AGO2 promotes tumor progression in KRAS-driven mouse models of non-small cell lung cancer.

Lung cancer is the deadliest malignancy in the United States. Non-small cell lung cancer (NSCLC) accounts for 85% of cases and is frequently driven by activating mutations in the gene encoding the KRAS GTPase (e.g., KRASG12D). Our previous work demonstrated that Argonaute 2 (AGO2)-a component of the RNA-induced silencing complex (RISC)-physically interacts with RAS and promotes its downstream signaling. We therefore hypothesized that AGO2 could promote KRASG12D-dependent NSCLC in vivo. To test the hypothesis, we evaluated the impact of Ago2 knockout in the KPC (LSL-KrasG12D/+;p53f/f;Cre) mouse model of NSCLC. In KPC mice, intratracheal delivery of adenoviral Cre drives lung-specific expression of a stop-floxed KRASG12D allele and biallelic ablation of p53 Simultaneous biallelic ablation of floxed Ago2 inhibited KPC lung nodule growth while reducing proliferative index and improving pathological grade. We next applied the KPHetC model, in which the Clara cell-specific CCSP-driven Cre activates KRASG12D and ablates a single p53 allele. In these mice, Ago2 ablation also reduced tumor size and grade. In both models, Ago2 knockout inhibited ERK phosphorylation (pERK) in tumor cells, indicating impaired KRAS signaling. RNA sequencing (RNA-seq) of KPC nodules and nodule-derived organoids demonstrated impaired canonical KRAS signaling with Ago2 ablation. Strikingly, accumulation of pERK in KPC organoids depended on physical interaction of AGO2 and KRAS. Taken together, our data demonstrate a pathogenic role for AGO2 in KRAS-dependent NSCLC. Given the prevalence of this malignancy and current difficulties in therapeutically targeting KRAS signaling, our work may have future translational relevance.

Targeting transcriptional regulation of SARS-CoV-2 entry factors ACE2 and TMPRSS2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for COVID-19, employs two key host proteins to gain entry and replicate within cells, angiotensin-converting enzyme 2 (ACE2) and the cell surface transmembrane protease serine 2 (TMPRSS2). TMPRSS2 was first characterized as an androgen-regulated gene in the prostate. Supporting a role for sex hormones, males relative to females are disproportionately affected by COVID-19 in terms of mortality and morbidity. Several studies, including one employing a large epidemiological cohort, suggested that blocking androgen signaling is protective against COVID-19. Here, we demonstrate that androgens regulate the expression of ACE2, TMPRSS2, and androgen receptor (AR) in subsets of lung epithelial cells. AR levels are markedly elevated in males relative to females greater than 70 y of age. In males greater than 70 y old, smoking was associated with elevated levels of AR and ACE2 in lung epithelial cells. Transcriptional repression of the AR enhanceosome with AR or bromodomain and extraterminal domain (BET) antagonists inhibited SARS-CoV-2 infection in vitro. Taken together, these studies support further investigation of transcriptional inhibition of critical host factors in the treatment or prevention of COVID-19.

Transcriptome Analysis Reveals the Immunoregulatory Activity of Rice Seed-Derived Peptide PEP1 on Dendritic Cells.

Some food-derived bioactive peptides exhibit prominent immunoregulatory activity. We previously demonstrated that the rice-derived PEP1 peptide, GIAASPFLQSAAFQLR, has strong immunological activity. However, the mechanism of this action is still unclear. In the present study, full-length transcripts of mouse dendritic cells (DC2.4) treated with PEP1 were sequenced using the PacBio sequencing platform, and the transcriptomes were compared via RNA sequencing (RNA-Seq). The characteristic markers of mature DCs, the cluster of differentiation CD86, and the major histocompatibility complex (MHC-II), were significantly upregulated after the PEP1 treatment. The molecular docking suggested that hydrogen bonding and electrostatic interactions played important roles in the binding between PEP1, MHC-II, and the T-cell receptor (TCR). In addition, the PEP1 peptide increased the release of anti-inflammatory factors (interleukin-4 and interleukin-10) and decreased the release of pro-inflammatory factors (interleukin-6 and tumor necrosis factor-α). Furthermore, the RNA-seq results showed the expression of genes involved in several signaling pathways, such as the NF-κB, MAPK, JAK-STAT, and TGF-ß pathways, were regulated by the PEP1 treatment, and the changes confirmed the immunomodulatory effect of PEP1 on DC2.4 cells. This findings revealed that the PEP1 peptide, derived from the byproduct of rice processing, is a potential natural immunoregulatory alternative for the treatment of inflammation.

