Royan Institute

Teratozoospermia, characterized by abnormal sperm morphology, is a major contributor to male infertility. Kinases, enzymes that catalyze the transfer of phosphate groups to proteins, are crucial regulators of cellular signaling pathways and play significant roles in sperm development and maturation. The purpose of the current study was to identify differentially expressed genes (DEGs) between teratozoospermia and normozoospermia samples and to investigate the role of kinases in these expression changes. An integrated analysis of transcriptome data was conducted from teratozoospermia and normozoospermia samples using publicly available gene expression omnibus (GEO) datasets. Three gene expression series (GSE) profiles of teratozoospermia from one superseries were selected and combined. Differential expression analysis was performed using the limma package in R, applying linear modeling and empirical Bayes statistics to identify DEGs with a threshold of adjusted p<0.05. A comprehensive list of human kinase genes was obtained from the KinHub database, and differentially expressed kinases between the two conditions were identified. Functional enrichment analyses including gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) pathways were conducted. Additionally, receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic potential of identified kinases. Our analysis identified 1,292 DEGs. Among these, 34 kinases were identified (10 upregulated and 24 downregulated). ROR1 and STK39 showed the most significant changes. ROC analysis demonstrated strong diagnostic values for these kinases. This study is the first comprehensive analysis integrating transcriptomic data and kinase-focused gene expression profiling specifically in teratozoospermia, suggesting that kinase dysregulation may contribute to teratozoospermia and male infertility.

Prenatal prediction of CD34+ adequacy supports cord-blood banking by reducing expenses on low-yield units and reserving capacity for clinically promising grafts. We have developed and evaluated a prenatal machine learning model that predicts whether an umbilical cord blood (UCB) unit will meet a clinically supported adequacy threshold (≥1.5×105 CD34+ cells/kg recipient) before collection for single-unit grafts. In this retrospective study, we analysed 126,406 records from the Royan Stem Cell Technology Company (RSCT; Tehran, Iran), which included routinely available maternal, neonatal, and family-history variables. A pipeline of imputation (IterativeImputer numeric; SimpleImputer+OrdinalEncoder categorical), feature selection (Extra Trees), and hyperparameter tuning using Bayesian optimisation with model training/evaluation was performed within cross-validation folds. Decision Tree (DT), K-Nearest Neighbours (KNN), Random Forest (RF), Support Vector Machine (SVM), and Multilayer Perceptron (MLP) classifiers were tuned via Bayesian optimisation. Models were ranked by the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS). Majority voting (MV) ensembles were constructed from the top-k models. Model interpretability used SHapley Additive exPlanations (SHAP). The MV (top-4: RF, KNN, DT, MLP) ensemble achieved an area under the receiver operating characteristic curve (ROC-AUC)=0.808 and an area under the precision-recall curve (PR-AUC)=0.744 on the held-out test set, with an accuracy=0.757, precision=0.726, recall/sensitivity=0.804, F1=0.762, specificity=0.716, and Brier score=0.181. SHAP highlighted history of hepatitis C, birth place, hyperthyroidism, history of anaemia, oral fungus, and rheumatism among the most influential features. Prenatal prediction of UCB CD34+ adequacy using an interpretable MV ensemble is feasible and accurate to support pre-collection triage and can potentially improve banking efficiency. The resultant model offers a non-invasive tool to enhance the efficiency of cord blood banking by prioritising units with higher transplantation potential.

Previous studies by the authors have shown that human amniotic membrane extract (AME) potentially improves epithelial damage in corneal and skin lesions. However, questions about its role in diabetic foot ulcer (DFU) treatment remain unanswered. In this study, patients with hard-to-heal DFUs were randomly selected after screening and, after confirming the safety of the AME in an initial selection of patients, the remaining participants were divided into two groups. The treatment group received an AME product (DiAMX, Royan Stem Cell Technology, Iran) (concentration 1mg/ml) with standard of care (SoC), topically, every 48 hours for the first week and then every 72 hours until complete closure. The control group received SoC. Patients were visited weekly, and any improvement, reduction in the wound area and any side-effects were recorded. The results of the safety phase (without the control group) showed that the AME had no adverse events. In the efficacy phase, there was no significant difference in baseline characteristics between the treatment and control groups. The results showed that the wound healing rate was 100% in the treatment group and 77.5±3.12% in the control group (p<0.0001) at week 6. All wounds (n=15) were closed in the treatment group and seven wounds had closed in the control group (p<0.0001). The findings of this study showed that the topical use of DiAMX, in addition to SoC, was patient-friendly, inexpensive, safe and effective in healing hard-to-heal DFUs.

