Cardiac injury from lipid overload features cardiomyocyte death, myocardial remodeling, and reduced contractile function. Although L-menthol has reported cardioprotective effects, its mechanisms are poorly defined. We therefore examined the role of transient receptor potential melastatin 8 (TRPM8) in lipid overload-induced cardiac dysfunction and tested whether L-menthol acts via TRPM8. After 12 weeks of high-fat diet, mouse ventricular myocytes showed a marked reduction in TRPM8 protein expression. In vitro, L-menthol reduced cardiomyocyte injury caused by lipid overload, and in vivo it mitigated cardiac injury in high-fat diet fed male mice. These protective effects were largely abolished by TRPM8 knockdown, indicating a TRPM8-dependent mechanism. Mechanistic studies indicate that L-menthol preserves mitochondrial Ca2+ homeostasis via a TRPM8/GRP75/VDAC1 associated pathway, which limits mitochondrial dysfunction and apoptosis during lipid overload. We also found that lipid overload decreased TRPM8 S-palmitoylation at the C707 regulatory site and reduced TRPM8 protein stability. Downregulation of zDHHC13 may contribute to this loss of S-palmitoylation, whereas L-menthol helped maintain TRPM8 S-palmitoylation and protein expression. Together, these results support a TRPM8-dependent protective effect of L-menthol against lipid overload-induced cardiac injury and suggest that TRPM8-related signaling may represent a potential therapeutic target.

The NLRP3 inflammasome is a multiprotein complex that primes cytokine production in the innate immune system. The inflammasome activation involves the cage-to-disk transition of NLRP3 oligomers, facilitated by the co-factor NEK7 protein. While NEK7's role in promoting cage disassembly has been reported, its involvement in the large conformational changes of the NLRP3 monomer during activation remains elusive. Here, by using multi-scale simulations, we uncovered a stage-dependent role of NEK7 in the inactive-to-active transition. In the early stage, NEK7 reshapes the dynamics of the highly unstable inactive NLRP3 monomer to resemble active state, priming the conformational transition. In the middle stage, NEK7 impedes progression by populating an intermediate state farther from the active conformation than the NEK7-free counterpart, and structures in this state exhibit reduced allosteric potential toward activation. In the late stage, NEK7 has negligible impact, as the active conformation remains inherently isolated by a high energy barrier regardless of NEK7 presence. This highlights the critical role of oligomeric assembly in enabling monomeric NLRP3 to complete its conformational transition, in agreement with experiment observations. Our work suggests a multilayered activation mechanism where oligomer-level assembly and monomeric conformational changes are coupled, providing new mechanistic insights into this physiologically essential macromolecular process.

Biofilms, formed within the necrotic tissue of chronic infected wounds, create a shielded microenvironment that promotes bacterial persistence and disrupts the healing process. This initiates a self-perpetuating cycle of bacterial infection and dysregulated host inflammation, making wound healing profoundly challenging. Here, we designed and fabricated a near-infrared (NIR)-driven Janus nanomotor capable of penetrating biofilms via autonomous propulsion to simultaneously deliver anti-inflammatory and photothermal therapy (PTT) for effective wound healing. Using a spatially selective super assembly strategy, we fabricated the self-thermophoresis Janus nanomotors with Au nanostars (AuNS) as the fundamental photothermal agent, enabling propulsion under NIR light irradiation. Combined with the spiky surface on the AuNS, the nanomotor can efficiently penetrate bacterial membranes. Furthermore, the Au was functionalized with humic acids (HAs) to introduce immunomodulatory functionality. We demonstrated that the Janus HAs-AuNS-PAA/mSiO2 nanomotor exhibits excellent biofilm penetration ability and efficacy in eliminating bacteria through a combination of anti-inflammatory and photothermal effects. In vitro and in vivo experiments confirmed that these nanomotors possess nanomolar-level bactericidal and anti-inflammatory efficacy, and they significantly accelerated wound healing in a mouse model. This platform combines potent, antibiotic-free antibacterial action with targeted immunomodulation, offering a comprehensive solution for managing persistent infectious wounds.

