Controlling stem cell differentiation is a longstanding goal in biomedical research. Here we explore how cell fate is influenced by RNA condensates, specifically P-bodies, which modulate gene expression posttranscriptionally. We profiled the transcriptomes of biomolecular condensates in diverse developmental contexts spanning multiple vertebrate species. Our analyses revealed conserved, cell type-specific sequestration of untranslated RNAs encoding cell fate regulators. P-body RNA contents do not reflect active gene expression in each cell type but are enriched for translationally repressed transcripts characteristic of the preceding developmental stage. Mechanistically, P-body contents are controlled by microRNAs and can be profoundly reshaped by perturbing AGO2 or polyadenylation site usage. Applying these insights to stem cell differentiation, we show that manipulating P-body assembly or microRNA activity can direct naive mouse and human pluripotent stem cells toward totipotency or primed human embryonic cells toward the germ cell lineage. Our findings link cell fate decisions to RNA condensates across vertebrates and provide a means of controlling cell identity. Stem cell differentiation is controlled by manipulating RNA condensates.

Germline-competent embryonic stem (ES) cells have been successfully derived from mice and rats, but not from other species. Here we report the development of culture conditions for deriving ES cells from chickens and seven other avian species. Chicken ES cells express core pluripotency markers and can differentiate into cells of all embryonic germ layers, as well as extra-embryonic lineages. Notably, chicken ES cells contribute to high rates of chimerism when injected into chicken embryos and give rise to germ cells both in vitro and in ovo, confirming their germline competence. In addition, we demonstrated that ES cell self-renewal pathways are conserved among avian species, allowing ES cells from multiple avian species to be established using optimized chicken ES cell culture conditions. The establishment of authentic avian ES cells lays the groundwork for future applications in genetic engineering and the conservation of avian biodiversity. Embryonic stem cells for chicken and seven other avian species are derived.

Glycogen synthase kinase 3 (GSK3) is a crucial regulator of cellular processes, including stem cell maintenance and differentiation. Although the roles of the two GSK3 isozymes, GSK3α and GSK3β, are well documented, their specific interactions remain less understood. In this study, we explored the regulatory interplay between GSK3α and GSK3β in mouse embryonic stem cells (mESCs). Using genetic manipulation, small-molecule inhibitors, and biochemical analysis, we found that inhibition of GSK3α kinase activity increases GSK3β protein levels and activity, whereas overexpression of GSK3α reduces GSK3β protein levels and activity. Domain-swapping experiments between the two isozymes identified the glycine-rich region at the N terminus of GSK3α as the key sequence responsible for downregulating GSK3β protein levels. Our findings reveal a novel interaction between GSK3 isozymes, with GSK3α modulating GSK3β activity to maintain the balance between stem cell pluripotency and neural differentiation. This insight may open new pathways for understanding stem cell fate mechanisms and developing GSK3-targeted therapeutic strategies in regenerative medicine.

BackgroundDoxorubicin (DOX) is a highly effective and widely used cytotoxic agent with application for various malignancies, but it’s clinically limited due to its cardiotoxicity Oxidative stress and inflammation were reported to take part in DOX-induced cardiotoxicity. Tirzepatide, a dual glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) receptor agonist has been approved to treat type 2 diabetes. However, its role in DOX-induced cardiotoxicity and the underlying mechanisms has not been explored.MethodsThe cardioprotective properties of Tirzepatide against DOX-induced cardiotoxicity are examined in this work both in vivo and in vitro. For four weeks, an intraperitoneal injection of 4 mg/kg DOX was used to cause cardiotoxicity in C57BL/6 mice. To ascertain the cardioprotective function and underlying mechanisms of Tirzepatide against DOX-induced cardiotoxicity, mice and H9c2 cells were treated with and without Tirzepatide.ResultsTirzepatide treatment significantly inhibited DOX-induced oxidative stress, inflammation and cardiac injury. Mechanistically, PI3K/Akt signaling pathway contributes to the protective effect of Tirzepatide against DOX-induced cardiotoxicity and inhibited PI3K/Akt signaling pathway with LY294002 almost blocked its therapeutic effect.ConclusionsCollectively, Tirzepatide could alleviate DOX-induced oxidative stress, inflammation and cardiac injury via activating PI3K/Akt signaling pathway and Tirzepatide may be a novel therapeutic target for DOX-induced cardiotoxicity.

