Multifunctional polymeric nanocapsules with enhanced cartilage penetration and retention for osteoarthritis treatment.

Osteoarthritis (OA) is a prevalent joint disease characterized by cartilage degeneration and subchondral bone homeostasis imbalance. Effective topical OA therapy is challenging, as therapeutic drugs often suffer from insufficient penetration and rapid clearance. We develop miniature polydopamine (PDA) nanocapsules (sub-60 nm), which are conjugated with collagen-binding polypeptide (CBP) and loaded with an anabolic drug (i.e., parathyroid hormone, PTH) for efficient OA treatment. Such multifunctional polymeric nanocapsules, denoted as PDA@CBP-PTH, possess deformability when interacting with the dense collagen fiber networks, enabling the efficient penetration into 1 mm cartilage in 4 h and prolonged retention within the joints up to 28 days. Moreover, PDA@CBP-PTH nanocapsules exhibit excellent reactive oxygen species scavenging property in chondrocytes and enhance the anabolism in subchondral bone. The nanosystem, as dual-mode treatment for OA, demonstrates rapid penetration, long-lasting effects, and combinational therapeutic impact, paving the way for reversing the progression of OA for joint health care.

Interfacial modification of recombinant protein for immunoglobulin G adsorption with spindle-shaped MOF as nano molecular containers.

Development of fresh solid phase extractant is critical for selective separation and purification of special proteins. Herein, we demonstrated a recombinant Staphylococcal Protein G (rSPG) with a His-tag modified the novel single-metal organic framework (rSPG@Ni-MOF-74). The proposed solid-phase extraction material possessed a uniform spindle-shaped structure, large surface area (709.60 m2 g-1) and pore volume (0.08 m3 g-1), high metal content (22.57 wt%), which facilitated the interaction between host and guest. As results, the composite displayed outstanding selective recognition and adsorption of IgG, due to synergistic effect of the binding ability of rSPG with the Fc region of IgG, maintained through hydrogen bonding and electrostatic attraction, as well as hydrophobic interaction. The adsorption performance and mechanism of rSPG@Ni-MOF-74 have been fully investigated. Additionally, the rSPG@Ni-MOF-74 composite could effectively separate IgG from serum obtained from healthy humans, with the purity of the separated IgG verified through SDS-PAGE analysis. Furthermore, LC-MS/MS analysis identified a high content of IgG (55.3 %) in the eluate from rSPG@Ni-MOF-74, suggesting the great potential of rSPG@Ni-MOF-74 in IgG separation and enrichment from complex matrix.

In-situ activation of biomimetic single-site bioorthogonal nanozyme for tumor-specific combination therapy.

Copper-catalyzed click chemistry offers creative strategies for activation of therapeutics without disrupting biological processes. Despite tremendous efforts, current copper catalysts face fundamental challenges in achieving high efficiency, atom economy, and tissue-specific selectivity. Herein, we develop a facile "mix-and-match synthetic strategy" to fabricate a biomimetic single-site copper-bipyridine-based cerium metal-organic framework (Cu/Ce-MOF@M) for efficient and tumor cell-specific bioorthogonal catalysis. This elegant methodology achieves isolated single-Cu-site within the MOF architecture, resulting in exceptionally high catalytic performance. Cu/Ce-MOF@M favors a 32.1-fold higher catalytic activity than the widely used MOF-supported copper nanoparticles at single-particle level, as first evidenced by single-molecule fluorescence microscopy. Furthermore, with cancer cell-membrane camouflage, Cu/Ce-MOF@M demonstrates preferential tropism for its parent cells. Simultaneously, the single-site CuII species within Cu/Ce-MOF@M are reduced by upregulated glutathione in cancerous cells to CuI for catalyzing the click reaction, enabling homotypic cancer cell-activated in situ drug synthesis. Additionally, Cu/Ce-MOF@M exhibits oxidase and peroxidase mimicking activities, further enhancing catalytic cancer therapy. This study guides the reasonable design of highly active heterogeneous transition-metal catalysts for targeted bioorthogonal reactions.

Colorimetric sensing for translational applications: from colorants to mechanisms.

