The chromatin remodeling amine oxidase lysine-specific demethylase 1 (LSD1) has become an attractive target for the design of specific inhibitors with therapeutic potential. We, and others, have described LSD1 inhibitors that have potential as antitumor agents. Many of the currently known LSD1 inhibitors are poor drug candidates, or are structurally based on the tranylcypromine backbone, thus increasing the potential for off-target effects mediated by other amine oxidases. We now describe a series of potent LSD1 inhibitors based on a novel 1,2,4-triazole scaffold; these inhibitors show a high degree of specificity for LSD1 in vitro, and cause increases in cellular histone 3 dimethyllysine 4 (H3K4me2), a gene transcription activating mark. Importantly, these inhibitors are not toxic to mammalian cells in vitro, and thus they may show utility in the treatment of epigenetically-based diseases where cell death is not a desired endpoint Figure 1. Structures of LSD1 inhibitors 1, verlindamycin 2, (bis)thioureas 3, amidoxime 4, cyclic peptide 5, N3-(2-chloro-6-phenoxybenzyl)-4H-1,2,4-triazole-3,5-diamine 6 and N3,N5-bis(2-methoxybenzyl)-1H-1,2,4-triazole-3,5-diamine 7.

Methylation at specific histone lysine residues is a critical post-translational modification that alters chromatin architecture, and dysregulated lysine methylation/demethylation is associated with the silencing of tumor suppressor genes. The enzyme lysine-specific demethylase 1 (LSD1) complexed to specific transcription factors catalyzes the oxidative demethylation of mono- and dimethyllysine 4 of histone H3 (H3K4me and H3K4me2 respectively). We have previously reported potent (bis)urea and (bis)thiourea LSD1 inhibitors that increase cellular levels of H3K4me and H3K4me2, promote the re-expression of silenced tumor suppressor genes and suppress tumor growth in vitro. Here we report the design additional (bis)urea and (bis)thiourea LSD1 inhibitors that feature 3-5-3 or 3-6-3 carbon backbone architectures. Three of these compounds displayed single-digit IC50 values in a recombinant LSD1 assay. In addition, compound 6d exhibited an IC50 of 4.2 μM against the Calu-6 human lung adenocarcinoma line, and 4.8 μM against the MCF7 breast tumor cell line, in an MTS cell viability assay. Following treatment with 6b–6d, Calu-6 cells exhibited a significant increase in the mRNA expression for the silenced tumor suppressor genes SFRP2, HCAD and p16, and modest increases in GATA4 message. The compounds described in this paper represent the most potent epigenetic modulators in this series, and have potential for use as antitumor agents.

Ischemia-reperfusion (IR) injury comprises a significant portion of morbidity and mortality from heart and brain diseases worldwide. This enduring clinical problem has inspired myriad reports in the scientific literature of experimental interventions seeking to elucidate the pathology of IR injury. Elective cardiac surgery presents perhaps the most viable scenario for protecting the heart and brain from IR injury due to the opportunity to condition the organs prior to insult. The physiological parameters for the preconditioning of vital organs prior to insult through mechanical and pharmacological maneuvers have been heavily examined. These investigations have revealed new insights into how preconditioning alters cellular responses to IR injury. However, the promise of preconditioning remains unfulfilled at the clinical level, and research seeking to implicate cell signals essential to this protection continues. Recent discoveries in molecular biology have revealed that gene expression can be controlled through posttranslational modifications, without altering the chemical structure of the genetic code. In this scenario, gene expression is repressed by enzymes that cause chromatin compaction through catalytic removal of acetyl moieties from lysine residues on histones. These enzymes, called histone deacetylases (HDACs), can be inhibited pharmacologically, leading to the de-repression of protective genes. The discovery that HDACs can also alter the function of non-histone proteins through posttranslational deacetylation has expanded the potential impact of HDAC inhibitors for the treatment of human disease. HDAC inhibitors have been applied in a very small number of experimental models of IR. However, the scientific literature contains an increasing number of reports demonstrating that HDACs converge on preconditioning signals in the cell. This review will describe the influence of HDACs on major preconditioning signaling pathways in the heart and brain.

