Pharmacological Research

Phosphodiesterase 3A (PDE3A) modulators are emerging anticancer agents for tumors that co-express PDE3A and Schlafen 12 (SLFN12), promoting the formation of a cytotoxic PDE3A-SLFN12 complex. The effect correlates with PDE3A and SLFN12 expression levels. In this study, we conducted a high-throughput screening of 526 compounds in three sarcoma cell lines to identify synergistic or antagonistic interactions with PDE3A modulators. Synergistic mechanisms were further investigated in cell lines and in patient-derived xenograft models of leiomyosarcoma and myxofibrosarcoma in mice. Several drug classes showed potential synergy, and PDE3A modulators sensitized cells to otherwise ineffective Bcl-2/xL inhibitors. The combination of a PDE3A modulator and a Bcl-xL inhibitor induced tumor regression in a patient-derived leiomyosarcoma model in vivo. Our findings reveal that PDE3A modulators synergized with several inhibitors of key cancer pathways, offering promising potential for future treatments. With further research, PDE3A modulators and combination therapies could provide effective targeted treatment options for tumors expressing both PDE3A and SLFN12, paving the way for innovative and hopeful advancements in cancer treatment.

Chemotherapy-induced cardiotoxicity has emerged as an important focus in oncology, driven by the growing number of cancer survivors. Intensive chemotherapies (iCT) used in the treatment of acute myeloid leukemia (AML) often lead to significant adverse cardiac events, which can reduce therapeutic benefits, limit treatment options, or even necessitate discontinuation--particularly for patients with pre-existing cardiac conditions, resulting in a loss of therapeutic opportunity. This study shows that the iCT triggers severe mitochondrial dysfunction in cardiac tissue, mirroring effects seen in ischemia-reperfusion models. Specifically, iCT results in succinate accumulation and elevated reactive oxygen species production, consistent with reverse electron transport phenomenon. Importantly, these effects were entirely prevented with RET inhibitors such as malonate or S1QEL1.1. In vivo, we demonstrate that malonate administration successfully prevents iCT-induced cardiotoxicity, maintaining left ventricular ejection fraction and fibrosis levels comparable to controls. Additionally, in an MLL-AF9-driven AML model, malonate sensitized leukemic cells to iCT. These findings support the dual potential of malonate: as an OXPHOS metabolism inhibitor to overcome chemoresistance in AML, while also reducing cardiotoxic risk for already vulnerable patients. One Sentence Summary: This study demonstrates that malonate prevents chemotherapy-induced cardiotoxicity by inhibiting mitochondrial reverse electron transport (RET), preserving cardiac function, and simultaneously sensitizing AML cells to intensive chemotherapy.

Ca2+/calmodulin-dependent protein kinase II alpha (CaMKII) is a major contributor to physiological and pathological glutamate-mediated Ca2+ signals, and its involvement in various critical cellular pathways demands specific pharmacological strategies. We recently presented GHB ligands as the first small molecules selectively targeting the CaMKII hub, a domain primarily responsible for holoenzyme oligomerisation, with an emerging functional role. Here, we report that the GHB ligand, HOCPCA, improves sensorimotor function after experimental stroke in mice when administered at clinically relevant time and in combination with alteplase. We observed that hub modulation by HOCPCA results in differential effects on distinct CaMKII pools, ultimately alleviating aberrant CaMKII signalling after cerebral ischemia. As such, HOCPCA normalised cytosolic Thr286 autophosphorylation after ischemia in mice and downregulated the ischemia-specific expression of a constitutively active CaMKII kinase fragment. Previous studies suggest holoenzyme stabilisation as a potential mechanism, yet a causal link to in vivo findings requires further studies. HOCPCAs selectivity and absence of effects on physiological CaMKII signalling highlight pharmacological modulation of the CaMKII hub domain as an attractive neuroprotective strategy.

