Journal of Cellular and Molecular Medicine最新文献

Childhood obesity has led to an increase in type 2 diabetes (T2D) among youth, with adolescent-onset T2D showing a rapid decline in β-cell function compared to adult-onset cases. While the disease progression is more aggressive in early life, treatment can lead to recovery or remission more often at younger ages. SGLT2i have proven multiple health benefits when prescribed to adults with T2D but may have a greater potential in improving insulin production and β-cell mass in youth. In our study, TallyHO mice, which develop severe early-onset T2D, were treated with canagliflozin (cana) while on a 10-week diet. Results showed a significant reduction in blood glucose levels and improved β-cell function, indicated by higher C-peptide, islet insulin content, and HOMA-B index compared to untreated mice. Cana treatment restored the islet area and β to α-cell ratio, while also decreasing apoptosis. Notably, cana promoted the transient appearance of endocrine bihormonal cells and small clusters of insulin-positive cells, suggesting a possible transdifferentiation process and β-cell neogenesis. Furthermore, cana stabilised β-cell phenotype, restoring the expression of key identity markers while reducing abnormal cell types and the dedifferentiation to precursors and mesenchymal cells. These findings suggest that canagliflozin can promote the regeneration of pancreatic islets and mitigate β-cell dedifferentiation in the early onset of β-cell deficiency.

Cancer cells rely on citrate for multiple metabolic processes, suggesting that limiting the availability of extracellular citrate may represent a novel therapeutic strategy. The plasma membrane citrate transporter (pmCiC) has been implicated in the pathogenesis of several cancers before, and its activity can be inhibited by gluconate. Tissue samples from patients were stained, and pmCiC expression was analysed and correlated with clinical course. Melanoma cells were treated with or without citrate in physiological concentration and with or without gluconate, a pmCiC inhibitor. Cell proliferation rates were subsequently measured. pmCiC expression was observed in 58.2% of primary melanomas and 76.5% of melanoma metastases, but only in 22.2% of benign nevi. However, pmCiC expression did not correlate with the response to novel melanoma-specific therapies. In the presence of pmCiC, melanoma cells exhibited significantly increased proliferation when exposed to extracellular citrate. This effect was blocked by the addition of gluconate. Extracellular citrate uptake via pmCiC appears to contribute to the pathogenesis of malignant melanoma. Notably, inhibition of pmCiC by gluconate effectively suppressed citrate-induced proliferation.

Although studies support disrupted bone remodelling within geriatric populations, mid-life changes are understudied. To investigate this, we performed bone marrow transplantation assays using either 8- or 40-week-old mice. Micro-CT analyses of lethally irradiated 8-week-old mice transplanted with 40-week-old bone marrow exhibited increased mid-shaft femoral cortical bone mass and thickness. Intensive bone marrow regeneration mirrors hematopoietic development in that erythro-myeloid progenitors (EMPs) first expand to support blood production before definitive hematopoietic stem cell (HSC) production. We hypothesized that reduced HSC capacity of 40-week-old bone marrow and compensatory expansion of EMPs may facilitate gains in cortical bone. Flow cytometry analyses revealed greater EMP to HSC ratios when mice were reconstituted with increasing percentages of middle-aged bone marrow. To identify cell types mediating these effects, we performed comparative scRNA-Seq analyses and identified CD11B⁺CD36⁺ myeloid cells exhibiting enriched expression of bone anabolic cytokines. Elevated levels of Wnt ligands, especially Wnt6, characterized these cells. In lineage tracing assays, CD11B⁺CD36⁺ cells were donor-derived myeloid cells. In functional assays, we demonstrate that soluble factors produced by CD11B⁺CD36⁺ cells enhance osteogenesis. Moreover, CD11B/CD36/Wnt6 exquisitely mark anabolic macrophages within human bone marrow. These findings reveal a myeloid population present during midlife that enhances cortical bone.

Oxidised 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (oxPAPC), dendritic cells (DCs), and long non-coding RNAs (lncRNAs) play crucial roles in atherosclerosis (AS). This study aimed to determine whether oxPAPC-induced DC-derived lncRNAs contribute to AS and to elucidate the underlying regulatory mechanisms. DCs were treated with increasing oxPAPC concentrations to assess transcriptomic changes. RNA sequencing was used to identify differential expression of lncRNAs. ChIP-Seq and RNA pull-down assays were used to assess direct binding between lncRNA CYP1B1-AS1 and NFATC2. The association between CYP1B1-AS1 and CYP1B1 was assessed using Pearson's correlation analysis. Elevated serum oxPAPC levels were confirmed in patients with coronary heart disease. In vitro, sustained oxPAPC stimulation activated the TLR4-MD2 pathway in DCs. CYP1B1-AS1 was identified as the key oxPAPC-induced DC-derived lncRNA, with Gm33055 as its murine homologue. RNA sequencing revealed oxPAPC-driven alterations in DC chemotaxis, differentiation, and lymphocyte activation. Analysis of human atherosclerotic plaque-derived DCs showed significant CYP1B1-AS1 upregulation. Gm33055 enhanced Cyp1b1 expression in murine DCs. Mechanistically, oxPAPC promoted NFATC2 nuclear translocation. NFATC2 binds to the CYP1B1-AS1 promoter, whereas CYP1B1-AS1 directly interacts with NFATC2, forming a positive regulatory loop. Adoptive transfer of m-CYP1B1-AS1-expressing DCs into Apoe^-/- mice accelerated AS progression. These findings identify a DC-derived lncRNA-mediated regulatory axis that promotes AS and suggest potential therapeutic targets.

