Macroautophagy (which we will call autophagy hereafter) is a critical intracellular bulk degradation system that is active at basal rates in eukaryotic cells. This process is embedded in the homeostasis of nutrient availability and cellular metabolic demands, degrading primarily long-lived proteins and some organelles, preserving cell viability. Autophagy is perturbed in many pathological conditions, and its manipulation to either enhance or inhibit this pathway therapeutically has received considerable attention. Although better probes are being developed for a more precise readout of autophagic activity in vitro and increasingly in vivo, many questions remain. These center in particular around the accurate measurement of autophagic flux and its translation from the in vitro to the in vivo environment as well as its application in the clinic. In this review, we highlight some of the key aspects that appear to contribute to stumbling blocks on the road towards clinical translation and discuss points of departure for reaching some of the desired goals. We discuss techniques that may be well aligned with achieving the desired spatiotemporal resolution to gather data on autophagic flux in a multi-scale fashion, so as to better apply the existing tools that are based on single-cell analysis and to use them in the living organism. We assess how current techniques may be used for the establishment of autophagic flux standards or reference points and consider strategies for a conceptual approach on titrating autophagy inducers based on their effect on autophagic flux and potency to achieve a favorable protein degradation rate. Finally, we discuss potential solutions for inherent controls for autophagy analysis that are accessible, so as to better discern systemic and tissue-specific autophagic flux in future clinical applications.

巨自噬（以下称为自噬）是一种关键的细胞内整体降解系统，在真核细胞中以基础速率活跃。此过程嵌入营养物可利用性和细胞代谢需求的动态平衡中，主要降解长寿蛋白和一些细胞器，从而保持细胞活力。自噬在许多病理条件下均受到干扰，其在治疗上增强或抑制该途径的操作受到了广泛的关注。尽管正在开发更好的探针以更精确地读出体外和体内越来越多的自噬活性，但仍然存在许多问题。这些中心尤其围绕自噬通量的准确测量及其从体外到体内环境及其在临床中的应用。在这篇综述中，我们重点介绍了一些关键方面，这些方面似乎有助于实现临床翻译的绊脚石，并讨论了实现某些预期目标的出发点。我们讨论的技术可能与实现所需的时空分辨率很好地吻合，以多尺度方式收集自噬通量的数据，以便更好地应用基于单细胞分析的现有工具并将其用于生活中。生物。我们评估如何将当前的技术用于建立自噬通量标准或参考点，并根据对自噬通量和对自噬通量的影响以及达到理想蛋白质降解率的能力，考虑概念化滴定自噬诱导剂的策略。最后，