Ligand-modified rAAV6 vectors with nanoblades allow high-level gene knockin in HSPCs without compromising cell survivalGutierrez-Guerrero, Périan, Leray
et alMol Ther Nucleic Acids (2025) 36 (2), 102495
Abstract: Nanoblades are viral particles loaded with the Cas9 protein complexed with gRNA, which allowed efficient gene editing in hematopoietic stem and progenitor cells (HSPCs). Combined with recombinant adeno-associated vector (rAAV) 6 containing two homologous arms to a gene locus resulted in 50% of expression cassette knockin into HSPCs. However, high effective doses of rAAV6 induced HSPC cell death. Here, we demonstrated that, at high doses, rAAV2 was much less toxic for template DNA delivery and allowed transduction levels in HSPCs equivalent to rAAV6. To improve donor template delivery, rAAV2 and rAAV6 were chemically bio-conjugated with a mannose ligand, via the lysine or tyrosine amino acid residues exposed at the adeno-associated vector (AAV) capsid surface. High-level transduction of HSPCs with mannose-coupled rAAV6 vectors accompanied by a remarkable lower toxicity was achieved as compared to control rAAV6 in correlation with highly reduced p53 pathway activation. Mannose-conjugated rAAV6 combined with nanoblades allowed efficient gene knockin and increased survival of HSPCs from 10% to 80% as compared to the unmodified rAAV6 even in the most immature CD34+CD38lowCD90+ hematopoietic stem cell (HSC) population. Summarizing, mannose-conjugated rAAV6 maintained high-level donor mediated gene knockin when combined with nanoblades without inducing significant toxicity for the HSPCs, an important feature for clinical translation of HSPC gene-editing strategies.© 2025 The Author(s).
Temperature modulates 17β-estradiol regulation of oogenesis protein expression in the liver of the eurythermal pupfish Cyprinodon nevadensisLema, Krayeva, Dale
et alGen Comp Endocrinol (2025) 365, 114707
Abstract: Female fish experiencing atypically high or prolonged elevations in temperature during oogenesis can suffer impaired oocyte development with fewer or smaller eggs, eggs with reduced yolk content or thinner envelopes, and lower egg viability. These changes in oocyte quality and quantity are in part caused by diminished liver synthesis of egg yolk (vitellogenin, Vtg) and egg envelope (choriogenin) proteins at anomalously high temperatures. Those declines in liver Vtg and choriogenin production are commonly paralleled by reduced blood concentrations of 17β-estradiol (E2). However, it is unclear whether declines in liver vitellogenin and choriogenin production at elevated temperatures result solely from lower circulating E2 or if other aspects of E2 signaling are also altered to diminish liver synthesis of oogenesis proteins. In this study, adult female Amargosa River pupfish (Cyprinodon nevadensis amargosae), a species with asynchronous follicular development, were maintained at 20 °C, 28 °C, or 36 °C and then administered E2 or vehicle solution. Ovarian gonadosomatic index (GSI) values and plasma E2 were lower in females at 36 °C compared to those at cooler temperatures. Females at 36 °C also had reduced plasma Vtg protein, lower liver abundances for mRNAs encoding vitellogenin genes (vtgAa, vtgAb, vtgc), choriogenin genes (cgh, cghm, cgl), and estrogen receptor α (esr1). Supplemental E2 increased plasma E2 in females at all temperatures, but only upregulated liver vitellogenin and choriogenin mRNAs at 36 °C, despite E2 upregulation of hepatic esr1 receptor transcripts at all temperatures. Females at 36 °C also exhibited higher liver mRNA abundances for sex hormone-binding globulin (shbg) and cytochrome P450 family 1 subfamily A member 1 (cyp1a1), an estrogen-metabolizing monooxygenase enzyme that converts E2 to 2-hydroxyestradiol. Together, these findings indicate elevated temperatures diminish E2 stimulation of liver Vtg and choriogenin expression in pupfish via effects on several aspects of E2 signaling including circulating E2 concentrations and liver esr1 expression as well as shbg and cyp1a1 expression, which may result in changes to free:bound E2 and the rate of hepatic E2 inactivation. These results also demonstrate that E2 replacement can help compensate for high temperature-induced declines in hepatic oogenesis gene expression in female pupfish.Copyright © 2025 The Author(s). Published by Elsevier Inc. All rights reserved.
The Alpha-coronavirus E protein inhibits the JAK-STAT pathway signaling by triggering STAT2 degradation through OPTN- and NBR1-mediated selective autophagyHuang, Gao, Huang
et alAutophagy (2025)
Abstract: The zoonotic transmission of coronaviruses continues to pose a considerable threat to humans. Swine acute diarrhea syndrome coronavirus (SADS-CoV), a bat coronavirus related to HKU2, causes severe economic losses in the pig industry and has the potential to trigger outbreaks in humans. However, our understanding of how SADS-CoV evades the host's innate immunity remains limited, hindering effective responses to potential human outbreaks. In this study, we demonstrate that the SADS-CoV envelope protein (E) inhibits type I interferon (IFN-I) signaling by inducing the degradation of STAT2 via the macroautophagy/autophagy-lysosome pathway. Mechanistically, the E protein evades host innate immunity by promoting STAT2 degradation through autophagy, mediated by the NBR1 and OPTN receptors. Notably, ubiquitination of E protein is required for the autophagic degradation of STAT2. Additionally, lysine residue K61 of the E protein is crucial for its stable expression; however, it is not involved in its ubiquitination. In conclusion, our study reveals a novel mechanism by which the E protein disrupts IFN-I signaling by targeting STAT2 via autophagy, enhancing our understanding of SADS-CoV's immune evasion strategies and providing potential drug targets for controlling viral infections.
Dengue virus-host interactions: Structural and mechanistic insights for future therapeutic strategiesKhanra, Ghosh, Khatun
et alJ Struct Biol (2025) 217 (2), 108196
Abstract: Dengue pathogen, transmitted by mosquitoes, poses a growing threat as it is capable of inflicting severe illness in humans. Around 40% of the global population is currently affected by the virus, resulting in thousands of fatalities each year. The genetic blueprint of the virus comprises 10 proteins. Three proteins serve as structural components: the capsid (C), the precursor of the membrane protein (PrM/M), and the envelope protein (E). The other proteins serve as non-structural (NS) proteins, consisting of NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5. The virus relies on these NS proteins to expropriate host proteins for its replication. During their intracellular replication, these viruses engage with numerous host components and exploit the cellular machinery for tasks such as entry into various organs, propagation, and transmission. This review explores mainly the relationship between dengue viral protein and host proteins elucidating the development of viral-host interactions. These relationships between the virus and the host give important information on the processes behind viral replication and the etiology of disease, which in turn facilitates the creation of more potent treatment strategies.Copyright © 2025 Elsevier Inc. All rights reserved.
膜杰作
Star Staining
