The progression from cirrhosis to hepatocellular carcinoma (HCC) is a key outcome in the management of chronic liver disease. This process has a long incubation period and significant individual differences, making early warning still difficult. Clinical follow-up mainly relies on imaging examinations and alpha fetoprotein, but the ability to identify high risk precancerous states is limited. The imbalance of gut microbiota and its metabolites may occur earlier than the visible stage of tumors. They can affect barrier integrity, chronic inflammation, immune surveillance, and metabolic homeostasis through the gut liver axis, and participate in the formation of a pro tumor microenvironment. Therefore, such changes may provide more upstream risk stratification clues for the population with cirrhosis. This article summarizes previous research evidence and summarizes the common microbiome and metabolite characteristics of cirrhosis and high-risk populations, including a decrease in short chain fatty acid (SCFA) related symbiotic bacteria, an increase in inflammation related bacteria, bile acid spectrum shift, and other intestinal derived metabolite abnormalities. This article also outlines the key mechanisms that these features may correspond to, such as barrier damage and microbial translocation, immune suppression, etc. There are still significant uncertainties at present. The effect of SCFA is context dependent. Different etiologies, diets, medications, and complications can lead to significant confounding and affect cross cohort consistency. Subsequent research requires longitudinal cohort validation and the promotion of multi omics integration and the construction of interpretable predictive models to support clinical translation.

BackgroundAlport syndrome (AS) is a common hereditary kidney disease, mainly characterized by hematuria, progressive renal dysfunction, sensorineural hearing loss, and ocular symptoms, which significantly impacts patients the quality of life patients’ quality of life and lifespan. However, due to its atypical and heterogeneous clinical features, the relationship between genotype and phenotype remains complex, posing AS diagnostic challenges.MethodGenetic variants were screened by whole exome sequencing (WES) followed by verification with Sanger sequencing. Genotype-phenotype analysis was also conducted, and a novel variant (COL4A3 c.3203G>A) was selected for in vitro functional studies.ResultsWe identified seven variants in six families, including autosomal dominant (COL4A3 c.352G>A, COL4A4 c.71 + 1G>C), autosomal recessive (COL4A3 c.2736dupA, c.4235G>T), X-linked (COL4A5 c.512del,COL4A5 c.3053del), and one spontaneous variant (COL4A3 c.3203G>A). Functional studies on the novel variants (COL4A3 c.3203G>A) demonstrated a significantly decrease significant decrease in the mRNA expression level in HEK293 T cells and the weakened cell migration ability.ConclusionWe identified four novel pathogenic changes causing AS, revealing the genetic heterogeneity of AS and expanding its genotype phenotype spectrum, holding significant implications for prenatal diagnosis.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies, largely due to its highly complex and immunosuppressive tumor microenvironment (TME). In recent years, non-coding RNAs (ncRNAs), including microRNAs, long non-coding RNAs, circular RNAs, and other regulatory RNA species, have emerged as critical mediators of cellular communication and functional remodeling within the PDAC TME. These molecules regulate tumor-stroma interactions, immune evasion, angiogenesis, metabolic reprogramming, and therapeutic resistance, thereby shaping disease progression. However, the pleiotropy of ncRNAs, together with the profound cellular and molecular heterogeneity of the PDAC TME, continues to hinder mechanistic interpretation and clinical translation. In this review, we summarize current knowledge on the multifaceted roles of ncRNAs across major cellular compartments of the TME, highlight key barriers that limit their diagnostic and therapeutic potential, and discuss emerging strategies, including single-cell and spatial transcriptomics, organoid and PDX models, and nanomedicine-based delivery systems, that may accelerate the development of ncRNA-guided precision medicine in PDAC. A deeper understanding of ncRNA-mediated TME regulation is essential for improving prognostic assessment and designing more effective therapeutic interventions for this devastating disease.

