Background/Objectives: Contemporary drug-eluting stents (DES) differ in antiproliferative drug, strut geometry, polymer, alloy, and drug release kinetics, and the independent association between the eluted drug and clinical outcomes remains uncertain. We compared outcomes among contemporary sirolimus-, zotarolimus-, and everolimus-eluting stents. Methods: This single-center retrospective cohort included 1940 patients who underwent percutaneous coronary intervention (PCI) between 2012 and 2021 with sirolimus- (n = 537), zotarolimus- (n = 400), or everolimus-eluting stents (n = 1003). The primary endpoint was target vessel failure (TVF), a composite of cardiac death, target vessel myocardial infarction (MI), or target vessel revascularization (TVR). The secondary endpoint was major adverse cardiovascular events (MACE), a composite of all-cause death, MI, or any revascularization. Stabilized inverse probability of treatment weighting (IPTW) based on pretreatment covariates and PCI era was used to address baseline and temporal differences; follow-up was censored at two years. Results: During a median follow-up of 731 days, weighted TVF rates were 5.4%, 7.7%, and 6.5% in the sirolimus, zotarolimus, and everolimus groups, respectively. Compared with sirolimus, TVF did not differ with zotarolimus (hazard ratio [HR] 1.38; 95% confidence interval [CI] 0.74-2.58) or everolimus (HR 1.19; 95% CI 0.70-2.02; overall p = 0.458). MACE also did not differ (overall p = 0.517). In complex PCI, TVF was higher with zotarolimus and everolimus than with sirolimus; however, event counts were small and the analyses were exploratory. Conclusions: In this observational cohort, no significant differences in mid-term outcomes were observed among the three drug-defined groups. Because drug type is closely intertwined with stent platform design, these findings should not be extrapolated to individual stent platforms. The exploratory complex-PCI subgroup finding requires confirmation in larger studies.

Philadelphia chromosome-positive (Ph+) B-cell acute lymphoblastic leukemia (ALL), driven by the BCR-ABL1 fusion, remains highly aggressive, with poor prognosis despite tyrosine kinase inhibitor (TKI) therapy. Aberrant PI3K/Akt/mTOR pathway activation contributes to resistance, warranting identification of predictive kinase biomarkers. To identify and validate critical phospho-kinase markers within the PI3K/Akt/mTOR axis as candidate adaptive signaling nodes and exploratory prognostic markers in Ph+ B-ALL. Network pharmacology using STRING and Cytoscape mapped the protein-protein interactions, identifying high-centrality nodes. Functional enrichment analyses (GO/KEGG) were then performed via ShinyGO. Experimentally, phospho-kinase arrays profiled 39 kinase phosphorylation sites in Ph+ B-ALL cells (SUPB-15) treated with PI3K/Akt/mTOR inhibitors (Rapamycin, GDC-0941, GSK690693, Perifosine). Gene expression was validated by RT-qPCR in 100 Ph+ B-ALL patients and protein validation employed Western blot in 50 patient samples with 15 and five healthy controls, respectively. Network analysis identified CREB1 and GSK3 isoforms as central hub regulators. Phospho-kinase profiling revealed p-CREB (Ser133) phosphorylation upregulation following Perifosine, GDC-0941, and Rapamycin treatment, with no statistically significant change after GSK690693. p-GSK3α/β (Ser21/9) phosphorylation showed a statistically significant increase after Perifosine, with non-significant trends after GDC-0941, and a significant reduction following Rapamycin. Hierarchical clustering then divided proteins into four distinct clusters positioning CREB1 in cluster 3 and GSK3α/β in cluster 2 as candidate adaptive response markers based on their phosphorylation profiles. RT-qPCR demonstrated marked downregulation of GSK3α (approximately 71%), GSK3β (63%), and CREB1 (65%) transcripts in patients versus controls, with even greater suppression seen in cell lines. Conversely, Western blot revealed a significant 2.49 fold elevated p-CREB (Ser133) (β = 1.317, p = 0.000347) and a phospho-GSK3α/β (Ser21/9) level that did not differ significantly between patients and controls (β = 0.467, p = 0.466), revealing mRNA-protein discordance indicative of compensatory post-transcriptional and post-translational regulation. These findings represent treatment-associated and disease-state associations between altered phosphorylation states and the Ph+ B-ALL context; functional validation remains an essential next step. mRNA-protein discordance with transcriptional downregulation (63-71%) contrasting with elevated p-CREB levels (2.49-fold, p = 0.000347) suggests post-translational compensatory mechanisms under therapeutic pressure. CREB1 and GSK3 isoforms emerge as central network nodes that might potentially mediate adaptive resistance, although functional validation is required. It should be noted that the evidence presented is associative rather than causal and a direct link between the observed phosphorylation changes and clinical drug resistance has not been established in this study. Sustained CREB phosphorylation despite PI3K/Akt/mTOR inhibition implicates alternative kinase pathways (MAPK/ERK, CaMK), supporting rational combination therapy strategies. These findings position p-CREB and p-GSK-3α/β as candidate biomarkers, highlighting the value of phospho-proteomics over transcriptomics alone in capturing pathway activation. Multi-omics integration may prove essential for overcoming adaptive resistance in ALL patients.