BackgroundGlioblastoma is the most invasive and malignant brain tumor, for which no effective treatment is currently available. Heat shock protein 90 (HSP90) is a potential target for the treatment of different types of cancer. Herein, we investigated the effect of gedunin, an HSP90 inhibitor, in murine GL261 glioblastoma in vitro and in vivo models. MethodsGedunin effects were assessed by in vitro cell viability assay (MTT), apoptosis (PI/Annexin staining), proliferation (CFSE), invasion (transmigration, scratch assay and zymography), protein detection (western blot) and in vivo orthotopic glioblastoma model (brain computed tomography and histology). ResultsGedunin treatment decreased GL261 cell proliferation and triggered apoptosis in a concentration-dependent manner. Gedunin also reduced GL261 cell migration and metalloproteinase-2 activity, suggesting that it impairs glioblastoma cell invasion. Despite the reduction of total protein content in gedunin-treated cells, the phosphorylation of STAT3 and ERK1/2 pathways was enhanced within 24 h. In situ treatment with gedunin did not significantly reduce tumor volume. Still, it reduced the tumor central area and the presence of vascular structures in xenograft glioblastoma in C57BL/6 mice, a clinical feature associated with morbidity and mortality. ConclusionGedunin inhibits murine glioblastoma cell growth and proliferation by inducing apoptosis. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC="FIGDIR/small/543101v4_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@15ea03corg.highwire.dtl.DTLVardef@169e5b1org.highwire.dtl.DTLVardef@16705bdorg.highwire.dtl.DTLVardef@a9c74b_HPS_FORMAT_FIGEXP M_FIG C_FIG

Endometriosis is a common and debilitating neuro-inflammatory disorder that is associated with chronic pain. Definitive diagnosis is based on the presence of endometrial-like tissue (lesions) in sites outside the uterus. Kynurenine monooxygenase (KMO) is a mitochondrial enzyme of tryptophan metabolism that regulates inflammation and immunity. Here, we show that KMO is expressed in epithelial cells in human endometriosis tissue lesions and in corresponding lesions in a mouse model of endometriosis. In mice, oral treatment with the potent KMO inhibitor KNS898 induced a biochemical state of KMO blockade with accumulation of kynurenine, diversion to kynurenic acid and ablation of 3-hydroxykynurenine production. In the mouse model of endometriosis, KMO inhibition improved histological outcomes and endometriosis pain-like behaviours, even when KNS898 treatment commenced one week after initiation of lesions. Taken together, these results suggest that KMO blockade is a promising new non-hormonal therapeutic modality for endometriosis.

As the third-most common type of cancers in infants and young children, neuroblastoma accounts for nearly 10% of all childhood cancers. Despite remarkable advances in tumor diagnosis and management during the past decades, the five-year survival rates for patients with high-risk neuroblastoma remain below 50%. Developing new therapies for this devastating type of childhood cancer is an urgent unmet need. Cyclooxygenase (COX) via synthesizing prostaglandin E2 (PGE2) promotes tumor cell proliferation, survival, migration and invasion, and fosters an inflammation-enriched microenvironment that can facilitate angiogenesis, immune evasion and treatment resistance. However, which downstream PGE2 receptor subtype - namely EP1, EP2, EP3 and EP4 - is directly involved in COX activity-promoted neuroblastoma growth remains elusive. Analyzing five major neuroblastoma patient datasets: Versteeg-88, Kocak-649, SEQC-498, Primary NRC-283, and Oberthuer-251, we show that COX-1/PGE2/EP2 signaling axis is highly associated with the aggressiveness of human neuroblastoma. A time-resolved fluorescence resonance energy transfer (TR-FRET) method reveals EP2 as the key Gs-coupled receptor that mediates PGE2-initiated cAMP signaling in neuroblastoma cells with various risk factors. Taking advantage of novel, selective and bioavailable small-molecule antagonists that we recently developed to target the PGE2/EP2 signaling in vivo, we have demonstrated that pharmacological inhibition of the peripheral EP2 receptor substantially impairs the growth of human neuroblastoma xenografts and the associated angiogenesis in mice. Collectively, our results suggest that the PGE2/EP2 pathway contributes to the growth and malignant potential of human neuroblastoma cells; pharmacological inhibition on EP2 receptor by our drug-like compounds might provide a novel therapeutic strategy for this deadly pediatric cancer.