Background The current treatment strategies for myocardial infarction (MI) mainly focus on recanalization of infarct-related arteries to alleviate myocardial damage. In contrast, the role of promoting microangiogenesis has not received sufficient attention. Lysine crotonylation (Kcr), a novel posttranslational protein modification, has an unclear role in revascularization following MI. Purpose This study investigated the role of TOM70 crotonylation in angiogenesis following MI and elucidated its potential mechanisms. Study design We first established a mouse model of MI and an in vitro model of hypoxia in hypoxic human umbilical vein endothelial cells (HUVECs). Using crotonylation-based sequencing, we identified the key gene TOM70 and its modification sites. Subsequently, we generated a specific antibody against TOM70 K199 crotonylation to validate this modification. To explore the underlying mechanism, we constructed an adeno-associated virus (AAV) vector carrying a mutation at the TOM70 locus. Finally, we performed high-throughput drug screening to identify compounds that potentially bind to the TOM70 K199 crotonylated protein. Results In this study, we found that the crotonylation of TOM70 was significantly increased in MI mice and hypoxic HUVECs. Quantitative analysis of crotonylation further identified K199 as the critical modification site. In vivo studies have shown that the TOM70 K199R mutant virus can improve cardiac function in mice with MI and promote angiogenesis. In vitro, transfection with the TOM70 K199R plasmid mitigated the hypoxia-induced reduction in tube-forming ability of HUVECs, whereas the K199Q plasmids exacerbated the damage. Mechanistically, acyl-coA synthetase short-chain family member 2 (ACSS2) serves as an upstream regulatory factor, upregulating TOM70 K199cr, leading to mitochondrial morphological abnormalities and dysfunction by inhibiting the mitochondrial import of MIC19. This inhibition ultimately worsens myocardial remodeling and impedes the revascularization process. Based on this mechanism, we identified parishin as specific inhibitor of TOM70 K199cr, which demonstrated efficacy in improving cardiac function and promoting revascularization. Conclusion We found that ACSS2 upregulates TOM70 K199cr, inhibits mitochondrial import of MIC19 protein, causes mitochondrial structural and functional damage, and suppresses angiogenesis, thereby aggravating the progression of myocardial infarction. Administration of parishin can inhibit this process, improve angiogenesis, and enhance cardiac function.

The VP4 protein of enteroviruses, such as Coxsackievirus B3, is a small, intrinsically disordered protein essential for perforating the host cell membrane during viral entry. A key feature of VP4 is its N-terminal myristoylation, which is required for infectivity in some enteroviruses but dispensable in others, suggesting a complex and context-dependent role that is not fully understood. The precise biophysical mechanisms by which this lipid anchor enables a disordered protein to breach a membrane remain unresolved. Here, using Coxsackievirus B3 VP4 as a model system and integrating multi-scale molecular dynamics simulations with confocal microscopy, we demonstrate that myristoylation is not a simple membrane tether but a multi-functional regulator that orchestrates VP4 activity through distinct, hierarchical roles. First, it provides the necessary hydrophobic anchor to recruit the disordered VP4 to the membrane interface. Second, the myristoyl group acts as a key molecular driver that promotes the liquid-liquid phase separation of VP4, leading to the formation of dynamic condensates on the membrane surface. These condensates actively remodel the membrane, generating substantial curvature that, in turn, lowers the free energy barrier for VP4 penetration. Furthermore, we find evidence that the myristoyl group plays a third role in stabilizing the final transmembrane pore. Our findings establish a novel paradigm where a single lipid modification empowers a disordered viral protein to form a functionally potent condensate that mechanically primes and physically breaches the target membrane, a mechanism that may explain the conditional myristoylation requirement across enteroviruses.

