Platelet factor 4(PF4) and its multiple roles in diseases.

Platelet factor 4 (PF4) combines with heparin to form an antigen that could produce IgG antibodies and participate in heparin-induced thrombocytopenia (HIT). PF4 has attracted wide attention due to its role in novel coronavirus vaccine-19 (COVID-9)-induced immune thrombotic thrombocytopenia (VITT) and cognitive impairments. The electrostatic interaction between PF4 and negatively charged molecules is vital in the progression of VITT, which is similar to HIT. Emerging evidence suggests its multiple roles in hematopoietic and angiogenic inhibition, platelet coagulation interference, host inflammatory response promotion, vascular inhibition, and antitumor properties. The emerging pharmacological effects of PF4 may help deepen the exploration of its mechanism, thus accelerating the development of targeted therapies. However, due to its pleiotropic properties, the development of drugs targeting PF4 is at an early stage and faces many challenges. Herein, we discussed the characteristics and biological functions of PF4, summarized PF4-mediated signaling pathways, and discussed its multiple roles in diseases to inform novel approaches for successful clinical translational research.

A review on artificial water channels incorporated polyamide membranes for water purification: Transport mechanisms and performance.

While thin-film composite (TFC) polyamide (PA) membranes are advanced for removing salts and trace organic contaminants (TrOCs) from water, TFC PA membranes encounter a water permeance-selectivity trade-off due to PA layer structural characteristics. Drawing inspiration from the excellent water permeance and solute rejection of natural biological channels, the development of analogous artificial water channels (AWCs) in TFC PA membranes (abbreviated as AWCM) promises to achieve superior mass transfer efficiency, enabling breaking the upper bound of water permeance and selectivity. Herein, we first discussed the types and structural characteristics of AWCs, followed by summarizing the methods for constructing AWCM. We discussed whether the AWCs acted as the primary mass transfer channels in AWCM and emphasized the important role of the AWCs in water transport and ion/TrOCs rejection. We thoroughly summarized the molecular-level mechanisms and structure-performance relationship of water molecules, ions, and TrOCs transport in the confined nanospace of AWCs, which laid the foundation for illustrating the enhanced water permeance and salt/TrOCs selectivity of AWCM. Finally, we discussed the challenges encountered in the field of AWCM and proposed future perspectives for practical applications. This review is expected to offer guidance for understanding the transport mechanisms of AWCM and developing next-generation membrane for effective water treatment.

Virulence and transmission characteristics of clade 2.3.4.4b H5N6 subtype avian influenza viruses possessing different internal gene constellations.

Clade 2.3.4.4 H5N6 avian influenza virus (AIV) has been predominant in poultry in China, and the circulating haemagglutinin (HA) gene has changed from clade 2.3.4.4h to clade 2.3.4.4b in recent years. In 2021, we isolated four H5N6 viruses from ducks during the routine surveillance of AIV in China. The whole-genome sequencing results demonstrated that the four isolates all belonged to the currently prevalent clade 2.3.4.4b but had different internal gene constellations, which could be divided into G1 and G2 genotypes. Specifically, G1 possessed H9-like PB2 and PB1 genes on the H5-like genetic backbone while G2 owned an H3-like PB1 gene and the H5-like remaining internal genes. By determining the characteristics of H5N6 viruses, including growth performance on different cells, plaque-formation ability, virus attachment ability, and pathogenicity and transmission in different animal models, we found that G1 strains were more conducive to replication in mammalian cells (MDCK and A549) and BALB/c mice than G2 strains. However, G2 strains were more advantageously replicated in avian cells (CEF and DF-1) and slightly more transmissible in waterfowls (mallards) than G1 strains. This study enriched the epidemiological data of H5 subtype AIV to further understand its dynamic evolution, and laid the foundation for further research on the mechanism of low pathogenic AIV internal genes in generating novel H5 subtype reassortants.

The Crosstalk between Nephropathy and Coagulation Disorder: Pathogenesis, Treatment, and Dilemmas.