JM-20 is a novel multifunctional benzodiazepine molecule with potent neuroprotective effects in rat focal cerebral ischemia. To confirm previous results obtained in single laboratories with small sample sizes, and to provide a robust preclinical evidence base for potential clinical development in stroke, we have performed a two-center preclinical trial with sufficiently large group sizes to detect relevant effects, minimizing biases in experimental design as much as possible (randomization, blinding, predefined in- and exclusion criteria) and increasing external and construct validities by performing experimental focal cerebral ischemia by different surgeons in two different laboratories on two continents, including two species (480 mice and 55 rats), different suppliers, young, young adult, and mature adult animals (range 2 -16 months) as well as comorbid animals (diabetes). While JM-20 improved functional outcomes after middle cerebral artery occlusion in young adult mice at day 7 and appeared to reduce mortality (not statistically significant), it had no effect in mature adult or comorbid (STZ-induced diabetes) mice. Effect sizes, where statistically significant, were modest, and much lower than those reported in the previous studies. Meta-analysis of all individual mouse data did not reveal statistically significant different functional outcomes or mortalities between vehicle- and JM-20-treated animals, although neuroscores and survival were slightly better in JM-20-treated animals. In the less severe model of permanent cortical focal cerebral ischemia in rats, JM-20 significantly reduced brain infarction. We conclude that we were able to confirm the neuroprotective potential of JM-20. However, effect sizes were substantially lower as previously described in small, monocentric trials. Further study is needed to determine whether JM-20 could be effective in less severe cases of focal cerebral ischemia or when used in combination with thrombolysis.

Ischemic stroke is a prominent cause of death and disability, demanding innovative therapeutic strategies. Accordingly, extracellular vesicles (EVs) released from mesenchymal stem cells are promising tools for stroke treatment. In this study, we evaluated the potential neuroprotective properties of EVs released from human adipose tissue stem cells (hAT-MSC), which were obtained from a healthy individual submitted to liposuction. A single intranasal EVs administration was performed 24 h after the ischemic stroke in rats. The EVs brain penetration and the tropism to brain zone of ischemia was observed 18 h after administration. Thus, we measured EVs neuroprotection against the ischemic stroke-induced impairment on long-term motor and behavioral performance. Indeed, one single intranasal EVs administration reversed the stroke damages on: i) front paws symmetry; ii) working memory, short- and long-term memory; iii) anxiety-like behavior. These findings highlight hAT-MSC-derived EVs as a promising therapeutic strategy in stroke.

BackgroundEndoplasmic reticulum (ER) stress signaling pathways have pivotal roles in atherosclerosis progression. Recently, we have developed Kyoto University Substance (KUS) 121, which selectively inhibits ATPase activities of valosin-containing protein (VCP) and consequently saves intracellular ATP consumption and mitigates ER stress. Methods and ResultsWe assessed the efficacy of KUS121 against atherosclerosis by its daily injection into Apoe-/- mice fed with Western Diet (WD) for 8 weeks. Consequently, KUS121 treatment reduced atherosclerosis progression by approximately 40% in atherosclerotic plaques. Interestingly, we found that C/EBP homologous protein (CHOP), an established ER stress marker, was mainly expressed in plaque endothelium. Therefore, we assessed the action of KUS121 in endothelial cells using the human endothelial cell line (EA.hy926 cells). As a result, KUS121 prevented ER stress-induced apoptosis and downregulated the IRE1 (Inositol-requiring enzyme) -associated inflammatory pathways. Consistent with these in vitro findings, KUS121 treatment also significantly reduced endothelial apoptosis assessed by TUNEL staining and inflammation examined by immunostaining of Nuclear factor kappa B (NF-{kappa}B) and Intercellular adhesion molecule (ICAM) 1 at plaque endothelium. We also demonstrated that KUS121 maintained ATP levels in EA.hy926 cells and atherosclerotic plaque lesions using the single-wavelength or the FRET-based fluorescent ATP sensor. Supplementation of intracellular ATP by Methyl pyruvate (MePyr) attenuated ER stress-induced apoptotic and inflammatory pathways in endothelial cells, which could be the main mechanism how KUS121 reduces ER stress. ConclusionsKUS121 can be a new therapeutic option for atherosclerotic diseases by maintaining intracellular ATP levels and attenuating ER stress-induced apoptosis and inflammation in plaque endothelium.

