Label-Free Optical Redox Imaging for Pretreatment Prognosis of Early-Stage Triple Negative Breast Cancer

无标记光学氧化还原成像对早期三阴性乳房治疗前预后的评价 癌 摘要 三阴性乳腺癌(TNBC)在肿瘤内微环境和转移方面存在显著差异。 LISM，它可以很大程度上影响疾病的进展和临床结果。TNBC肿瘤可以含有hy- 缺氧区和常氧区，表现为糖酵解、氧化或混合代谢亚型。这些新陈代谢 在临床表现的乳腺肿瘤中，亚型通常在亚毫米尺度上，低于 目前的临床代谢成像方法，突出了对新的高分辨率代谢成像的需求 方法临床应用。光学氧化还原成像(ORI)，一种在 氧化还原成像的布里顿·钱斯实验室可以检测肿瘤内代谢亚型三种- 尺寸(3D)，分辨率低至25微米。ORI测量Re-Re的本征荧光信号 诱导烟酰胺腺嘌呤二核苷酸(NADH)和氧化黄素蛋白(FP)，并测定氧化 氧化还原代谢状态降低。我们的初步结果发现了氧化还原热点，高度氧化的Re- 治疗中的DOX亚型--来自早期TNBC患者试点队列的幼稚样本。值得注意的是，在 研究队列，ORI氧化还原热点比传统临床更好地预测疾病进展的风险 指标(例如，肿瘤大小、分期、分级和结节状态)。我们研究未经处理的TNBC异种移植和 细胞培养还表明，氧化还原热点是由Warburg效应(糖酵解开关)支撑的， 证实了ORI对进展风险的预测。基于这些结果，这一项目的长期目标是 计划是通过扩大在TNBC临床中的观察来建立预处理ORI的预后价值 标本和临床前模型。目标1将是一项全面的冷冻未处理手术的ORI研究- 早期TNBC患者的CAL标本。我们将测试假设，肿瘤内氧化还原热点 预测进展风险增加，并为常规预后指标增加价值。目标2将评估 应用进展法评价ORI在人-TNBC-小鼠移植瘤和培养细胞模型中的预后价值 并确认激活的糖酵解开关是更高的氧化还原热点和风险的基础 进步。该项目的成功完成将使体外ORI成为一种新的前处理方法 以亚毫米分辨率指示瘤内糖酵解开关的工具，并告知有利于- 对于早期TNBC患者的个体化治疗。我们的项目将帮助解决未得到满足的临床需求 准确的预后生物标志物，以改善TNBC的风险分层和个性化治疗。

Label-Free Optical Redox Imaging for Pretreatment Prognosis of Early-Stage Triple Negative Breast Cancer Abstract Triple negative breast cancer (TNBC) has significant intratumor variations in microenvironments and metabo- lism, which can substantially affect disease progression and clinical outcomes. A TNBC tumor can contain hy- poxic and normoxic regions and exhibit glycolytic, oxidative, or mixed metabolic subtypes. These metabolic subtypes are typically on a submillimeter scale in clinically presented breast tumors, below the resolution of current clinical metabolic imaging methods, highlighting the need of new high-resolution metabolic imaging methods in the clinic. Optical Redox Imaging (ORI), a label-free fluorescence imaging technique developed at the Britton Chance Laboratory of Redox Imaging, can detect intratumor metabolic subtypes three- dimensionally (3D) with a resolution down to 25 µm. ORI measures the intrinsic fluorescence signals of re- duced nicotinamide adenine dinucleotide (NADH) and oxidized flavoproteins (Fp) and determines the oxidized and reduced status of redox metabolism. Our preliminary results found redox hotspots, the highly oxidized re- dox subtype in treatment-naïve specimens, from a pilot cohort of early-stage TNBC patients. Notably, in the studied cohort, ORI redox hotspots predicted the risk of disease progression better than conventional clinical indicators (e.g., tumor size, stage, grade, and nodal status). Our data studying untreated TNBC xenografts and cell cultures also suggested that the redox hotspots are underpinned by the Warburg effect (glycolytic switch), corroborating the prediction of risk of progression by ORI. Based on these results, the long-term goal of this program is to establish the prognostic value of pretreatment ORI by expanding observations in TNBC clinical specimens and preclinical models. Aim 1 will be a full-scale retrospective ORI study of frozen untreated surgi- cal specimens from early-stage TNBC patients. We will test the hypothesis that the intratumor redox hotspots predict an increased progression risk and add value to conventional prognostic indicators. Aim 2 will assess the prognostic value of ORI in human TNBC mouse xenografts and cultured cell models using progression endpoints and confirm that the activated glycolytic switch is the basis for the higher redox hotspot and risk of progression. The successful accomplishment of this project will establish ex vivo ORI as a novel pretreatment tool that is indicative of the intratumor glycolytic switch at submillimeter resolution and inform the risks of pro- gression for individual patients with early-stage TNBC. Our project will help resolve the unmet clinical need for accurate prognostic biomarkers to improve risk stratification and personalized treatment in TNBC.