Receptor tyrosine kinases (RTKs) are validated anticancer targets, and targeting their DFGout conformations represents a mainstream strategy for developing highly selective type II inhibitors. RTKs can adopt various DFGout conformations, but only the classical conformations with a fully formed back pocket are structurally validated to accommodate type II inhibitors. However, experimentally determined structures of kinases in a classical DFGout conformation are scarce, which presents a significant obstacle for the structure-based design of selective RTK inhibitors. Recently, a conserved disordered motif N-terminal to the kinase domain of RTKs, called the juxtamembrane (JM) motif, has been reported to regulate the binding of inhibitors to the DFGout conformation of VEGFR2, an RTK involved in angiogenesis. In this study, we performed extensive MD simulations to explore the impact of the disordered JM motif on the conformational space of the DFG motif in RTKs and to investigate how that influence may affect inhibitor binding to VEGFR2. We revealed that in VEGFR2, the disordered JM is highly dynamic and forms transient contacts with the kinase domain and consequently fine-tunes the DFGout subconformational space to shift populations from nonclassical to classical DFGout conformations. This dynamic model provides alternative structural explanations underpinning the reported regulatory effects of the JM motif on binding of the inhibitor to VEGFR2. Additionally, we demonstrated that in other RTKs beyond VEGFR2, the disordered JM similarly promotes classical DFGout conformations. Such a role of JM is particularly favorable for creating druggable DFGout conformations that can be exploited for designing high-selectivity type II inhibitors.

Combining glucose oxidase (GOx) with peroxidase-like (POD) nanozymes is a well-established strategy for synergistic starvation and chemodynamic therapy (CDT). However, the low catalytic efficacy of most nanozymes compared to GOx often leads to wasteful accumulation of hydrogen peroxide (H2O2), which can inhibit GOx activity and limit the therapeutic effect. Therefore, enhancing the H2O2 consumption efficiency is critical to unlocking the full potential of this approach. Here, we designed a cobalt-doped mixed-valence copper oxide (Co-CuxO) nanozyme with metals uniformly dispersed using a polyacrylic acid (PAA) scaffolding method. This nanozyme tightly couples the redox cycles that drive the valence state interconversions of cobalt (Co) and copper (Cu) elements, enabling highly efficient consumption of H2O2 and glutathione (GSH). While Co provides multi-valent characteristics ideal for Fenton-like reactions, Cu acts not only as a co-catalyst but also triggers cuproptosis. We further installed GOx onto the nanozyme surface, allowing H2O2 produced by GOx channels to Co-CuxO active sites, thereby further accelerating the catalytic cascade. In vitro experiments demonstrated effective glucose and H2O2 depletion with abundant hydroxyl radical (·OH) generation. When functionalized with hyaluronic acid (HA) for tumor targeting, in vivo experiments demonstrated robust multimodal cell death, confirming the high efficacy of our integrated dual-enzyme platform.

Abstract Predicting protein–protein interaction (PPI) affinities from structural data remains a challenge. Although binding funnel theory describes the formation of native complexes, the topography of the funnel bottom and its influence on affinity are often overlooked. Using two well-controlled nanobody-antigen datasets as model systems, we demonstrate that PPI can exist in both static and dynamic binding paradigms. These two nanobody series adopt nearly identical binding poses toward their respective antigens yet exhibit diverse affinities, each representing a distinct paradigm: a static paradigm in which affinity can be ranked by Rosetta scoring of co-crystal structures alone, and a dynamic paradigm in which affinity can only be ranked using molecular dynamics–sampled ensembles. The two paradigms differ in interfacial dynamics. In the dynamic series, the relative motion between binding partners (Δ RMSF ) is temperature-sensitive, with optimal affinity correlation at 298 K. In the static series, Δ RMSF is minimal and insensitive to temperature. Local frustration analysis establishes a mechanistic bridge between interfacial dynamics and landscape topology. Dynamic interfaces exhibit increased frustration upon thermal sampling, facilitating sampling of functionally relevant microstates, whereas static interfaces show minimal frustration changes across temperatures. The temperature-dependent frustration difference mirrors Δ RMSF sensitivity, confirming local frustration as a determinant of interfacial dynamics. Furthermore, static interfaces are characterized by a higher density of canonical hotspot residues, while dynamic interfaces utilize interfacial ruggedness to modulate affinity. Together, these results demonstrate that conserved binding modes can encode different energy landscapes and provide a practical framework for determining when ensemble-based sampling is required for accurate affinity prediction.