The identification of somatic drivers has turned a field once based on morphology into a set of molecular pathways that can potentially be targeted with drugs. Vascular lesions of the eye and ocular adnexa used to be classified by their appearance; many of them are now defined by a post-zygotic variant in a signaling cascade for which an inhibitor is already available. This review groups these lesions by signaling axis and examines how close each pathway is to clinical application. Belzutifan seems to be the most advanced. In a single-arm phase 2 subgroup with prospectively graded retinal endpoints, HIF-2α inhibition improved all 16 evaluable eyes in 12 participants, without a comparator arm. PI3K and mTOR inhibition is supported by clinical studies in predominantly non-ocular venous and lymphatic malformations, with more consistent evidence for symptomatic benefit than for volume reduction. Direct orbital evidence remains limited to small series. MEK inhibition treats RAS-driven disease in other locations, but it can also cause a retinopathy of its own, which makes it both a therapy and an ophthalmic concern. Gq/11 signaling accounts for most capillary malformations, Sturge-Weber syndrome and both choroidal hemangiomas, which are separated by the mutated codon rather than by histology, and it remains a preclinical target. Junctional GJA4 signaling defines the orbital cavernous venous malformation, probably the clearest example of a disease defined by its driver. Genotyping of ocular lesions is limited by tissue access and by variant allele frequencies that are commonly missed by standard panels.

Ovarian cancer remains a major cause of gynecological cancer-related mortality, highlighting the need for novel therapeutic strategies capable of enhancing antitumor activity while targeting multiple mechanisms involved in tumor progression. Everolimus, an inhibitor of the mammalian target of rapamycin pathway, and carboplatin are established antineoplastic agents with distinct mechanisms of action, whereas astaxanthin is a bioactive carotenoid with antioxidant, anti-inflammatory, and potential anticancer properties. This study compared the individual and pairwise combination effects of carboplatin, everolimus, and astaxanthin on cell viability, migratory capacity, and inflammatory and tumor-associated biomarkers in SKOV3 ovarian cancer cells. Human SKOV3 ovarian cancer cells were cultured under standard conditions and exposed to different concentrations of carboplatin, everolimus, and astaxanthin, either individually or as pairwise two-drug combinations, for 24-72 h. Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay. Cell migration was evaluated using a scratch wound-healing assay. Based on the experimental treatment protocol, 100 nM everolimus, 100 μM carboplatin, and 100 μM astaxanthin, alone and in combination, were further evaluated for their effects on β-defensin 1, β-defensin 2, α-defensin 1, tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) levels. Biomarker concentrations were determined by ELISA. Experiments were independently performed in triplicate. Everolimus, carboplatin, and astaxanthin produced concentration- and time-dependent alterations in SKOV3 cell viability, with more pronounced cytotoxic effects generally observed following prolonged exposure. The pairwise combination treatments also substantially affected cell viability, particularly at later experimental time points. Assessment of migratory capacity demonstrated significant differences in wound closure among treatment groups at 24, 48, and 72 h (p = 0.0224, p = 0.0155, and p = 0.0028, respectively), indicating a time-dependent inhibitory effect on SKOV3 cell migration. Treatment with carboplatin, everolimus, and astaxanthin, individually or in pairwise combinations, also significantly modulated β-defensin 1, β-defensin 2, α-defensin 1, TNF-α, and IL-1β levels compared with control cells, with the magnitude and direction of these changes varying according to treatment regimen and exposure duration. Everolimus, carboplatin, and astaxanthin exerted significant time- and treatment-dependent effects on the viability and migratory behavior of SKOV3 ovarian cancer cells and markedly modulated defensin and pro-inflammatory cytokine responses. The findings demonstrate distinct treatment- and time-dependent responses to the individual agents and their pairwise combinations in SKOV3 ovarian cancer cells. These observations represent preliminary comparative in vitro effects rather than evidence of pharmacological synergy or therapeutic benefit. Further mechanistic studies using additional ovarian cancer cell lines, formal drug interaction analyses, and in vivo models are required to establish the biological and potential therapeutic relevance of these treatment combinations.

