Profiling chemical tumor microenvironment: application for diagnostics & therapy

描述（由申请人提供）：如今，人们已经广泛认识到，恶性细胞通常会根据其特定的组织微环境（TME）表现不同。在这里，我们假设肿瘤TME的双面特性，提出TME氧合、细胞外pH （pHe）、无机磷酸盐（Pi）、氧化还原和谷胱甘肽（GSH）稳态的特定模式利用一种协调机制促进癌细胞存活，同时对正常细胞具有高毒性和致突变性。该项目的总体目标是使创新的磁共振方法能够用于化学TME的体内多功能分析，以提高对恶性转移的早期诊断的预测能力，并为未来合理设计针对TME的抗癌治疗方法。具体目标是：（SA1）推进磁共振技术和顺磁探针在体内实时组织微环境分析。利用电子顺磁共振（EPR）和质子-电子双磁共振成像（PEDRI）技术进行多功能测量的顺磁探针将被推进到进行体内实时TME分析。即优化用于同时监测pO2、pHe和Pi的三烷基探针，以及用于测量pH、GSH和还原容量的氮氧化物探针。SA 2：在肿瘤向恶性发展时进行体内肿瘤微环境分析。来自（SA1）的探针阵列将用于使用我们的PyMT转基因小鼠群体进行化学TME分析，当乳腺肿瘤进展为恶性肿瘤时，这些小鼠会自发发展为乳腺癌并模拟人类肿瘤分期。除了l波段EPR光谱测量外，PEDRI功能作图将用于评估肿瘤TME的探针分布和功能异质性。动态对比增强（DCE） MRI将用于量化肿瘤的空间异质性。目的3：确定预后因素并评估组织微环境操作的疗效。化学TME的正常化可能为肿瘤TME靶向治疗干预提供新的依据。在SA 3A中，我们将单独进行TME操作，在SA 3B中，我们将联合标准化疗。我们期望旨在预防/补偿组织缺氧和酸中毒的程序也会影响组织氧化还原，Pi和GSH，并改善癌前乳腺病变小鼠模型的结果。在Aim 3B中，我们将验证我们的假设，即TME化学正常化将增加使用标准化疗多西紫杉醇的治疗效果，以解决肿瘤解毒和化疗耐药性的重要临床问题。

DESCRIPTION (provided by applicant): Today it is becoming widely recognized that malignant cells very often behave differently depending on their specific tissue microenvironment (TME). Here we hypothesized Janus-faced properties of tumor TME proposing that specific patterns of TME oxygenation, extracellular pH (pHe), inorganic phosphate (Pi), redox and glutathione (GSH) homeostasis act to utilize an orchestrated mechanism to promote cancer cell survival while at the same time being highly toxic and mutagenic for normal cells. The overall goal of this project is to enable innovative magnetic resonance methods for in vivo multifunctional profiling of chemical TME to improve prediction power of early diagnostics for the malignant transition and for future rational design of TME-targeted anticancer therapeutics. The specific aims are: (SA1) To advance magnetic resonance technology and paramagnetic probes for in vivo real-time tissue microenvironment profiling. The paramagnetic probes for multifunctional measurements using Electron Paramagnetic Resonance (EPR) and Proton-Electron Double- Resonance imaging (PEDRI) techniques will be advanced to perform in vivo real-time TME profiling. Namely, trityl probes for concurrent monitoring of pO2, pHe and Pi, and nitroxide probes for pH, GSH and reducing capacity measurements will be optimized. SA 2: To perform in vivo tumor microenvironment profiling as the tumor progresses to malignancy. The array of the probes from (SA1) will be used to perform chemical TME profiling as the mammary tumors progress to malignancy using our colony of PyMT transgenic mice which spontaneously develop breast cancer and emulate human tumor staging. In addition to L-band EPR spectroscopic measurements, PEDRI functional mapping will be used to assess probe distribution and functional heterogeneity of tumor TME. Dynamic contrast-enhanced (DCE) MRI will be used to quantify tumor spatial heterogeneity. SA 3: To identify prognostic factors and evaluate efficacy of tissue microenvironment manipulation. Normalizing chemical TME may provide new basis for anticancer TME-targeted therapeutic interventions. In SA 3A we will perform TME manipulation alone, and in combination with standard chemotherapy in SA 3B. We expect that the procedures aimed to prevent/compensate tissue hypoxia and acidosis will also affect tissue redox, Pi and GSH, and improve outcome in a mouse model of pre-cancer breast lesions. In Aim 3B, we will test our hypothesis that TME chemical normalization will augment therapeutic effectiveness using the standard chemotherapy, docetaxel, to address the important clinical problem of tumor detoxification and chemotherapy resistance.