ABSTRACT: The interaction between the kidney and the coagulation system greatly affects each other because of the abundant vessel distribution and blood perfusion in the kidney. Clinically, the risks of complicated thrombosis and bleeding have become important concerns in the treatment of nephropathies, especially nephrotic syndrome, CKD, ESKD, and patients with nephropathy undergoing RRTs. Adverse effects of anticoagulant or procoagulant therapies in patients with nephropathy, especially anticoagulation-related nephropathy, heparin-induced thrombocytopenia, and bleeding, seriously worsen the prognosis of patients, which have become challenges for clinicians. Over the decades, the interaction between the kidney and the coagulation system has been widely studied. However, the effects of the kidney on the coagulation system have not been systematically investigated. Although some coagulation-related proteins and signaling pathways have been shown to improve coagulation abnormalities while avoiding additional kidney damage in certain kidney diseases, their potential as anticoagulation targets in nephropathy requires further investigation. Here, we review the progression of research on the crosstalk between the coagulation system and kidney diseases and systematically analyze the significance and shortcomings of previous studies to provide new sight into future research. In addition, we highlight the status of clinical treatment for coagulation disorder and nephropathy caused by each other, indicating guidance for the formulation of therapeutic strategies or drug development.

[Inositol 1,4,5-triphosphate receptor 3 promotes renal cyst development in autosomal dominant polycystic kidney disease].

The purpose of the present study was to determine the role of inositol 1,4,5-trisphosphate receptor 3 (IP3R3) in renal cyst development in autosomal dominant polycystic kidney disease (ADPKD). 2-aminoethoxy-diphenyl borate (2-APB) and shRNA were used to suppress the expression of IP3R3. The effect of IP3R3 on cyst growth was investigated in Madin-Darby canine kidney (MDCK) cyst model, embryonic kidney cyst model and kidney specific Pkd1 knockout (PKD) mouse model. The underlying mechanism of IP3R3 in promoting renal cyst development was investigated by Western blot and immunofluorescence staining. The results showed that the expression level of IP3R3 was significantly increased in the kidneys of PKD mice. Inhibiting IP3R3 by 2-APB or shRNA significantly retarded cyst expansion in MDCK cyst model and embryonic kidney cyst model. Western blot and immunofluorescence staining results showed that hyperactivated cAMP-PKA signaling pathway in the growth process of ADPKD cyst promoted the expression of IP3R3, which was accompanied by a subcellular redistribution process in which IP3R3 was translocated from endoplasmic reticulum to intercellular junction. The abnormal expression and subcellular localization of IP3R3 further promoted cyst epithelial cell proliferation by activating MAPK and mTOR signaling pathways and accelerating cell cycle. These results suggest that the expression and subcellular distribution of IP3R3 are involved in promoting renal cyst development, which implies IP3R3 as a potential therapeutic target of ADPKD.

The challenges and opportunities of αvß3-based therapeutics in cancer: From bench to clinical trials.

Integrins are main cell adhesion receptors serving as linker attaching cells to extracellular matrix (ECM) and bidirectional hubs transmitting biochemical and mechanical signals between cells and their environment. Integrin αvß3 is a critical family member of integrins and interacts with ECM proteins containing RGD tripeptide sequence. Accumulating evidence indicated that the abnormal expression of integrin αvß3 was associated with various tumor progressions, including tumor initiation, sustained tumor growth, distant metastasis, drug resistance development, maintenance of stemness in cancer cells. Therefore, αvß3 has been explored as a therapeutic target in various types of cancers, but there is no αvß3 antagonist approved for human therapy. Targeting-integrin αvß3 therapeutics has been a challenge, but lessons from the past are valuable to the development of innovative targeting approaches. This review systematically summarized the structure, signal transduction, regulatory role in cancer, and drug development history of integrin αvß3, and also provided new insights into αvß3-based therapeutics in cancer from bench to clinical trials, which would contribute to developing effective targeting αvß3 agents for cancer treatment.

Targeting integrin pathways: mechanisms and advances in therapy.