Background: Pulmonary vein stenosis (PVS) is a progressive disease characterized by neointimal hyperplasia with poor outcomes. Clinical observations show that adjunctive therapy with imatinib mesylate and sirolimus is associated with improved survival and reduced rates of reintervention. In this study, we aimed to investigate the cellular mechanisms underlying PVS development and progression and to elucidate how these targeted therapies modulate these processes. Methods: We retrospectively analyzed cardiac catheterization and computed tomography, and lung scan data from three patients with PVS to estimate wall shear stress (WSS) at multiple time points. Tissue specimens resected during anatomical repair surgery were analyzed for tissue composition and fibrosis using hematoxylin and eosin staining, Masson's trichrome staining, and flow cytometric analysis of endothelial cell (ECs) and fibroblast markers to determine endothelial-to-mesenchymal transition (EndMT). The safety and efficacy of imatinib mesylate and sirolimus were evaluated in healthy and PVS-derived ECs using cellular viability and proliferation assays, and by assessing fibrotic marker expression indicative of EndMT. Results: Patients exhibited severely elevated WSS estimates at multiple time points throughout disease progression. Disproportionate fibrotic remodeling driven by ECs transitioning into mesenchymal cells through EndMT was the cellular substrate of resected tissue in PVS. Targeted therapy with clinically relevant concentrations of imatinib mesylate and sirolimus was safe and significantly inhibited fibroproliferation and EndMT. At follow-up, all patients were alive and free of surgical and stent-based reintervention after anatomic repair surgery and adjunctive imatinib mesylate and sirolimus therapy. Conclusions: Fibroproliferative ECs undergoing EndMT are likely the cellular substrate underlying PVS development and progression. A multimodal approach consisting of anatomically focused surgical repair with adjunctive imatinib mesylate and sirolimus therapy may attenuate fibroproliferative remodeling.

Physiologically based pharmacokinetic (PBPK) models are increasingly used in pediatric drug development and matching such models to the target population is an important consideration. A US pediatric PBPK model was developed incorporating four sub-groups describing physiology and enzymatic/transporter levels for White, African American, Hispanic/Latino and Asian subjects. Virtual populations could then be created reflecting the demographics of the clinical study design. Ten different drugs (14 studies) were used to verify the pediatric PBPK model performance. Predicted PK parameters showed reasonable agreement with observed data, 50% of the predicted: observed values were within 0.8-1.25-fold, 74% within 0.67-1.5-fold, and 97% within 0.5-2-fold. There was minimal difference for most drugs in performance between the mixed US and North European pediatric populations. This was partly due to reported studies having low numbers of subjects and under-representation of some ethnicities. Power calculations showed that to detect a pharmacokinetic difference for the CYP3A4/5 substrates midazolam, sirolimus and tacrolimus between African American and White American children with a power of 80%, 25, 18 and 12 subjects respectively, in each group would be needed. Developing adult and pediatric PBPK models to closely match study populations is important due to their increased regulatory use and with the move to their application in personalized medicine. Such models may also help to bridge some of the ethnic differences not captured in pediatric clinical studies due to the low numbers of subjects.