Colorimetric sensing offers instant reporting via visible signals. Versus labor-intensive and instrument-dependent detection methods, colorimetric sensors present advantages including short acquisition time, high throughput screening, low cost, portability, and a user-friendly approach. These advantages have driven substantial growth in colorimetric sensors, particularly in point-of-care (POC) diagnostics. Rapid progress in nanotechnology, materials science, microfluidics technology, biomarker discovery, digital technology, and signal pattern analysis has led to a variety of colorimetric reagents and detection mechanisms, which are fundamental to advance colorimetric sensing applications. This review first summarizes the basic components (e.g., color reagents, recognition interactions, and sampling procedures) in the design of a colorimetric sensing system. It then presents the rationale design and typical examples of POC devices, e.g., lateral flow devices, microfluidic paper-based analytical devices, and wearable sensing devices. Two highlighted colorimetric formats are discussed: combinational and activatable systems based on the sensor-array and lock-and-key mechanisms, respectively. Case discussions in colorimetric assays are organized by the analyte identities. Finally, the review presents challenges and perspectives for the design and development of colorimetric detection schemes as well as applications. The goal of this review is to provide a foundational resource for developing colorimetric systems and underscoring the colorants and mechanisms that facilitate the continuing evolution of POC sensors.

Activable Nano-Immunomodulator Assembled from π-Extended Naphthalenediimide for Precision Photothermal Immunotherapy.

A nano-immunomodulator (R-NPT NP) comprising a tumor microenvironment (TME) activable resiquimod (R848) and a π-extended NIR-absorbing naphthophenanthrolinetetraone (NPT) has been engineered for spatiotemporal controlled photothermal immunotherapy. R-NPT NP demonstrated excellent photostability, while R848 promoted synergistic immunity as a toll-like receptor 7/8 (TLR7/8) agonist. Upon accumulation at the tumor site, R-NPT NP released R848 in response to redox metabolite glutathione (GSH), triggering dendritic cell (DC) activation. The photothermal effect endowed by R-NPT NP can ablate tumors directly and trigger immunogenic cell death to augment immunity after photoirradiation. The synergistic effect of GSH-liable TLR7/8 agonist and released immunogenic factors leads to a robust evocation of systematic immunity through promoted DC maturation and T cell infiltration. Thus, R-NPT NP with photoirradiation achieved 99.3 % and 98.2 % growth inhibition against primary and distal tumors, respectively.

Accelerating carrier separation to boost the photocatalytic CO2 reduction performance of ternary heterojunction Ag-Ti3C2Tx/ZnO catalysts.

Developing low-cost and efficient photocatalyst/co-catalyst systems that promote CO2 reduction remains a challenge. In this work, Ag-Ti3C2Tx composites were made using a self-reduction technique, and unique Ag-Ti3C2Tx/ZnO ternary heterojunction structure photocatalysts were created using an electrostatic self-assembly process. The photocatalyst's close-contact heterogeneous interface increases photogenerated carrier migration efficiency. The combination of Ti3C2Tx and Ag improves the adsorption active sites and reaction centers for ZnO, making it a key site for CO2 adsorption and activation. The best photocatalysts had CO and CH4 reduction efficiencies of 11.985 and 0.768 µmol g-1 h-1, respectively. The CO2 conversion was 3.35 times better than that of pure ZnO, which demonstrated remarkable stability even after four cycle trials with no sacrificial agent. Furthermore, in situ diffuse reflectance infrared Fourier transform spectroscopy (in situ DRIFTS) and valence band spectroscopy were utilized to propose the photocatalytic reaction mechanism and electron transfer channels of the Ag-Ti3C2Tx/ZnO system, confirming that CHO* and CO* are the important intermediates in the generation of CH4 and CO. This study introduces a novel method for the development of new and efficient photocatalysts and reveals that Ti3C2Tx MXene is a viable co-catalyst for applications.

A Cancer Nanovaccine Based on an FeAl-Layered Double Hydroxide Framework for Reactive Oxygen Species-Augmented Metalloimmunotherapy.