OBJECTIVE:Early diagnosis of preeclampsia (PE) remains challenging. We previously linked the long non-coding (lnc) RNA MIR210HG to pathological placental development. Here, we investigated its differential expression, pathophysiological phenotypic specificity, and non-invasive diagnostic potential for PE in the peripheral blood and placental tissue. METHODS:We enrolled 88 PE patients (case group) and 81 normal pregnant women (control group) from the Affiliated Hospital of Qingdao University during January 2020-December 2021. MIR210HG expression was detected via RT-PCR and western blotting, and was modulated using siRNA/overexpression plasmid transfection in human trophoblasts and vascular endothelial cells. RESULTS:MIR210HG expression was specifically upregulated in the peripheral blood of patients with early-onset PE, with superior diagnostic efficacy for the PE-fetal growth restriction subtype. Additionally, CDHR5 was upregulated in PE-associated placental tissues (P = 0.004). Cellular assays confirmed that MIR210HG knockdown reduced CDHR5 expression in trophoblasts and endothelial cells (Bewo: P = 0.019; JEG3: P = 0.006), and CDHR5 knockdown also downregulated MIR210HG expression (Bewo: P = 0.023). CONCLUSION:Therefore, MIR210HG may contribute to placental pathological injury by regulating CDHR5, though the pathogenic mechanism remains unclear. In contrast to the invasive and lagging placental-derived lncRNA biomarkers reported in previous studies, peripheral blood MIR210HG allows for non-invasive monitoring during pregnancy, making it more clinically applicable for early screening and phenotypic stratification of PE.

Objective This study reports the rare co-occurrence of CYP4V2 (causing Bietti crystalline dystrophy, BCD) and LRTOMT (causing nonsyndromic hearing loss) variants within a single family and analyzes their clinical correlation. Methods Exome sequencing was performed on three siblings with distinct clinical phenotypes. Sanger sequencing was used for variant confirmation. Although parental data was unavailable, the inheritance pattern was analyzed through co-segregation within the family. Results The proband, affected by both conditions, carried compound heterozygous variants in both CYP4V2 and LRTOMT genes. This correlated with more severe hearing loss compared to siblings carrying only LRTOMT variants. Siblings with isolated BCD or hearing loss carried variants only in the respective single gene. The findings suggest a double-compound heterozygous state in the proband. Conclusion This case highlights the genetic complexity of multisensory disorders, where the co-inheritance of variants in distinct genes may lead to a more severe phenotype. It underscores the need for comprehensive genetic testing and counseling in affected families.

Aims Warsaw Breakage Syndrome (WABS) is a rare autosomal recessive disorder caused by biallelic variants in DDX11. This study aims to elucidate genotype-phenotype correlations and pathogenic mechanisms of novel DDX11 variants. Methods We present a detailed case report of a proband with WABS. Whole exome sequencing was performed to identify DDX11 variants. Functional validation included minigene splicing assays, analysis of mRNA/protein expression, cellular localization, protein stability, and assessments of proliferation, apoptosis, cell-cycle progression, and neuronal migration. RNA-seq was conducted on a transfected neural cell model, and findings were further validated using fetal abortion tissue from a subsequent pregnancy. Results Novel compound heterozygous variants in DDX11 were identified: a maternally inherited c.1949-3C>T variant and a paternally inherited c.2120delT variant. The c.1949-3C>T variant induced aberrant intron retention, while the c.2120delT variant led to reduced mRNA/protein levels, nuclear damage aggregation, accelerated protein degradation, impaired neuronal migration, suppressed proliferation, elevated apoptosis, and S-phase cell-cycle arrest. RNA-seq revealed marked upregulation of SERF1B, which was validated in fetal tissue. Mechanistically, SERF1B overexpression may accelerate SNCA protein aggregation and exacerbate neurotoxicity. Conclusions Pathogenic DDX11 variants aggravate neurodevelopmental disorder phenotypes through dysregulation of the SERF1B-SNCA pathway. These findings provide novel mechanistic insights into the pathophysiology of WABS.

Autism spectrum disorder (ASD) exhibits significant genetic heterogeneity, and a large number of risk genes may eventually converge on a limited number of common pathways. Among them, SCN2A, which encodes the Nav1.2α subunit of the voltage-gated sodium channel, is one of the important risk genes. This article reports a case of ASD caused by a novel mutation in SCN2A. The patient is a 6-year-old female, with the main clinical manifestations being language development delay and social communication disorders, but without epilepsy. Whole-exome sequencing revealed that she carried a heterozygous variant in the SCN2A c.4023_4077del (p.Val1343Alafs*17). This case enriches the ASD phenotype spectrum related to the SCN2A, especially providing a clinical example without comorbid epilepsy.

[This corrects the article DOI: 10.3389/fimmu.2024.1511824.].

