Activation of non-classical Wnt signaling pathway effectively enhances HLA-A presentation in acute myeloid leukemia.

Objective: The escape from T cell-mediated immune surveillance is an important cause of death for patients with acute myeloid leukemia (AML). This study aims to identify clonal heterogeneity in leukemia progenitor cells and explore molecular or signaling pathways associated with AML immune escape. Methods: Single-cell RNA sequencing (scRNA-seq) was performed to identified AML-related cellular subsets, and intercellular communication was analyzed to investigate molecular mechanisms associated with AML immune escape. Bulk RNA sequencing (RNA-seq) was performed to screen differentially expressed genes (DEGs) related to hematopoietic stem cell progenitors (HSC-Prog) in AML, and critical ore signaling pathways and hub genes were found by Gene Set Enrichment Analysis (GSEA), Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The mRNA level of the hub gene was verified using quantitative real-time PCR (qRT-PCR) and the protein level of human leukocyte antigen A (HLA-A) using enzyme-linked immuno sorbent assay (ELISA). Results: scRNA-seq analysis revealed a large heterogeneity of HSC-Prog across samples, and the intercellular communication analysis indicated a strong association between HSC-Prog and CD8+-T cells, and HSC-Prog also had an association with HLA-A. Transcriptome analysis identified 1748 DEGs, enrichment analysis results showed that non-classical wnt signaling pathway was associated with AML, and 4 pathway-related genes (RHOA, RYK, CSNK1D, NLK) were obtained. After qRT-PCR and ELISA validation, hub genes and HLA-A were found to be down-regulated in AML and up-regulated after activation of the non-classical Wnt signaling pathway. Conclusion: In this study, clonal heterogeneity of HSC-Prog cells in AML was identified, non-classical wnt signaling pathways associated with AML were identified, and it was verified that HLA-A could be upregulated by activation of non-classical wnt signaling, thereby increasing antigen presentation.

Clinical efficacy of recombinant human thrombopoietin combined with glucocorticoids in the treatment of immune thrombocytopenia.

OBJECTIVE: Herein, we aimed to determine the clinical efficacy of recombinant human thrombopoietin (rhTPO) combined with glucocorticoids for treating immune thrombocytopenia (ITP). METHODS: Clinical data of 87 patients with ITP admitted to our hospital were retrospectively analyzed, and patients were divided into two groups according to the treatment employed: 42 patients in the control group (CG) were prescribed glucocorticoids, and 45 patients in the study group (SG) received rhTPO combined with glucocorticoids. RESULTS: The total effective treatment rate in the SG (95.56%) was higher than that in the CG (76.19%) (P < 0.05). The SG achieved a platelet (PLT) count > 50 × 109/L faster and required fewer PLT transfusions than the CG (P < 0.05). At 1, 7, and 14 days after treatment, the PLT count increased in both groups and was higher in the SG than in the CG (P < 0.05). After treatment, CD3+, CD4+, and CD4+/CD8+ T cells increased, whereas CD8 + decreased in both groups, with the SG exhibiting a superior improvement to the CG (P < 0.05). Considering prothrombin time, activated partial thromboplastin time, and fibrinogen, differences between the two groups were not statistically significant, both before and after treatment (P > 0.05). CONCLUSION: rhTPO combined with glucocorticoids for treating ITP can effectively enhance the therapeutic effect, regulate the T lymphocyte subpopulation, rapidly increase the PLT level, and induce no significant effect on the coagulation function of patients, with good safety and high clinical promotion value.

[Effects of PTPN11 on the Biological Characteristics of AML Cells].

OBJECTIVE: To detect the expression of PTPN11 gene in acute myeloid leukemia (AML) cell line and to explore the effects of PTPN11 over expressing on proliferation and apotosis of AML cell lines. METHODS: The expression of PTPN11 in AML cell lines（HEL,U937, K562, KG-1, HL -60） was detected by RT-PCR, Q-PCR and Western blot. The PTPN11 gene was amplified by RT-PCR. PTPN11 DNA fragement and the lentiviral vector PCDH-CD513B were digested by BamHI and EcoRI, and then ligated by T4 DNA ligase. Recombinant lentivirus was generated by co-transfection of three-plasmids into 293FT cells using lipofectamine 2000. Then Q-PCR and Western blot were used to detect the expression of PTPN11 in the lentivirus infected HEL and U937 cells. The CCK-8 and Annexin V/7-AAD assays were performed to evaluate effects of PTPN11 on proliferation, apoptosis of HEL and U937 cells. RESULTS: All 5 AML cell lines expressed the PTPN11 gene, restriction analysis and gene sequencing confirmed that recombinant lentiviral vector was successfully constructed. After transfection of cells with the lentivirus, the recombinant plasmid could stably up-regulate the expression of PTPN11. Analysis of the proliferation and apoptosis of transfected AML cells indicated that as compared with the control group, the OD values of over-expression group were significantly higher and the apoptotic rates were significantly lower (P<0.05). CONCLUSION: PTPN11 is expressed in all the 5 AML cell lines. The lentiviral expression vector carrying human PTPN11 and the engineered HEL and U937 cell lines stably up-regulating PTPN11 gene expression are successfully obtained. Over-expression of PTPN11 promotes the proliferation of AML cell lines and inhibit then apoptosis.