Porphyrin NanoMOFs as a catalytic label in nanozyme-linked immunosorbent assay for Aflatoxin B1 detection.

Food safety is a global problem, and methods to reliably and sensitivity detect contaminants could be transformative. Herein, we synthesized a novel Porphyrin NanoMoFs (NanoPCN-223(Fe)) with excellent dispersion and peroxidase-like activity. The Km value of NanoPCN-223(Fe) was 2.0 × 10-4 M toward the H2O2 substrate during the catalytic process. We use the NanoPCN-223(Fe) to construct an enhanced dispersion MOF-linked immunosorbent assay (Ed-MOFLISA) to sensitive detect AFB1 in milk. The optimized Ed-MOFLISA displayed a broad quantitative range from 0.05 to 10 ng/mL and a limit of detection of 0.003 ng/mL. Spiked peanut and soy milk recovery ranged from 91.22% to 97.63%. Intra-assay and inter-assay coefficient of variation values ranged from 0.78 to 3.85%, demonstrating the outstanding reproducibility and accuracy of Ed-MOFLISA. We applied the Ed-MOFLISA assay to test the milk samples, demonstrating its potential use for monitoring food quality.

In Vitro Anti-Inflammatory Activity of Three Peptides Derived from the Byproduct of Rice Processing.

Inflammation is a contributing factor to the initiation and progression of many diseases, and some food-derived biofunctional peptides show high anti-inflammatory activity. In our previous study, we demonstrated that peptides derived from trypsin hydrolysis of rice protein show good immunological activity. In the present study, proteins of broken rice were extracted and identified by macroporous resin fractionation and liquid chromatography/tandem mass spectrometry (LC-MS/MS). Subsequently, a bioinformatics prediction and in silico simulation approach was used to screen for peptides showing anti-inflammatory activity, including inhibition of the production of nitric oxide and proinflammatory cytokines (interleukin-1ß, interleukin-6, and tumor necrosis factor-α) by lipopolysaccharide-stimulated RAW264.7 mice macrophages. Three peptides (DNIQGITKPAIR, IAFKTNPNSMVSHIAGK, and IGVAMDYSASSKR) that demonstrated the highest binding affinity were synthesized, and their in vitro anti-inflammatory activity was investigated. This is the first study that integrates LC-MS/MS identification and bioinformatics prediction for reporting the anti-inflammatory activity of anti-inflammatory peptides derived from broken rice protein. The study findings revealed that the peptides derived from the byproduct of rice milling could be potentially used as natural anti-inflammatory alternativities.

Three-dimensional assembly and disassembly of Fe3O4-decorated porous carbon nanocomposite with enhanced transversal relaxation for magnetic resonance sensing of bisphenol A.

