Uncovering the cellular and omics characteristics of natural killer cells in the bone marrow microenvironment of patients with acute myeloid leukemia.

BACKGROUND: Acute myeloid leukemia (AML) is a highly heterogeneous hematologic malignancy and the most frequently acute leukemia of stem cell precursors and the myeloid derivatives in adult. Longitudinal studies have indicated the therapeutic landscape and drug resistance for patients with AML are still intractable, which largely attribute to the deficiency of detailed information upon the pathogenesis. METHODS: In this study, we compared the cellular phenotype of resident NK cells (rAML-NKs, rHD-NKs) and expanded NK cells (eAML-NKs, eHD-NKs) from bone marrow of AML patients (AML) and healthy donors (HD). Then, we took advantage of the co-culture strategy for the evaluation of the in vitro cytotoxicity of NK cells upon diverse tumor cell lines (e.g., K562, Nalm6, U937). With the aid of RNA-sequencing (RNA-SEQ) and bioinformatics analyses (e.g., GOBP analysis, KEGG analysis, GSEA, volcano plot), we verified the similarities and differences of the omics features between eAML-NKs and eHD-NKs. RESULTS: Herein, we verified the sharp decline in the content of total resident NK cells (CD3-CD56+) in rAML-NKs compared to rHD-NKs. Differ from the expanded eHD-NKs, eAML-NKs revealed decline in diverse NK cell subsets (NKG2D+, CD25+, NKp44+, NKp46+) and alterations in cellular vitality but conservations in cytotoxicity. According to transcriptomic analysis, AML-NKs and HD-NKs showed multifaceted distinctions in gene expression profiling and genetic variations. CONCLUSIONS: Collectively, our data revealed the variations in the cytobiological and transcriptomic features between AML-NKs and HD-NKs in bone marrow environment. Our findings would benefit the further development of novel biomarkers for AML diagnosis and NK cell-based cytotherapy in future.

Characterization of the biological and transcriptomic signatures of natural killer cells derived from cord blood and peripheral blood.

Longitudinal studies have indicated the pivotal role of natural killer cells (NKs) in the elimination of certain infections and malignancies. Currently, perinatal blood (PB) and cord blood (CB) have been considered with promising prospective for autogenous and allogeneic NKs transplantation, yet the similarities and differences at the biological and molecular levels are largely obscure. We isolated mononuclear cells (MNCs) from PB and CB, and compared the biological phenotypes of resident NKs by flow cytometry and cell counting. Then, we turned to our well-established "3ILs" strategy and co-culture for NK cell activation and cytotoxicity analyses, respectively. Finally, with the aid of transcriptomic analyses, we further dissected the signatures of PB-NKs and CB-NKs. CB-NKs revealed superiority in cellular vitality over PB-NKs, together with variations in subpopulations. CB-NKs showed higher cytotoxicity over PB-NKs against K562 cells. Furthermore, we found both NKs revealed multifaceted conservations and differences in gene expression profiling and genetic variations, together with gene subsets and signaling pathway. Collectively, both NKs revealed multifaceted similarities and diverse variations at the cellular and transcriptomic levels. Our findings would benefit the further exploration of the biological and transcriptomic properties of CB-NKs and PB-NKs, together with the development of NK cell-based cytotherapy.

[Clinical Analysis of 208 Patiets with BCR/ABL Negative Myeloproliferative Neoplasms].

