异常的缺氧应激将引发在肿瘤发生中显着观察到的一系列病理后果。了解缺氧在肿瘤发生中的作用对于开发有效的治疗方法非常重要，这需要可靠的工具来特异性地区分缺氧肿瘤细胞（或组织），并将其动态与复杂生活环境中的疾病状态相关联，以进行精确的治疗诊断。到目前为止，不同的缺氧响应探针分子和前药是通过化学或酶促反应设计的，但由于不受限制的扩散和对环境响应的选择性较低，它们实时报告发病机制发展的能力常常受到损害。在此，我们提出了一种氧不敏感硝基还原酶 (NTR) 可激活的聚糖代谢报告基因 (pNB-ManNAz)，能够共价标记缺氧肿瘤细胞和组织。在病理生理缺氧环境下，笼状的不可代谢前体 pNB-ManNAz 对细胞 NTR 表现出独特的响应性，最终导致结构自焚，所得的 ManNAz 可以整合到细胞表面糖蛋白上，从而促进通过生物正交化学进行荧光标记。这种 NTR 响应代谢报告基因表现出对多细胞缺氧标记的广泛适用性，特别是在动态监测原位肿瘤发生和体内靶向肿瘤光疗方面。我们预计这种方法有望用于研究缺氧相关的病理进展，为准确的诊断和治疗提供有价值的见解。© 2024 Wiley‐VCH GmbH。

Aberrant hypoxic stress will initiate a cascade of pathological consequence observed prominently in tumorigenesis. Understanding of hypoxia's role in tumorigenesis is  highly essential for developing effective therapeutics, which necessitates reliable tools to specifically distinguish hypoxic tumor cells (or tissues) and correlate their dynamics with the status of disease in complex living settings for precise theranostics. So far, disparate hypoxia-responsive probe molecules and prodrugs were designed via chemical or enzymatic reactions, yet their capability in real-time reporting pathogenesis development is often compromised due to unrestricted diffusion and less selectivity towards the environmental responsiveness. Herein we present an oxygen-insensitive nitroreductase (NTR)-activatable glycan metabolic reporter (pNB-ManNAz) capable of covalently labeling hypoxic tumor cells and tissues. Under pathophysiological hypoxic environments, the caged non-metabolizable precursor pNB-ManNAz exhibited unique responsiveness to cellular NTR, culminating in structural self-immolation and the resultant ManNAz could incorporate onto cell surface glycoproteins, thereby facilitating fluorescence labeling via bioorthogonal chemistry. This NTR-responsive metabolic reporter demonstrated broad applicability for multicellular hypoxia labeling, particularly in the dynamic monitoring of orthotopic tumorigenesis and targeted tumor phototherapy in vivo. We anticipate that this approach holds promise for investigating hypoxia-related pathological progression, offering valuable insights for accurate diagnosis and treatment.© 2024 Wiley‐VCH GmbH.