Purpose:Neovascular age-related macular degeneration (nAMD) is a prevalent cause of blindness in the elderly. Standard treatment includes anti-vascular endothelial growth factor (anti-VEGF) drugs, such as aflibercept. However, anti-VEGF drugs may have limited efficacy and cause drug resistance. This study explores whether Kavain, an anti-inflammatory molecule from Piper methysticum, can treat choroidal neovascularization (CNV). Methods:Various experiments were conducted to assess the Kavain's toxicity. The impact of Kavain on in vitro cultured endothelial cells was examined through 5-ethynyl-20-deoxyuridine (EdU) assays, transwell migration assays, and tube formation assays. The therapeutic effects of Kavain on CNV were investigated using a laser-induced CNV mice model. To elucidate the mechanism of Kavain, network pharmacology analysis, molecular docking, and western blots were performed. Results:Kavain exhibited no apparent toxicity both in vitro and in vivo. Kavain significantly decreased endothelial cell viability, proliferation, migration, and tube formation ability in a dose-dependent manner compared to the hypoxia groups (P<0.05). Kavain alleviated CNV in the laser-induced CNV mouse model compared to the control groups (P<0.05). These effects were statistically significantly enhanced in the Kavain plus aflibercept groups (P<0.05). Following Kavain administration, the expression levels of various inflammatory factors were markedly reduced in retinal pigment epithelium (RPE)/choroid complexes (P<0.05). Mechanistically, Kavain decreased the activity of the hypoxia-inducible factor 1α (HIF-1α)/VEGF-A/ VEGF receptor 2 (VEGFR2) signaling pathway. Conclusion:Our study is the first to demonstrate Kavain's potential as a promising treatment for nAMD, owing to its dual effects of anti-inflammation and anti-angiogenesis.

BackgroundTo investigate the pharmacovigilance (PV) and make pairwise comparisons on reporting proportion, seriousness, and severity of outcomes of major adverse cardiovascular events (MACE) among poly(ADP-ribose) polymerase-inhibitors (PARPis) in treating ovarian cancer, fallopian tube carcinoma, and primary peritoneal cancer (collectively named EOC) from the US Food and Drug Administration Adverse Event Reporting System (FAERS).Research design and methodsData on adverse cardiovascular events reports related to EOC treatment submitted to FAERS from the first quarter of 2015 to the second quarter of 2023 were harvested. Three PARPis were identified: olaparib, niraparib, and rucaparib.ResultsEventually, a total of 258,596 eligible records were enrolled with 12,331 reports including 5,292 reports of MACE and 7,039 reports of other cardiovascular events. For the primary composite endpoint, a PV signal associated with MACE was detected in niraparib (ROR = 1.12; IC025 = 0.03), whereas it was not detected in olaparib and rucaparib; For the secondary endpoint, PV signals associated with other cardiovascular events were detected in niraparib (ROR = 1.17;IC025 = 0.04), but not in olaparib and rucaparib.ConclusionsFor EOC patients, close monitoring of blood pressure, heart rate, and coagulation function should be conducted when selecting niraparib for treatment.

BACKGROUND:We aimed to demonstrate the regulatory effect of long non-coding RNA (lncRNA) ENAH-202 on oral squamous cell carcinoma (OSCC) development as well as its molecular mechanism.METHODS:We detected ENAH-202 expression in OSCC tissues and cell lines by quantitative real-time PCR (qPCR). The biological function of ENAH-202 was assessed in vitro and in vivo using CCK-8, colony formation assays, transwell assays, xenograft formation, and tail vein injection. The further molecular mechanism by which ENAH-202 promoted OSCC progression was identified using RNA pull-down, LS-MS/MS analysis, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (ChIP) assays.RESULTS:ENAH-202 was significantly upregulated in OSCC tissues and cells. ENAH-202 promoted OSCC cell proliferation, migration, and invasion in vitro and in vivo. The expression of enabled homolog (ENAH) and epithelial-to-mesenchymal transition (EMT)-related proteins was changed with the expression of ENAH-202. Moreover, ENAH-202 promoted the transcription of Vimentin (VIM) by binding with ZNF502, which can help ENAH-202 promote OSCC progression.CONCLUSIONS:ENAH-202 facilitated OSCC cell proliferation and metastasis by regulating ZNF502/VIM axis, which played an important role in OSCC progression.