The design and construction of a novel magnetic resonance sensor (MRS) is presented for bisphenol A (BPA) detection. The MRS has been built based on the core component of magnetic Fe3O4 nanoparticles (~ 40 nm), which were uniformly distributed in nanoporous carbon (abbreviated as Fe3O4@NPC). The synthesis was derived from the calcination of the metal organic framework (MOF) precursor of Fe-MIL-101 at high temperature. Fe3O4@NPC was confirmed with enhanced transversal relaxation with r2 value of 118.2 mM-1 s-1, which was around 1.7 times higher than that of the naked Fe3O4 nanoparticle. This enhancement is attributed to the excellent proton transverse relaxation rate of Fe3O4@NPC caused by the reduced self-diffusion coefficient of water molecules in the vicinity of Fe3O4 nanoparticles in the nanoporous carbon. BPA antibody (Ab) and antigen (Ag)-ovalbumin (OVA) were immobilized onto the Fe3O4@NPC to form Ab-Fe3O4@NPC and Ag-Fe3O4@NPC, respectively. These two composites can cause the three-dimensional assembly of Fe3O4@NPC via immunological recognition. The presence of BPA can compete with antigen-OVA to combine with Ab-Fe3O4@NPC, thereby breaking the assembly process (disassembly). The difference in the change of the T2 value before and after adding BPA can thus be used to monitor BPA. The proposed MRS not only revealed a wide linear range of BPA concentration from 0.05 to 50 ng mL-1 with an extremely low detection limit of 0.012 ng mL-1 (S/N = 3), but also displayed high selectivity towards matrix interferences. The recoveries of BPA ranged from 95.6 to 108.4% for spiked tea π, and 93.4 to 104.7% for spiked canned oranges samples, respectively, and the RSD (n = 3) was less than 4.4% for 3 successive assays. The versatility of Fe3O4@NPC with customized relaxation responses provides the possibility for the adaptation of magnetic resonance platforms for food safety development. The magnetic Fe3O4 nanoparticles are uniformly dispersed in the nanoporous carbon (Fe3O4@NPC), which derived from the calcinating of the metal organic framework (MOF) precursor of Fe-MIL-101. And the magnetic Fe3O4@NPCs are adopted for the construction of magnetic resonance sensor (MRS) for bisphenol A (BPA) detection.

Peroxidase-mimetic activity of a nanozyme with uniformly dispersed Fe3O4 NPs supported by mesoporous graphitized carbon for determination of glucose.

A Fe3O4/mesoporous graphitized carbon (Fe3O4/m-GC) composite was prepared through a facile calcination method with iron-based metal-organic frameworks (Fe-MOFs) as a sacrificial template. After carbonization, the Fe3O4 nanoparticles were uniformly dispersed in the mesoporous carbon support, resulting in spatial structural stability. The mesoporous carbon support obtained was highly graphitized and exhibited eminent electrical conductivity, which accelerated the electron transfer between the Fe3O4 nanoparticles by Fe(II)/Fe(III) redox cycles and m-GC by C = Csp2/C-Csp3 redox cycles, leading to the excellent peroxidase-mimetic activity of Fe3O4/m-GC. Km values for tetramethylbenzidine (TMB) and H2O2 were 26.8 and 15.8 times lower than that of natural horseradish peroxidase, respectively. Taking advantage of the peroxidase-mimetic activity of Fe3O4/m-GC, a colorimetric assay was fabricated for detecting glucose in the range 0.5 ~ 200 µM, with a limit of detection of 0.24 µM. Fig 1 A Schematic illustration of the preparation process of Fe3O4/m-GC, B schematic illustration of a proposed synergistic catalytic mechanism of TMB oxidation by Fe3O4/m-GC.

Recent Advances in Porphyrin-Based Materials for Metal Ions Detection.

Porphyrins have planar and conjugated structures, good optical properties, and other special functional properties. Owing to these excellent properties, in recent years, porphyrins and their analogues have emerged as a multifunctional platform for chemical sensors. The rich chemistry of these molecules offers many possibilities for metal ions detection. This review mainly discusses two types of molecular porphyrin and porphyrin composite sensors for metal ions detection, because porphyrins can be functionalized to improve their functional properties, which can introduce more chemical and functional sites. According to the different application materials, the section of porphyrin composite sensors is divided into five sub-categories: (1) porphyrin film, (2) porphyrin metal complex, (3) metal-organic frameworks, (4) graphene materials, and (5) other materials, respectively.

Metal-Organic Frameworks of MIL-100(Fe, Cr) and MIL-101(Cr) for Aromatic Amines Adsorption from Aqueous Solutions.

