The KRASG12D mutation, which occurs in nearly half of pancreatic ductal adenocarcinoma (PDAC) and colorectal cancer (CRC) cases, has demonstrated considerable promise as a target for cancer therapy. In this study, we report a series of VHL-recruiting proteolysis targeting chimeras (PROTACs) targeting KRASG12D based on the analogues of MRTX1133. Through extensive structure–degradation relationship (SDR) studies involving the optimization of linker parameters, E3-binding ligand, and KRASG12D warhead, ZJK-366 is obtained as a representative compound that can selectively, effectively, and sustainably degrade KRASG12D in a dose-, time- and ubiquitin–proteasome system-dependent manner. ZJK-366 also inhibits the growth of KRASG12D mutant cell lines. Moreover, ZJK-366 is capable of overcoming resistance induced by secondary mutations. ZJK-366 shows attractive pharmacokinetic properties and effectively and safely inhibits tumor growth in the cell line-derived xenograft model in a dose-dependent manner. These results underscore the potential of PROTAC-mediated KRASG12D degradation as a strategy for treating KRAS mutation-driven cancers.
Stable cnidarian–dinoflagellate symbioses provide the trophic foundation of coral reef ecosystems. Understanding nutrient exchange underpinning this symbiosis, and the need for interventions to improve coral health via aquaculture, grow increasingly urgent as reefs face accelerating threats from climate change. Liposomes and lipid nanoparticles, widely used as delivery vehicles in biomedical science, are emerging as promising tools to supplement coral nutrition. However, physiological impacts of liposomes on cnidarians, including their uptake, nutritional value, and holobiont response, remain largely unexplored. Here, we delivered “empty” phosphatidylcholine liposomes to both symbiotic and aposymbiotic Exaiptasia diaphana, an anemone model for corals, and analyzed the host proteomic response via mass spectrometry. Liposome supplementation elicited broad proteome shifts, with notable overlap between symbiosis- and liposome-induced protein expression. Proteins involved in lipid catabolism, lipid transport, β-oxidation, lysosomal function, and protein translation were significantly more abundant, consistent with enhanced lipid processing and metabolic activity. Liposome supplementation, like symbiosis, suppressed both asexual reproduction and the expression of a suite of predation- and digestion-associated venom proteins and proteases, suggesting a conserved “sated” phenotype in response to lipid supply. Variations in feeding frequency with Artemia had minimal effect on the proteome. These data suggest that adult Exaiptasia may be capable of direct uptake of liposomes, and we propose a model offering a potential tool for probing lipid metabolism, signaling, and symbiotic function in cnidarians. Moreover, the ability to manipulate host physiology using defined lipid formulations holds potential for advancing coral aquaculture and stress resilience, including for reef restoration strategies.
Limited carbohydrate utilization is a common metabolic challenge in carnivorous fish, including largemouth bass (Micropterus salmoides). This study integrated in vivo feeding trials, in vitro primary hepatocyte experiments, and comparative genomic analyses to investigate the mechanisms underlying limited carbohydrate utilization in this species. Prolonged high-carbohydrate feeding decreased serum insulin levels and increased hepatic glycogen levels, accompanied by alterations in hepatic insulin signaling and glucose metabolism. A high-carbohydrate diet suppressed hepatic insulin signaling, including components of the IR/PI3K/AKT pathway, downregulated glycolytic genes (gk and pk), and differentially regulated gluconeogenic genes (pepck, fbp1, and g6pc), indicating limited glucose utilization and altered regulation of gluconeogenesis. Comparative genomic analysis across 18 teleost species revealed that key insulin signaling genes (ins, ir, irs, pi3k, and akt) were highly conserved in largemouth bass, although ins exhibited greater evolutionary divergence, suggesting lineage-specific diversification. In in vitro, insulin supplementation partially ameliorated high-glucose-induced alterations by upregulating irs and pik3r1 expressions and restoring akt1 expression, indicating that insulin pathway responsiveness was retained. Western blot further confirmed insulin-mediated modulation of INSR, PI3K, and AKT1 phosphorylation. Meanwhile, the selective recovery of glycolytic and gluconeogenic gene expression indicated pathway-specific sensitivity to insulin. These findings indicate that largemouth bass exhibit reduced serum insulin levels and widespread suppression of hepatic insulin signaling under high-carbohydrate conditions, whereas exogenous insulin partially restores the associated metabolic alterations.
Cryptocaryon irritans is a devastating marine parasite, necessitating the development of effective control strategies. While copper–zinc alloy (CZA) exhibits potent antiparasitic activity, its underlying mechanism of action remains unclear. Here, we demonstrate that CZA eliminates C. irritans primarily by inducing ferredoxin reductase 1 (fdx1)-dependent cuproptosis. Pharmacological inhibition assays revealed that blocking cuproptosis significantly rescued parasite viability (survival rate: 90
As dominant components of microzooplankton, tintinnid ciliates act as a vital link between the microbial food web and the traditional planktonic food chain in marine ecosystems. However, polymorphism of the lorica and the lack of ciliature information and gene sequence data have significantly complicated taxonomic analysis and restricted the development of tintinnid-related research fields such as ecology and physiology. In the present study, the morphological and molecular characterization of three poorly described species—Tintinnopsis estuariensis, T. angusta, and Leprotintinnus nordqvisti—and one novel species, Metacylis parajorgensenii sp. nov., were revealed, and the evolutionary relationships of tintinnids were discussed based on all currently known information on lorica morphology, ciliary pattern, and molecular phylogeny. The ciliary patterns of this group of ciliates were grouped into eight types—VO-type, S-type, A-type, N-type, E-type, F-type, M-type, and C-type—from which an evolutionary hypothesis was proposed. In combination with the SSU rRNA gene phylogeny tree, six types (corresponding gene sequence data were unavailable for species of VO-type and N-type), from simple to complex, were found to be consistent with the evolutionary direction of the order Tintinnida. However, the lorica type did not agree with the topology, suggesting that the ciliary pattern can be more reliable than the lorica morphology for classifying tintinnids. The interspecific comparison of known ciliary patterns demonstrated that the numbers of somatic kineties and kinetids could distinguish these species well. In addition, the population distributions of the genera Metacylis, Tintinnopsis, and Leprotintinnus, based on reliable data, revealed large differences in lorica opening diameters between inshore species, suggesting niche differentiation caused by competition for food resources.