Aging is a major risk factor for atherosclerosis and is accompanied by profound changes in the immune system, including effector T cell expansion, senescent cell accumulation, and increased pro-inflammatory signaling. Rapamycin, a promising rejuvenation therapeutic that inhibits mTORC1 and modulates immune aging, was investigated for its potential to attenuate pro-atherogenic immune responses in aged atherosclerotic mice. 80- to 90-week-old male Ldlr[-/-] mice with established atherosclerotic lesions were treated intraperitoneally with rapamycin or control triweekly for 8 weeks. Systemic and plaque immunity was assessed by flow cytometry or single-cell RNA-sequencing. Plaque morphology was evaluated by histology. Rapamycin treatment reduced plaque macrophage content (0.73 ± 0.07 × 10[5] μm[2]) compared to control (ctrl: 0.95 ± 0.09 × 10[5] μm[2], p < 0.05) and decreased total T cell numbers in vascular and lymphoid tissues. This was accompanied by a shift from effector to central memory phenotypes, particularly within the CD8[+] compartment, and a relative enrichment of regulatory CD4[+] T cells (Tregs). Single-cell transcriptomics of aortic Tregs revealed increased expression of suppressive genes (Foxp3, Tgfb1) upon rapamycin. Rapamycin also reduced T follicular helper cells, IL-21 expression, germinal center B cells, plasma cells, and total and oxidation-specific epitope-specific immunoglobulin levels. Additionally, age-associated B cells were diminished in spleen and lymph nodes, reflecting a broader anti-inflammatory effect on B cell immunity. Finally, aortic leukocytes displayed profoundly reduced senescent enrichment scores. Rapamycin remodels the immune landscape in aged atherosclerotic mice, promoting regulatory lymphocyte subsets and suppressing antigen-driven and inflammatory immune responses. These findings highlight the potential of immune-targeted rejuvenation to mitigate age-related atherogenic inflammation.

Poor aqueous solubility can limit drug release from biodegradable implants. We developed a multifunctional dual-jet electrospun drug-delivery system (DDS) designed to accelerate the release of poorly water-soluble bioactive substances. Its principle is based on introducing the drug-carrying matrix and release-rate modifier as separate, interlaced fractions. Poly(D,L-lactide-co-glycolide) (PDLGA) nonwovens or poly(ε-caprolactone-co-trimethylene carbonate) (PCLTMC) membranes served as sirolimus carriers, whereas poly(vinyl alcohol) nanofibres loaded with lactic acid (PVA(LA)) were designed to increase incubation-medium accessibility, lower the pH of the aqueous environment and catalyse early-stage matrix hydrolysis. Both copolymers had defined chain microstructures with relatively short comonomer sequences, intended to limit changes in physical properties and molar composition during incubation. Drug release and early-stage hydrolytic degradation were evaluated over 28 days in deionised water and phosphate-buffered saline. Dissolution of the PVA(LA) fraction increased incubation-medium uptake and was associated with accelerated sirolimus release in water. After 28 days, cumulative release increased from 12.76 to 29.32% for PDLGA and from 22.03 to 59.03% for PCLTMC. LA release reduced bulk-medium pH to 3.14-3.21 after one day. Modest hydrolytic changes were detected for PDLGA, whereas the effect on PCLTMC was limited. Both carriers remained amorphous, showed only small changes in glass-transition temperature (Tg) and retained stable molar compositions. Release was diffusion-dominated, with possible contributions from medium uptake and structural reorganisation. This approach integrates a biodegradable drug carrier with an interlaced, dissolving modifier fraction, allowing the release environment to be modified and drug release to be accelerated without altering the chemical composition of the matrix.