Park Jaena, Kamerer Rebecca L, Marjanovic Marina, Sorrells Janet E, You Sixian, Barkalifa Ronit, Selting Kimberly A, Boppart Stephen A
Department of Bioengineering, University of Illinois at Urbana-Champaign IL, USA.
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign IL, USA.
Am J Cancer Res. 2022 May 15;12(5):2068-2083. eCollection 2022.
Extracellular vesicles (EVs) have been studied for their potential applications in cancer screening, diagnosis, and treatment monitoring. Most studies have focused on the bulk content of EVs; however, it is also informative to investigate their metabolic status, and changes under different physiological and environmental conditions. In this study, noninvasive, multimodal, label-free nonlinear optical microscopy was used to evaluate the optical redox ratio of large EVs (microvesicles) isolated from the urine of 11 dogs in three cohorts (4 healthy, 4 transitional cell carcinoma (TCC) of the bladder, and 3 prostate cancer). The optical redox ratio is a common metric comparing the autofluorescence intensities of metabolic cofactors FAD and NAD(P)H to characterize the metabolic profile of cells and tissues, and has recently been applied to EVs. The optical redox ratio revealed that dogs with TCC of the bladder had a more than 2-fold increase in NAD(P)H-rich urinary EVs (uEVs) when compared to healthy dogs, whereas dogs with prostate cancer had no significant difference. The optical redox ratio values of uEVs kept at -20°C for 48 hours were significantly different from those of freshly isolated uEVs, indicating that this parameter is more reliable when assessing freshly isolated uEVs. These results suggest that the label-free optical redox ratio of uEVs, indicating relative rates of glycolysis and oxidative phosphorylation of parent cells and tissues, may act as a potential screening biomarker for bladder cancer.
细胞外囊泡（EVs）因其在癌症筛查、诊断和治疗监测中的潜在应用而受到研究。大多数研究集中在EVs的总体内容上；然而，研究它们的代谢状态以及在不同生理和环境条件下的变化也很有意义。在本研究中，使用无创、多模态、无标记非线性光学显微镜来评估从三个队列的11只狗的尿液中分离出的大型EVs（微囊泡）的光学氧化还原比。这三个队列分别为4只健康狗、4只膀胱移行细胞癌（TCC）狗和3只前列腺癌狗。光学氧化还原比是一种常用指标，用于比较代谢辅因子FAD和NAD(P)H的自发荧光强度，以表征细胞和组织的代谢特征，最近已应用于EVs。光学氧化还原比显示，与健康狗相比，患有膀胱TCC的狗富含NAD(P)H的尿EVs（uEVs）增加了两倍多，而患有前列腺癌的狗则没有显著差异。在-20°C保存48小时的uEVs的光学氧化还原比值与新鲜分离的uEVs的光学氧化还原比值有显著差异，这表明在评估新鲜分离的uEVs时，该参数更可靠。这些结果表明，uEVs的无标记光学氧化还原比，表明母细胞和组织的糖酵解和氧化磷酸化的相对速率，可能作为膀胱癌的潜在筛查生物标志物。
