The Association Between Annexin A5 c.-302G > T Variant and the Risk of Recurrent Pregnancy Loss: Systematic Review and Meta-analysisZhou, Zhu, Liu
et alReprod Sci (2025)
Abstract: This study aimed to conduct a comprehensive meta-analysis of the relationship between Annexin A5 (ANXA5) c.-302G > T variant and susceptibility to recurrent pregnancy loss (RPL). We conducted a comprehensive systematic search of the literature published before June 17, 2023 using PubMed, Embase, Web of Science, and Cochrane Library. The Newcastle-Ottawa Scale (NOS) was used to evaluate the quality of the included studies. The odds ratio was used to estimate the correlation of the ANXA5 c.-302 T > G genetic alteration with RPL susceptibility. The I squared (I2) statistic and Q statistic were used to assess the heterogeneity among the included studies. And Begg's test and Egger's regression were then conducted to evaluate publication bias. In this meta-analysis, we included seven studies comprising 1535 RPL cases and 1328 healthy pregnant women to investigate the relationship between ANXA5 c.-302G > T variants and the susceptibility of RPL. No significant publication bias was detected across different comparison models. In the overall population analysis, the ANXA5 c.-302G > T variant was positively associated with the risk of RPL under the dominant, allelic, heterozygote and homozygous models. Meta-regression analysis was performed to identify the potential source of heterogeneity, revealing that neither publication year nor country contributed to heterogeneity. Our findings provided robust evidence that the ANXA5 c.-302G > T variant was significantly associated with an increased risk of RPL, highlighting its potential as a genetic marker for identifying women at high risk of developing RPL. These results emphasized the importance of genetic screening in improving the understanding and clinical management of RPL.© 2025. The Author(s), under exclusive licence to Society for Reproductive Investigation.
Structural insights into the fusion of annexin A5 and fluorescent proteins generating hundredfold differentiated binding affinities to phosphatidylserineGao, Tang, Wang
et alProtein Sci (2025) 34 (4), e70086
Abstract: Fluorescent proteins (FPs) are an indispensable part of modern biology. Numerous studies utilize FPs for protein labeling and cell tracking purposes. They are commonly fused with proteins to aid in their visualization. It is generally assumed that these FP tags have minimal impact on the properties of the fusion proteins. Do the FP types affect the function and characteristics of target proteins on earth? So far, there is no definite answer. Fluorescent annexin A5 (AnxA5) has been extensively employed as apoptosis probes. However, except for chemically labeled AnxA5, there are few developed FP-based AnxA5 probes. Therefore, it is essential to screen out suitable FPs for developing high-affinity AnxA5 probes. Here, various fusion proteins (AnxA5-FPs) were developed. The fusion of AnxA5 did not change the chromophore environments of FPs, while the fusion of FPs led to over a 100-fold difference in AnxA5's affinity for phosphatidylserine (PS). We found that polymeric AnxA5-FPs had higher PS-affinity. Remarkably, although the structures of FPs were similar, they fused with AnxA5 in different modes, generating fusion proteins with different spatial conformations. The difference in conformation resulted in variations in the PS-binding pattern of AnxA5, leading to differing levels of PS-affinity. More importantly, we found five high-affinity (Kd > 10-7 M) FP-based AnxA5 probes with different excitation wavelengths. Together, these observations suggested that differences in the fusion modes of AnxA5 and FPs provided a robust mechanism for modulating PS-affinity of AnxA5. We anticipate that our findings can provide a guideline to develop highly sensitive AnxA5 probes.© 2025 The Protein Society.
