Combination antiretroviral therapy (ART) is currently the standard-of-care to control HIV replication in infected individuals. However, since this treatment mainly targets viral enzymes, the presence of latently-infected reservoirs and the emergence of drug resistance remain major barriers to viral eradication. To overcome this, a “Shock and Kill” approach has been developed, using “latency-reversing agents” (LRAs) to activate latent proviruses (“shock”), leading to viral elimination by ART and immune-mediated clearance (“kill”). We previously screened a small-molecule library on monocytic and CD4+ T cell lines latently infected with HIV, and identified two novel LRA candidates (compounds #C1 and #C3). Based on their structural features, we now identified two related derivatives, compounds #C2 and #C4. Compounds #C1 and #C2 possess diimide structures, whereas #C3 and #C4 have hydrazone structures. All four compounds synergistically enhanced LRA activity when combined with JQ1 or PKC activators in J-Lat 10.6 cells. Notably, combining #C4 with JQ1 further enhanced HIV reactivation showing limited responsiveness to JQ1 alone in J-Lat 10.6 model cells. In ex vivo purified CD4+ T cells from ART-suppressed individuals, #C4 elicited a statistically significant, donor-matched increase in HIV-1 transcription (ratio paired t-test, P = 0.0137, n = 8), whereas JQ1 did not reach significance under the same paired analysis despite a comparable mean magnitude. Notably, #C4 additionally induced measurable reactivation in donors with limited responsiveness to JQ1 alone. RNA-sequencing analyses indicated that compound #C4, JQ1 and PKC activators mediated LRA activity through distinct molecular mechanisms. Collectively, these findings delineate a structurally novel class of LRA candidates that engage a latency-reversal mechanism distinct from that of JQ1 and canonical PKC signaling, and thereby support further investigation of their utility within combinatorial strategies aimed at HIV-1 reservoirs.
Background Chronic kidney disease (CKD) causes millions of deaths and is a known risk factor for the development of other diseases. CKD has been associated with lung injury characterized by neutrophil infiltration and edema. Despite evidence that the lungs are distant organs affected in CKD, there is no treatment to impair lung injury. Cannabinoid receptors type 2 (CB2) activation has been shown to reduce renal dysfunction and organ damage in sepsis. Objective Therefore, this study aimed to evaluate the effects of β-caryophyllene (βC), a CB2 agonist, on CKD-induced pulmonary alterations. Methods Male Swiss mice were subjected to 5/6th partial nephrectomy (PNx) model and randomly assigned to three experimental groups: sham, PNx + vehicle and PNx+βC (10 mg/kg/day). Treatment was administered from days 7 to 14 post-surgery. On day 15, bronchoalveolar lavage, blood, and organs were obtained. Results PNx animals exhibited increased pulmonary leukocyte infiltration, MPO activity, and protein content. Histological analysis of lung tissue showed signs of injury and excessive superoxide production. Increased expression of ICAM-1 was also detected by Western blotting in the lungs of PNx animals. βC treatment attenuated inflammatory cell infiltration and protein extravasation, and reduced ICAM-1 expression in lungs of CKD mice. βC improved renal function parameters, which may have contributed, at least in part, to the reduction in lung inflammation. CB2 receptors expression did not differ among the groups. Conclusion In conclusion, βC attenuated PNx-induced pulmonary changes, supporting its potential as an adjuvant therapy for lung injury in CKD.
Idiopathic pulmonary fibrosis (IPF) is driven by persistent fibroblast activation, excessive extracellular matrix (ECM) deposition, and progressive tissue stiffening, yet the mechanosensitive signaling programs that sustain fibrotic fibroblast states remain incompletely defined. Here, we investigated the association between a periostin (POSTN)-enriched fibrotic program and PIEZO1-dependent Yes-associated protein (YAP)/TEA domain transcription factor (TEAD) activation in IPF. Integrated analysis of public bulk lung transcriptomic datasets identified a reproducible POSTN-enriched ECM remodeling program in IPF lungs. Single-cell and donor-level pseudobulk analyses further showed that a PIEZO1/YAP activity-related signature was co-enriched in POSTN-positive fibrotic fibroblast and myofibroblast compartments. In bleomycin-induced pulmonary fibrosis, Postn upregulation, Piezo1 induction, and increased Yap nuclear localization were recapitulated in fibrotic lung regions. Verteporfin, used as a pharmacological inhibitor of Yap/TEAD-associated transcription, attenuated collagen deposition, hydroxyproline accumulation, profibrotic gene expression, and Yap target protein induction in vivo. In transforming growth factor-β1 (TGF-β1)-stimulated lung fibroblasts, verteporfin suppressed fibroblast migration, Yap nuclear localization, and profibrotic marker expression at non-cytotoxic concentrations. Knockdown of Piezo1 or Yap1 reduced TGF-β1-induced fibroblast activation, while Piezo1 knockdown decreased Yap nuclear localization and Yap/TEAD reporter activity. Active YAP partially restored profibrotic gene expression after Piezo1 knockdown, supporting Yap/TEAD as an important downstream effector of Piezo1. Collectively, these findings identify an association between a POSTN-enriched fibrotic fibroblast state and enhanced PIEZO1-YAP/TEAD signaling, while functional experiments support a role for PIEZO1-YAP/TEAD in fibroblast activation and fibrotic lung remodeling.
Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies, characterized by poor survival rates and limited therapeutic options. Oncogenic KRAS mutations are frequent in pancreatic cancer patients and are key drivers of tumorigenesis. Recent studies have underscored the essential role of extracellular signal-regulated kinase (ERK) in sustaining KRAS-mutant tumors and mediating resistance to KRAS-ERK-targeted therapies. However, the specific prognostic value of KRAS-ERK-associated genes in KRAS-mutant pancreatic cancer remains largely undefined. In this study, we performed a comprehensive analysis of molecular stratification based on KRAS-ERK-associated genes and established a five-gene KRAS-ERK prognosis signature (KEPS) comprising ARNTL2, ZWILCH, KIAA0513, KIAA0913, and C4orf43. The KEPS model effectively stratified patients into high- and low-risk groups, with significantly poorer prognosis observed in the high-risk group. External validation using independent KRAS-mutant pancreatic cancer cohorts validated the prognostic performance of KEPS. Further analyses revealed that the high-risk group was enriched in tumor progression-related pathways and exhibited elevated expression of immune checkpoint molecules, including PD-L1, PD-L2, and CD276. Among the KEPS signature genes, ARNTL2 was prioritized for further functional characterization. In two anti-PD-L1-treated cohorts, ARNTL2 correlated with immune checkpoint expression and therapeutic response, suggesting potential relevance to immune checkpoint blockade. Functional assays confirmed that ARNTL2 depletion suppressed malignant phenotypes of KRAS-mutant PDAC cells in vitro and impaired tumor growth in vivo. In conclusion, this work establishes KEPS as a novel molecular tool for risk stratification in KRAS-mutant pancreatic cancer and highlights that ARNTL2 may represent a potential personalized therapeutic target.
Age-associated cardiac remodeling, characterized by hypertrophy and fibrosis, is a major contributor to cardiovascular morbidity in the elderly and is often exacerbated by metabolic stressors such as hypercholesterolemia. Given the central role of mitochondrial dysfunction in these processes, we investigated whether mitochondria-targeted esculetin (Mito-Esc) could mitigate pathological cardiac remodeling associated with aging and hypercholesterolemia. To model this condition, we utilized 12-month-old Apoe−/− mice, which exhibit chronic hypercholesterolemia and develop age-related cardio-metabolic structural alterations. The efficacy of Mito-Esc was compared with metformin, a putative geroprotective agent. Treatment with Mito-Esc or metformin significantly attenuated cardiac hypertrophy, fibrosis, inflammation, and oxidative stress in aged Apoe−/− mice. These protective effects were further validated in vitro using H9c2 cardiomyocytes and primary adult mouse cardiac fibroblasts. Mechanistically, both Mito-Esc and metformin exerted cardioprotective effects through activation of the AMPK-SIRT1 signaling axis. Depletion of either AMPK or SIRT1 markedly reduced the ability of these compounds to suppress cellular senescence, hypertrophic responses, and fibrotic signaling, indicating a critical role for this pathway. AMPK functioned upstream of SIRT1, as SIRT1 activation was attenuated following AMPK depletion. In parallel, both compounds significantly suppressed p38 MAPK activation. While depletion of AMPK or SIRT1 did not affect p38 MAPK activity, pharmacological inhibition of p38 MAPK enhanced AMPK and SIRT1 activation, supporting a model in which p38 MAPK activation negatively regulates the AMPK-SIRT1 axis. Collectively, these findings demonstrate that Mito-Esc alleviates pathological cardiac remodeling in the setting of aging and hypercholesterolemia through coordinated activation of AMPK-SIRT1 signaling and inhibition of p38 MAPK.
