Background: Beta-thalassemia is a hereditary blood disorder characterized by reduced synthesis of the beta-globin chain. MicroRNAs (miRs) are small RNA molecules that regulate gene expression and have been implicated in beta-thalassemia. To explore dysregulated miR-222 and miR-15a expression in transfusion-dependent beta-thalassemia and assess their potential associations with Torque Teno Virus and cytomegalovirus infections. Materials and Methods: This study included 57 TDT patients registered at the Thalassemia Clinic affiliated with the Hematology Research Center, Shiraz, Iran. The expression levels of miR-222 and miR-15a were analyzed using the real-time SYBR Green PCR method. TTV and CMV infections were detected by analyzing the presence of their genomic DNA using an in-house semi-nested PCR protocol. Results: The expression level of miR-222 was significantly upregulated (47.5-fold, P≤0.001) in TDT patients compared to healthy controls. However, the expression of miR-15a in TDT patients was slightly decreased compared to healthy controls, but the difference was not statistically significant (P=0.193). TTV infection was observed in 21.1% of TDT patients, while CMV infection was detected in 5.2% of the patients. Although miR-222 and miR-15a gene expression levels were higher in TTV-positive patients compared to TTV-negative patients, the differences were not statistically significant (P=0.926 and P=0.243, respectively). Conclusion: MiR-222 was upregulated in TDT patients, but miR-15a did not show a significant difference. TTV and CMV infections were detected, but their association with miR expression was not significant, possibly due to the small sample size. Larger studies are needed for a more comprehensive evaluation.

Background: Iron homeostasis is a complex process involving multiple factors. Erythroblasts secrete erythroferrone (ERFE), which affects hepcidin production, thereby enhancing iron uptake. This study aimed to investigate the expression of the ERFE-encoding FAM132B gene in Iranian patients with beta-thalassemia major (BTM) and beta-thalassemia intermedia (BTI). Materials and Methods: A total of 40 BTM and BTI patients and 20 healthy blood donors as a control group were recruited. Total RNA was extracted from whole blood samples, and cDNA was synthesized. Gene expression was quantified using a SYBR Green-based real-time PCR (RT-PCR) assay. Data analysis was performed by the GraphPad Prism program using the one-way ANOVA test. Results: The expression of the FAM132B gene was upregulated in BTI patients compared to BTM patients (p = 0.0151). Despite the higher mRNA fold change in BTI patients, no significant difference was observed in the FAM132B gene expression between beta-thalassemia patients (major and intermedia) and the control group. Conclusion: The expression of the FAM132B gene was different between beta-thalassemia major and intermedia patients. Further studies should be conducted to better elucidate the role of erythroferrone as a potential therapeutic target in patients with beta-thalassemia.

β-hemoglobinopathies rank among the most prevalent inherited blood disorders globally. Traditional management strategies are primarily palliative and often associated with significant challenges, including iron overload and limited long-term efficacy. Allogeneic hematopoietic stem cell transplantation (HSCT) is a potentially curative option for transfusion-dependent patients, but its broader applicability is constrained by factors that limit its use. Utilizing viral vectors and gene-editing tools, particularly CRISPR-Cas9 technology, researchers have developed therapies that target the root causes of these disorders. These innovative approaches have demonstrated substantial therapeutic potential, accompanied by favorable safety profiles, in clinical settings. Since the initial investigations, the genome editing tool has rapidly advanced for genetic abnormalities, particularly monogenic blood diseases, including β-hemoglobinopathies. This method suggests an approach with lower concerns in viral gene integration and insertional mutagenesis issues. This review comprehensively surveys the therapeutic strategies for β-thalassemia and sickle cell disease (SCD) currently in preclinical and clinical development, with a focus on the evolving treatment paradigm. Looking forward, critical research priorities include optimizing the efficiency and specificity of gene-editing platforms and pioneering novel delivery systems to guarantee both therapeutic efficacy and clinical safety.