N7-methylguanosine (m7G) modification is closely related to the occurrence of tumors. However, the m7G modification of circRNAs in oral squamous cell carcinoma (OSCC) remains to be investigated. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) was used to measure the methylation levels of m7G and identify m7G sites in circRNAs in human OSCC and normal tissues. The host genes of differentially methylated and differentially expressed circRNAs were analyzed by Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, and circRNA-miRNA-mRNA networks were predicted using the miRanda and miRDB databases. The analysis identified 2348 m7G peaks in 624 circRNAs in OSCC tissues. In addition, the source of m7G-methylated circRNAs in OSCC was mainly the sense overlap region compared with normal tissues. The most conserved m7G motif in OSCC tissues was CCUGU, whereas the most conserved motif in normal tissues was RCCUG (R = G/A). Importantly, GO enrichment and KEGG pathway analysis showed that the host genes of differentially methylated and differentially expressed circRNAs were involved in many cellular biological functions. Furthermore, the significantly differentially expressed circRNAs were analyzed to predict the circRNA-miRNA-mRNA networks. This study revealed the whole profile of circRNAs of differential m7G methylation in OSCC and suggests that m7G-modified circRNAs may impact the development of OSCC.

Abstract Background Despite surgical and medical efforts in the upfront treatment, about 70% of OC patients have a disease relapse within 2–3 years after diagnosis. ROC is still an incurable condition; targeted agents as maintenance therapies have been shown to improve survival, but the most appropriate maintenance regimens are still controversial and opaque.Objective This Bayesian network meta-regression analysis provides a head-to-head comparison of maintenance therapy PARP-Inhibitors and Angiogenesis Inhibitors for Platinum-Sensitive Recurrent Ovarian Cancer(PSROC) using the BRCA genes status as a covariate.Methods To identify relevant studies, the databases of Pubmed, Scopus, Embase, Web of Science, and the Cochrane Central Register of Controlled Trials were searched until January 31, 2023. Comparing progression-free survival (PFS) and overall survival (OS) of different interventions at the same time node by NMA. NMR was performed on the hazard ratios (HR) associated with PFS and OS as the primary endpoints, with the BRCA genes status as the covariate. The ability of each treatment was ranked using the surface under the cumulative ranking (SUCRA) curve.Results Eventually,16 studies with 5696 patients and 8 maintenance therapy were enrolled.From a horizontal perspective,bevacizumab(HR = 1.83, 95%CI: 1.47 to 2.33), niraparib(HR = 2.9, 95%CI: 2.15 to 3.95), olaparib (HR = 2.74, 95%CI: 2.05 to 3.6) ,rucaparib (HR = 2.61, 95%CI: 1.81 to 3.57) and fuzuloparib (HR = 3.99, 95%CI: 2.41 to 6.6) were significantly superior to the placebo on PFS of AC.Niraparib(HR = 4.08, 95%CI: 1.49 to 11.33), olaparib (HR = 4.29, 95%CI: 1.9 to 9.75) ,rucaparib (HR = 4.16, 95%CI: 1.04 to 16.49) and fuzuloparib(HR = 7.16, 95%CI: 1.63 to 30.82) were significantly superior to the placebo on PFS of BRCAm.Niraparib(HR = 2.33, 95%CI: 1.17 to 4.7) was significantly superior to the placebo on PFS of BRCAwt.From a longitudinal perspective, the results for PFS are similar to the above; Olaparib significantly increased OS rates from the 3rd to the 36th month(HR from4.43 to26.55). Meta-regression analyses support the above conclusions.Conclusions Considering the efficacy, durability, safety, and compliance, one of the standard maintenance therapy, Olaparib, should be recommended as the best choice for PSROC. For BRCAm and BRCAwt patients,Olaparib and niraparib may be the first choice for improving PFS.Furthermore, more head-to-head trials are needed to confirm those findings.