Understanding how disease-causing missense mutations (DCMMs) affect protein function is fundamental. As protein dynamics is increasingly recognized as a key regulator of protein function, it is essential to incorporate dynamics into the once heavily emphasized structure-function framework to explain the effects of DCMMs. Although research in this area is emerging, evidence supporting a definitive role of dynamics in mediating the DCMM effects on protein function remains limited. Here, we used Menin─a mutation-prone scaffold protein involved in various pathologies─as a model system to explore the DCMMs' effects on Menin's dynamics and on Menin's interaction with a disordered JunD peptide. By performing MD simulations on 24 clinically confirmed DCMMs coupled with Rosetta conformation energy calculations, we showed that DCMMs do not necessarily destabilize protein stability. Instead, they induce similar dynamic changes in the protein. Using multireplica umbrella sampling to compare binding affinities, we show that DCMMs reduce Menin-JunD affinity by disrupting the conserved dissociation pathway observed in wild-type Menin. The underlying mechanism was revealed through allosteric analysis, which showed that, despite being located far from the JunD binding site, DCMMs uniformly disturbed the coupling between Menin residue E179 and the binding pocket, and forced maintenance of E197-pocket coupling restored the impaired Menin-JunD interactions in DCMMs. Together, these data demonstrate that DCMMs affect Menin-JunD interactions via changing protein dynamics, with allostery playing a crucial role, necessitating the incorporation of dynamics to better explain the effects of DCMMs.

Ferroptosis is a promising anticancer strategy, yet its efficacy is limited by insufficient cellular hydrogen peroxide (H2O2) levels and intrinsic antioxidant defenses. Meanwhile, mild photothermal therapy (mPTT) enhances ferroptosis by accelerating lipid peroxidation. Herein, we designed a Janus nanoplatform that specifically arrests cells in thermo-vulnerable S-phase to maximize the ferroptosis-sensitizing capacity of mPTT. A Janus architecture with a unique morphology was engineered to incorporate three functional modules: (1) A photothermal half-bowl composed of polydopamine (PDA) that generates localized hyperthermia under near-infrared (NIR) laser irradiation; (2) a drug-loading module utilizing π-π stacking to deliver irinotecan (CPT-11), which arrests the cell cycle at the heat-sensitive S-phase and creates a favorable environment for mPTT; and (3) a ferroptosis-inducing Fe-zinc sulfide (ZnS) core anchored to the other half-bowl, which initiates ferroptosis via Fe2+ release and amplifies it through the ZnS component. This amplification operates via two mechanisms: hydrogen sulfide (H2S)-mediated catalase (CAT) inhibition elevates H2O2 for the Fenton reaction, while Zn2+-triggered p53 activation suppresses SLC7A11, disrupting the glutathione (GSH)-glutathione peroxidase 4 (GPX4) antioxidant axis. The resulting nanoplatform demonstrated excellent antitumor efficacy, with ferroptosis confirmed as critical to its performance. This Janus nanoplatform presents a novel multi-mechanism cooperative strategy for anticancer therapy centered on ferroptosis.

Circular RNAs (circRNAs) play a crucial role in the initiation and development of cancers. Understanding circRNAs’ functions and molecular mechanisms in tumor development is expected to reveal new diagnostic indicators and therapeutic targets of prostate cancer (PCa). In our study, we identified a new circRNA hsa-circ-0057553 (circSLC39A10) in PCa from a bioinformatic microarray analysis. The levels of circSLC39A10 were observed to be markedly elevated in both prostate cancer cells and tissues. This increased expression was associated with multiple clinicopathological features, suggesting its potential as a new diagnostic indicator for PCa. CircSLC39A10 exhibited oncogenic effects on the proliferation, migration, invasion, and metastasis of prostate cancer cells both in vivo and in vitro. CircSLC39A10 was identified as a factor that promoted the malignant progression of PCa cells through the miR-936/PROX1/β-catenin pathway, ultimately leading to the activation of Wnt signaling. Overall, circSLC39A10 is an oncogenic circRNA with potential as a biomarker for PCa. The identified circSLC39A10/miR-936/PROX1/β-catenin axis shows promise as an innovative therapeutic target for PCa.