Liver fibrosis, characterized by the excessive deposition of extracellular matrix (ECM) driven by hepatic stellate cells (HSCs) activation, remains a critical challenge due to its progression to cirrhosis and hepatocellular carcinoma (HCC). This review clarifies the complex crosstalk between the immune system and HSCs, highlighting key cellular players including macrophages, natural killer (NK) cells, regulatory T cells (Tregs), and their cytokine‐mediated signaling pathways that regulate fibrogenesis and fibrosis resolution. We describe pivotal molecular mechanisms such as transforming growth factor (TGF)‐β, platelet‐derived growth factor (PDGF), Wnt/β‐catenin, and NF‐κB signaling in HSCs modulation, emphasizing their interplay with immune responses. Novel therapeutic strategies targeting this complex immune–HSCs interaction, ranging from immunomodulatory agents, macrophage polarization, and NK cell‐based therapies, to stem cell‐derived exosomes, offer promising opportunities for preventing and reversing fibrosis. We further discuss innovative combination therapies integrating immunotherapies with antifibrotic agents, personalized strategies based on immune profiling, and the challenges of immune heterogeneity in fibrosis management. This review discusses recent advances in molecular interplay of immune system and HSCs, highlighting novel therapeutic targets, and future perspectives for managing chronic liver diseases.

Solid tumors remain difficult to treat due to antigen heterogeneity, physical barriers that limit immune-cell trafficking, and a profoundly immunosuppressive tumor microenvironment (TME). Over the past decade, cancer immunotherapy advanced considerably through innovative strategies, including macrophage reprogramming and CAR-macrophages, dendritic-cell (DC) vaccines, natural killer (NK) and natural killer T (NKT) cell approaches, tumor-infiltrating lymphocyte (TIL) therapy, TCR-engineered and CAR-T cells, emerging B-cell engineering, and cell-derived extracellular vesicles (EVs). Here we summarize how each modality interacts with the TME, highlight key clinical milestones (e.g., FDA approval of a TIL product for melanoma in 2024), and outline bioengineering strategies—multi-antigen targeting, cytokine armoring, trafficking cues, and safety switches—that aim to overcome resistance and toxicity. We also review EV-based, cell-free strategies that retain tumor specificity with potentially improved safety and manufacturability. Finally, we discuss remaining barriers—standardized manufacturing, on-target/off-tumor effects, limited persistence—and propose rational combinations with checkpoint blockade, radiotherapy, and targeted agents. This overview positions immune cell–based therapy as a rapidly maturing, transformative approach for solid tumors.Graphical abstract

ABSTRACTN,N‐dimethylacetamide (DMA) is an organic solvent, used in busulphan, iv‐formulation, (Busulfex). DMA is metabolized primarily to N‐methylacetamide (MMA) via CYP2E1. In the present study, we investigated the pharmacokinetics and toxicity of DMA and MMA in patients, mice and cell lines. In pediatric patients (6 months to 18 years) undergoing hematopoietic stem cell transplantation, the pharmacokinetics of DMA and MMA were followed during the 4‐days Busulfex conditioning (3.2–4 mg/kg, b.i.d). No accumulation of DMA was found; however, a significant increase in clearance and a significant decrease (p < 0.005) in half‐lives by the end of treatment were observed which might indicate CYP2E1 autoinduction. Furthermore, continuous increases in plasma concentration of MMA were observed during treatment and a terminal half‐life of 15.2 ± 1.7 h was detected. Moreover, ALT was significantly (p = 0.04) increased in > 61% of the patients after conditioning. Additionally, mice were treated with either dimethyl sulfoxide (DMSO), DMA, busulphan in DMSO, Busulfex or saline for 4 days. DMA‐treated mice showed elevated ALT and AST values. Interestingly, Busulfex administration did not alter mice liver enzymes. Busulphan in DMA showed higher cytotoxicity compared to busulphan in DMSO in HepG‐2, Huh‐7 and HL‐60 cells. The combination Bu‐DMA‐MMA exhibited increased cytotoxicity in a concentration‐dependent manner. In conclusion, Busulfex administration to pediatric patients resulted in an accumulation of MMA during the 4‐days treatment. Busulfex is less toxic compared to Bu dissolved in DMA. MMA addition to Bu in DMA showed higher cytotoxicity. Therefore, MMA with a relatively long half‐life may induce hepatotoxicity and/or interaction with subsequently administered drugs; thus further investigations are urgently warranted.

Tau is a microtubule-associated protein. Hyperphosphorylation of tau at neurotoxic sites, particularly at Thr231 within the Thr231-Pro motif, is a pathological hallmark of Alzheimer's disease (AD) and other tauopathies. Phosphorylated tau at Thr231 exists in two distinct conformations: cis and trans. The Cis pThr231-Pro Tau confomer is neurotoxic and promotes neurodegeneration. Furthermore, tau is subject to O-linked N-acetylglucosamine (O-GlcNAc) modification, and it has been suggested that O-GlcNAcylation of tau can influence tau phosphorylation. In this study, we utilized Thiamet G, an O-GlcNAcase (OGA) inhibitor, to elevate tau O-GlcNAcylation levels. Our findings demonstrate that treatment of nutrient-deprived primary cortical neurons with this OGA inhibitor increased tau O-GlcNAcylation, inhibited the formation of the neurotoxic Cis p-Tau conformation, and reduced neuronal cell loss. Additionally, we observed that the Trans p-Tau conformation represents a normal conformer under physiological conditions. Collectively, our data support tau O-GlcNAcylation as a promising therapeutic strategy for Alzheimer's disease and other tauopathies.