Background: Androgen receptor pathway inhibitors (ARPIs) have transformed the treatment of high risk and metastatic prostate cancer, though are associated with increased cardiovascular risk. Glucagon-like peptide-1 receptor agonists (GLP-1 RAs) have been shown to reduce cardiovascular events in non-cancer populations, but their role in patients receiving ARPIs is unclear. Methods: Retrospective analysis of 120 men with PC treated with ARPIs between 2015-2025 with any GLP-1 RA exposure. The time of GLP-1 RA use was categorized relative to ARPI initiation (pre- vs post-ARPI). Cumulative incidences for major adverse cardiac events (MACE) any grade 2 or greater cardiac common terminology criteria for adverse events (CTCAE) were estimated. Fine-Gray regressions were performed (non-cardiac death as a competing risk). Results: The median follow-up was 2.3 years (interquartile range [IQR] 1.3-3.7). The median age was 72 years (IQR 66-78). Atherosclerotic cardiovascular disease (ASCVD) was present in 45.0% (n=54). Overall, 55.0% (n=66) initiated GLP-1 RA therapy prior to ARPI and 45.0% (n=54) after ARPI initiation, with a median duration of GLP-1 RA use of 4.0 years (IQR, 2.3-7.0) and 1.3 years (IQR, 0.6-2.1), respectively. Four patients experienced MACE, including three coronary revascularizations and one ischemic stroke. 25 patients experienced at least one grade 2 or greater cardiac event, most commonly arrhythmia (n=20) and thromboembolic disease (n=11). The 2-year cumulative incidence of MACE and grade [≥]2 cardiac events was 1.7% and 16.1%, respectively. Adjusting for pre-existing cardiovascular risk, GLP-1 RA duration, and pre-ARPI androgen deprivation therapy use, GLP-1RA use prior to ARPI initiation (vs. after ARPI start) was associated with reduced risk of grade [≥]2 cardiac events (subdistribution hazard ratio 0.26, 95% CI 0.08-0.91; p=0.036). Conclusion: GLP-1 RA use prior to ARPI initiation was associated with reduced risk of cardiac events, suggesting that earlier metabolic optimization may influence cardiovascular outcomes. These hypothesis-generating findings support investigation of early GLP-1 RA initiation as a potential cardiovascular risk mitigation strategy during ARPI therapy.

ATP-sensitive potassium (KATP) channel opener levcromakalim is a potent inducer of vasodilation, headache, and migraine attacks in humans and tactile hypersensitivity in mice. Other migraine-inducing agents such as nitric oxide (NO) donors, CGRP, and PACAP are thought to activate second messengers leading to KATP opening. Yet, how KATP channel opening leads to migraine remains unclear. Here, we investigated the contribution of nitric oxide synthase (NOS) isoforms and downstream signaling cascades in a mouse model of migraine-relevant tactile hypersensitivity induced by repeated administration of levcromakalim. The non-selective NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) effectively prevented levcromakalim-induced hypersensitivity. Gene expression analysis in the dura mater suggested contributions from endothelial NOS (eNOS) and inducible NOS (iNOS). Semi-selective nNOS inhibition with S-methyl-L-Thiocitrulline (SMTC) or genetic deletion of neuronal NOS (nNOS) had minimal effect on hypersensitivity and no effect on vasodilation. In contrast, eNOS-/- mice were partially protected from levcromakalim-induced hypersensitivity and exhibited impaired vascular response, highlighting eNOS as a key mediator. Inhibition of iNOS with S-methylisothiourea (SMT) revealed a possible contribution from iNOS as well. Surprisingly, inhibition of soluble guanylate cyclase (sGC) had no effect, while the peroxynitrite decomposition catalyst FeTPPS partially attenuated hypersensitivity, implicating nitrosative stress--rather than classical NO-sGC-cGMP signaling--as the critical downstream pathway. We propose that levcromakalim induces both coupled and uncoupled eNOS activity, enhanced NO production and generation of reactive nitrogen species, including peroxynitrite. Our findings reveal a pivotal role for eNOS and peroxynitrite in KATP channel-induced migraine-relevant hypersensitivity and support targeting nitrosative stress as a potential therapeutic strategy.