Integrins are considered the main cell-adhesion transmembrane receptors that play multifaceted roles as extracellular matrix (ECM)-cytoskeletal linkers and transducers in biochemical and mechanical signals between cells and their environment in a wide range of states in health and diseases. Integrin functions are dependable on a delicate balance between active and inactive status via multiple mechanisms, including protein-protein interactions, conformational changes, and trafficking. Due to their exposure on the cell surface and sensitivity to the molecular blockade, integrins have been investigated as pharmacological targets for nearly 40 years, but given the complexity of integrins and sometimes opposite characteristics, targeting integrin therapeutics has been a challenge. To date, only seven drugs targeting integrins have been successfully marketed, including abciximab, eptifibatide, tirofiban, natalizumab, vedolizumab, lifitegrast, and carotegrast. Currently, there are approximately 90 kinds of integrin-based therapeutic drugs or imaging agents in clinical studies, including small molecules, antibodies, synthetic mimic peptides, antibody-drug conjugates (ADCs), chimeric antigen receptor (CAR) T-cell therapy, imaging agents, etc. A serious lesson from past integrin drug discovery and research efforts is that successes rely on both a deep understanding of integrin-regulatory mechanisms and unmet clinical needs. Herein, we provide a systematic and complete review of all integrin family members and integrin-mediated downstream signal transduction to highlight ongoing efforts to develop new therapies/diagnoses from bench to clinic. In addition, we further discuss the trend of drug development, how to improve the success rate of clinical trials targeting integrin therapies, and the key points for clinical research, basic research, and translational research.

Aquaporins in Urinary System.

There are at least eight aquaporins (AQPs) expressed in the kidney. Including AQP1 expressed in proximal tubules, thin descending limb of Henle and vasa recta; AQP2, AQP3, AQP4, AQP5, and AQP6 expressed in collecting ducts; AQP7 expressed in proximal tubules; AQP8 expressed in proximal tubules and collecting ducts; and AQP11 expressed in the endoplasmic reticulum of proximal tubular epithelial cells. Over years, researchers have constructed different AQP knockout mice and explored the effect of AQP knockout on kidney function. Thus, the roles of AQPs in renal physiology are revealed, providing very useful information for addressing fundamental questions about transepithelial water transport and the mechanism of near isoosmolar fluid reabsorption. This chapter introduces the localization and function of AQPs in the kidney and their roles in different kidney diseases to reveal the prospects of AQPs in further basic and clinical studies.

Rational design of trimetallic AgPt-Fe3O4 nanozyme for catalyst poisoning-mediated CO colorimetric detection.

Carbon monoxide (CO) is not only a highly poisonous gas that brings great health risk, but also a significant signaling molecule in body. However, it is still challengeable for development of alternative colorimetric probes to traditional organic chromophores for simple, sensitive and convenient CO sensing. Here, for the first time, we rationally design a novel hydrophilic AgPt-Fe3O4 nanozyme with a unique heterodimeric nanostructure for colorimetric sensing of CO based on the excellent peroxidase-like catalytic activity as well as highly poisonous effect of CO on the nanozyme's catalytic activity. Both experimental evidence and theoretical calculations reveal the trimetallic AgPt-Fe3O4 nanozyme is susceptible to poisoning with the strongest affinity towards CO compared to individual Fe3O4 or Ag-Fe3O4, which is attributed to the adequate exposure of the active metallic sites and efficient interfacial synergy of unique heterodimeric nanostructure. Accordingly, a novel nanozyme-based colorimetric strategy is developed for CO detection with a low detection limit of 5.6 ppb in solution. Furthermore, the probe can be prepared as very convenient test strips and integrated with the portable smartphone platforms for detecting CO gas samples with a low detection limit of 8.9 ppm. Overall, our work proposes guidelines for the rational design of metallic heterogeneous nanostructure to expand the analytical application of nanozyme.

1-Indanone retards cyst development in ADPKD mouse model by stabilizing tubulin and down-regulating anterograde transport of cilia.

Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited kidney disease. Cyst development in ADPKD involves abnormal epithelial cell proliferation, which is affected by the primary cilia-mediated signal transduction in the epithelial cells. Thus, primary cilium has been considered as a therapeutic target for ADPKD. Since ADPKD exhibits many pathological features similar to solid tumors, we investigated whether targeting primary cilia using anti-tumor agents could alleviate the development of ADPKD. Twenty-four natural compounds with anti-tumor activity were screened in MDCK cyst model, and 1-Indanone displayed notable inhibition on renal cyst growth without cytotoxicity. This compound also inhibited cyst development in embryonic kidney cyst model. In neonatal kidney-specific Pkd1 knockout mice, 1-Indanone remarkably slowed down kidney enlargement and cyst expansion. Furthermore, we demonstrated that 1-Indanone inhibited the abnormal elongation of cystic epithelial cilia by promoting tubulin polymerization and significantly down-regulating expression of anterograde transport motor protein KIF3A and IFT88. Moreover, we found that 1-Indanone significantly down-regulated ciliary coordinated Wnt/ß-catenin, Hedgehog signaling pathways. These results demonstrate that 1-Indanone inhibits cystic cell proliferation by reducing abnormally prolonged cilia length in cystic epithelial cells, suggesting that 1-Indanone may hold therapeutic potential to retard cyst development in ADPKD.

Achyranthes bidentata polysaccharides alleviate endoplasmic reticulum stress in osteoarthritis via lncRNA NEAT1/miR-377-3p pathway.

Endoplasmic reticulum stress (ERS) has been identified to be an important factor leading to chondrocyte apoptosis in osteoarthritis (OA). Previous studies have confirmed that Achyranthes bidentata polysaccharides (ABPS) can inhibit chondrocyte apoptosis; however, the mechanism of action of ABPS on chondrocyte ERS remains unclear. Thus in this study, we aim to investigate whether ABPS could inhibit OA-associated chondrocyte apoptosis by regulating ERS, especially by observing the relationship between the lncRNA NEAT1 and miR-377-3p, to explore further the protective mechanism of ABPS in OA. In vitro and in vivo experiments showed that ABPS inhibited chondrocyte ERS by regulating the expression of lncRNA NEAT1 and miR-377-3p. Moreover, both lncRNA NEAT1 silencing and miR-377-3p inhibition could attenuate the therapeutic effect of ABPS on ERS. Dual-luciferase results indicated that miR-377-3p targets the lncRNA NEAT1 gene in mouse chondrocytes. Therefore, we concluded that ABPS could inhibit thapsigargin (TG)-induced chondrocyte ERS through the lncRNA NEAT1/miR-377-3p axis.

Protective role of Achyranthes bidentata polysaccharides against chondrocyte extracellular matrix degeneration through lncRNA GAS5 in osteoarthritis.

Achyranthes bidentata polysaccharides (ABPS) is an active ingredient of the flowering plant Achyranthes bidentata that has been previously reported to be effective for the treatment of osteoarthritis (OA). However, the underlying molecular mechanism remain to be fully clarified. Emerging studies have shown that the long non-coding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) is involved in the pathogenesis of OA. Therefore, the present study aimed to investigate the potential mechanism of ABPS by focusing on its effects on the regulation of chondrocyte extracellular matrix (ECM) homeostasis, with particular emphasis on lncRNA GAS5. In the present study, the modified Hulth method was used to construct OA rats, which were gavaged with 400 mg/kg ABPS for 8 weeks. Histopathological changes in cartilage and subchondral bone were evaluated by hematoxylin-eosin staining and Safranin O-fast green staining. In in vitro experiments, IL-1ß-treated chondrocytes were infected with Lenti-lncRNA GAS5. Fluorescence in situ hybridization assay was performed to measure the expression of the lncRNA GAS5 in chondrocytes. Moreover, the relative expression level of lncRNA GAS5 in cartilage tissue and chondrocytes was detected using reverse transcription-quantitative PCR. Western blot analysis was used to detect protein expression levels of MMP-9, MMP-13, TIMP-1, TIMP-3 and type II collagen in cartilage tissue and chondrocytes. The results indicated that ABPS delayed the degradation of the ECM by chondrocytes in addition to reducing lncRNA GAS5 expression both in vivo and in vitro. Furthermore, silencing of lncRNA GAS5 expression in IL-1ß-treated chondrocytes downregulated the protein expression of MMP-9 and MMP-13 whilst upregulating the expression of tissue inhibitor matrix metalloproteinase (TIMP)-1, TIMP-3 and type II collagen. To conclude, the present study provides evidence that ABPS can inhibit the expression of lncRNA GAS5 in chondrocytes to regulate the homeostasis of ECM, which in turn may delay the occurrence of cartilage degeneration during OA.