OBJECTIVE: To analyze the incidence, hemogram, genetics, clinical manifestations, therapeutic efficacy and outcome of patients with myeloproliferative neoplasms(MPN) so as to provide much more therapeutic basis for clinically studying the pathogenesis, diagnosis, and treatment as well as evaluating the prognosis of MPN patients. METHODS: The clinical data and related laboratory test results in 208 cases of BCR/ABL fusion gene regative MPN were collected and analyzed retrospectively. RESULTS: The MPN could occur at any age, but the highest incidence was observed in patients aged 40-79. Among 208 patients with MPN, the patients with essential thrombocythemia(ET) accounted for 48.56%(101/208), the patients with polycythemia vera(PV) accounted for 25.96%(54/208), and the patients with primary myelofibrosis(PMF) accounted for 25.48(53/208). The clinical manifestation of MPN varied, the first manifestations was no-specific, onset of disease presented slow. The JAK2V617F gene mutation existed in 130 out of 208 patients with MPN, total mutation rate was 62.5%;JAK2V617F mutation rate in PV patients was 81.5%(44/54), while that in ET and PMF patients was 58.4%(59/101) and 50.9%(27/53) respectively, the detected rate of this mutation in PV patients was significantly higher than that in ET and PMF patients (P<0.05), while there was no significant difference between ET and PMF patients(P>0.05). In PV group, the WBC count of JAK2V617F positive patients was significantly enhanced (P<0.01), while there were no statistical differences of hemoglobin level and platelet count (P>0.05); in ET and PMF groups, the JAK2V617F positive patients had a higher WBC count and hemoglobin level(P<0.05), while the difference of platelet count was no significant(P>0.05). The most common vascular event in patients with MPN was ischemic cerebrovascular disease. The JAK2V617F mutation related with risk of thrombosis (OR=2.222, 95% CI=1.101 to 4.486). The difference in the incidence of vascular event between ET and PV patients was no statistically significant (P>0.05), but the incidence of vascular event in ET and PV patients was higher than that in PMF patients(P<0.05). The disease conversion much more easily happened in JAK2V617F positive patients. After treatment, the MPN could be controlled, yet the maintained treatment is needed. CONCLUSION: The MPN can occur almost at any age, but more commonly occures in middle-aged and elderly persons. The onset of MPN varies, the clinical manifestation was similar, a high detected rate of JAK2V617F mutation is observed in MPN patients and relates closely with onset of MPN; moreover, JAK2V617F mutation rate relates with type of MPN. The MPN patients with JAK2V617F mutation have higher WBC count and higher incidence of thrombosis. After treatment, the MPN can be better controlled, and need maintenance treatment. So as to avoid the reccurence of disease, control the complications and obtain the longterm survival.

[Clinical Significance of the Serum EPO Level in Patients with Iron Deficiency Anemia].

OBJECTIVE: To investigate the expression level of erythropoietin (EPO) and ferritin before and after treatment of patients with iron deficiency anemia (IDA) so as to explore their clinical significance in diagnosis and discrimination. METHODS: The EPO and ferritin levels in serum of 37 patients with IDA were determined by using chemiluminescence analysis (CLIA method) and electrical chemiluminescence analysis (ECLIA method), 30 healthy people were randomly selected as normal controls. RESULTS: (1) the sEPO level in IDA patients of group before treatment, group treated for 1 month and group treated for 2 months was higher than that in normal control group (P < 0.05). The level of sEPO of IDA patients in different groups after treatment was lower than that in IDA patients of groups before treatment, along with improvement of anemia status, the level of EPO was gradually reduced, and the level of sEPO in patients of group treated for 3 months was not statistical significant in comparison with that in normal control (P > 0.05). The level of ferritin in IDA patients before and after treatment was lower than that in normal control group (P < 0.05). The level of ferritin in IDA patient of groups after treatment was all higher than that in patients of groups before treatment, but comparision of serum ferritin level in patients of groups after treatment did not show statistical significance. (2) The level of logEPO in IDA patient before and after treatment was negatively related with level of Hb, but the level of ferritin in IDA patients was positively related with the level of Hb before treatment (r = 0.449, P = 0.005), the level of ferritin in patients of different group after treatment and in normal group did not related with level of HB. (3) The level of serum EPO in patients of severe anemia group was obviously higher than that in patients of moderate and mild anemia groups, and along with aggravation of anemia, the EPO level was gradually arised. CONCLUSION: The serum EPO is involved in the process of erythrocyte hematopoiesis, and can indicate the level of anemia, its sensitivity for anemia is higher than that of ferritin, and has important clinical value for evaluating status of diseases, observing therapeutic efficacy and judging prognosisi of IDA.