The complexity and heterogeneity of individual tumors have hindered the efficacy of existing therapeutic cancer vaccines, sparking intensive interest in the development of more effective in situ vaccines. Herein, we introduce a cancer nanovaccine for reactive oxygen species-augmented metalloimmunotherapy in which FeAl-layered double hydroxide (LDH) is used as a delivery vehicle with dihydroartemisinin (DHA) as cargo. The LDH framework is acid-labile and can be degraded in the tumor microenvironment, releasing iron ions, aluminum ions, and DHA. The iron ions contribute to aggravated intratumoral oxidative stress injury by the synergistic Fenton reaction and DHA activation, causing apoptosis, ferroptosis, and immunogenic cell death in cancer cells. The subsequently released tumor-associated antigens with the aluminum adjuvant form a cancer nanovaccine to generate robust and long-term immune responses against cancer recurrence and metastasis. Moreover, Fe ion-enabled T1-weighted magnetic resonance imaging can facilitate real-time tumor therapy monitoring. This cancer-nanovaccine-mediated metalloimmunotherapy strategy has the potential for revolutionizing the precision immunotherapy landscape.

Ultrahigh-Ionic-Conductivity, Antifreezing Poly(amidoxime)-Grafted Polyzwitterion Hydrogel for Facile Integrated into High-Performance Stretchable Flexible Supercapacitor.

Developing wearable supercapacitors (SCs) with high stretchability, arbitrary deformability, and antifreezing ability is still a challenge. In the present work, an ultrahigh-ionic-conductivity, antifreezing poly(amidoxime)-graft-polyzwitterion (PAO-g-PSBMA) hydrogel electrolyte is fabricated by grafting PSBMA in PAO. Owing to the abundant hydrophilic and high ionic adsorption capacity of amidoxime groups in PAO and zwitterion groups in PSBMA, the as-prepared PAO-g-PSBMA hydrogel can facilitate the dissociation of lithium salt and exhibit an ultrahigh ionic conductivity of 29.8 S m-1 at 25 °C and 3.4 S m-1 even at -30 °C. Employing mATi3C2Tx and mSTi3C2Tx, which contain small amounts of PAO-AGE and PAO-g-PSBMA dispersions, respectively, coated onto both sides of the PAO-g-PSBMA hydrogel, we followed a thermal treatment to facilely form integrated stretchable flexible SCs. The as-prepared SCs show an outstanding recoverable tensile stain of 80% and an excellent electrochemical stability under many types and times of arbitrary deformation. More importantly, as-prepared mATi3C2Tx- and mSTi3C2Tx-based SCs present fantastic antifreezing ability and excellent stability with 74.6 and 78.3% retention of the initial capacitance, respectively, even after 1000 times of stretching to 60% at -30 °C. This work offers a new strategy of using PAO-grafted polyzwitterion for obtaining an antifreezing stretchable SC, which shows a high potential for application in next-generation integrated stretchable devices in various fields.

Triptolide induces apoptosis and cytoprotective autophagy by ROS accumulation via directly targeting peroxiredoxin 2 in gastric cancer cells.

Triptolide, a natural bioactive compound derived from herbal medicine Tripterygium wilfordii, has multiple biological activities including anti-cancer effect, which is being tested in clinical trials for treating cancers. However, the exact mechanism by which Triptolide exerts its cytotoxic effects, particularly its specific protein targets, remains unclear. Here, we show that Triptolide effectively induces cytotoxicity in gastric cancer cells by increasing reactive oxygen species (ROS) levels. Further investigations reveal that ROS accumulation contributes to the induction of Endoplasmic Reticulum (ER) stress, and subsequently autophagy induction in response to Triptolide. Meanwhile, this autophagy is cytoprotective. Interestingly, through activity-based protein profiling (ABPP) approach, we identify peroxiredoxins-2 (PRDX2), a component of the key enzyme systems that act in the defense against oxidative stress and protect cells against hydroperoxides, as direct binding target of Triptolide. By covalently binding to PRDX2 to inhibit its antioxidant activity, Triptolide increases ROS levels. Moreover, overexpression of PRDX2 inhibits and knockdown of the expression of PRDX2 increases Triptolide-induced apoptosis. Collectively, these results indicate PRDX2 as a direct target of Triptolides for inducing apoptosis. Our results not only provide novel insight into the underlying mechanisms of Triptolide-induced cytotoxic effects, but also indicate PRDX2 as a promising potential therapeutic target for developing anti-gastric cancer agents.