Combing magnetic resonance imaging (MRI) and cancer therapy modalities within one nanoplatform holds great potential for imaging-guided cancer theranostics. However, simultaneously achieving optimal MRI performance and satisfactory anti-cancer effects is still a challenge. Here, by manipulating the position of a Fe3O4 core that has MRI capability within a porous shell, this work synthesizes a series of core-shell structured Fe3O4@carbon/copper oxide nanoparticles (Fe3O4@C/CuOx NPs) in which the Fe3O4 core has different degrees of exposure. This work demonstrates that in an eccentric structure with maximally exposed Fe3O4 core, the NPs exhibits optimal MRI capability. Additionally, using polymer as scaffold blended with carbon-doped copper oxide, the shell of the NPs can integrate chemotherapy, photothermal therapy (PTT) and cuproptosis to exert synergistic anti-cancer effect in a pH/NIR dual responsive manner. The porous shell allows efficient DOX loading for chemotherapy, the copper oxide component serves as photothermal agents for PTT and triggers for cuproptosis. Last, the Fe3O4 core is also a nanozyme that possesses peroxidase (POD)-like properties, which enhances the efficacy of above-mentioned anti-cancer effects via promoting reactive oxygen species (ROS) generation. This work evaluates the MRI-guided anti-cancer efficacy of the NPs both in vitro and in vivo, and demonstrates NPs' superior performance in imaging-guided cancer theranostics.

The heart adapts to cardiac demand via chemical modifications of contractile myofilament proteins. Many of these modifications, such as phosphorylation, occur in proteins’ intrinsically disordered regions (IDRs). These IDRs, though challenging to study, are recognized as dynamic, tunable regulators of protein function. Since cardiac dysfunction often involves altered posttranslational modifications (PTMs) in myofilament proteins, understanding how IDR changes affect protein and myofilament behavior is crucial. We hypothesized that PTMs, primarily phosphorylation, regulate ABLIM1 (a myofilament protein) by altering its IDR conformational ensemble, thereby modulating its binding to other myofilament proteins. We tested this using multiscale modeling (including molecular dynamics simulations) to predict ABLIM1’s conformational ensembles pre- and postphosphorylation at sites altered in a canine model of heart failure with reduced GSK3β activity. A state-based contraction model then rationalized the physiological consequences. Our data show that local physicochemical alterations from phosphorylation in ABLIM1’s IDRs significantly affect its conformational ensemble. This ensemble change subsequently influences the ability of its LIM domains to interact with titin. Furthermore, using the contraction model, we show that a reduced ability to recruit myosin heads for cross-bridge formation, resulting from the modified LIM domain/titin interactions, provides a mechanism that elucidates previous findings of diminished length-dependent activation. These findings offer critical molecular insights, reframing IDRs not merely as structural noise but as key, tunable elements that control protein interactions and ultimately impact mechanical behavior in the sarcomere. This work bridges molecular disorder and biomechanical function, providing a new perspective to understand dynamic control and dysfunction in cardiomyocyte contraction.

Prostate cancer (PCa) remains a major challenge in oncology, driving the need for continuous exploration and development of innovative treatment strategies. NCAPH plays a critical role in tumorigenesis and progression across multiple cancer types; however, its specific role in PCa has yet to be fully understood. This study aims to elucidate the biological functions of NCAPH in PCa. Our findings reveal that NCAPH gene expression is upregulated in PCa patients and correlates with poor prognosis. Enrichment analysis, flow cytometry, and correlation analysis demonstrate that NCAPH promotes the PI3K/AKT/mTOR pathway and facilitates cell cycle transition in PCa cells. Additionally, we identified E2F1 as a novel downstream target of NCAPH in PCa cells. Mechanistically, ChIP analysis showed that NCAPH regulates E2F1 transcription by binding to the proximal promoter of E2F1, subsequently stimulating the PI3K/AKT/mTOR pathway and activating downstream targets for cell cycle progression in PCa cells. Notably, combining NCAPH knockdown with an mTOR inhibitor (Everolimus) or a cyclin-dependent kinase inhibitor (Flavopiridol) demonstrated promising anti-tumor effects both in vitro and in vivo. This study highlights the significant pro-tumor role of NCAPH in PCa and suggests its potential as a therapeutic target.