MIL-100(Fe, Cr) and MIL-101(Cr) were synthesized by the hydrothermal method and applied to the adsorptions of five aromatic amines from aqueous solutions. These three metal-organic frameworks (MOFs) were well characterized by powder X-ray diffraction (PXRD), scanning electron microscope (SEM), transmission electron microscope (TEM), thermogravimetric analysis (TGA) and surface area analysis. The adsorption mechanism of three MOFs and the effects of the structures of MOFs on the adsorption of aromatic amines were discussed. The results show that the cavity system and suitable hydrogen bond acceptor were important factors for the adsorption for five aromatic amines of aniline, 1-naphthalamine, o-toluidine, 2-amino-4-nitrotoluene and 2-nitroaniline: (a) the saturated adsorption capacity of aniline, 1-naphthylamine and o-toluidine on MIL-100(Fe) were 52.0, 53.4 and 49.6 mg/g, respectively, which can be attributed to the intermolecular hydrogen bond interaction and cavity system diffusion. (b) The adsorption capacity of 2-nitroaniline and 2-amino-4-nitrotoluene on MIL-101(Cr) were 54.3 and 25.0 mg/g, respectively, which can be attributed to the more suitable pore size of MIL-101(Cr) than that of MIL-100(Fe, Cr). The MOFs of MIL-100(Fe) and MIL-101(Cr) can be potential materials for removing aromatic amines from aqueous solutions.

MicroRNA expression profile and functional analysis reveal their roles in contact inhibition and its disruption switch of rat vascular smooth muscle cells.

Contact inhibition and its disruption of vascular smooth muscle cells (VSMCs) are important cellular events in vascular diseases. But the underlying molecular mechanisms are unclear. In this study we investigated the roles of microRNAs (miRNAs) in the contact inhibition and its disruption of VSMCs and the molecular mechanisms involved. Rat VSMCs were seeded at 30% or 90% confluence. MiRNA expression profiles in contact-inhibited confluent VSMCs (90% confluence) and non-contact-inhibited low-density VSMCs (30% confluence) were determined. We found that multiple miRNAs were differentially expressed between the two groups. Among them, miR-145 was significantly increased in contact-inhibited VSMCs. Serum could disrupt the contact inhibition as shown by the elicited proliferation of confluent VSMCs. The contact inhibition disruption accompanied with a down-regulation of miR-145. Serum-induced contact inhibition disruption of VSMCs was blocked by overexpression of miR-145. Moreover, downregulation of miR-145 was sufficient to disrupt the contact inhibition of VSMCs. The downregulation of miR-145 in serum-induced contact inhibition disruption was related to the activation PI3-kinase/Akt pathway, which was blocked by the PI3-kinase inhibitor LY294002. KLF5, a target gene of miR-145, was identified to be involved in miR-145-mediated effect on VSMC contact inhibition disruption, as it could be inhibited by knockdown of KLF5. In summary, our results show that multiple miRNAs are differentially expressed in contact-inhibited VSMCs and in non-contact-inhibited VSMCs. Among them, miR-145 is a critical gene in contact inhibition and its disruption of VSMCs. PI3-kinase/Akt/miR-145/KLF5 is a critical signaling pathway in serum-induced contact inhibition disruption. Targeting of miRNAs related to the contact inhibition of VSMCs may represent a novel therapeutic approach for vascular diseases.

Phospholipase D2-dependent inhibition of the nuclear hormone receptor PPARgamma by cyclic phosphatidic acid.

Cyclic phosphatidic acid (1-acyl-2,3-cyclic-glycerophosphate, CPA), one of nature's simplest phospholipids, is found in cells from slime mold to humans and has a largely unknown function. We find here that CPA is generated in mammalian cells in a stimulus-coupled manner by phospholipase D2 (PLD2) and binds to and inhibits the nuclear hormone receptor PPARgamma with nanomolar affinity and high specificity through stabilizing its interaction with the corepressor SMRT. CPA production inhibits the PPARgamma target-gene transcription that normally drives adipocytic differentiation of 3T3-L1 cells, lipid accumulation in RAW264.7 cells and primary mouse macrophages, and arterial wall remodeling in a rat model in vivo. Inhibition of PLD2 by shRNA, a dominant-negative mutant, or a small molecule inhibitor blocks CPA production and relieves PPARgamma inhibition. We conclude that CPA is a second messenger and a physiological inhibitor of PPARgamma, revealing that PPARgamma is regulated by endogenous agonists as well as by antagonists.