Modification of ultrafiltration membrane with antibacterial agent intercalated layered nanosheets: Toward superior antibiofouling performance for water treatment.

Membrane fouling, especially biofouling induced by biofilm formation on membranes, can result in frequent cleaning or even replacement of membranes. Fabrication of membrane with excellent antibiofouling property is quite attractive due to its effectiveness and low-impact on the operation of membrane-based process. Herein, a cationic antibacterial agent, quaternary ammonium compound (QAC), was intercalated into the interlayer spaces of the MgAl layered double hydroxide (QAC/LDH) by self-assembly. The QAC/LDH composite was incorporated into polyethersulfone (PES) ultrafiltration (UF) membrane (PES-QLDH). The QAC/LDH enhanced the hydrophilicity, water flux, and resistance to organic fouling for the PES-QLDH membrane. The PES-QLDH membrane exhibited superior antibiofouling performance than the control PES membrane, with deposition of a thinner biofilm layer consisted of almost dead cells. The superior antibacterial activity inhibits the adhesion and growth of bacteria on the membrane surface, effectively retarding the formation of biofilms. Importantly, the synergistic effect of QAC and LDH in the PES-QLDH membrane resulted in a high biocidal activity based on both direct and indirect killing mechanisms. The PES-QLDH membrane maintained a stable and high antibacterial activity after several fouling-cleaning cycles. These results imply that the PES-QLDH membrane provides an effective and promising strategy for its long-term application in wastewater treatment.

Highly Specific Colorimetric Probe for Fluoride by Triggering the Intrinsic Catalytic Activity of a AgPt-Fe3O4 Hybrid Nanozyme Encapsulated in SiO2 Shells.

Current colorimetric probes for fluoride (F-) primarily rely on organic chromophores that often suffer from unsatisfactory selectivity, complex organic synthesis, and low aqueous compatibility. Herein, we proposed a highly specific colorimetric method for F- with 100% aqueous compatibility by triggering the intrinsic peroxidase-like activity of a AgPt-Fe3O4 nanozyme encapsulated in SiO2 shells. The excellent catalytic performance of the AgPt-Fe3O4 nanozyme serves as an ideal platform for sensitive colorimetric sensing. After being encapsulated in SiO2, the enzyme-like activity of AgPt-Fe3O4 is inhibited and only F- can exclusively etch the SiO2 shell to expose the active site of the nanozyme, thereby inducing color changes via oxidation of the chromogenic substrate. The limit of detection of the proposed method can reach as low as 13.73 µM in aqueous solution, which is lower than the maximum allowable concentration (79 µM) stipulated in the World Health Organization drinking water regulation. More importantly, this method is highly specific toward F- over other types of anions commonly found in environmental water, making it capable of analyzing sewage samples with very complex matrices. Finally, the nanoprobe is embedded into a test strip by electrostatic spinning to enable the rapid, visual, and on-site detection of F-.

[Clinical features and genetic analysis of a Chinese pedigree affected with X-linked adrenoleukodystrophy].