The residual risks of advanced atherosclerosis remain substantial despite current preventive strategies and pharmacotherapy. Circulating branched-chain amino acids are biomarkers of cardiovascular disease risk. However, the mechanism of leucine in atherosclerosis progression remains unclear. Leucine transporter-SLC7A5-mediated leucine intake that promotes advanced atherosclerosis in mice, increasing apoptotic macrophages and lipids accumulation within plaques. Multi-omics analyses showed that leucine deprivation enhanced macrophage mitochondrial function and increased plaque CD5Lhi macrophages, under SLC7A5-deficiency-mediated leucine deprivation, these cells exhibited stronger oxidative phosphorylation and lipid metabolism. Mechanistically, leucine deficiency reduced SLC7A5-PGAM5 binding in macrophages, promoting PGAM5-NDUFV1 interaction and enhancing mitochondrial function, which attenuates atherosclerosis progression. Collectively, these findings elucidate the function and mechanism of SLC7A5 in Cd5lhi macrophages, highlighting it as a potential therapeutic target. Strategies aimed at improving mitochondrial function also offer a promising approach for advanced atherosclerosis treatment.

1 The heart adapts to cardiac demand through various mechanisms, including chemical modifications of myofilament proteins responsible for cell contraction. Many of these modifications, such as phosphorylation, occur in unstructured, or intrinsically disordered, regions (IDRs) of proteins. Although often challenging to study, these IDRs are increasingly recognized as dynamic, tunable regulators of protein function. Given that cardiac dysfunction can involve changes in the post-translational modification (PTM) status of myofilament proteins, it is critical to assess how alterations within these disordered regions impact intact protein and myofilament behavior. We hypothesized that the function of ABLIM1, a myofilament protein containing an important IDR, is regulated by altering its IDR conformational ensemble through PTMs, primarily phosphorylation. We proposed that this conformational change would modulate its ability to bind to other myofilament proteins. To evaluate this hypothesis, we employed a multiscale modeling approach including molecular dynamics simulations. This was used to predict the conformational ensembles of ABLIM1 before and after phosphorylation, at sites known to be altered in a canine model of heart failure with reduced GSK3 β activity. We then used a state-based model of contraction to rationalize the physiological consequences of the molecular-scale predictions. Based on our data, we observed that local physicochemical alterations induced by phosphorylation in ABLIM1's intrinsically disordered regions significantly affect its overall conformational ensemble properties. This ensemble change subsequently influences the ability of its LIM domains to interact with titin. Furthermore, using the contraction model, we show that a reduced ability to recruit myosin heads for cross-bridge formation, resulting from the modified LIM domain/titin interactions, provides a mechanism that elucidates previous findings of diminished length-dependent activation. These findings offer crucial molecular insights, reframing IDRs not merely as structural noise but as key, tunable elements that control protein interactions and ultimately impact mechanical behavior in the sarcomere. This work bridges molecular disorder and biomechanical function, providing a new lens to understand dynamic control and dysfunction in cardiomyocyte contraction.

We present a comprehensive review of biomolecular binding kinetics and their modeling via Brownian dynamics simulations, with particular emphasis on enzyme-substrate interactions in cellular environments. We outline the theoretical foundations of Brownian dynamics (BD) and its application to modeling association and dissociation processes in both homogeneous and heterogeneous media. We further examine the role of BD in relation to emerging machine learning (ML) approaches for directly predicting binding kinetics. Finally, we propose that BD simulations can serve as a critical bridge between molecular-scale models and continuum, cell-level descriptions, offering a pathway toward a multiscale understanding of in vivo kinetic phenomena.

