Transcriptomic analysis on prostate tissues uncover the diverse gene expression features induced by Lonp1. Transcriptomic analysis on prostate tissues uncover the diverse gene expression features induced by Lonp1

Lon protease 1 (LONP1) is an ATP-dependent protease located in the mitochondrial matrix and plays a crucial role in regulating mitochondrial proteostasis, metabolism and cellular stress responses. Here we show that overexpression of LONP1 is closely related to adverse clinicopathological features and poor prognosis in prostate cancer (PCa) patients. Mechanistically, the findings reveal that LONP1 is implicated in modulating the metabolic switch from oxidative phosphorylation (OXPHOS) to aerobic glycolysis, thereby promoting tumor proliferation, invasion and metastasis both in vitro and in vivo. Meanwhile, we prove that LONP1 as a protease directly targets mitochondrial pyruvate carrier 1 (MPC1), a key metabolic protein in the process of glycolysis, and enhances its degradation, which in turn suppresses tricarboxylic acid (TCA) cycle and ultimately promotes the progression of PCa. Furthermore, using PCa in cancer-prone mice homozygous for a prostate-targeted conditional Pten knockout and Lonp1 knockin, we integrate transcriptomic and proteomic analyses of prostate tumors, upon which reveals that Lonp1 overexpression results in a significant downregulation of NADH: ubiquinone oxidoreductase activity, consequently impeding the electron transfer process and mitochondrial ATP synthesis, associated with metastasis of PCa. Collectively, our results highlight that metabolic reprogramming induced by LONP1 in PCa is closely coupled with disease progression, suggesting that targeting the LONP1-mediated cascade in the mitochondrial may provide therapeutic potential for PCa disease. Overall design: To investigate whether lonp1 overexpression has a causal role in PCa metastasis, genetically engineered mouse was used to generate a PCa model. Firstly, we crossed Lonp1KI mice withProbasin-Cre mice to induce Lonp1 expression specifically in the prostate epithelium. Subsequently, we generated Pten-/-; Lonp1KI mice by crossing Lonp1KI mice with Ptenfloxed/floxed mice. Transcriptomic analysis was then performed on prostates extracted from 40-week-old wild-type (WT), Lonp1KI, Pten-/- and Pten-/-; Lonp1KI mice.

Lon蛋白酶1（Lon protease 1, LONP1）是一种定位于线粒体基质的ATP依赖性蛋白酶，在调控线粒体蛋白质稳态、代谢及细胞应激反应中发挥关键作用。本研究发现，LONP1过表达与前列腺癌（prostate cancer, PCa）患者的不良临床病理特征及不良预后密切相关。机制层面，研究结果揭示LONP1参与调控从氧化磷酸化（oxidative phosphorylation, OXPHOS）到有氧糖酵解的代谢转换，进而在体内外均能促进肿瘤增殖、侵袭与转移。同时，本研究证实，作为蛋白酶的LONP1可直接靶向糖酵解过程中的关键代谢蛋白线粒体丙酮酸载体1（mitochondrial pyruvate carrier 1, MPC1）并增强其降解，由此抑制三羧酸循环（tricarboxylic acid cycle, TCA），最终推动前列腺癌的进展。此外，本研究利用前列腺靶向条件性Pten敲除且纯合Lonp1敲入的前列腺癌易感小鼠模型，对前列腺肿瘤开展转录组与蛋白质组整合分析，结果显示LONP1过表达会显著下调NADH:泛醌氧化还原酶活性，进而阻碍电子传递过程与线粒体ATP合成，该现象与前列腺癌转移密切相关。总体实验设计：为探究LONP1过表达在前列腺癌转移中是否具有因果作用，本研究采用基因工程小鼠构建前列腺癌模型。首先，将Lonp1敲入（Lonp1KI）小鼠与Probasin-Cre小鼠杂交，使LONP1特异性在前列腺上皮细胞中表达；随后，将Lonp1KI小鼠与Pten floxed/floxed小鼠杂交，获得Pten-/-; Lonp1KI小鼠。最后，对40周龄的野生型（wild-type, WT）、Lonp1KI、Pten-/-及Pten-/-; Lonp1KI小鼠的前列腺组织进行转录组分析。