Background Non-small cell lung cancers (NSLC) are aggressive cancers that are insensitive to chemotherapies and accounts for nearly 33% of all cancer deaths in the United States. Two hallmarks of cancer that allow cells to invade and metastasize are sustained proliferation and enhanced motility. In this study we investigate the relationship between urokinase plasminogen activator (uPA)/uPA receptor (uPAR) signaling and Na+/H+ exchanger isoform 1 (NHE1) expression and activity. Methods and Results The addition of 10nM uPA increased the carcinogenic potential of three NSCLC cell lines, NCI-H358, NCI-H460, and NCI-H1299. This included an increase in the rate of cell proliferation 1.6 to 1.9 fold; an increase in the percentage of cells displaying stress fibers 3.05 to 3.17 fold; and an increase in anchorage-independent growth from 1.64 to 2.0 fold. In each of these cases the increase was blocked when the experiments were performed with NHE1 inhibited by 10 μM EIPA (ethylisopropyl amiloride). To further evaluate the role of uPA/uPAR and NHE1 in tumor progression we assessed signaling events using full-length uPA compared to the uPA amino terminal fragment (ATF). Comparing uPA and ATF signaling in H460 cells, we found that both uPA and ATF increased stress fiber formation approximately 2 fold, while uPA increased matrix metalloproteinase 9 (MMP9) activity 5.44 fold compared to 2.81 fold for ATF. To expand this signaling study, two new cell lines were generated, one with reduced NHE1 expression (H460 NHE1 K/D) and one with reduced uPAR expression (H460 uPAR K/D). Using the K/D cell lines we found that neither uPA nor ATF could stimulate stress fiber formation or MMP9 activity in cells with dramatically decreased NHE1 or uPAR expression. Finally, using in vivo tumor formation studies in athymic mice we found that when mice were injected with H460 cells 80% of mice formed tumors with an average volume of 390 mm3. This was compared to 20% of H460 uPAR K/D injected mice forming tumors with an average volume of 15 mm3 and 10% of H460 NHE1 K/D injected mice forming tumors with an average volume of 5 mm3. Conclusion Taken together, these data demonstrate that uPA/uPAR-mediated tumor progression and metastasis requires NHE1 in NSCLC cells and suggests a potential therapeutic approach to blocking cancer progression.

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INTRODUCTION: The U.S. Navy experienced a series of physiological events in aircrew involving primarily the F/A-18 airframe related to rapid decompression of cabin pressures, of which aviation decompression sickness (DCS) was felt to contribute. The underlying pathophysiology of aviation DCS is the same as that of diving-related. However, based on the innate multifactorial circumstances surrounding hypobaric DCS, in clinical practice it continues to be unpredictable and less familiar as it falls at the intersect of aerospace and hyperbaric medicine. This retrospective study aimed to review the case series diagnosed as aviation DCS in a collaborative effort between aerospace specialists and hyperbaricists to increase appropriate identification and treatment of hypobaric DCS.METHODS: We identified 18 cases involving high-performance aircraft emergently treated as aviation DCS at a civilian hyperbaric chamber. Four reviewers with dual training in aviation and hyperbaric medicine retrospectively reviewed cases and categorized presentations as “DCS” or “Alternative Diagnosis”.RESULTS: Reviewers identified over half of presenting cases could be attributed to an alternative diagnosis. In events that occurred at flight altitudes below 17,000 ft (5182 m) or with rapid decompression pressure changes under 0.3 atm, DCS was less likely to be the etiology of the presenting symptoms.CONCLUSIONS: Aviation physiological events continue to be difficult to diagnose. This study aimed to better understand this phenomenon and provide additional insight and key characteristics for both flight physicians and hyperbaric physicians. As human exploration continues to challenge the limits of sustainable physiology, the incidence of aerospace DCS may increase and underscores our need to recognize and appropriately treat it.Kutz CJ, Kirby IJ, Grover IR, Tanaka HL. Aviation decompression sickness in aerospace and hyperbaric medicine. Aerosp Med Hum Perform. 2023; 94(1):11–17.

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Non‐small cell lung cancer (NSCLC) is an aggressive cancer where 10% of patients survive beyond five years of initial diagnosis. One of the reasons for the poor survival rate is the robust invasion and metastatic properties of NSCLC. Urokinase plasminogen activator (uPA) and its receptor are directly associated with a poor prognosis in NSCLC. Na+/H+ transporter (NHE1) has been implicated in regulation of cell motility in a number of cancer cells. We wished to identify the role NHE1 plays in uPA activation of tumor progression in NSCLC. We used both uPA and the inactive‐receptor binding amino terminal fragment of uPA (ATF) to stimulate human lung H460 carcinoma cells. 10 nM uPA or ATF stimulated proliferation 1.4 ‐ 1.2 fold over non‐treated cells. Inhibition of NHE1 with 10 μM EIPA abrogated agonist‐stimulated proliferation. Tumor formation size and number was significantly increased by addition of either uPA or ATF, while NHE1 inhibition blocked the ability of either agonist to induce tumors in soft agar assays. Migration of H460 cells was inhibited in the presence of EIPA. Both uPA and ATF enhanced MMP9 activity but not in cells treated with EIPA. Only ATF increased MMP9 expression. We also show the effect of uPA in NHE1 and uPAR knockdown cell lines where we investigated MMP9 activity, and the location of NHE1 and uPAR in lipid rafts. Our results indicate that uPAR activation of cancer cell properties requires NHE1. Support from NIH‐1‐R15‐CA135616‐01