Core A: Bioanalysis

项目摘要 生物分析核心旨在提供集中的生物分析支持。这一核心将促进 并提供可共享和比较基本信息 项目中的三个项目。这一新的、重组后的核心将为以下方面提供支持： 用于临床的现有光动力敏化剂（PS）的临床和临床前组织和生物流体分布 以及临床前研究和任何新开发的用于光动力疗法（PDT）的PS。这些数据至关重要 用于解释患者和动物模型中的PDT活性。项目2中描述的PS吸收了 可见光谱的红色或NIR区域，并且是强荧光的。因此，标准化检测 已经开发了利用这些化合物的物理化学性质的方法。生物分析 核心将评估选定化合物在荷瘤小鼠和患者中的肿瘤组织保留。 使用荧光分光光度法（并且，如果可能的话，液体闪烁计数， 检测放射性同位素标记的[14C]光敏剂）。荧光分光光度法将用于 测定人血清和溶解的活检和手术标本中的光敏剂水平。核心A将 测量SADA（血清碱性DNA酶）作为循环生物标志物，有可能预测PDT 头部和颈部患者的反应。这将包括从患者中采集常规血液， SADA活性标准分光光度分析。核心也会收集血液， 我们的头颈部病人的免疫细胞进行分析。具体而言，各个项目已确定 为临床和转化科学研究提供多个领域的支持。核心的具体目标 是：1）确定患者组织和生物体液中的光敏剂水平，2）收集、分离和 储存来自患者的免疫细胞用于后续分析，3）测量非侵入性循环生物标志物 （血清碱性DNA酶）和4）测量光敏剂在带有异种移植头部的小鼠中的保留 颈部肿瘤。

PROJECT SUMMARY The purpose of the Bioanalysis Core is to provide centralized bioanalysis support. This Core will promote standardization and efficiency of analysis and provide essential information that can be shared and compared across all three Projects in the Program. This new, reorganized Core will provide support for all aspects of clinical and preclinical tissue and biofluid distribution of existing photodynamic sensitizers (PS) used in clinical and preclinical studies and any newly developed PS for photodynamic therapy (PDT). These data are essential for interpreting PDT activity in patients and animal models. PSs described in Project 2 have absorption in the red or NIR regions of the visible spectrum and are strongly fluorescent. As such, standardized detection methods have been developed to exploit the photophysical properties of these compounds. The Bioanalysis Core will evaluate the tumor tissue retention of select compounds in mice bearing established and patient- derived human tumors using fluorescence spectrophotometry (and, if possible, liquid scintillation counting to detect radioisotopically-labeled [14C] photosensitizers). Fluorescence spectrophotometry will be used to determine photosensitizer levels in human serum and solubilized biopsy and surgical specimens. Core A will measure SADA (serum alkaline DNase) as a circulating biomarker that has the potential to predict PDT response in head and neck patients. This will involve routine blood collection from patients followed by standard spectrophotometric analysis of SADA activity. The Core will also collect blood and isolate and store immune cells from our head and neck patients for analysis. Specifically, the individual Projects have identified a number of areas of support for both clinical and translational science studies. The specific aims of the Core are: 1) to determine photosensitizer levels in tissues and biological fluids of patients, 2) to collect, isolate and store immune cells from patients for subsequent analysis, 3) to measure noninvasive, circulating biomarker (serum alkaline DNase) and 4) to measure the retention of photosensitizers in mice bearing xenografted head and neck tumors.