LAMP2-FLOT2 interaction enhances autophagosome-lysosome fusion to protect the septic heart in response to ILC2Shao, Liu, Feng
et alAutophagy (2025)
Abstract: Cardiac dysfunction is a serious complication of sepsis-induced multiorgan failure in intensive care units and is characterized by an uncontrolled immune response to overwhelming infection. Type 2 innate lymphoid cells (ILC2s), as a part of the innate immune system, play a crucial role in the inflammatory process of heterogeneous cardiac disorders. However, the role of ILC2 in regulating sepsis-induced cardiac dysfunction and its underlying mechanism remain unknown. The present study demonstrated that autophagic flux blockage exacerbated inflammatory response and cardiac dysfunction, which was associated with mortality of sepsis. Using a cecal ligation and puncture (CLP) mouse sepsis model, we observed an expansion of ILC2s in the septic heart. Furthermore, IL4 derived from ILC2 mitigated cardiac inflammatory responses and improved cardiac function during sepsis. Additionally, IL4 enhanced LAMP2 (lysosomal associated membrane protein 2) expression through STAT3 (signal transducer and activator of transcription 3) activation to stabilize lysosomal homeostasis and rescue the impaired autophagic flux during sepsis. Notably, LAMP2 was preferentially bound to FLOT2 (flotillin 2) after IL4 exposure, and the interaction enhanced autophagosome-lysosome fusion in cardiac endothelial cells. Loss of FLOT2 reversed the regulatory effects of LAMP2 on autophagy mediated by IL4, leading to autophagosome accumulation and suppressed autophagosome clearance. Conclusively, these findings provide novel insights that ILC2 regulates incomplete autophagic flux to protect septic heart and expand our understanding of immunoregulation for sepsis.Abbreviation: ACTB: actin beta; ACTN: actinin, alpha; ADGRE1/F4/80: adhesion G protein-coupled receptor E1; ANXA5/annexin V: annexin A5; AO: acridine orange; BECN1/Beclin1: beclin 1, autophagy related; CKM: creatine kinase, muscle; CKB: creatine kinase, brain; CLP: cecal ligation and puncture; CO: cardiac output; CQ: chloroquine; CTS: cathepsin; DAPI: 4'6-diamidino-2-phenylindole; EC: endothelial cell; EF: ejection fraction; ELISA: enzyme-linked immunosorbent assay; FLOT: flotillin; FS: fractional shortening; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GATA3: GATA binding protein 3; GLB1/β-Gal: galactosidase, beta 1; HCMEC: human cardiac microvascular endothelial cell; IL: interleukin; ILC: innate lymphoid cell; IL1RL1/ST2: interleukin 1 receptor-like 1; IL4c: IL4 complex; IL7R/CD127: interleukin 7 receptor; KEGG: Kyoto Encyclopedia of Genes and Genomes; LAMP: lysosomal-associated membrane protein; LDH: lactate dehydrogenase; LMP: lysosome membrane permeabilization; LPS: lipopolysaccharide; LVEDd: left ventricular end-diastole diameter; LVEDV: left ventricular end-diastole volume; LVESd: left ventricular end-systolic diameter; LVESV: left ventricular end-systole volume; MAN: mannosidase alpha; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MS: mass spectrometry; PECAM1/CD31: platelet/endothelial cell adhesion molecule 1; PTPRC/CD45: protein tyrosine phosphatase receptor type C; RORC/RORγt: RAR related orphan receptor gamma; SQSTM1/p62: sequestosome 1; TBX21/T-bet: T-box 21; TEM: transmission electron microscopy; THY1/CD90.2: thymus cell antigen 1, theta; TNF/TNF-α: tumor necrosis factor; V-ATPase: vacuolar-type H+-translocating ATPase; VIM: vimentin.
Long circulating XTEN864-HGV-Apoptin fusion protein for selective cancer therapyYang, Haeckel, Beindorff
et alInt J Biol Macromol (2025) 306 (Pt 4), 141679
Abstract: The virus protein CAV-Apoptin and its homologue HGV-Apoptin selectively kill cancer cells but are not suitable for systemic treatment. The aim was to develop Apoptin-based fusion proteins for intravenous application in cancer therapy, which also contain the hydrophilic polypeptide XTEN, a cleavage site for MMP-2/9, and a TAT peptide for cell penetration. Expression of XTEN864-HGV-Apoptin in E. coli and purification using XTEN as a tag yielded 100 mg protein/L tissue culture. The expression of XTEN864-CAV-Apoptin did not generate a sufficient yield. Cytotoxic effects were assessed using MTT and Annexin A5 assays, whereas cellular uptake was visualized using Cy3.5-XTEN864-HGV-Apoptin. Blood half-life and biodistribution were evaluated with 99mTc-XTEN864-HGV-Apoptin using SPECT-CT and gamma counting. The fusion protein significantly reduced cancer cell growth and induced apoptosis with minimal effects on non-cancerous cells. It accumulates in the nucleus and associates with F-actin. In mice, the protein showed a blood half-life of 0.68 h (fast phase) and 17 h (slow phase), with a tumor/muscle ratio of 9.36 ± 6.22 (SD). In a 4T1 mouse tumor model, it effectively inhibited tumor growth. The cancer-specific cytotoxicity and prolonged circulation of XTEN864-HGV-Apoptin suggest its potential for systemically applicable, biodegradable, and E. coli-producible antitumor drugs.Copyright © 2025 The Authors. Published by Elsevier B.V. All rights reserved.
膜杰作
Star Staining