Iron overload is associated with significant health risks, underscoring the importance of understanding its pathophysiology as well as establishing accurate diagnostic and monitoring methods. Chronic iron overload is associated with either genetic disorders characterized by excessive iron accumulation (hereditary hemochromatosis), or is secondary to diseases of ineffective erythropoiesis and/or requiring regular blood transfusions (like thalassemia, sickle cell disease, myelodysplastic syndromes). Diagnosis is based on clinical suspicion, corroborated by laboratory findings such as elevated serum ferritin and transferrin saturation, along with imaging techniques (magnetic resonance imaging) which facilitate non-invasive assessment of iron levels in parenchymal organs. Elevated ferritin and transferrin saturation and exclusion of secondary causes should prompt genetic evaluation for hereditary disorders predisposing iron overload. Chronic systemic iron overload causes progressive tissue iron accumulation, leading to severe clinical implications, including myocardial dysfunction, liver cirrhosis, and increased risk of hepatocellular carcinoma. Monitoring includes evaluating iron overload indices (serum ferritin, transferrin saturation, liver and heart iron concentration) along with serum and urine indices of parenchymal organ damage at different timepoints regarding the type of disease, patient's age, severity and response to treatment, and aims in improving disease progression and preventing complications. This article provides a comprehensive overview of the pathophysiologic mechanisms and the diagnostic and monitoring techniques of iron overload, in order to revise current knowledge and to raise clinical awareness for effective management.

Recent advances in Artificial Intelligence (AI) have profoundly transformed the field of genome editing, particularly through integration with the Clustered Regularly Inter-spaced Short Palindromic Repeats (CRISPR) technology. This review highlights how AI-driven computational models are reshaping guide RNA (gRNA) design, off-target prediction, and editing precision in CRISPR-Cas systems. A PRISMA-informed literature survey was conducted using PubMed, Scopus, EMBASE, and Google Scholar databases to identify studies exploring AI-assisted CRISPR applications in gene therapy and biomedical research. The results demonstrate that deep learning, machine learning, and reinforcement learning approaches significantly enhance prediction accuracy, algorithmic efficiency, and translational potential across genetic diseases such as β-thalassemia, muscular dystrophy, and cancer. Moreover, ethical challenges, algorithmic bias, and data security concerns remain critical barriers to clinical adoption. This review also discusses the emerging landscape of AI-assisted CRISPR research in Iran, emphasizing national progress, infrastructural constraints, and future opportunities. Overall, the convergence of AI and CRISPR technologies promises to advance precision medicine by accelerating the development of personalized, efficient, and ethically responsible genome-editing solutions.

Background: Iron overload and its associated complications are major concerns in patients with transfusion-dependent β-thalassaemia (TDT). Iron chelation is an important part of TDT therapy with monotherapy or dual iron chelation being the most commonly used strategies. Evidence regarding the efficacy and safety of triple iron chelation therapy remains limited. Case presentation: We present the case of a 21-year-old immigrant from the Middle East with TDT and a history of irregular transfusion management without chelation therapy, leading to clinically significant iron overload. She was successfully treated with the combination of deferoxamine, deferasirox and deferiprone over a course of 8 years. Triple chelation therapy led to sustained reductions in serum ferritin levels and improvement in hepatic and cardiac iron burden on follow-up MRI, with good tolerability. Conclusions: This case highlights the potential role of triple iron chelation therapy as a therapeutic strategy in TDT patients with severe iron overload. Further studies are needed to establish optimal dosing, eligible patients and long-term safety.