Background and purpose: Oral squamous cell carcinoma (OSCC) is the most common subtype of head and neck squamous cell carcinoma (HNSCC), and its pathogenesis is unclear. Nucleolar protein 8 (NOL8), as one of the RNA-binding protein (RBP), plays a key role in the occurrence and development of many kinds of tumors, however its role in OSCC is not clear. This study aimed to investigate the expression level of NOL8 in OSCC and its effects on the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of OSCC. Methods: The expression of NOL8 in HNSCC was analyzed online by gene expression profiling interactive analysis 2 (GEPIA2), tumor immune estimation resource (TIMER), the University of Alabama at Birmingham cancer data analysis portal (UALCAN) and the encyclopedia of RNA interactomes (ENCORI). The mRNA expression level of NOL8 in OSCC cells was detected by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR). siRNA interference technique was used to knock down the expression of NOL8 in CAL-27 cells to form NOL8 knockdown group (si-NOL8-1, si-NOL8-2) and negative control group. CAL-27 and HN6 cells were overexpressed with NOL8 by lentivirus transfection technique to form NOL8 overexpression group and negative control group. Cell counting kit-8 (CCK-8) assay, scratch healing assay and transwell assay were used to detect the effect of NOL8 expression on the proliferation, migration and invasion of OSCC cells. Western blot assay was used to detect the effect of NOL8 on the expression of EMT-related genes including E-cadherin, vimentin and N-cadherin. The effect of NOL8 on the proliferation of OSCC cells in vivo was examed by xenograft formation assays. Results: The online analysis of GEPIA2, TIMER, UALCAN and ENCORI showed that the expression of NOL8 was higher in HNSCC than in normal tissues, and the expression of NOL8 in OSCC was significantly higher than in normal control cells. The relative expression of NOL8 in CAL-27 cells transfected with si-NOL8-1 and si-NOL8-2 was significantly lower compared with the negative control group. The results of CCK-8 assay, scratch healing assay and transwell assay showed that the proliferative ability, cell migration rate and invasion number of CAL-27 cells were significantly decreased after knockdown of NOL8 expression. The relative expression of NOL8 in CAL-27 and HN6 cells transfected with NOL8 was significantly higher compared with the control group. The proliferative ability, cell migration rate and invasion number of CAL-27 and HN6 cells in the overexpression NOL8 group were significantly higher compared with the negative control group. The results of Western blot showed that in CAL-27 cells, the expression of E-cadherin increased and the expressions of N-cadherin and vimentin decreased after NOL8 knockdown, while in CAL-27 and HN6 cells, the expression of E-cadherin decreased and the expressions of N-cadherin and vimentin increased after NOL8 overexpression. The xenograft formation assays showed that the weight of tumor was significantly higher in NOL8 overexpression group than in NOL8 control group. Conclusion: NOL8 is highly expressed in OSCC and can promote the proliferation, migration and invasion of OSCC, which may be related to the process of EMT.

Background Despite conspicuous advances in innovating novel drugs and combination regimens in multiple myeloma (MM) in recent decades, the most appropriate maintenance regimens after inductive therapy are still controversial and opaque.Objective We aimed to identify the most effective maintenance treatment for newly diagnosed multiple myeloma (NDMM) patients via network meta-analysis.Method We searched PubMed, Embase, Cochrane Library, Scopus, and Google Scholars from inception to April, 2022. Odds ratios (ORs) were generated for dichotomous variants. The primary endpoint was overall survival (OS).Results Eventually a total of 19 trials, including 11 treatments and 8337 patients, were included in this analysis. For OS, lenalidomide (OR ranged from 1.61 to 1.99) and daratumumab (OR ranged from 1.83 to 2.41) showed significant efficacy over placebo. Maintenance therapy comprising lenalidomide-carfilzomib (OR ranged from 3.19 to 6.95), lenalidomide-prednisone (OR ranged from 2.62 to 4.44), bortezomib-thalidomide (OR ranged from 2.48 to 3.64), daratumumab (OR ranged from 2.0 to 2.98), lenalidomide (OR ranged from 1.4 to 3.19), ixazomib (OR ranged from 1.36 to 2.05), thalidomide (OR ranged from 1.5 to 1.86) demonstrated significant effects in prolonging PFS compared with placebo; Among the efficient therapies, lenalidomide-carfilzomib was significantly superior to lenalidomide (OR ranged from 2.18 to 2.20), daratumumab (OR ranged from 1.49 to 2.66) and ixazomib (OR ranged from 2.75 to 3.57).Conclusion Considering OS and PFS, lenalidomide-carfilzomib should be recommended as the best therapy. In clinical practice, this must be weighed against the increased risk of adverse events and financial burden. However, more head-to-head studies are needed to confirm these findings.