Protective effects of a wheat germ peptide (RVF) against H2O 2-induced oxidative stress in human neuroblastoma cells.

RVF (Arg-Val-Phe), a peptide derived from wheat germ, shows antioxidant properties. Here, the neuroprotective efficacies of RVF were investigated in human neuroblastoma cells (SH-SY5Y) that were pretreated with RVF (150-250 µM, 4 h) and exposed to H2O2 (200 µM). RVF increased viable cell numbers by 37 % and reduced the release of lactate dehydrogenase. Pretreatment with RVF also inhibited H2O2-induced accumulation of reactive oxygen species and maintained the mitochondrial transmembrane potential as well as preventing intracellular Ca(2+) dysregulation during H2O2 exposure. Furthermore, pretreatment with RVF increased the Bcl-2/Bax ratio and blocked cleavage poly(ADP-ribose) polymerase by inhibiting caspase-3 activation, thus decreasing apoptosis.

Comprehensive Review of EGCG Modification: Esterification Methods and Their Impacts on Biological Activities.

Epigallocatechin gallate (EGCG), the key constituent of tea polyphenols, presents challenges in terms of its lipid solubility, stability, and bioavailability because of its polyhydroxy structure. Consequently, structural modifications are imperative to enhance its efficacy. This paper comprehensively reviews the esterification techniques applied to EGCG over the past two decades and their impacts on bioactivities. Both chemical and enzymatic esterification methods involve catalysts, solvents, and hydrophobic groups as critical factors. Although the chemical method is cost-efficient, it poses challenges in purification; on the other hand, the enzymatic approach offers improved selectivity and simplified purification processes. The biological functions of EGCG are inevitably influenced by the structural changes incurred through esterification. The antioxidant capacity of EGCG derivatives can be compromised under certain conditions by reducing hydroxyl groups, while enhancing lipid solubility and stability can strengthen their antiviral, antibacterial, and anticancer properties. Additionally, esterification broadens the utility of EGCG in food applications. This review provides critical insights into developing cost-effective and environmentally sustainable selective esterification methods, as well as emphasizes the elucidation of the bioactive mechanisms of EGCG derivatives to facilitate their widespread adoption in food processing, healthcare products, and pharmaceuticals.

Influence of substrate aggregation state on the enzymatic-induced crosslinking of soy protein isolate.

Transglutaminase (TGase) induced-crosslinking of soy protein isolate (SPI) was markedly influenced by the substrate aggregation state. Results showed that appropriate heating significantly accelerated the TGase crosslinking, and the 7S and 11S acidic subunits were more susceptible to the enzyme than the 11S basic proteins. The content of ε-(γ-glutamyl)-lysine isopeptide bonds increased from 4.74 to 8.61 µmol/g protein when the heating intensity was increased from 75 °C for 15 min to 95 °C for 30 min, due to sufficient unfolding of the protein structure. Rheological data indicated that the gel formed from the SPI heated at 95 °C for 30 min exhibited the best properties, with a 60 % increase in the storage modulus compared with the unheated sample. However, excessive heating (95 °C, 60-120 min) caused severe aggregation of SPI and formation of insoluble aggregates, resulting in poor crosslinking efficiency and weaker gel properties.

Functionalized magnetic metal organic framework nanocomposites for high throughput automation extraction and sensitive detection of antipsychotic drugs in serum samples.