Allogeneic hematopoietic cell transplantation (HCT) has been used for decades to treat certain malignant and non-malignant hematological conditions, but challenges remain. Increased understanding of disease mechanisms and recent developments in genome editing have enabled alternative strategies utilizing gene-edited autologous HCT and many of these have progressed to the clinic. We present here a comprehensive review of clinical trials of gene-edited autologous hematopoietic stem cells for the treatment of hemoglobinopathies and immunodeficiencies. Searches of major international clinical trial registries were carried out using specific key words. In total, 44 interventional clinical trials investigating gene-edited autologous stem cell therapies were identified, with CASGEVY (exagamglogene autotemcel) being the only product approved to date. Hemoglobinopathies were the most common indication (n = 37) followed by immunodeficiencies (n = 4), with single trials in HIV-1 infection, pyruvate kinase deficiency and limb-girdle muscular dystrophy. Gene-editing strategies fall into three categories: disruption of the BCL11A erythroid enhancer, editing of the γ-globin promoter and direct correction or disruption of disease-relevant genes. CD34[+] hematopoietic stem and progenitor cells are the most common cell types edited, and CRISPR-Cas9 is the most widely used gene-editing modality. While results are encouraging, efficient intracellular delivery of gene-editing tools, editing efficiencies and off-target editing remain challenges for the field.

Thalassemia is a hereditary hemoglobinopathy characterized by ineffective erythropoiesis, chronic hemolysis, and transfusion-related iron overload, which collectively contribute to oxidative stress and organ dysfunction. The present study aimed to investigate the relationships between iron metabolism, oxidative stress biomarkers, and immune cell function across different clinical conditions. Peripheral blood samples were obtained from healthy individuals and patients with iron deficiency anemia, obesity, thalassemia trait (TT), β-thalassemia HbE (BTE), and β-thalassemia major (BTM). Hematological parameters were measured using automated hematology analyzers, while biochemical indicators, including liver enzymes and bilirubin, were determined using clinical chemistry assays. Iron overload was evaluated using serum iron parameters and T2*-weighted magnetic resonance imaging. Oxidative stress biomarkers, including reduced glutathione, thiobarbituric acid-reactive substances, and total antioxidant capacity, were assessed spectrophotometrically. Flow cytometric analysis was used to measure reactive oxygen species, redox-active iron, and lipid peroxide levels in granulocytes and lymphocytes. Thalassemia patients exhibited severe anemia, elevated liver enzymes, increased bilirubin levels, and significant alterations in iron metabolism compared with healthy controls. Hepatic iron accumulation was more common than cardiac iron deposition, particularly in BTE patients. Granulocyte oxidative burst activity was significantly reduced in thalassemia patients, whereas lymphocyte responses remained relatively preserved. Increased variability in glutathione levels suggested activation of intracellular antioxidant defense mechanisms in response to chronic oxidative stress. These findings highlight the complex interplay between iron overload, oxidative stress, and the immune cell dysfunction associated with thalassemia, thereby providing insights into improved monitoring and therapeutic strategies.

Thalassemia is a relatively common monogenic inherited blood disorder in southern China. This study aims to evaluate the additional diagnostic yield of targeted sequencing for thalassemia gene compared to traditional mainstream methods, providing evidence to support clinical decision-making and application. A total of 449 specimens with the negative common thalassemia test result were randomly selected for next-generation sequencing(NGS) testing. For rare thalassemia genotypes and abnormal hemoglobin variants detected by NGS, hematological information such as mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and hemoglobin electrophoresis was summarized and statistically analyzed. Through targeted sequencing for thalassemia, an additional 42 cases of rare thalassemia genotypes and 10 cases of abnormal hemoglobin variants were detected in the traditional test-negative groups (Neg_70 and Neg_80). Compared with traditional gene testing, the overall additional positive rate for NGS was 9.35% (42/449). In the traditional test-positive group (Pos_80), 2 cases of abnormal hemoglobin variants were identified, with an additional positive rate of 0.88% (2/226). The most frequently detected rare genotypes were the α-thalassemia variant -THAI and the δβ-thalassemia variant Chinese(Gγ + (Aγδβ)⁰), found in 20 and 7 cases, respectively. Additionally, a novel heterozygous variant of the HBA2 gene was identified: NM_000517.6:c.16delG. NGS demonstrates superior detection efficacy and higher detection rates in individuals with low mean corpuscular volume, particularly in those with severe microcytic hypochromic anemia. It effectively identifies rare mutations and reduces the misdiagnosis rate of thalassemia.