HighlightsCas9 RNA functions as a miRNA sponge.Let-7 is the dominant regulated miRNA by Cas9 RNA.RNA sequence optimization of Cas9 by synonymous mutation improves its safety.

Background and ObjectivesPancreatic cancer (PC) is one of the deadliest cancers worldwide although substantial advancement has been made in its comprehensive treatment. The development of artificial intelligence (AI) technology has allowed its clinical applications to expand remarkably in recent years. Diverse methods and algorithms are employed by AI to extrapolate new data from clinical records to aid in the treatment of PC. In this review, we will summarize AI's use in several aspects of PC diagnosis and therapy, as well as its limits and potential future research avenues. MethodsWe examine the most recent research on the use of AI in PC. The articles are categorized and examined according to the medical task of their algorithm. Two search engines, PubMed and Google Scholar, were used to screen the articles. ResultsOverall, 66 papers published in 2001 and after were selected. Of the four medical tasks (risk assessment, diagnosis, treatment, and prognosis prediction), diagnosis was the most frequently researched, and retrospective single-center studies were the most prevalent. We found that the different medical tasks and algorithms included in the reviewed studies caused the performance of their models to vary greatly. Deep learning algorithms, on the other hand, produced excellent results in all of the subdivisions studied. ConclusionsAI is a promising tool for helping PC patients and may contribute to improved patient outcomes. The integration of humans and AI in clinical medicine is still in its infancy and requires the in-depth cooperation of multidisciplinary personnel.

Objective To explore the effects of the expression level of miR-520-5p/PPP5C in pancreatic cancer cells and exosomes on cell viability, angiogenesis, autophagy, which involved in the mechanism of gemcitabine resistance in pancreatic cancer. Methods APSC-1 cell line was treated with gemcitabine, after which its exosomes were extracted for NTA assay. Subsequently, the drug resistance of APSC-1 cells was assayed using CCK8, as well as the activity of HUVEC cells treated with exosomes from each group of APSC-1 cells after drug resistance treatment as well as overexpression treatment. Five groups of HUVEC cells treated with exosomes were subjected to in vitro tubule formation assay. levels of PPP5C in each group of ASPC-1 cells and their exosomes, levels of overexpressed PPP5C, and related exosomal proteins were examined by WB. mRNA expression levels of PPP5C and levels of miR-520a were examined by qPCR The relationship between miR-520a-5p and PPP5C was investigated. After that, the autophagy of PPP5C was detected. Finally, it was analyzed by TCGA database for survival prognosis analysis. Results APSC-1 cells had an IC50 value of 227.1 μM for gemcitabine, elevated PPP5C expression, drug resistance, and enhanced HUVEC cell activity; exosomes CD9, CD63, and CD81 were significantly expressed in all groups; meanwhile, enhanced PPP5C expression not only promoted in vitro tubule formation but also increased autophagy levels; meanwhile, its relationship with miR-520-5p and There was a targeted inhibitory relationship between its level and miR-520-5p and PPP5C, and its elevated level also led to a decrease in the survival level of patients over 3-5 years. Conclusion PPP5C has a prognostic role in pancreatic cancer by promoting the value-added and invasion of pancreatic cancer cells, and a targeted inhibitory relationship between miR-520-5p and PPP5C was found.

