The FASEB Journal最新文献

Gestational diabetes mellitus (GDM) increases the risks for fetal complications such as large-for-gestational-age infants and stillbirth, and disposes both the mother and fetus to develop type 2 diabetes in later life. The placenta, which is crucial for fetal development, exhibits structural and functional changes in GDM, but the impact of these alterations on placental trophoblast function remains unclear. During their differentiation, villous cytotrophoblast display several characteristics of senescent cells; however, the role of senescence pathways in placental function remains unexplored in normoglycemic as well as GDM pregnancies. Here, we investigate placental senescence pathways utilizing term villous tissue and primary trophoblasts to assess senescence pathways and determine fetal sex-based differences in normoglycemic and GDM pregnancies. Our data suggest that both p21- and p16-mediated senescence pathways are activated during trophoblast differentiation and are dysregulated in the GDM placenta in a sexually dimorphic manner. We also provide evidence for increased activation of p53-Lamin A/C and p16-RB pathways in trophoblasts from GDM placentas. Reduced expression of p21 and its downstream effects on GCM1 expression and βhCG secretion outline how altered physiological senescence can affect trophoblast differentiation and function. This is an important study highlighting the disruption of placental senescence pathways in pregnancies complicated by GDM.

Kidney injury is a common complication of deep hypothermic circulatory arrest (DHCA) in patients undergoing cardiac surgery. However, the role of competitive endogenous RNA (ceRNA) networks in mediating DHCA-induced kidney injury has not been fully elucidated. In the present study, we aimed to systematically analyze ceRNA networks and identify a novel lncRNA, MSTRG.16386.1, that promotes DHCA-induced kidney injury by regulating the miR-466b-5p/Sprouty RTK signaling antagonist 2 (Spry2) axis. Kidney injury induced by DHCA was confirmed using a rat model of cardiopulmonary bypass (CPB) with or without DHCA. In addition, lncRNA and mRNA sequencing of kidney tissues revealed 309 specific shared differentially expressed mRNAs (DEGs) and 439 differentially expressed lncRNAs (DELs) in the kidneys of CPB+DHCA rats compared with those of SHAM and CPB rats. Differentially expressed miRNAs (DEMs) were predicted by coexpression and binding site analysis. A ceRNA network consisting of 12 DEGs, 6 DELs, and 11 DEMs was constructed, and the top ranked RNAs were Sprouty RTK Signaling Antagonist 2 (Spry2), MSTRG.16386.1, and miR-466b-5p, which were verified using qRT–PCR. We found that MSTRG.16386.1 overexpression resulted in NRK-52E cell apoptosis, which was suppressed by the interaction with miR-466b-5p. In addition, we demonstrated that Spry2 is a key target of MSTRG.16386.1 and miR-466b-5p. Rescue experiment results revealed that the downregulated expression of Spry2 protected NRK-52E cells against apoptosis mediated by MSTRG.16386.1 overexpression or an miR-466b-5p inhibitor. Our findings provide novel insights into ceRNA regulation of the MSTRG.16386.1/miR-466b-5p/Spry2 axis in DHCA-induced kidney injury by the induction of apoptosis.

Brucella is a significant pathogen in the livestock industry, causing Brucellosis, a zoonotic disease that leads to considerable health and economic losses in both humans and animals. Current diagnostic methods for Brucella, including culture, serological assays, and PCR/qPCR, are valuable tools but have inherent limitations. These include the requirement for BSL-3 laboratories, trained personnel, complex procedures, expensive equipment, issues with sensitivity and specificity, and the time-consuming nature of assays, making them unsuitable for large-scale epidemiological screening. Therefore, there is a critical need to develop a rapid, portable, and cost-effective diagnostic method with high specificity and sensitivity. In this study, we established a rapid, portable, reliable, and inexpensive detection method for Brucella genus identification based on RPA-CRISPR/Cas12a technology. Specific RPA primers and crRNA sequences were designed targeting the bcsp31 gene of Brucella. Subsequently, both a fluorescence assay and a lateral flow strip (LFS) assay were developed after optimizing the conditions using the RPA-CRISPR/Cas12a system. The limit of detection (LoD) was 1 copy/μL for RPA-CRISPR/Cas12a-F and 10 copies/μL for RPA-CRISPR/Cas12a-LFS and the entire assay was completed in less than 30 min. This method demonstrated excellent specificity in distinguishing Brucella from other closely related pathogens. Moreover, the RPA-CRISPR/Cas12a assay showed high concordance with classical quantitative real-time PCR when testing diverse clinical samples (blood, serum, milk, semen, vaginal secretions). Together, these findings make this method a promising tool for Brucella detection, with potential applications in both field surveillance and clinical diagnostics.

Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder characterized by the development of fluid-filled cysts in the kidneys. Tolvaptan is the only FDA-approved drug to treat ADPKD, which has significant side effects, prompting the need for safer novel treatments. Given the pathophysiological similarities between ADPKD and malignant tumors, repurposing anti-cancer compounds signifies a promising strategy. In this study, we explored the therapeutic potential of a novel spirocyclic compound 4l with anti-cancer effects in ADPKD. Our preliminary results demonstrated that 4l reduced cell viability in a dose-dependent manner and inhibited cyst growth in 3D culture. Transcriptomic analysis indicated hypoxia-related pathways, mitophagy, and necroptosis (Ripk1 and Mlkl activation) as key mechanisms, alongside cell cycle arrest via Cdkn1a-Rb1 pathway and inhibition of fibrotic markers (Tgfb2 and Col4a6). To strengthen the translational relevance, we established an ADPKD iPSC-derived renal epithelial model, confirming successful differentiation into proximal tubule/collecting duct-like cells (LTL/DBA staining and upregulation of renal markers AQP1, LRP2, and GATA3). Differentiated ADPKD cells exhibited hyperproliferation, reflecting the cystic epithelium observed in vivo. 4l treatment significantly reduced cell viability without immediate cytotoxicity, suggesting a cytostatic effect. Our findings emphasize 4l as a multi-target agent, proficient in reducing cystogenesis through mitophagy induction, modulation of necroptosis, cell cycle inhibition, and suppression of fibrosis. 4l's efficiency across 2D, 3D, and iPSC-derived models highlights its therapeutic potential. This study also recognizes mitophagy and necroptosis as novel targets in ADPKD and corroborates iPSC-derived renal cells as a powerful platform for drug screening.

Skin-derived precursors (SKPs) isolated from the dermis are poorly immunogenic and capable of modulating the allogeneic immune responses. Previous studies implied mouse SKPs-derived exosomes (mSKPs-exo) might possess immunomodulatory functions. In the present study we collected mSKPs-exo using ultracentrifugation method and characterized them. We then explored mSKPs-exo's impact on acne inflammation with Cutibacterium acnes (C. acnes)-challenged murine monocyte-macrophages RAW264.7 and Sprague Dawley (SD) rats auricular acne model. The in vitro studies demonstrated mSKPs-exo reduced nitric oxide, TNF-α, and IL-6 concentration in C. acnes-challenged RAW264.7 cells supernatant. mSKPs-exo also ameliorated NF-κB p65 nuclear translocation and CD86 and iNOS antigens expression in C. acnes-challenged RAW264.7 cells. The RT-PCR and Western blots revealed mSKPs-exo alleviated inflammation by regulating key genes (TLR2, TNF-α, MyD88, and IL-6) and proteins (TLR2, MyD88, IκBa, and NF-κBp65) in the TLR2/MyD88/NF-κB pathway. The anti-inflammatory effects could be synergistically enhanced by IκB/IKK inhibitor. The in vivo studies validated mSKPs-exo's efficacy on acne inflammation and TLR2/MyD88/NF-κB pathway. Topical dermal injection of mSKPs-exo alleviated auricular inflammatory manifestations in SD rats. Immunohistochemistry and Western blot results indicated mSKPs-exo reduced the expression of TLR2, MyD88, MMP3, p-p38 MAPK, and NF-κB p65. The effect was comparable with topical adapalene gel application. The current study initiated mSKPs-exo research and provided theoretical support and experimental data for their potential application in inflammation regulation, acne treatment, and novel therapeutic targets identification.