Covalently surface-grafting α­zirconium phosphate nanoplatelets enables collagen fiber matrix with ultraviolet barrier, antibacterial, and flame-retardant properties.

Manipulating the dispersibility and reactivity of two-dimensional nanomaterials in collagen fibers (CFs) matrix has aroused attention in the fabrication of multifunctional collagen-based nanocomposites. Here, α­zirconium phosphate nanoplatelets (ZrP NPs) were surface-functionalized with gallic acid (GA) to afford ZrP-GA NPs for engineering CFs matrix. The influence of ZrP-GA NPs on the ultraviolet barrier, antibacterial, and flame-retardant properties of resultant CFs matrix were investigated. Microstructural analysis revealed that ZrP-GA NPs were dispersed and bound within the collagen fibrils and onto the collagen strands in the CFs matrix. The resultant CFs matrix also maintained typical D-periodic structures of collagen fibrils and native branching and interwoven structures of CFs networks with increased porosity and enhanced ultraviolet barrier properties. Inhibition zone testing presented excellent antibacterial activities of the CFs matrix owing to surface grafting of antibacterial GA. Thanks to enhanced dispersion and binding of ZrP NPs with the CFs matrix by surface-functionalization with GA, the resultant CFs matrix reduced the peak heat release rate and the total heat release by 42.9 % and 39.0 %, respectively, highlighting improved flame-retardant properties. We envision that two-dimensional nanomaterials possess great potential in developing reasonable collagen-based nanocomposites towards the manufacture of emergent multifunctional collagen fibers-based wearable electronics.

PSMC5 regulates microglial polarization and activation in LPS-induced cognitive deficits and motor impairments by interacting with TLR4.

Luteolin is a flavonoid found in high concentrations in celery and green pepper, and acts as a neuroprotectant. PSMC5 (proteasome 26S subunit, ATPase 5) protein levels were reduced after luteolin stimulation in activated microglia. We aimed to determine whether regulating PSMC5 expression could inhibit neuroinflammation, and investigate the underlying mechanisms.BV2 microglia were transfected with siRNA PSMC5 before the addition of LPS (lipopolysaccharide, 1.0 µg/ml) for 24 h in serum free DMEM. A mouse model of LPS-induced cognitive and motor impairment was established to evaluate the neuroprotective effects of shRNA PSMC5. Intracerebroventricular administration of shRNA PSMC5 was commenced 7 days prior to i.p. injection of LPS (750 µg/kg). Treatments and behavioral experiments were performed once daily for 7 consecutive days. Behavioral tests and pathological/biochemical assays were performed to evaluate LPS-induced hippocampal damage. Molecular dynamics simulation was used to confirm the interaction between PSMC5 and TLR4 (Toll-like receptor 4) in LPS-stimulated BV2 microglia. SiRNA PSMC5 inhibited BV2 microglial activation, and suppressed the release of inflammatory factors (IL-1ß, COX-2, PGE2, TNF-α, and iNOS) upon after LPS stimulation in BV2 microglia. LPS increased IκB-α and p65 phosphorylation, which was attenuated by siRNA PSMC5. Behavioral tests and pathological/biochemical assays showed that shRNA PSMC5 attenuated LPS-induced cognitive and motor impairments, and restored synaptic ultrastructure and protein levels in mice. ShRNA PSMC5 reduced pro-inflammatory cytokine (TNF-α, IL-1ß, PGE2, and NO) levels in the serum and brain, and relevant protein factors (iNOS and COX-2) in the brain. Furthermore, shRNA PSMC5 upregulated the anti-inflammatory mediators interleukin IL-4 and IL-10 in the serum and brain, and promoted a pro-inflammation-to-anti-inflammation phenotype shift in microglial polarization. Mechanistically, shRNA PSMC5 significantly alleviated LPS-induced TLR4 expression. The polarization of LPS-induced microglial pro-inflammation phenotype was abolished by TLR4 inhibitor and in the TLR-4-/- mouse, as in shRNA PSMC5 treatment. PSMC5 interacted with TLR4 via the amino sites Glu284, Met139, Leu127, and Phe283. PSMC5 site mutations attenuated neuroinflammation and reduced pro-inflammatory factors by reducing TLR4-related effects, thereby reducing TLR4-mediated MyD88 (myeloid differentiation factor 88)-dependent activation of NF-κB. PSMC5 could be an important therapeutic target for treatment of neurodegenerative diseases involving neuroinflammation-associated cognitive deficits and motor impairments induced by microglial activation.