Sugemalimab is a full-length, fully human PD-L1 targeted immunoglobin G4 (IgG4, s228p) mAb. GEMSTONE-302 is a randomized, double-blind, phase 3 study to evaluate the efficacy and safety of sugemalimab or placebo in combination with chemotherapy as first-line treatment in metastatic squamous (sq) or non-squamous (nsq) NSCLC. The PFS interim analysis data as of 08 June 2020 showed that sugemalimab plus chemotherapy demonstrated a clinically meaningful and statistically significant prolongation of PFS with a well-tolerated safety profile in metastatic NSCLC patients irrespective of tumor pathology and PD-L1 expression.

Background Postmenopausal osteoporosis is a chronic metabolic bone disease caused by excessive osteoclast activation, and osteoclasts are considered to be the sole participants in the degeneration and resorption of bone matrix for controlling bone integrity and continuity. The biological functions of osteoclasts depend critically on the number and activity of fused polykaryon. Hence, targeting osteoclast differentiation and activity can modulate bone resorption and alleviate osteoporosis. Alpinetin is widely used for excellent anti-inflammatory activities and little side-effect, but its role in osteoporosis remains unknown. Results In this study, we investigated for the first time the ability of alpinetin to inhibit estrogen deficiency-induced bone loss. Alpinetin significantly reduced the expression levels of NFATc1 and its downstream genes, thereby inhibiting osteoclast differentiation in a concentration- and time-dependent manner. Additionally, alpinetin inhibited F-actin ring formation and bone resorption, as well as reduced the activation levels of NF-κB, ERK, and AKT signaling cascades. In mature osteoclasts, alpinetin remarkably inhibited integrin-mediated migration and lysosomal biogenesis and trafficking by modulating the PKCβ/TFEB and ATG5/LC3 axes. Importantly, alpinetin treatment in mice alleviated ovariectomy-induced bone volume loss. Conclusion Our findings strongly suggest that alpinetin plays a significant role in the regulation of NFATc1 production for the differentiation of osteoclasts and inhibits integrin-mediated cell migration and lysosomal function in mature osteoclasts, thus weaken the increased osteolytic ability due to estrogen deficiency. Alpinetin may represent a promising agent for the treatment of osteoporosis and other metabolic bone diseases.

During embryonic development, neural stem cells (NSCs) emerge as early as the neural plate stage and give rise to the nervous system. Early-stage NSCs express Sry-related-HMG box-1 (Sox1) and are biased towards neuronal differentiation. However, long-term maintenance of early-stage NSCs in vitro remains a challenge. Here, we report development of a defined culture condition for the long-term maintenance of Sox1-positive early-stage mouse NSCs. The proliferative ability of these Sox1-positive NSCs was confirmed by clonal propagation. Compared to the NSCs cultured using the traditional culture condition, the long-term self-renewing Sox1-positive NSCs efficiently differentiate into neurons and exhibit an identity representative of the anterior and midbrain regions. These early-stage Sox1-positive NSCs could also be switched to late-stage NSCs by being cultured with bFGF/EGF, which can then differentiate into astrocytes and oligodendrocytes. The long-term self-renewing Sox1-positive NSCs were defined as naïve NSCs, based on their high neuronal differentiation capacity and anterior regional identity. This culture condition provides a robust platform for further dissection of the NSC self-renewal mechanism and promotes potential applications of NSCs for cell-based therapy on nervous system disorders.

Glycogen synthase kinase 3 (GSK3) plays a central role in diverse cellular processes. GSK3 has two mammalian isozymes, GSK3α and GSK3β, whose functions remain ill-defined because of a lack of inhibitors that can distinguish between the two highly homologous isozymes. Here, we show that GSK3α and GSK3β can be selectively inhibited in mouse embryonic stem cells (ESCs) using a chemical-genetic approach. Selective inhibition of GSK3β is sufficient to maintain mouse ESC self-renewal, whereas GSK3α inhibition promotes mouse ESC differentiation toward neural lineages. Genome-wide transcriptional analysis reveals that GSK3α and GSK3β have distinct sets of downstream targets. Furthermore, selective inhibition of individual GSK3 isozymes yields distinct phenotypes from gene deletion, highlighting the power of the chemical-genetic approach in dissecting kinase catalytic functions from the protein’s scaffolding functions. Our study opens new avenues for defining GSK3 isozyme-specific functions in various cellular processes.