OBJECTIVE: To analyze the clinical features and variants of ABCD1 gene in a Chinese pedigree affected with X-linked adrenoleukodystrophy. METHODS: Clinical data of the proband were collected and analyzed. Potential variant of the ABCD1 gene were analyzed by PCR and Sanger sequencing of the proband, his parents and 100 unrelated healthy individuals. RESULTS: The prominent features of the proband included cerebellar and brainstem lesions, along with increased serum level of very-long chain fatty acids. He was found to harbor a hemizygous c.1509delG (p.L504Sfs*54) variant of the ABCD1 gene, for which his mother was heterozygous. The same variant was not detected in his father and 100 healthy controls. CONCLUSION: X-linked adrenoleukodystrophy has a variety of clinical manifestations. Discovery of the c.1509delG (p.L504Sfs*54), as a novel pathogenic variant of the ABCD1 gene, has enabled diagnosis and genetic counseling for this pedigree.

Pd-Fe3O4 Janus nanozyme with rational design for ultrasensitive colorimetric detection of biothiols.

Although nanozyme-based colorimetric assays have been broadly used for biosensing, some limitations such as low catalytic activity of nanozyme, poor sensitivity to analytes and lack of understanding the structure-activity relationship remain unsolved. In this work, we developed an ultrasensitive colorimetric method for biothiols detection based on density functional theory-assisted design of janus Pd-Fe3O4 nanozyme. The Pd-Fe3O4 dumbbell-like nanoparticles (DBNPs) prepared by seed-mediated approach shows a uniform heterodimeric nanostructure. Ultrasensitive biothiols detection is achieved from two aspects. On one hand, due to the synergistic effect between Pd and Fe3O4 in the dumbbell structure, Pd-Fe3O4 DBNPs show enhanced peroxidase-mimic activity compared to the individual components. On the other hand, when the target biothiols molecule is present, its inhibition effect on the janus Pd-Fe3O4 nanozyme is also significantly enhanced. The above results are confirmed both in experiment and theoretical calculation. Based on the rational design, a simple, highly selective and urtrasensitive colorimetric and quantitative assay for biothiols is developed. The limit of detection (LOD) can reach as low as 3.1 nM in aqueous solution. This assay is also successfully applied to the detection of biothiols in real urine samples. Moreover, the Pd-Fe3O4 nanozyme is used to discriminate biothiols levels in normal and cancer cells with high sensitivity at the cell density of 15,000/mL, which demonstrates its great potential in biological and clinical analysis. This work not only shows the great promise of janus bimetallic nanozymes' excellent functionalities but also provides rational guidelines to design high-performance nanozymes for biosensing and biomedical applications.

Construction of lignin-based nano-adsorbents for efficient and selective recovery of tellurium (IV) from wastewater.

Tellurium is massively used as the main light-absorbing layer component in the manufacturing of CdTe thin-film solar cells, a critical component in the photovoltaic industry. However, the process of manufacturing and renewing components has produced large amounts of tellurium-containing wastewater that is difficult to degrade and poses a serious threat to the aquatic ecosystem and human health. Hence, to achieve the recovery of tellurium resources for reducing their damages, a win-win approach was employed to utilize waste lignin to construct functional copper-doped activated lignin (CAL) adsorbents for selective separation and recovery of tellurium from wastewater. CAL exhibited superior adsorption properties towards tellurium (248.45 mg/g), mainly attributed to the adsorption mechanism of coordination interactions. Kinetic and isotherm results elucidated that monolayer chemisorption dominated CAL adsorption process. Besides, CAL had a satisfactory regeneration capability with minimal loss adsorption capacity after six consecutive cycles, which also exhibited high antifouling properties. Meanwhile, CAL achieved high selectivity for tellurium adsorption under the simulated wastewater, revealing the potential of CAL for practical application in wastewater. Therefore, this work provides a promising environmental strategy for exploring the application of lignin-based materials for tellurium recovery from wastewater.

Ganoderic Acids Prevent Renal Ischemia Reperfusion Injury by Inhibiting Inflammation and Apoptosis.