Due to the complexity of biological sample matrix, the automated and high-throughput pretreatment technology is urgently needed for monitoring the antipsychotic drugs for mental patients. In this study, functionalized magnetic zirconium-based organic framework nanocomposites (Fe3O4@SiO2@Zr-MOFs) were successfully designed and synthesized by the layer-by-layer growth. Among them, Fe3O4@SiO2@UiO-67-COOH showed the best adsorption performance, and at the same time it exhibited excellent water dispersibility, high thermal stability, chemical stability and high hydrophobicity. Results of adsorption kinetics, isotherm and FT-IR showed that the adsorption process was dominated by chemical adsorption (hydrogen bond, electrostatic interaction, π-π interaction) and monolayer adsorption. Moreover, the smaller pore size improved the protein exclusion rate which reached 98.9-99.8%. Based on the above result, the synthesized magnetic nanoparticles were introduced to 96-well automatic extractor, antipsychotic drugs in 96 serum samples were automatically extracted within 9 min, which most greatly saved the time and labor costs and avoided artificial errors. By further integrating with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), antipsychotic drugs can be detected in the range of 0.2-3.0 ng mL-1 with a quantitative limit of 0.06-0.9 ng mL-1. The recoveries of antipsychotic drugs and their metabolites in serum ranged from 95.7% to 112.3% within 1.4-6.5% of RSD. These features indicate that the proposed method is promising for high throughput and sensitively monitoring of drugs and other hazardous substances.

Nano­selenium functionalized chitosan gel beads for Hg(II) removal from apple juice.

The presence of potentially toxic elements and compounds poses threats to the quality and safety of fruit juices. Among these, Hg(II) is considered as one of the most poisonous heavy metals to human health. Traditional chitosan-based and selenide-based adsorbents face challenges such as poor adsorption capacity and inconvenient separation in juice applications. In this study, we prepared nano­selenium functionalized chitosan gel beads (nanoSe@CBs) and illustrated the synergistic promotions between chitosan and nanoSe in removing Hg(II) from apple juice. The preparation conditions, adsorption behaviors, and adsorption mechanism of nanoSe@CBs were systematically investigated. The results revealed that the adsorption process was primarily controlled by chemical adsorption. At the 0.1 % dosage, the adsorbent exhibited high uptake, and the maximum adsorption capacity from the Langmuir isotherm model could reach 376.5 mg/g at room temperature. The adsorbent maintained high adsorption efficiency (> 90 %) across a wide range of Hg(II) concentrations (0.01 to 10 mg/L) and was unaffected by organic acids present in apple juice. Additionally, nanoSe@CBs showed negligible effects on the quality of apple juice. Overall, nanoSe@CBs open up possibilities to be used as a safe, low-cost and highly-efficient adsorbent for the removal of Hg(II) from juices and other liquid foods.

Length-controlled hydrophobic CF3-COF as a highly efficient absorbent coating for dual-mode solid-phase microextraction of sixteen polycyclic aromatic hydrocarbons in water samples.

Polycyclic aromatic hydrocarbons (PAHs), a group of seriously hazardous environmental contaminants, have attracted extensive attention due to their carcinogenicity, genotoxicity, mutagenicity, and ubiquity. In this work, the excellent hydrophobic trifluoromethyl-enriched covalent organic framework (CF3-COF) was designed and synthesized as coating of solid-phase microextraction (SPME). The CF3-COF offered a high adsorption selectivity for PAHs, which could be attributed to the multiple interactions between the CF3-COF and PAHs, including hydrophobicity interaction, π-π and H bond interactions. Furthermore, headspace (HS) and direct immersion (DI) dual-mode solid-phase microextraction (HS/DI-SPME) were innovatively integrated as a dual-mode extraction by varying the length of SPME coating on stainless-steel, which could simultaneously and efficiently extract 16 PAHs with different volatile. Amazingly, the proposed strategy achieved fast adsorption for PAHs and shortened the adsorption equilibrium time to 15 min. By further integrating with gas chromatography tandem mass spectrometry (GC-MS/MS), PAHs could be detected in the range of 0.008-0.16 ng mL-1 with a quantitative limit of 0.029-0.47 ng mL-1, respectively. The recoveries of PAHs in water samples ranged from 80.84 to 117.67 %. This work indicates that the dual-mode CF3-COF-SPME is a promising candidate for the enrichment of multiple hazardous substances in complicated samples.

A novel magnetic resonance tuning-magnetic relaxation switching sensor based on Gd-MOF/USPIO assembly for sensitive and convenient aflatoxin B1 detection.

