将其光谱与模型化合物的光谱进行比较来确定。通过将因子图像与免疫组织化学和骨组织学图像进行比较，可以获得额外的指导。 该项目完成后，将开发一种定量光谱评分系统，当与拉曼光谱结合使用时，该系统将能够对转移瘤中的骨质量进行原位评估。这将为她建立独立的骨转移研究提供PI工具。这一结果将作为国际和平协会未来申请外部资金的初步数据。

DESCRIPTION (provided by applicant): The PI's long term career goal is to become an independent investigator in quantitative cancer research specializing in bone metastasis. The main research interest is to develop novel optical methodologies for cancer diagnosis or surgical guidance, and to develop/evaluate therapeutic treatments for bone metastasis. The immediate goals for the PI over the next four years are to obtain necessary trainings, gain experience in cancer photonics, establish independent research and build collaborations with the fellow colleagues. These goals will be achieved by realizing the following objectives 1) to enrich her knowledge in cancer biology, 2) to expand her specialty in the field of biomedical optics, 3) to pursue a serious research project from which she will gain experience in biomedical research and obtain preliminary data for future extramural funding, and 5) to establish collaborations and obtain other necessary trainings in career development through the research and educational resources at Vanderbilt University. The K25 award will allow the PI protected time to accomplish these goals by following a detailed training plan including hands on laboratory training in biological skills, several courses/workshop over the first two years, and a mentored research plan. For the proposed research project, the PI seeks to develop a statistical model that can be used in combination with Raman fiber optic probe, providing real-time quantitative assessment on the quality of metastatic bone in situ.This goal will be realized in three aims. Aim 1) Characterize the temporal alterations of metastatic bone in breast and prostate cancers using Raman spectroscopy. This aim will prove the overall hypothesis of the study that that the alterations in the quality of metastatic bone can be detected by Raman spectroscopy. Both lyitc and blastic metastases will be studied in an intra-cardiac and an intra-tibial injection mouse model for breast and prostate cancers, respectively. The structural and compositional properties of the bone matrix will be assessed for intact non-tumor and tumor-bearing bones using a fiber optic probe interfaced Raman spectroscopy. These Raman-derived bone quality measurements will be correlated with bone structure (by X-ray imaging and Micro-CT analysis), tumor burden (assessed by bioluminescence imaging in vivo), bone turnover (by serum biomarkers) and bone cell activities (by bone histomorphometry). Statistics evaluation will test whether bone quality is changed with the progression of tumor, and how the osteoblast/osteoclast activities and pathological bone turnover are related to the alterations in the quality of metastatic bone. Aim 2) Develop a statistical model that can provide real-time automated assessment on the quality of metastatic bone in situ. The full range of spectral data collected in Aim 1 will be analyzed using statistical pattern recognition method for extracting the diagnostic features and classifying them into corresponding categories of bone quality. Aim 3) Correlate Raman spectral biomarkers to the microenvironment of metastatic bone. This aim seeks to identify the biological contributors of the apparent spectral signatures (biomarkers) for metastatic bone. Two-Dimensional Raman maps will be acquired at the marked sites where Raman probe measurements are performed using conventional confocal Raman micro-spectroscopy with a spatial resolution at the cellular level. Multivariate factor analysis on the Raman maps can extract the spectrum and spatial distribution of each component (factor) in the sample. The molecular nature of each component will be identified by comparing its spectrum with that of model compounds. Additional guidance is available by comparing the image of factors with the immunohisto chemistry and bone histology images. Upon completion of this project, a quantitative spectral scoring system will be developed, which will enable in situ assessment for bone quality in metastasis when used in combination with Raman spectroscopy. This will provide the PI tools to establish her independent research in bone metastasis. The results will serve as preliminary data for the PI's future extramural funding application.