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骨转移定量癌症研究的独立研究者。主要研究兴趣是开发用于癌症诊断或手术指导的新光学方法，并开发/评估骨转移的治疗方法。PI在未来四年的直接目标是获得必要的培训，获得癌症光子学的经验，建立独立的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）追求一个严肃的研究项目，从中她将获得生物医学研究的经验，并为未来的校外资助获得初步数据，5）通过范德比尔特大学的研究和教育资源，建立合作关系，并获得职业发展方面的其他必要培训。K25奖将允许PI保护的时间来完成这些目标，通过以下详细的培训计划，包括生物技能的实验室实践培训，前两年的几个课程/研讨会，以及指导研究计划。 对于拟议的研究项目，PI寻求开发一种统计模型，该模型可与拉曼光纤探针结合使用，提供原位转移性骨质量的实时定量评估。这一目标将通过三个目标实现。目的1）应用拉曼光谱技术研究乳腺癌和前列腺癌骨转移灶的时间变化。这一目的将证明本研究的总体假设，即转移性骨质量的改变可以通过拉曼光谱检测。将分别在乳腺癌和前列腺癌的心脏内和胫骨内注射小鼠模型中研究lyitc和母细胞转移。将使用光纤探针接口拉曼光谱法评估完整非肿瘤骨和荷瘤骨的骨基质的结构和组成特性。这些拉曼衍生的骨质量测量值将与骨结构（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（通过血清生物标志物）和骨细胞活性（通过骨组织形态计量学）相关。统计学评价将检验骨质量是否随肿瘤进展而改变，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改变的关系。目的2）建立一个能实时自动评估骨转移瘤骨质量的统计模型。将使用统计模式识别方法分析目标1中收集的全范围光谱数据，以提取诊断特征并将其分类为相应的骨质类别。目的3）将拉曼光谱生物标志物与骨转移瘤的微环境相关联。该目的旨在确定转移性骨的表观光谱特征（生物标志物）的生物学贡献者。将在标记部位采集二维拉曼图，其中使用具有细胞水平空间分辨率的常规共焦拉曼显微光谱进行拉曼探针测量。对拉曼图谱进行多元因子分析，可以提取样品中各组分（因子）的光谱和空间分布。将通过将其光谱与模型化合物的光谱进行比较来鉴定每种组分的分子性质。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，可获得额外的指导。 本项目完成后，将开发一种定量光谱评分系统，当与拉曼光谱结合使用时，该系统将能够原位评估转移中的骨质。这将为PI提供工具，以建立她在骨转移方面的独立研究。研究结果将作为PI未来校外资助申请的初步数据。 公共卫生相关性：在该项目中，PI寻求开发一种新的拉曼光谱方法，该方法可以在真实的时间内非破坏性地评估转移性疾病的骨质。在临床上，骨质量的正确评估对于帮助骨科医生评估骨转移引起的骨损伤至关重要，而对于治疗开发，骨质量是评估骨对治疗的反应的重要决定因素。因此，从长远来看，该项目将有助于改善骨转移的诊断和预后，并通过避免不必要的测试来限制治疗成本。

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. PUBLIC HEALTH RELEVANCE: In this project, the PI seeks to develop a novel Raman spectroscopy method that can assess bone quality in metastatic disease in real time non-destructively. In clinics the correct assessment of bone quality is crucial to help orthopaedic surgeons evaluate bone damage caused by bone metastasis, while for treatment development, bone quality is an important determinant to evaluate the response of bone to therapy. Thus this project in the long run will help improve the diagnosis and prognosis of bone metastasis and limit treatment costs by avoidance of unnecessary tests.