Aflatoxin B1 (AFB1) can accumulate in different organs or tissues and seriously harm humans. Traditional magnetic relaxation switching (MRS) sensors have relatively low sensitivity, but are complex to use. Rapid small-trace molecule analysis in complex samples is challenging. In this study, we used a gadolinium-based metal-organic framework (Gd-MOF) and ultra-small superparamagnetic iron oxide (USPIO) assembly to develop a magnetic resonance tuning-magnetic relaxation switching (MRET-MRS) sensor to improve conventional MRS sensor sensitivity and simplify operational steps in complex samples. Importantly, the local magnetic field generated by USPIO interfered with Gd-MOF electron spin fluctuation and directly affected dipole-dipole interactions between Gd electrons and water molecules, thus rendering relaxation signal changes more sensitive. The sensitivity (0.54 pg mL-1) was 833 times more sensitive than that of a conventional MRS sensor (0.45 ng mL-1). Finally, a convenient one-step detection approach can be achieved by mixing antigen/antibody functionalized Gd-MOF/USPIO and target samples.

Novel Immunosensor Based on Metal Single-Atom Nanozymes with Enhanced Oxidase-Like Activity for Capsaicin Analysis in Spicy Food.

Capsaicin (CAP) is a primary indicator for assessing the level of pungency. Herein, iron-based single-atom nanozymes (SAzymes) (Fe/NC) with exceptional oxidase-like activity were used to construct an immunosensor for CAP analysis. Fe/NC could imitate oxidase actions by transforming O2 to •O2- radicals in the absence of hydrogen peroxide (H2O2), which could avoid complex operations and unstable results. By regulating the Fe atom loads, an optimal Fe0.7/NC atom usage rate could improve the catalytic activity (Michaelis-Menten constant (Km) = 0.09 mM). Fe0.7/NC was integrated with goat antimouse IgG by facile mix incubation to develop a competitive enzyme-linked immunosorbent assay (ELISA). Our Fe0.7/NC immunosensing platform is anticipated to outperform the conventional ELISA in terms of stability and shelf life. The proposed immunosensor provided color responses across 0.01-1000 ng/mL CAP concentrations, with a detection limit of 0.046 ng/mL. Fe/NC may have potential as nanozymes for CAP detection in spicy foods, with promising applications in food biosensing.

p300/CBP degradation is required to disable the active AR enhanceosome in prostate cancer.

Prostate cancer is an exemplar of an enhancer-binding transcription factor-driven disease. The androgen receptor (AR) enhanceosome complex comprised of chromatin and epigenetic coregulators assembles at enhancer elements to drive disease progression. The paralog lysine acetyltransferases p300 and CBP deposit histone marks that are associated with enhancer activation. Here, we demonstrate that p300/CBP are determinant cofactors of the active AR enhanceosome in prostate cancer. Histone H2B N-terminus multisite lysine acetylation (H2BNTac), which is exclusively reliant on p300/CBP catalytic function, marked active enhancers and was notably elevated in prostate cancer lesions relative to the adjacent benign epithelia. Degradation of p300/CBP rapidly depleted acetylation marks associated with the active AR enhanceosome, which was only partially phenocopied by inhibition of their reader bromodomains. Notably, H2BNTac was effectively abrogated only upon p300/CBP degradation, which led to a stronger suppression of p300/CBP-dependent oncogenic gene programs relative to bromodomain inhibition or the inhibition of its catalytic domain. In vivo experiments using an orally active p300/CBP proteolysis targeting chimera (PROTAC) degrader (CBPD-409) showed that p300/CBP degradation potently inhibited tumor growth in preclinical models of castration-resistant prostate cancer and synergized with AR antagonists. While mouse p300/CBP orthologs were effectively degraded in host tissues, prolonged treatment with the PROTAC degrader was well tolerated with no significant signs of toxicity. Taken together, our study highlights the pivotal role of p300/CBP in maintaining the active AR enhanceosome and demonstrates how target degradation may have functionally distinct effects relative to target inhibition, thus supporting the development of p300/CBP degraders for the treatment of advanced prostate cancer.