Covalent JNK Inhibitor, JNK-IN-8, Suppresses Tumor Growth in Triple-Negative Breast Cancer by Activating TFEB- and TFE3-Mediated Lysosome Biogenesis and Autophagy.

The heterogeneity and aggressiveness of triple-negative breast cancer (TNBC) contribute to its early recurrence and metastasis. Despite substantial research to identify effective therapeutic targets, TNBC remains elusive in terms of improving patient outcomes. Here, we report that a covalent JNK inhibitor, JNK-IN-8, suppresses TNBC growth both in vitro and in vivo. JNK-IN-8 reduced colony formation, cell viability, and organoid growth in vitro and slowed patient-derived xenograft and syngeneic tumor growth in vivo. Cells treated with JNK-IN-8 exhibited large, cytoplasmic vacuoles with lysosomal markers. To examine the molecular mechanism of this phenotype, we looked at the master regulators of lysosome biogenesis and autophagy transcription factor EB (TFEB) and TFE3. JNK-IN-8 inhibited TFEB phosphorylation and induced nuclear translocation of unphosphorylated TFEB and TFE3. This was accompanied by an upregulation of TFEB/TFE3 target genes associated with lysosome biogenesis and autophagy. Depletion of both TFEB and TFE3 diminished the JNK-IN-8-driven upregulation of lysosome biogenesis and/or autophagy markers. TFEB and TFE3 are phosphorylated by a number of kinases, including mTOR. JNK-IN-8 reduced phosphorylation of mTOR targets in a concentration-dependent manner. Knockout of JNK1 and/or JNK2 had no impact on TFEB/TFE3 activation or mTOR inhibition by JNK-IN-8 but inhibited colony formation. Similarly, reexpression of either wildtype or drug-nonbinding JNK (C116S) in JNK knockout cells did not reverse JNK-IN-8-induced TFEB dephosphorylation. In summary, JNK-IN-8 induced lysosome biogenesis and autophagy by activating TFEB/TFE3 via mTOR inhibition independently of JNK. Together, these findings demonstrate the efficacy of JNK-IN-8 as a targeted therapy for TNBC and reveal its novel lysosome- and autophagy-mediated mechanism of action.

Most YouTube Videos About Carpal Tunnel Syndrome Have the Potential to Reinforce Misconceptions.

BACKGROUND: Studies of online health information have addressed completeness and adherence to evidence, which can be difficult because current evidence leaves room for debate about etiology, diagnosis, and treatment. Fewer studies have evaluated whether online health information can reinforce misconceptions. It can be argued that information with the potential to harm health by reinforcing unhelpful misconceptions ought to be held to a higher standard of evidence. QUESTIONS/PURPOSES: (1) What is the prevalence and nature of health information in YouTube videos with the potential to reinforce common misconceptions about symptoms and treatment associated with carpal tunnel syndrome (CTS)? (2) What factors (such as the number of views, likes, and subscribers) are associated with Potential Reinforcement of Misconception scores of YouTube videos about CTS? METHODS: After removing all personalized data, we searched for the term "carpal tunnel syndrome" on YouTube, reviewed the first 60 English-language videos that discussed the diagnosis and treatment of CTS, and collected available metrics. The primary outcome was the number of statements that could reinforce misconceptions about CTS, rated by two authors using a checklist. As a secondary outcome, we counted the number of statements that could help patients by reorienting or balancing common misconceptions, providing agency, and facilitating decisions, and we subtracted the number of potential misconceptions from this count. A modified version of the DISCERN instrument (a validated scoring system designed to gauge the quality and reliability of health information) was used to evaluate each video. We sought factors associated with the Potential Reinforcement of Misconception score-in both the negative-only and combined (positive and negative) variations-accounting for various YouTube metrics (such as the number of views, number of likes and dislikes, and duration) and the modified DISCERN score. The interrater reliability was excellent for both the Potential Reinforcement of Misconceptions checklist (ICC = 0.97; Pearson correlation [r] = 0.97) and DISCERN information quality score (ICC = 0.96; r = 0.97). RESULTS: Seventy-eight percent of the YouTube videos (47 of 60 videos) contained at least one statement that could reinforce common misconceptions about CTS. The median number of potentially misconception-reinforcing statements was two (range one to three), with the most common statements being that CTS is caused by hand use (38%; 23 of 60 videos) and that splints can alter the natural history of the disease (37%; 22 videos). Videos that were more popular (higher number of views or likes) did not contain less potential reinforcement of misconceptions. In the multivariable analysis, we found a strong association between the DISCERN score and the CTS Potential Reinforcement of Misconceptions score (regression coefficient = 0.67; 95% CI 0.22-1.2; partial r2 = 0.13; p = 0.004) and a lower number of subscribers (calculated per one million subscribers: regression coefficient = -0.91; 95% CI -1.8 to -0.023; p = 0.045). CONCLUSION: Potential reinforcement of misconceptions is prevalent in YouTube videos about CTS, more so in videos with lower information quality scores. CLINICAL RELEVANCE: Online health information should be held to a standard of accuracy (alignment with best evidence), and where such evidence leaves room for debate, it should be held to a standard by which unhealthy misconceptions are not reinforced.