Mixed-valence gold-porphyrin two-dimensional coordination networks for repurposing of chrysotherapy.

Catalytic gold nanomaterials typically exhibit antibacterial properties, albeit significantly weaker than ionic gold in chrysotherapy. The inherent stability of gold nanoparticles prevents the release of gold ions, limiting their ability to achieve efficient antibacterial therapy. To address this limitation, we propose a novel sustained ionic gold release strategy through the construction of a mixed-valence gold-porphyrin coordination network (Au-Por). By adjusting the ratio of Au to porphyrin molecule, an ultrathin two-dimensional Au-Por nanosheet was successfully synthesized, which contains 85.9 % of Au (III). In addition, the remaining gold existed in the form of uniformly distributed ultrasmall nanoclusters on the Au-Por nanosheet. Notably, the Au-Por nanosheet exhibited a sustained release of gold ions. Thus, a multimodal antibacterial therapy was achieved by integrating the direct bactericidal action of ionic gold and lethal reactive oxygen species (ROS) generated through the peroxidase (POD)-like activity of gold nanoclusters and photodynamic therapy (PDT) using porphyrins. The innovative Au-Por exerted broad-spectrum bactericidal activity against both Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus bacteria mediated by bacterial membrane disruption and DNA damage. Moreover, in vivo studies demonstrated the synergistic effect of Au-Por on combating skin wound infections and facilitating wound healing. Comprehensive safety evaluations proved that Au-Por exhibited no hematotoxicity or hepatorenal toxicity, and it also displayed rapid renal clearance after treatment, indicating favorable biocompatibility. The repurposing of chrysotherapy has revolutionized the antibacterial strategy of nanoscale gold, resulting in a dramatic boost in antibacterial activity and valuable insights for designing highly efficient nanoscale antibacterial agents.

Potent gene delivery from fluorinated poly(ß-amino ester) in adhesive and suspension difficult-to-transfect cells for apoptosis and ferroptosis.

Tremendous efforts have been made to improve polymeric property in gene delivery performances, especially when obstacle of transferring gene construct into difficult-to-transfect cells occurs. Innovations in the area of fluorination and fluorinated compounds with biomedical potential in medicinal chemistry are believed to assist in the development of new therapeutics. Fluorine modified polymers have shown to navigate the gene transfection cellular barriers and promoted the transfection outcomes. Gene transfer into some liver cancer cells and human leukemia cells has always been a challenge. Here, by facile incorporation of a fluorine containing amine monomer, 1H,1H-undecafluorohexylamine, fluorinated poly(ß-amino ester) (FPAE) was synthesized to significantly improve the transfection performance, achieving high transfection efficiency of 87% and 55% in two representative difficult-to-transfect cells, HepG2 and Molt-4, which were cultured in adhesive and suspension condition, respectively. However, the potency of Lipofectamine 3000 was very limited. More importantly, functional studies revealed that FPAE can dramatically outperform Lipofectamine 3000 in delivering Bcl-xL and PKCßII to either provide the protection against apoptosis or promote the ferroptosis in HepG2 cells. This work facilitates gene therapies by overcoming biological barriers for targeting difficult-to-transfect cells and disease models when medically necessary.

Cationic Carbon Monoxide-Releasing Polymers as Antimicrobial and Antibiofilm Agents by the Synergetic Activity.