BackgroundAccumulating evidence substantiates the capacity of targeted radionuclide therapy (TRT) to potentiate tumor immunotherapy responses. However, combining TRT and immunotherapies exhibits substantial varying efficacy, underscoring the importance of investigating mechanisms through which TRT remodels the tumor microenvironment (TME). Despite recent advancements, the specific signaling molecules mediated by TRT that regulate the functions of cytotoxic T lymphocytes (CTLs) remain unclear. MethodsGiven the demonstrated potential of -radionuclides targeting the fibroblast activation protein (FAP) in TRT, this study employed [225Ac]Ac-FAPI-04 to investigate the mechanisms of TRT-induced TME remodeling. The cytotoxic effects were first evaluated in the B16F10 cell line, followed by an assessment of its in vivo therapeutic efficacy in tumor-bearing mice. RNA-sequencing analysis was utilized to elucidate underlying signaling pathways involving immune responses. Results[225Ac]Ac-FAPI-04 significantly induced DNA double-strand breaks, further elevated ROS levels, and apoptosis in tumor cells. Transcriptomic profiling revealed that [225Ac]Ac-FAPI-04 activates the IL-27/JAK-STAT pathway to amplify interferon-mediated responses, indicating IL-27 serves as a critical regulatory signal for CTLs during TRT. A concomitant observation was the downregulation of Tnfaip3 following [225Ac]Ac-FAPI-04 treatment. Tnfaip3 is implicated in modulating CTLs behavior and intratumoral infiltration, while its downregulation was associated with the upregulation of cytotoxic effector molecules, as confirmed in this study. ConclusionThis study identifies a TRT-induced pattern of synergistic signaling mediated by IL-27 and Tnfaip3, which collectively enhances CTL recruitment and potentiates their cytotoxic function. These findings provide a detailed mechanistic rationale for strategies aimed at augmenting the therapeutic efficacy of TRT and immunotherapy combinations.

Currently, there are a lack of FDA-approved tocolytics for the management of preterm labor. We previously observed that the isoflavones mundulone and mundulone acetate (MA) inhibit intracellular Ca2+-regulated myometrial contractility. Here, we further probed the potential of these natural products to be small molecule leads for discovery of novel tocolytics by: (1) examining uterine-selectivity by comparing concentration-response between human primary myometrial cells and a major off-target site, aortic vascular smooth muscle cells (VSMCs), (2) identifying synergistic combinations with current clinical tocolytics to increase efficacy or and reduce off-target side effects, (3) determining cytotoxic effects and (4) investigating the efficacy, potency and tissue-selectivity between myometrial contractility and constriction of fetal ductus arteriosus (DA), a major off-target of current tocolytics. Mundulone displayed significantly greater efficacy (Emax = 80.5% vs. 44.5%, p=0.0005) and potency (IC50 = 27 M and 14 M, p=0.007) compared to MA in the inhibition of intracellular-Ca2+ from myometrial cells. MA showed greater uterine-selectivity, compared to mundulone, based on greater differences in the IC50 (4.3 vs. 2.3 fold) and Emax (70% vs. 0%) between myometrial cells compared to aorta VSMCs. Moreover, MA demonstrated a favorable in vitro therapeutic index of 8.8, compared to TI = 0.8 of mundulone, due to its significantly (p<0.0005) smaller effect on the viability of myometrial (hTERT-HM), liver (HepG2) and kidney (RPTEC) cells. However, mundulone exhibited synergism with two current tocolytics (atosiban and nifedipine), while MA only displayed synergistic efficacy with only nifedipine. Of these synergistic combinations, only mundulone + atosiban demonstrated a favorable TI = 10 compared to TI=0.8 for mundulone alone. While only mundulone showed concentration-dependent inhibition of ex vivo mouse myometrial contractions, neither mundulone or MA affected mouse fetal DA vasoreactivity. The combination of mundulone and atosiban yielded greater tocolytic efficacy and potency on term pregnant mouse and human myometrial tissue compared to single-drugs. Collectively, these data highlight the difference in uterine-selectivity of Ca2+-mobilization, effects on cell viability and tocolytic efficacy between mundulone and MA. These natural products could benefit from medicinal chemistry efforts to study the structural activity relationship for further development into a promising single- and/or combination-tocolytic therapy for management of preterm labor. Chemical compounds studied in this articleatosiban (Pubchem CID: 5311010); indomethacin (Pubchem CID: 3715); mundulone (Pubchem CID: 4587968); mundulone acetate (Pubchem CID: 6857790); nifedipine (Pubchem CID: 4485); oxytocin acetate (Pubchem CID: 5771); U46619 (Pubchem CID: 5311493)