Renal ischemia reperfusion injury (RIRI) is one of the main causes of acute kidney injury (AKI), which can lead to acute renal failure. The development of RIRI is so complicated that it involves many factors such as inflammatory response, oxidative stress and cell apoptosis. Ganoderic acids (GAs), as one of the main pharmacological components of Ganoderma lucidum, have been reported to possess anti-inflammatory, antioxidant, and other pharmacological effects. The study is aimed to investigate the protective effect of GAs on RIRI and explore related underlying mechanisms. The mechanisms involved were assessed by a mouse RIRI model and a hypoxia/reoxygenation model. Compared with sham-operated group, renal dysfunction and morphological damages were relieved markedly in GAs-pretreatment group. GAs pretreatment could reduce the production of pro-inflammatory factors such as IL-6, COX-2 and iNOS induced by RIRI through inhibiting TLR4/MyD88/NF-kB signaling pathway. Furthermore, GAs reduced cell apoptosis via the decrease of the ratios of cleaved caspase-8 and cleaved caspase-3. The experimental results suggest that GAs prevent RIRI by alleviating tissue inflammation and apoptosis and might be developed as a candidate drug for preventing RIRI-induced AKI.

Inhibiting Focal Adhesion Kinase Ameliorates Cyst Development in Polycystin-1-Deficient Polycystic Kidney Disease in Animal Model.

BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is characterized by numerous cysts originating from renal tubules and is associated with significant tubular epithelial cell proliferation. Focal adhesion kinase (FAK) promotes tumor growth by regulating multiple proliferative pathways. METHODS: We established the forskolin (FSK)-induced three-dimensional (3D) Madin-Darby Canine Kidney cystogenesis model and 8-bromoadenosine-3`,5`-cyclic monophosphate-stimulated cyst formation in ex vivo embryonic kidney culture. Cultured human renal cyst-lining cells (OX-161) and normal tubular epithelial cells were treated with FAK inhibitors or transfected with green fluorescent protein-tagged FAK mutant plasmids for proliferation study. Furthermore, we examined the role of FAK in two transgenic ADPKD animal models, the kidney-specific Pkd1 knockout and the collecting duct-specific Pkd1 knockout mouse models. RESULTS: FAK activity was significantly elevated in OX-161 cells and in two ADPKD mouse models. Inhibiting FAK activity reduced cell proliferation in OX-161 cells and prevented cyst growth in ex vivo and 3D cyst models. In tissue-specific Pkd1 knockout mouse models, FAK inhibitors retarded cyst development and mitigated renal function decline. Mechanically, FSK stimulated FAK activation in tubular epithelial cells, which was blocked by a protein kinase A (PKA) inhibitor. Inhibition of FAK activation by inhibitors or transfected cells with mutant FAK constructs interrupted FSK-mediated Src activation and upregulation of ERK and mTOR pathways. CONCLUSIONS: Our study demonstrates the critical involvement of FAK in renal cyst development, suggests that FAK is a potential therapeutic target in treating patients with ADPKD, and highlights the role of FAK in cAMP-PKA-regulated proliferation.

Tumors exploit FTO-mediated regulation of glycolytic metabolism to evade immune surveillance.

The ever-increasing understanding of the complexity of factors and regulatory layers that contribute to immune evasion facilitates the development of immunotherapies. However, the diversity of malignant tumors limits many known mechanisms in specific genetic and epigenetic contexts, manifesting the need to discover general driver genes. Here, we have identified the m6A demethylase FTO as an essential epitranscriptomic regulator utilized by tumors to escape immune surveillance through regulation of glycolytic metabolism. We show that FTO-mediated m6A demethylation in tumor cells elevates the transcription factors c-Jun, JunB, and C/EBPß, which allows the rewiring of glycolytic metabolism. Fto knockdown impairs the glycolytic activity of tumor cells, which restores the function of CD8+ T cells, thereby inhibiting tumor growth. Furthermore, we developed a small-molecule compound, Dac51, that can inhibit the activity of FTO, block FTO-mediated immune evasion, and synergize with checkpoint blockade for better tumor control, suggesting reprogramming RNA epitranscriptome as a potential strategy for immunotherapy.