Recent studies indicate that carbon monoxide-releasing molecules (CORMs), a class of organometallic compounds, exert antibacterial activities through the delivery of carbon monoxide (CO) molecules. We developed a new-class CO-delivery system by conjugating classical low-molecular-weight CORMs (i.e., [Ru(CO)3Cl2]2 and Mn(CO)5Br) onto a positively charged carrier, polyimidazolium (PIM), giving cationic CO-releasing polymers Ru@PIM and Mn@PIM, respectively. Compared with low-molecular-weight CORMs, our polymeric CO vehicles showed improved water solubility, reduced cytotoxicity, significantly extended CO-releasing duration, and enhanced antimicrobial ability against both planktonic and biofilm microorganisms. Ru@PIM and Mn@PIM inhibited the growth of a broad spectrum of free Gram-positive and Gram-negative bacteria as well as fungus with the lowest minimum inhibitory concentration (MIC) at 8 µg/mL. They were effective in preventing pathogenic Pseudomonas aeruginosa biofilm formation with biofilm reduction by more than 92% at 16 µg/mL and 99% at 32 µg/mL. They also demonstrated potent dispersal efficacy on recalcitrant well-established biofilms through a synergetic activity with a biofilm log10 reduction of 2.5-3.2 ≥ 64 µg/mL and nearly 2.0 at the concentration of as low as 16 µg/mL. This CO-releasing system may retain long-time antimicrobial ability after the complete release of CO molecules owing to the cationic structure. The novel CO-releasing polymers have great potential as antimicrobial and antibiofilm agents in biomedical applications.

Correction: Polymers as advanced antibacterial and antibiofilm agents for direct and combination therapies.

[This corrects the article DOI: 10.1039/D1SC05835E.].

Oculocerebrorenal syndrome of Lowe (OCRL) controls leukemic T-cell survival by preventing excessive PI(4,5)P2 hydrolysis in the plasma membrane.

T-cell acute lymphoblastic leukemia (T-ALL) is one of the deadliest and most aggressive hematological malignancies, but its pathological mechanism in controlling cell survival is not fully understood. Oculocerebrorenal syndrome of Lowe is a rare X-linked recessive disorder characterized by cataracts, intellectual disability, and proteinuria. This disease has been shown to be caused by mutation of oculocerebrorenal syndrome of Lowe 1 (OCRL1; OCRL), encoding a phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] 5-phosphatase involved in regulating membrane trafficking; however, its function in cancer cells is unclear. Here, we uncovered that OCRL1 is overexpressed in T-ALL cells, and knockdown of OCRL1 results in cell death, indicating the essential role of OCRL in controlling T-ALL cell survival. We show OCRL is primarily localized in the Golgi and can translocate to plasma membrane (PM) upon ligand stimulation. We found OCRL interacts with oxysterol-binding protein-related protein 4L, which facilitates OCRL translocation from the Golgi to the PM upon cluster of differentiation 3 stimulation. Thus, OCRL represses the activity of oxysterol-binding protein-related protein 4L to prevent excessive PI(4,5)P2 hydrolysis by phosphoinositide phospholipase C ß3 and uncontrolled Ca2+ release from the endoplasmic reticulum. We propose OCRL1 deletion leads to accumulation of PI(4,5)P2 in the PM, disrupting the normal Ca2+ oscillation pattern in the cytosol and leading to mitochondrial Ca2+ overloading, ultimately causing T-ALL cell mitochondrial dysfunction and cell death. These results highlight a critical role for OCRL in maintaining moderate PI(4,5)P2 availability in T-ALL cells. Our findings also raise the possibility of targeting OCRL1 to treat T-ALL disease.

Membrane Potential-Dependent Uptake of Cationic Oligoimidazolium Mediates Bacterial DNA Damage and Death.

The treatment of bacterial infections is becoming increasingly challenging with the emergence of antimicrobial resistance. Thus, the development of antimicrobials with novel mechanisms of action is much needed. Previously, we designed several cationic main-chain imidazolium compounds and identified the polyimidazolium PIM1 as a potent antibacterial against a wide panel of multidrug-resistant nosocomial pathogens, and it had relatively low toxicity against mammalian epithelial cells. However, little is known about the mechanism of action of PIM1. Using an oligomeric version of PIM1 with precisely six repeating units (OIM1-6) to control for consistency, we showed that OIM1-6 relies on an intact membrane potential for entry into the bacterial cytoplasm, as resistant mutants to OIM1-6 have mutations in their electron transport chains. These mutants demonstrate reduced uptake of the compound, which can be circumvented through the addition of a sub-MIC dose of colistin. Once taken up intracellularly, OIM1-6 exerts double-stranded DNA breaks. Its potency and ability to kill represents a promising class of drugs that can be combined with membrane-penetrating drugs to potentiate activity and hedge against the rise of resistant mutants. In summary, we discovered that cationic antimicrobial OIM1-6 exhibits an antimicrobial property that is dissimilar to the conventional cationic antimicrobial compounds. Its killing mechanism does not involve membrane disruption but instead depends on the membrane potential for uptake into bacterial cells so that it can exert its antibacterial effect intracellularly.