Here we report the results of a single-center phase 2 clinical trial combining sorafenib tosylate, valproic acid, and sildenafil for the treatment of patients with recurrent high-grade glioma (NCT01817751). Clinical toxicities were grade 1 and grade 2, with one grade 3 toxicity for maculopapular rash (6.4%). For all evaluable patients, the median progression-free survival was 3.65 months and overall survival (OS) 10.0 months. There was promising evidence showing clinical activity and benefit. In the 33 evaluable patients, low protein levels of the chaperone GRP78 (HSPA5) was significantly associated with a better OS (p < 0.0026). A correlation between the expression of PDGFR and OS approached significance (p < 0.0728). Five patients presently have a mean OS of 73.6 months and remain alive. This is the first therapeutic intervention glioblastoma trial to significantly associate GRP78 expression to OS. Our data suggest that the combination of sorafenib tosylate, valproic acid, and sildenafil requires additional clinical development in the recurrent glioma population.

Stroke is a prevalent chronic disease, significantly contributing to mortality and long-term disability. The cerebrovascular recovery process after stroke can be complicated by certain comorbidities, including obstructive sleep apnea syndrome (OSA). However, the mechanisms underlying this deleterious impact of OSA on post-stroke recovery remain elusive. We conducted a preclinical study in rats submitted to stroke and intermittent hypoxia (IH), the main characteristic of OSA, to investigate the effects of IH on stroke lesion and to decipher the pathophysiological mechanisms of this stroke-OSA interaction. Using a malonate model of ischemic stroke on Sprague Dawley rats exposed to either normoxia or intermittent hypoxia for over 56 days, we monitored brain lesion size, microvascular plasticity and blood brain barrier (BBB) permeability using in vivo 4.7T-MRI. Finally, we assessed oxidative stress, inflammation, and angiogenesis-related gene expression by qPCR, and NeuN, GFAP, Collagen IV and ZO-1 expression by immunohistological staining. Our findings indicate that while IH does not significantly affect lesion volume reduction over time, it exacerbates ischemic injury-induced necrosis and neuronal loss. Additionally, IH amplifies post-stroke inflammation, as evidenced by increased IL-6 and TGF-{beta} expression, and induces oxidative stress by increasing DHE staining and decreasing Superoxide Dismutase (SOD1) level. Vascular assessment revealed IH-induced modifications in vessel radius and angiogenic factors expression (VEGF, Ang2), alongside increased BBB permeability and altered expression of aquaporin 1 and claudin 1, particularly in the acute phase post-stroke. These results suggest that IH alters cerebrovascular integrity and exacerbates inflammatory response following ischemic stroke, potentially contributing to poorer long-term functional outcomes. Understanding the mechanisms through which IH exacerbates post-stroke injury may guide the development of targeted neuroprotective strategies to improve stroke recovery in patients with OSA.