Adams-Oliver syndrome (AOS) is a rare developmental disorder, and the DOCK6 gene is an identified AOS gene. This report highlights the prenatal diagnosis of AOS-2 by ultrasonography and genetic testing. A growth-restricted fetus with bilateral ventriculomegaly, paraventricular calcifications, and ventricular septal defect underwent trio-whole-exome sequencing (trio-WES). Functional validation of the splice-altering variant was performed via minigene assays and protein structural modeling. Trio-WES revealed compound heterozygous DOCK6 variants: a paternal frameshift (c.3190_3191del; p. Leu1064Valfs60) and a maternal splice-site variant (c.3241-1G > T). Minigene assays demonstrated that c.3241-1G > T caused intron 26 retention (486 bp), introducing a premature termination codon (p. Val1081Glufs37). Structural modeling confirmed the loss of critical DHR2 domains in both truncated proteins. This study expands the mutational spectrum of DOCK6 and underscores the importance of combining prenatal imaging with functional genomics for early diagnosis of AOS2.

BACKGROUND:SIRT1 and SIRT6, key members of the Sirtuins (SIRTs) family, play crucial roles in regulating cell metabolism and survival, providing beneficial effects against aging and metabolic diseases, which may be involved in the pathogenesis of preeclampsia (PE) by modulating cell senescence phenotypes in the placenta through antioxidant and inflammatory responses. This study aimed to investigate the association of SIRT1 and SIRT6 polymorphisms with PE in Chinese Han women. METHOD:A total of 619 healthy pregnant women were enrolled as controls, and 699 pregnant women with PE as cases. DNA was extracted from peripheral blood samples and was used for genotyping rs12415800 in SIRT1 and rs350844 in SIRT6 via real-time quantitative polymerase chain reaction (PCR). The relationship between SIRT1 and SIRT6 polymorphisms and PE was assessed by comparing genotypic and allelic frequencies between the two groups. RESULTS:The results indicated that the SIRT1 rs12415800 polymorphism may be associated with PE risk and the AA genotypes was associated with a lower risk of developing PE compared to the AG and GG (genotype χ2 = 6.612, P = 0.037; allele χ2 = 1.279, P = 0.258; OR = 0.720, 95 % CI = 0.548 ∼ 0.951, P = 0.002). Subgroup analyses (early-onset vs. late-onset PE, mild vs. severe PE) further supported these findings, showing consistent or even more pronounced associations. CONCLUSION:Our findings suggest that the genetic variant rs12415800 in SIRT1 may be linked to PE risk in Chinese Han women, with the AA genotype conferring a reduced risk. Further research is warranted to validate these findings in other populations and larger prospective studies.

IntroductionGlucose-6-phosphate dehydrogenase (G6PD) deficiency has a distinct regional and ethnic heterogeneity in distribution, and information on the molecular characteristics of G6PD deficiencies in the Heze area, Shandong Province, China, is limited. We aimed to explore the incidence and genetic mutations characteristic of G6PD enzyme deficiencies in newborns in the Heze area to investigate the pathogenicity of new G6PD mutations.MethodsWe measured G6PD activity in 114,285 neonates born in the Heze area and identified 80 patients with G6PD deficiencies. The genetic mutations in G6PD in these patients were analyzed using Sanger sequencing. Functional studies were conducted by constructing eukaryotic expression vectors, transfecting them into HEK-293T and HELA cells, and measuring the mRNA and protein levels and G6PD enzymatic activity.ResultsThe incidence of G6PD deficiency in the study population was 0.07% (80/114,285). We identified 17 mutation types with a 100% G6PD mutation detection rate, with four of them being significant: c.479G>A, c.404A>T, and c.486-7C>G being globally novel mutations, while c.682G>A has never been reported in China before. Functional studies revealed that the heterozygous missense mutations c.479G>A/p.S160N and c.404A>T/p.N135I increased mRNA levels, decreased protein expression, and reduced G6PD activity.DiscussionThe incidence of neonatal G6PD deficiency in the Heze area is low, and the most commonly mutated loci were c.1388G>A, c.487G>A, and c.1376G>T. Among these mutations, c.479G>A/p.S160N, and c.404A>T/p.N135I are potentially pathogenic. These mutations may cause G6PD deficiency via different mechanisms, thereby requiring further experimental investigation.