Gastric cancer (GC) has a high incidence in China. There is a closed-loop structure in circRNAs, which is involved in various cellular biological processes such as tumor development. However, there is a lack of research on the function of circRNAs in GC. In this study, we aimed to explore the potential of circXPO1 as a diagnostic biomarker and the role of circXPO1 in the progression of GC. We screened out circXPO1 through circRNA sequencing. Using exonuclease digestion assay, agarose gel electrophoresis (AGE), Sanger sequencing, and gDNA experiment, we proved that circXPO1 contained a cyclic structure. Quantitative real-time fluorescent Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of circXPO1 in plasma and GC tissues. The receiver operating characteristic curve (ROC curve) was established to evaluate the diagnostic efficacy of circXPO1. The role of circXPO1 was assessed in vitro. The binding sites between circXPO1 and miRNAs were predicted by CircBank, Circinteractome, CircAtlas and miRanda databases. CircXPO1 was up-regulated in 67 GC tissues compared with the adjacent normal tissues (p = 0.0002). It was stable and hard to be degraded, which made it an ideal tumor biomarker. Compared with the patients in the normal control group, the expression level of circXPO1 in plasma was higher in GC patients (p < 0.001) and those with benign lesions (p = 0.0031) with statistically significant differences. CircXPO1 was proved to have satisfactory diagnostic efficacy in distinguishing GC patients from healthy donors (AUC = 0.813, 95% CI: 0.749–0.877). Besides, the diagnostic efficacy, sensitivity, and specificity could achieve 0.853, 78% and 86%, respectively, when circXPO1, CEA and CA199 were used together in diagnosis. In addition, in vitro experiments indicated that circXPO1 knockdown significantly weakened the proliferation, invasion and migration of GC cells. It was also predicted that circXPO1 could serve as a sponge of miR-1248 to regulate the progression of GC.

Thalidomide, a glutamate derivative with teratogenicity, possesses anti-inflammatory, immunomodulatory, and anti-angiogenic properties that enable its use in treating refractory diseases unresponsive to conventional therapies. Dry age-related macular degeneration (AMD), characterized by retinal pigment epithelium (RPE) degeneration and lacking effective therapies, represents a significant unmet medical need. Our findings demonstrated that thalidomide significantly restores mitochondrial function, alleviates G2/M phase cell cycle arrest, and suppresses sustained endoplasmic reticulum (ER) stress in oxidatively injured RPE cells. Mechanistically, these effects are coordinated through E2F2 activation, which subsequently regulates FBXO5 expression. Moreover, thalidomide was able to ameliorate oxidative stress-induced retinal structural disorders and RPE degeneration, and improve visual function in mice. In summary, this study elucidates that thalidomide synergistically regulates cell cycle progression and endoplasmic reticulum homeostasis through the E2F2-FBXO5 signaling pathway, providing a new drug candidate and therapeutic target for the prevention and treatment of dry AMD.

Ischemia–reperfusion injury (IRI) is one of the pathogenic factors of acute kidney injury (AKI). Anostosis, including apoptosis, proptosis, necroptosis, along with ferroptosis and autophagy, is involved in IRI. Based on bioinformatics analysis, we aimed to explore the effect of Ubiquitin Conjugating Enzyme E2 C/Pituitary Tumor-Transforming Gene 1 on renal IRI in the present study. The IRI model in vitro and in vivo was established; correlation of UBE2C with PTTG1, including ubiquitination through down and over-expression of UBE2C/PTTG1 construction in IRI-induced AKI was investigated. Our results indicated that IRI-induced increasing UBE2C/PTTG1 expression at 3 days reperfusion and decreased gradually at 7 days to 14 days after ischemia. Down expression of UBE2C/PTTG1 promoted apoptosis and ferroptosis while suppressing autophagy in the IRI-induced kidney. The effect of UBE2C may be reversed by intervention on PTTG1 expression, and UBE2C regulated PTTG1 expression by affecting its ubiquitination. Our results suggested that UBE2C might promote PTTG1 expression by enhancing its stability through decreasing ubiquitination to inhibit Panopolis and ferroptosis while facilitating autophagy to protect the kidney from IR-induced AKI.