Acquired chemoresistance, often linked to metabolic adaptation, severely limits therapeutic efficacy in non-small cell lung cancer (NSCLC). Strategies targeting resistance-associated metabolic plasticity are urgently needed. Here, we investigated the combination of Metformin (mitochondrial Complex I inhibitor) and Dichloroacetate (DCA; PDK inhibitor) in chemo-sensitive human A549 NSCLC cells and an acquired Doxorubicin-resistant subline (A549-DR). A549-DR cells displayed significant Doxorubicin resistance ([~]15.6-fold; IC50: 3.9 {micro}M vs 0.25 {micro}M in parental A549) and exhibited enhanced glycolysis (higher ECAR) while retaining substantial oxidative phosphorylation (OxPhos) capacity. While single-agent Metformin or DCA showed limited cytotoxicity, their combination induced potent synergistic cell death (Combination Index < 0.5) specifically in A549-DR cells compared to parental A549 (CI [~] 0.8-1.1). Mechanistically, Metformin inhibited OxPhos, while DCA promoted pyruvate entry into the TCA cycle, preventing compensatory glycolysis. This dual blockade precipitated a catastrophic metabolic collapse uniquely in A549-DR cells, marked by simultaneous suppression of OxPhos (OCR) and glycolysis (ECAR), leading to severe ATP depletion ([~]85% reduction), hyperactivation of the energy sensor AMPK, and robust induction of apoptosis ([~]58% Annexin V positive). Parental A549 cells were less susceptible, experiencing moderate metabolic inhibition and apoptosis ([~]32% Annexin V positive). Our findings highlight a powerful synergistic strategy that exploits the acquired metabolic vulnerabilities of chemoresistant NSCLC cells, inducing an irrecoverable energy crisis and offering a promising therapeutic approach to counteract treatment failure.

Metabolic dysfunction-associated steatohepatitis (MASH) is a progressive liver disease associated with liver-related complications and death. Kylo-0603 is a novel agonist for the thyroid hormone receptor {beta} (THR-{beta}) that has been developed by merging the structures of three acetylgalactosamine (GalNAc)-modified ASPGR ligands with a triiodothyronine (T3) analog. This unique design allows for both THR-{beta} activation and targeted delivery to hepatocytes, significantly reducing the risk of adversed effects related to increased systemic thyroid hormone-like effects. Additionally, it effectively lowers serum cholesterol by as much as 69.2% and low-density lipoprotein cholesterol (LDL-C) levels by up to 88.2% in the MASH mouse model. Meanwhile, Kylo-0603 was shown to reduce steatosis by up to 1.3 points (P < 0.001), inflammation by 1.8 points (P < 0.0001), and ballooning by 0.8 points (P < 0.01). The non-alcoholic steatohepatitis (NASH) activity score (NAS) decreased by up to 3.7 points (P < 0.0001), and the fibrosis score dropped by 0.6 points (P < 0.05). These findings suggest that Kylo-0603 is effective in enhancing liver tissue NASH status and inhibiting fibrosis progression. In summary, Kylo-0603, as a highly both tissue and target selective and low-toxicity THR-{beta} agonist, shows significant promise for treating MASH and is likely to emerge as a new therapeutic option for patients with this condition.

Immune checkpoint inhibitors, particularly T cell targeting anti-PD(L)1 therapies, have revolutionized the treatment landscape for solid malignancies, but challenges related to non-responsiveness and the development of treatment resistance continue to be observed. An additional immunosuppressive axis relates to prostaglandin signaling downstream of cyclooxygenase-2 (COX2), where COX2 inhibitors have shown clinical promise in re-engaging both T and non-T cell immune compartments, yet have suffered from toxicity concerns. We report here the preclinical characterization of OKN4395, a highly potent and specific first-in-class triple antagonist of EP2, EP4, and DP1, major tumor immunosuppressive receptors downstream of COX2. OKN4395 restores immune function on both T cells and NK cells in vitro. Additionally, OKN4395 acts synergistically with anti-PD1 to increase speed and depth of antitumor activity. Overall, these findings robustly support the clinical investigation of OKN4395 in an ongoing Phase 1 trial (NCT06789172) as an innovative cancer immunotherapy for solid tumors, as a single agent and in combination with anti-PD1 therapy. Statement of significanceOKN4395, a first-in-class oral EP2/EP4/DP1 antagonist, reverses prostanoid-driven immunosuppression to restore antitumor immunity. Integrated pharmacology defines mechanism, translational biomarkers as well as both monotherapy and anti-PD1 combination strategies. These data position prostanoid tri-receptor antagonism as a translatable strategy in solid tumors. A global Phase 1 study is underway (NCT06789172).