The interplay between keratinocyte (KC) hyperproliferation and T cell activation underpins psoriasis pathogenesis, posing significant challenges for effective treatment. Here, cell membrane‐coated nanoparticles (CNPs) are designed to simultaneously inhibit T cell activation and KC hyperproliferation for optimized psoriasis therapy. A customized cell membrane is genetically engineered to overexpress the immunoregulatory V‐domain immunoglobulin suppressor of T cell activation (VISTA) protein to enable potent T‐cell inhibition. The encapsulated core is designed as Janus nanoparticles (mSiO 2 &rPMO) for co‐delivery of hydrophilic 5‐aminolevulinic acid (5‐ALA) and hydrophobic tazarotene, facilitating synergistic sonodynamic therapy (SDT) and anti‐inflammatory effects. Mechanistically, under ultrasound stimulation, accumulated protoporphyrin IX (PpIX) derived from 5‐ALA metabolism generates reactive oxygen species (ROS) that inhibit epidermal proliferation, while tazarotene activates retinoic acid receptor gamma (RARγ) to exert anti‐inflammatory effects. Concurrently, the VISTA‐overexpressed membrane enhances anti‐inflammatory responses, further amplifying the anti‐psoriatic effect. CNP efficacy in a psoriatic mouse model is validated, demonstrating mitigated lesion severity, suppressed epidermal hyperplasia, and reduced inflammatory cytokine levels in skin lesions. Notably, CNPs outperformed non‐membrane counterparts in both cellular and animal models, highlighting the anti‐psoriatic amplification conferred by the engineered membrane. Overall, this research presents a novel therapeutic strategy for psoriasis management using engineered membrane‐coated mesoporous Janus nanoparticles to target keratinocyte proliferation and T cell activation.

Introduction Hyperlipidemia is a key contributor to cardiovascular diseases, underscoring the necessity for alternative lipid-lowering treatments beyond statins. Objectives This study aimed to synthesize and identify small, low-toxicity lipid-lowering compounds and investigate their mechanisms of action. Methods A series of tetrahydroisoquinoline compounds were synthesized, with HepG2 cells used to screen and identify B11 as a potent, low-toxicity lipid-lowering candidate. B11′s efficacy was tested in various hyperlipidemic animal models, including C57BL/6 mice, hamsters, and humanized PCSK9 transgenic (B6-hPCSK9) mice. Target interactions were investigated using various in vitro techniques, including molecular docking, cellular thermal shift assays (CETSA), drug affinity responsive target stability (DARTS), and surface plasmon resonance (SPR). Furthermore, we evaluated the synergistic effects of B11 combined with statins in C57BL/6 mice. Results A series of tetrahydroisoquinoline compounds was synthesized, identifying B11 as a potent lipid-lowering candidate with minimal toxicity. B11 significantly reduced total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and triglycerides (TG) in the plasma and liver of high fat diet (HFD) induced mice, hamsters, and B6-hPCSK9 mice, without causing any adverse effects. Mechanistically, B11 targets the 455–692 amino acid region of pro-protein convertase subtilisin/kexin type 9 (PCSK9), blocking its interaction with the low-density lipoprotein receptor (LDL-R) and inducing PCSK9 degradation via the ubiquitin–proteasome pathway. This process leads to increased LDL-R levels, enhancing LDL-C clearance. Notably, The unique mechanism of B11 enables combination therapy with atorvastatin, leading to stronger lipid-lowering effects and lower liver toxicity. Conclusions These findings demonstrate a small, non-statin compounds, and provide the potential alternative treatment approach for hyperlipidemic patients.

ASPP2 and iASPP bind to p53 through their conserved ANK-SH3 domains to respectively promote and inhibit p53-dependent cell apoptosis. While crystallography has indicated that these two proteins employ distinct surfaces of their ANK-SH3 domains to bind to p53, solution NMR data has suggested similar surfaces. In this study, we employed multi-scale molecular dynamics (MD) simulations combined with free energy calculations to reconcile the discrepancy in the binding modes. We demonstrated that the binding mode based solely on a single crystal structure does not enable iASPP's RT loop to engage with p53's C-terminal linker-a verified interaction. Instead, an ensemble of simulated iASPP-p53 complexes facilitates this interaction. We showed that the ensemble-average inter-protein contacting residues and NMR-detected interfacial residues qualitatively overlap on ASPP proteins, and the ensemble-average binding free energies better match experimental KD values compared to single crystallgarphy-determined binding mode. For iASPP, the sampled ensemble complexes can be grouped into two classes, resembling the binding modes determined by crystallography and solution NMR. We thus propose that crystal packing shifts the equilibrium of binding modes towards the crystallography-determined one. Lastly, we showed that the ensemble binding complexes are sensitive to p53's intrinsically disordered regions (IDRs), attesting to experimental observations that these IDRs contribute to biological functions. Our results provide a dynamic and ensemble perspective for scrutinizing these important cancer-related protein-protein interactions (PPIs).