Protein & Cell最新文献

Epithelial ovarian cancer (EOC) is an aggressive malignancy with limited therapeutic options. Poly(ADP-ribose) polymerase inhibitors (PARPi) have shown remarkable efficacy, especially in BRCA-mutant patients, and are approved as maintenance therapy to prevent recurrence after initial response to chemotherapy. However, the development of PARPi resistance poses a major clinical challenge. This study utilized a whole-genome CRISPR-Cas9 genetic screening to identify genes associated with PARPi sensitivity upon knockout. Based on the screening and validated through further experiments, we confirmed that CLK1 knockdown is synthetically lethal with PARPi in ovarian cancer. The combination of the PARPi Olaparib and CLK1 inhibitor TG003 exhibited potent anti-proliferative effects both in vitro and in vivo. Mechanistically, CLK1 inhibition downregulated the functional ERCC1-202 isoform, resulting in enhanced DNA damage and apoptosis. Our findings reveal a novel mechanism underlying PARPi sensitivity and suggest that targeting CLK1 in combination with PARPi may represent a promising therapeutic strategy for PARPi-resistant ovarian cancer.

Human tumor organoids represent a paradigm shift in cancer modeling, overcoming critical limitations of conventional systems by faithfully recapitulating genetic heterogeneity, three-dimensional architecture, and tumor microenvironment dynamics of patient tumors. Our review explores how human tumor organoids serve as a transformative preclinical platform, bridging the gap between basic research and clinical translations. We highlight recent advances in tumor organoid generation, spanning patient-derived organoids to genetically engineered platforms from normal tissue and human pluripotent stem cells, and their applications in deciphering carcinogenesis, clonal evolution, and metastatic mechanisms. We further examine technological innovations in culture systems that enhance the interpretability and translatability of tumor phenotypes and drug responses. We present an in-depth exploration of how integrated tumor microenvironment co-culture systems-combining immune cells, cancer-associated fibroblasts, and vascular components-enable novel investigations into tumor-stroma-immune crosstalk. Clinically, human tumor organoid biobanks have shown great promise in predicting personalized therapy responses. Emerging technologies like organoids-on-chip platforms, three-dimensionally bioprinting and artificial intelligence-driven analytics are enhancing high-throughput drug screening efficiency and biomarker identification. Despite advances, complete microenvironmental modeling remains challenging, particularly in replicating vascular complexity and systemic immune responses. Future advancements will demand convergence of synthetic biology, functional genomics, and machine learning to transform human tumor organoids from static avatars into dynamic "living biosensors". In summary, this review provides an in-depth exploration of the organoid field and presents a clear and actionable framework for positioning tumor organoids as indispensable tools in functional precision medicine-a strategy that ultimately bridges mechanistic discoveries with clinical translation.

Nipah virus (NiV) poses a significant public health threat due to its high mortality rate and the absence of approved treatments. Nonetheless, the host-virus interactions underlying its pathogenesis remain poorly understood. Here, we identified the 5-methylcytosine (m5C) methyltransferase NSUN2 as a critical host factor hijacked by NiV to facilitate replication via dual mechanisms. The viral matrix (M) protein stabilizes NSUN2 by inhibiting its proteasomal degradation. In turn, NSUN2 catalyzes m5C deposition on NiV RNAs, enhancing M RNA stability and protein expression. Simultaneously, NSUN2's noncatalytic domain engages GNB2 as an adaptor to facilitate the recruitment of the E3 ubiquitin ligase TRIM28 to M, promoting M ubiquitination and consequent nuclear export for virion assembly. Targeting both pathways using the proteasome inhibitor carfilzomib and the m5C inhibitor MY-1B suppressed NiV replication in vitro and in hamsters. Our findings uncover a dual epigenetic-posttranslational regulatory axis exploited by NiV and present a promising combinatorial therapeutic approach.

Genome-wide off-target effect poses a safety risk for clinical use of adenine base editor (ABEs), among which ABE8e is one of the most efficient. Two-cell embryo injection (GOTI) analysis showed that the rate of genome-wide single nucleotide variants (SNVs) in ABE8e-edited cells was ∼30-fold higher than that of spontaneous SNVs in control cells, indicating prevalent off-target effects of ABE8e, but no off-target effect for ABE7.10, from which ABE8e was derived. We performed saturation mutagenesis of eight amino acid sites of the deaminase (TadA8e) within ABE8e and obtained ABE8eY149V that exhibited high editing efficiency without detectable off-target effect. Furthermore, TadA8eY149V could be fused with other Cas homologs (PAM-relaxed SpRY, hypercompact SaKKH, or IscB) to expand its target range. Finally, ABE8eY149V editing of hydroxyphenylpyruvate dioxygenase (Hpd) gene prevented lethality in hereditary tyrosinemia type I mice. The high efficiency and fidelity of ABE8eY149V suggest its potential application in ABE-based gene therapies.

Esophageal squamous cell carcinoma (ESCC) remains a major health burden, particularly in Asia, with poor patient prognosis despite advancements in radiotherapy, chemotherapy, and immunotherapy. The marked inter-patient and intra-tumor heterogeneity of ESCC underscores the need for molecularly informed diagnostic and therapeutic strategies. Recent high-throughput omics technologies, including genomics, transcriptomics, proteomics, and metabolomics, have substantially advanced our understanding of ESCC biology. Genomic profiling has revealed recurrent alterations such as TP53 and NOTCH1 mutations, as well as actionable targets including PIK3CA, FGFR1, and SOX2 amplifications, which provide new opportunities for precision therapy. Epigenomic and transcriptomic analyses have identified methylation-based early detection markers (e.g., PAX9, SIM2) and immune-related transcriptomic subtypes associated with prognosis and immunotherapy responsiveness. Proteomic and metabolomic studies have further uncovered cell cycle and spliceosome pathway activation and altered lactate metabolism, offering additional biomarker and therapeutic insights. In this review, we synthesize these multi-omics advances and highlight how they collectively inform improved diagnostic, prognostic, and therapeutic strategies for ESCC. Despite these developments, the clinical translation of multi-omics findings remains limited due to the lack of standardized analytical pipelines, insufficient multi-center validation, and the high cost and technical complexity of integrating multi-omics data into routine clinical workflows. Future research integrating artificial intelligence with multi-omics data holds promise for enhancing diagnostic accuracy and enabling more precise therapeutic decision-making in ESCC.

Mouse extended pluripotent stem (EPS) cells have demonstrated significant potential for generating embryo models in vitro. However, their limited capacity for extraembryonic trophoblast development has hindered their use in constructing whole embryo models, particularly post-implantation embryoids. Here, we establish a stepwise induction protocol to generate trophectoderm-like cells from mouse EPS cells. These cells retain trophectoderm-specific transcriptomic features and can differentiate into trophoblast lineages in vivo. Moreover, combining these trophectoderm-like cells with EPS cell-derived primitive endoderm/epiblast bilineage structures enabled the robust generation of post-implantation embryoids in a transgene-free manner. EPS-derived embryoids recapitulate key developmental events of post-implantation mouse embryos, including the formation of the pro-amniotic cavity, anterior-posterior axis, primitive streak, gastrulation, and complex extraembryonic tissues. Notably, single-cell transcriptomic analysis revealed a high degree of transcriptional similarity between EPS-derived embryoids at day 6 and natural E7.5 mouse embryos. Our study presents a novel platform for modeling post-implantation mouse embryogenesis in vitro.