The efficacy of anthracycline-based chemotherapeutics, which include doxorubicin and its structural relatives daunorubicin and idarubicin, remains almost unmatched in oncology, despite a side effect profile including cumulative dose-dependent cardiotoxicity, therapy-related malignancies and infertility. Detoxification of anthracyclines while preserving their anti-neoplastic effects is arguably a major unmet need in modern oncology, as cardiovascular complications that limit anti-cancer treatment are now a leading cause of morbidity and mortality among the 17 million cancer survivors in the U.S.. To address this, we examined different clinically relevant anthracycline drugs with respect to a series of features including mode of action (chromatin and DNA damage), bio-distribution, anti-tumor efficacy and cardiotoxicity in pre-clinical models and patients. We show that different anthracycline drugs have surprisingly individual efficacy and toxicity profiles. In particular, aclarubicin stands out in pre-clinical models and clinical trials as it potently kills cancer cells, does not induce therapy-related malignancies or cardiotoxicity, and can be safely administered even after a maximum cumulative dose of either ida- or doxorubicin has been reached. Retrospective analysis of aclarubicin used in second-line treatment of relapsed/refractory AML patients showed similar survival effects to its use in first line, leading to an almost 25% increase in 5-year overall survival. Considering individual anthracyclines as different drugs provides new treatment options that strongly improve survival of cancer patients while limiting the toxic side-effects. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=188 SRC="FIGDIR/small/23298950v1_ufig1.gif" ALT="Figure 1"> View larger version (49K): org.highwire.dtl.DTLVardef@4d6483org.highwire.dtl.DTLVardef@101d8b7org.highwire.dtl.DTLVardef@3adf1aorg.highwire.dtl.DTLVardef@10d2c49_HPS_FORMAT_FIGEXP M_FIG C_FIG

Chemotherapy-induced peripheral neuropathy (CIPN) is a debilitating complication of cancer treatment. CB2 cannabinoid receptor activation reduces inflammation and is an attractive therapeutic target. Antibodies targeting G protein-coupled receptors (GPCRs), like CB2, offer high specificity and peripheral-restriction, thereby minimizing off-target activity. Here, we investigated the efficacy of first-in-class CB2-specific antibody agonists (AB110 and AB120) and an isotype control (AB100) on mechanical and cold hypersensitivity induced by paclitaxel in both tumor-free and mammary (4T1) tumor-bearing female mice. These CB2 antibody agonists exhibit biased G- signaling and also reduce macrophage markers and pro-inflammatory cytokines in vitro. Paclitaxel produced behavioral hypersensitivities to mechanical and cold stimulation, which were reduced by AB110 and AB120 for approximately 48 hours post-injection in female mice. Repeated daily dosing did not lead to tolerance to the anti-allodynic effects. Prophylactic treatment with AB110 and AB120 during paclitaxel treatment delayed, but did not prevent, the development of paclitaxel-induced behavioral hypersensitivities after termination of dosing with antibody agonists. AB100 had no effect under any conditions. The anti-allodynic effects of AB120 were absent in CB2 knockout mice, confirming pharmacological specificity via CB2 receptors. Furthermore, AB120 remained effective in paclitaxel-treated tumor-bearing mice. Neither AB110 nor AB120 affected locomotor activity in otherwise naive mice. The cytotoxic activity of paclitaxel on 4T1 tumor cell line was maintained in the presence of CB2 antibody agonists in vitro. Overall, our results suggest that CB2-specific antibody agonists are promising candidates for treating CIPN, providing lasting pain relief without tolerance, off target effects or unwanted CB1-mediated motor side effects.