Context: Dual oxidases (DUOXs) are essential for thyroid hormone synthesis. Rare DUOX variations have been detected in patients with congenital hypothyroidism (CH); however, their mode of inheritance and genotype-phenotype correlations remain unclear. Additionally, no study has determined whether common DUOX variants confer a risk of CH. Objective: To elucidate the molecular and clinical characteristics of CH caused by rare and common DUOX variants. Methods: Targeted next-generation sequencing was performed on 203 trios (parents and their child with CH) to screen for rare DUOX variants. For common variants, 8 tag single nucleotide polymorphisms (SNPs) were genotyped among 298 trios and 439 healthy controls. The association between these SNPs and CH risk was analyzed using a case-control study and a family-based transmission disequilibrium test. Results The genetic contribution of rare DUOX variants to CH was 16.3% (DUOX2 14.3% and DUOXA2 2.0%). Familial cosegregation analysis suggested that DUOX variants were transmitted by an autosomal recessive pattern. These patients exhibited dyshormonogenesis and were more likely to develop into transient CH with the lower requirement of levothyroxine dose. Regarding common variants, 5 SNPs distributed across DUOXs were significantly associated with CH in both the case-control and family-based study. DUOX1 rs16939752 C > T and DUOXA1 rs3784576 C > A protected against CH, whereas DUOX2 rs269868 A > G, rs2001616 A > G and DUOXA2 rs2252371 T > C were associated with increased susceptibility to CH. Conclusion: Our research confirmed that DUOX variants are inherited in an autosomal recessive manner. We present a comprehensive spectrum of rare and common DUOX variants that provides more accurate insights into the pathogenesis of CH associated with DUOX.

Background:The absence of universal diagnosis and treatment recommendations for congenital hypothyroidism (CH) has led to suboptimal diagnostic and therapeutic outcomes. This study aimed to provide immunologically relevant evidence for the diagnosis and treatment of CH. Methods:Datasets related to CH were selected. Differentially expressed genes were screened, followed by enrichment analysis, weighted gene coexpression network analysis (WGCNA), protein-protein interaction analysis, and machine learning for the identification of hub genes. The reliability of these hub genes was verified through least absolute shrinkage and selection operator (LASSO) regression, box plot comparison, and receiver operating characteristic (ROC) curve analysis. Through gene set enrichment analysis (GSEA) of coexpressed genes, the common pathways of the hub genes and the inflammatory factors involved were identified. Immunoinfiltration analysis was carried out to verify the immunological correlation. Inflammatory factors and immune cells were screened by batch Mendelian randomization. Finally, the reliability of the hub genes, their relationships with inflammatory factors, and their impacts on cell function and the synthesis of free thyroxine (FT4) were validated through real-time quantitative polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay (ELISA), and cell proliferation experiments. Results:Multiomics analysis confirmed that APP, DDB1, MRPS5, and MRPL33 were hub genes with low expression levels in CH. These genes negatively regulated IL-2, and subsequently, through the STAT5 and MTORC1 pathways, they positively regulated (CD27 on IgD- CD38+ B cells) and (CD27 on switched memory B cells)/CD244 and negatively regulated (CD33dim HLA DR+ CD11b- Absolute Count)/IL-18 and (CD28+ CD4-CD8- T-cell %T cell)/OPG, promoting the progression of CH. Increasing the expression levels of these hub genes could increase the activity of thyroid cells and promote the synthesis of FT4 through the abovementioned pathways. Conclusions:APP, DDB1, MRPS5, and MRPL33 regulate the expression of IL-2, act on relevant immune cell subtypes through the STAT5 and MTORC1 pathways, negatively regulate IL-18 and OPG, and positively regulate CD244, thereby influencing the activity of thyroid cells and the synthesis of FT4.