Biochemical mechanisms of tributyltin chloride-induced cell toxicity in Sertoli cells.

Tributyltin chloride (TBTCL) is a widely used fungicide and heat stabilizer in compositions of PVC. TBTCL has been detected in human bodies and potentially causes harmful effects on humans' thyroid, cardiovascular and other organs. As one of the first examples of endocrine disruptors, the toxicity effects of TBTCL on the male reproduction system have aroused concerns. However, the potential cellular mechanisms are not fully explored. In the current study, by using Sertoli cells, a critical regulator of spermatogenesis as a cell model, we showed that with 200 nM exposure for 24 h, TBTCL causes apoptosis and cell cycle arrest. RNA sequencing analyses suggested that TBTCL probably activates endoplasmic reticulum (ER) stress, and disrupts autophagy. Biochemical analysis showed that TBTCL indeed induces ER stress and the dysregulation of autophagy. Interestingly, activation of ER stress and inhibition of autophagy is responsible for TBTCL-induced apoptosis and cell cycle arrest. Our results thus uncovered a novel insight into the cellular mechanisms for TBTCL-induced toxicology in Sertoli cells.

Tributyltin chloride (TBTCL) induces cell injury via dysregulation of endoplasmic reticulum stress and autophagy in Leydig cells.

Tributyltin chloride (TBTCL), a commonly used antiseptic substance, is commonly found in the environment. Human exposure to TBTCL through the consumption of contaminated seafood, fish, or drinking water has aroused concern. It is well-characterized that TBTCL has multiple detrimental effects on the male reproductive system. However, the potential cellular mechanisms are not fully elucidated. Here, we characterized molecular mechanisms of TBTCL-induced cell injury in Leydig cells, a critical supporter for spermatogenesis. We showed that TBTCL induces apoptosis and cell cycle arrest in TM3 mouse Leydig cells. RNA sequencing analyses revealed that endoplasmic reticulum (ER) stress and autophagy were potentially involved in TBTCL-induced cytotoxicity. We further showed that TBTCL causes ER stress and inhibited autophagy flux. Notably, the inhibition of ER stress attenuates not only TBTCL-induces autophagy flux inhibition but also apoptosis and cell cycle arrest. Meanwhile, the activation of autophagy alleviates, and inhibition of autophagy exaggerates TBTCL-induced apoptosis and cell cycle arrest flux. These results suggest that TBTCL-induced ER stress and autophagy flux inhibition contributed to apoptosis and cell cycle arrest in Leydig cells, providing novel understanding into the mechanisms of TBTCL-induced testis toxicity.

An acquired phosphatidylinositol 4-phosphate transport initiates T-cell deterioration and leukemogenesis.

Lipid remodeling is crucial for malignant cell transformation and tumorigenesis, but the precise molecular processes involved and direct evidences for these in vivo remain elusive. Here, we report that oxysterol-binding protein (OSBP)-related protein 4 L (ORP4L) is expressed in adult T-cell leukemia (ATL) cells but not normal T-cells. In ORP4L knock-in T-cells, ORP4L dimerizes with OSBP to control the shuttling of OSBP between the Golgi apparatus and the plasma membrane (PM) as an exchanger of phosphatidylinositol 4-phosphate [PI(4)P]/cholesterol. The PI(4)P arriving at the PM via this transport machinery replenishes phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] and phosphatidylinositol (3,4,5) trisphosphate [PI(3,4,5)P3] biosynthesis, thus contributing to PI3K/AKT hyperactivation and T-cell deterioration in vitro and in vivo. Disruption of ORP4L and OSBP dimerization disables PI(4)P transport and T-cell leukemogenesis. In summary, we identify a non-vesicular lipid transport machinery between Golgi and PM maintaining the oncogenic signaling competence initiating T-cell deterioration and leukemogenesis.