Profiling chemical tumor microenvironment: Application for diagnostics & therapy

项目摘要/摘要 TME在癌症进展、治疗耐药以及作为治疗干预靶点方面的关键作用日益受到重视。基于体内EPR的组织氧合(Po{2})、细胞外pH(pH{e})、氧化还原、谷胱甘肽(GSH)和间质无机磷(PI)的无创波谱和成像为了解TME的生物学过程提供了独特的见解，并可作为临床前筛选抗癌药物和优化TME靶向治疗策略的工具。在这次竞争性续签申请中，我们的新方向建立在前一个资助期的知识和发现的基础上。我们的R01续订提案有两个目标。第一是进一步推进顺磁探针和磁共振技术，重点提高探针的生物兼容性、功能性，并开发多功能成像模式。第二，利用这些进展和在最初的R01项目期间使用光谱模式积累的知识，以获得对TME在癌症进展中的作用的进一步见解，并测试我们关于间质无机磷在肿瘤发生中的作用的假设。具体目标是：(SA1)推进基于EPR的分子多功能成像技术和顺磁探针。对细胞外组织微环境中的酸性(pH{e})、氧气(Po{2})和磷酸盐(PI)敏感的多功能三叔丁基HOPE探针的生物相容性衍生物，以及双功能氮氧化物pH和氧化还原探针，将使用尖端成像方式、快速扫描EPR成像和Overhaser增强磁共振成像进行优化，以同时进行多功能成像。(SA2)随着乳腺肿瘤的恶性进展，对化学物质TME进行分子成像。多功能空间分辨TME分析将在乳腺癌小鼠模型中验证基于形态和基因表达的分期，并将绘制具有不同表型的肿瘤区域。来自具有恶性表型TME化学特征的地区的组织样本将被分离，TME研究将通过免疫组织化学、生化和遗传组织分析以及血液中总磷和磷(PI)含量的测量来补充。总之，开发的多功能成像技术和探针将扩大临床前EPR的范围，允许在癌症和其他疾病的各种疾病模型中绘制生理相关的组织参数图。需要对肿瘤进展过程中特定阶段的TME进化的新知识来优化TME靶向抗癌治疗。这也将为唤起公众对加工食品中高含量的磷酸盐改性剂及其潜在的健康风险的认识提供科学依据。

PROJECT SUMMARY/ABSTRACT A key role of the TME in cancer progression, treatment resistance, and as a target for therapeutic intervention is increasingly appreciated. Noninvasive in vivo EPR-based spectroscopy and imaging of tissue oxygenation (pO{2}), extracellular pH (pH{e}), redox, glutathione (GSH) and interstitial inorganic phosphate (Pi) provide unique insights into biological processes in the TME, and may serve as a tool for preclinical screening of anticancer drugs and optimizing TME-targeted therapeutic strategies. In this competitive renewal application, our new directions are built on knowledge and discoveries made during the previous funding period. There are two goals of our R01 renewal proposal. First is to further advance paramagnetic probes and magnetic resonance technology with a focus on improving probes biocompatibility, functionality, and developing multifunctional imaging modalities. Second is to utilize these advances and knowledge accumulated using spectroscopic modalities in the initial R01 project period to obtain further insights into the role of TME in cancer progression, and to test our hypothesis on the roles of interstitial inorganic phosphate in tumorigenesis. The specific aims are: (SA1) To advance molecular multifunctional EPR-based imaging technology and paramagnetic probes. The biocompatible derivatives of multifunctional trityl HOPE probe sensitive to acidity (pH{e}), oxygen (pO{2}) and phosphate (Pi) in Extracellular tissue microenvironment, and dual function nitroxide pH & redox probes will be optimized for concurrent multifunctional imaging using cutting edge imaging modalities, rapid scan EPR imaging and Overhauser-enhanced magnetic resonance imaging. (SA2) To perform molecular imaging of chemical TME as the mammary tumors progress to malignancy. Multifunctional spatially-resolved TME profiling will validate morphological and gene expression-based staging in a mouse model of breast cancer and will map tumor regions with different phenotypes. Tissue samples from the areas with the chemical TME characteristics of a malignant phenotype will be isolated and the TME study will be complemented with immunohistochemical, biochemical, and genetic tissue analysis, and with measurements of total phosphorus and phosphate (Pi) contents in blood. In summary, the developed multifunctional imaging techniques and probes will broaden the scope of preclinical EPR allowing for mapping of physiologically relevant tissue parameters in various disease models in cancers and beyond. New knowledge on stage-specific TME evolution during tumor progression is required to optimize TME-targeted anticancer therapies. It will also provide a scientific basis to evoke public awareness of high content of the phosphate-based modifiers in the processed food and the potential health risk.