IntroductionCongenital hypothyroidism (CH) is a metabolic disorder in newborns due to insufficient synthesis, abnormal secretion, or defective action of thyroid hormones. While newborn screening enables early detection, the precise etiology remains elusive in most cases, with genetic factors playing a crucial but incompletely characterized role. This study comprehensively investigated the association of the rs9789446 polymorphism with CH risk, its interactions with biological sex and clinical subtypes, and its impact on thyroid function severity.MethodsA case-control study was conducted with 306 CH patients and 441 controls. Genotyping for rs9789446 was performed using SNPscan™. Association analyses included chi-square tests, logistic regression stratified by biological sex and clinical features, linear regression for thyroid parameters, and family-based validation in 201 trios using TaqMan™ assays.ResultsThe minor G allele frequency was significantly lower in CH patients (0.348) than in controls (0.407). A protective association was observed for the G allele against CH risk (OR = 0.78, p = 0.021), with a stronger effect in males under the dominant model (OR = 0.57, p = 0.008) but no association in females, highlighting a pronounced sex-specific effect. Stratification by permanent or temporary subtypes showed no significant association, while a modest effect was detected in the goitrous subgroup under the dominant model. Initial thyroid hormone levels exhibited no significant correlation. Importantly, family-based analyses robustly validated the case-control findings.DiscussionThe rs9789446 G allele confers a sex-specific protective effect against CH, particularly in males. This supports its potential utility in genetic risk assessment and personalized screening strategies for early intervention.

BACKGROUND:Congenital hypothyroidism (CH) is the most common neonatal disorder, primarily caused by thyroid dysgenesis (TD). While the genetic cause has been identified in less than 5% of TD cases, there is an urgent need to investigate additional gene mutations that may be responsible. In 2018, TUBB1 was identified as a novel candidate gene associated with TD. Nevertheless, further research is required to confirm the role of TUBB1 in TD pathogenesis and the association between TUBB1 mutations and TD in humans. Based on the previous genetic analysis of TUBB1 in 289 Chinese TD patients, this study aimed to further validate the association between TUBB1 and TD, and to explore the pathogenic mechanisms of TUBB1 c.952C>T at the cellular level. MATERIAL AND METHODS:We performed real-time polymerase chain reaction (RT-PCR), western blot, Cell Counting Kit 8 (CCK8), and wound healing assay to evaluate the effect of TUBB1 c.952C>T on gene expression, cell proliferation, and migration. RESULTS:The c.952C>T mutant decreased the expression of TUBB1 in both mRNA and protein level, and inhibited the proliferation of thyroid cells significantly. Also, c.952C>T mutant showed restrain effects on the migration, although there was no stistical significance. Notably, pathogenic TUBB1 variants were not detected in patients with dyshormonogenesis (DH). CONCLUSIONS:TUBB1 variants confer genetic susceptibility to TD but not DH. The pathogenic variant in TUBB1 was identified in 1.38% (4/289) of our Chinese TD patient cohort, and burden test analysis revealed an association between TUBB1 variants and TD. Functional experimental results indicated that the c.952C>T mutant dominantly affects gene expression and proliferation of thyroid cells.

BACKGROUND:Oral antioxidants especially L-carnitine (LC) and N-Acetylcysteine (NAC) are commonly used as the drug treatment method for idiopathic male infertility (IMI). METHODS:Randomized controlled trials (RCTs) of LC and NAC monotherapy for IMI were searched systematically by using MEDLINE, EMBASE and the Cochrane Controlled Trials Register. The reference lists of retrieved studies were also perused. We analyzed the sperm concentration, normal morphology, sperm motility, and ejaculation volume. RESULTS:Seven Randomized controlled trials were included. Four trials compared the efficacy of LC with placebo, and three trials compared the efficacy of NAC with placebo. In the efficacy analysis, LC increased sperm concentration (p < 0.001), normal morphology (p = 0.03), and sperm motility (p = 0.02); NAC improved the first three indicators while also increasing ejaculation volume (p = 0.002). In hormone level analysis, LC increased serum testosterone levels (p < 0.001), but the changes in other hormone levels were not statistically significant. CONCLUSIONS:Both LC and NAC can improve sperm motility, sperm concentration, and normal morphology, and increase serum testosterone concentration, but have no significant effect on other serum hormones. THE PROSPERO REGISTRATION:CRD42024552120.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy with limited early diagnostic methods and therapeutic options, contributing to its poor prognosis. Recent advances in high-throughput sequencing have highlighted the critical roles of noncoding RNAs (ncRNAs), particularly PIWI-interacting RNAs (piRNAs), in cancer biology. In this review, we systematically summarize the emerging roles of piRNAs and their associated PIWI proteins in PDAC pathogenesis, progression, and prognosis. We provide a comprehensive analysis of the molecular mechanisms by which piRNAs/PIWIs regulate gene expression and cellular signaling pathways in PDAC. Furthermore, we discuss their potential as novel biomarkers for early diagnosis and therapeutic targets. Importantly, this review identifies key piRNAs/PIWIs involved in PDAC and proposes innovative strategies for improving diagnosis and treatment outcomes. Our work not only consolidates current knowledge but also offers new perspectives for future research and clinical applications in PDAC management.