Hepatocellular carcinoma (HCC) is a primary malignant tumour that impacts patients' quality of life. Currently, clinical experience from The First Affiliated Hospital of Guangzhou University of Chinese Medicine suggests that Jianpi Jiedu Xiaozheng Fang (JPJDXZF) demonstrates promising efficacy in the treatment of HCC. We aimed to explore the mechanisms of JPJDXZF in HCC based on network pharmacology. The components and their relevant targets of JPJDXZF were identified using databases such as SymMap, TCMID, TCMSP, and TCM-ID. Following ADME screening, 1443 active components of JPJDXZF were identified, and 435 corresponding drug targets were predicted using the SwissTargetPrediction database. Subsequently, prognosis-related differentially expressed genes (DEGs) associated with HCC were analyzed using TCGA and GTEx datasets, and a gene expression matrix was derived. Key genes involved in HCC regulation were identified, and functional analyses were performed. Furthermore, we explored the regulatory effects of JPJDXZF at the cellular, organoid, and animal levels. We identified 18 intersecting genes between HCC prognosis-related genes and JPJDXZF-target genes. Venn diagram analysis successfully identified BIRC5 and CYP2E1 as two potential targets for JPJDXZF in treating HCC. Pathway enrichment analysis indicated that the core targets of JPJDXZF were enriched in multiple signalling pathways, including the Hippo pathway, in which BIRC5 is involved as a downstream regulatory gene. In in vitro experiments, JPJDXZF-containing serum significantly reduced the viability and migration of HepG2 and MHCC97-H cells, leading to a decrease in organoid diameter and ATP activity in HCC organoids. In in vivo experiments, tumours in nude mice treated with JPJDXZF exhibited reduced volume and weight, along with decreased expression of BIRC5 and Hippo pathway effectors YAP and TAZ. At the mechanistic level, JPJDXZF treatment was associated with altered Hippo pathway-related signalling, accompanied by reduced YAP/TAZ activity and changes in BIRC5 expression, together with effects on HCC cell proliferation and apoptosis. In addition, siMST1/2 interference and EMT inhibitor-1 treatment partially attenuated the effects of JPJDXZF on cell viability, migration, and apoptosis. JPJDXZF regulates BIRC5 expression in association with Hippo pathway activity in HCC. In vitro, in vivo, and molecular mechanism analyses support JPJDXZF as a potential therapeutic strategy for HCC by modulating key proteins in the Hippo pathway, thus affecting HCC cell proliferation, apoptosis, and migration.

Peroxiredoxins (PRDXs) are antioxidant enzymes that scavenge hydrogen peroxide and protect cells from reactive oxygen species (ROS). There are six genes encode different types of PRDXs (PRDX1-PRDX6) in humans and most of them are overexpressed in tumours; however, their expression patterns and prognostic value in bladder cancer (BLCA) remain unclear. In this study, we examined the aberrant expression of all six types of PRDX genes in BLCA and identified distinct clinical and immune associations. High expression of PRDX1 and PRDX6 was correlated with poor overall survival (OS), increased mutational burden and chromosomal instability. Overexpression of PRDX4 and PRDX6 was associated with advanced tumour stage, larger tumour size, higher immune scores, and increased immune cell infiltration. By contrast, PRDX2 overexpression showed only modest effects on OS and was associated with reduced immune signalling and diminished infiltration of anti-tumor immune cells. These findings highlight the differential roles of PRDX family members in shaping BLCA tumour immune microenvironment. PRDXs may serve as prognostic biomarkers for patients tratification and represent potential therapeutic targets to enhance immunotherapy response. Further in vitro and in vivo studies are required to confirm our in silico data and define their clinical relevance for BLCA prognosis.

Dent's disease, an X-linked recessive disorder predominantly affecting males, is characterized by nephrocalcinosis, nephrolithiasis, and a high risk of progression to end-stage renal disease. Dent's disease type 1, accounting for 60% of cases, caused by mutations in the CLCN5 gene encoding the chloride ion channel protein ClC-5, exhibits significant clinical heterogeneity and variability in disease progression. The lack of hotspot mutations poses challenges for genetic diagnosis and counselling, complicating the prediction of disease outcomes. This study systematically evaluated the functional and structural impacts of 181 CLCN5 missense mutations using computational tools, including PredictSNP, MAGPIE, and molecular dynamics simulations, to propose a robust method for improving genetic counselling and prognosis prediction. Our analysis identified mutations at the dimer interface and chloride selectivity filter as critical disruptors of ClC-5 function and stability. Notably, molecular dynamics simulations of L200R, P213L, and G512R mutations revealed that L200R significantly destabilized the protein structure. Clinical data from a multicentre cohort of Chinese patients with CLCN5 mutations corroborated our computational predictions, highlighting the essential role of helix O in ClC-5 function. By integrating bioinformatics analyses with clinical validation, this study provides molecular insights into Dent's disease heterogeneity and proposes a framework for enhancing genetic counselling and prognostic assessment for affected patients.