Purpose: Retinitis pigmentosa (RP) is an inherited heterogeneous neurodegenerative retinal disease leading to blindness eventually. Currently, a large number of studies have explored its heterogeneity, but the genotype-phenotype correlation remains unclear. The present study aimed to explore genetic mutations and the correlation between genotype-phenotype in three RP families from the Chinese Han population. Methods: Genomic DNA was obtained from peripheral blood samples of patients and their relatives and subjected to whole-exome and Sanger sequencing. The corresponding visual acuity and fundus examinations were also performed, including fundus photography and ophthalmologic examinations. Results: In this study, three novel variants, including CERKL c.1482delT (p.Val495fs*), RPRH2 c.-5_3dup (p.Ala2Glufs*6), and RPGR c.1539del(p.Lys513Asnfs*), and a heterozygous mutation c.239-2A>G from three families were identified from three inheritance formats. All above variants were cosegregated, with the PRPH2 variant inherited in an autosomal dominant pattern, the CERKL variants in an autosomal recessive pattern, and the RPGR variant in an X-chromosome-linked recessive pattern, respectively. Conclusions: This study laid the foundation for prenatal diagnosis of RP in three family pedigrees, offering a comprehensive understanding of the genetic and clinical characteristics of patients with RP, which provided theoretical support for addressing complex genetic heterogeneity to enable accurate prenatal screening and diagnosis, early detection, and treatment of RP.

BackgroundThis study aims to identify the hub genes and immune-related pathways in acute myeloid leukemia (AML) to provide new theories for immunotherapy.MethodsWe use bioinformatics methods to find and verify the hub gene. At the same time, we use the results of GSEA enrichment analysis to find immune-related mediators. Through Mendelian randomization(MR) analysis, on the one hand, we look for related immune cells, and on the other hand, we use it to determine the causal relationship among immune cells, immune mediators, and AML. Finally, in vitro experiments are conducted to further verify and improve the reliability and physiological functions of the hub gene and its immune-related pathways.ResultsComplement Factor D(CFD) gene is identified as the highly expressed hub gene and is positively correlated with IL-2. IL-2 is also positively correlated with CD27 on CD24+CD27+B cells, JAK/STAT, and PI3K/Akt. The latter three are positively correlated with the occurrence and development of AML.ConclusionWe conclude that CFD gene uses IL-2 as a mediator to promote the disease progression of AML by promoting the CD27 on CD24+CD27+B cells, JAK-STAT, and PI3K-Akt pathways.

BackgroundAplastic anemia (AA), myelodysplastic syndromes (MDS), and acute myeloid leukemia (AML) exhibit complex pathogenic mechanisms and interrelated characteristics. We aimed to identify the common hub genes, establishing a foundation for preventing disease progression.MethodsWe selected relevant datasets from the Gene Expression Omnibus(GEO) database for differential gene expression, gene set enrichment, and weighted gene co-expression network analyses to identify hub genes, and then validated them. Subsequent analyses included immune infiltration analysis, single-cell sequencing, and cell communication analysis. We performed Mendelian randomization to screen inflammatory factors and immune cells. We used RT-qPCR, Enzyme - Linked Immunosorbent Assay(ELISA), and cell proliferation assays to validate the identified hub genes, their relationship with cellular communication mediators and inflammatory factors, and their impact on cellular function.ResultsPOLG and MAP2K7 were identified as common hub genes, with low expression observed across AA, MDS, and AML. There were distinct immune differentials among these diseases, with an enhanced correlation between immune cells and hub genes as the disease progressed. Macrophage Migration Inhibitory Factor(MIF) emerged as a key mediator of cellular communication. We identified 20 regulatory pathways of immune cells and inflammatory factors across different disease stages. In vitro validation confirmed low expression of the hub genes, which were inversely correlated with MIF and inflammatory factors, though they showed no significant impact on cell proliferation or migration.ConclusionsPOLG and MAP2K7 demonstrate crucial roles in the progression from AA to MDS and, ultimately, to AML. These genes regulate more than 20 immune regulatory pathways through MIF-mediated communication, thereby influencing disease progression.