Effects of targeted inhibition of deubiquitinase USP7/USP47 on proliferation and apoptosis of acute myeloid leukemia cells with or without Flt3-ITD mutation / 中华微生物学和免疫学杂志

Objective:To investigate the effects of ubiquitin-specific protease (USP) 7/47 inhibitor (Cat. No. 1247825-37-1) on the proliferation and apoptosis of acute myeloid leukemia (AML) cells with or without internal tandem duplications of the Flt3 gene (Flt3-ITD). Methods:ATP assay was used to detect the effects of 1247825-37-1 on the cell viability of two AML cell lines (MOLM13 and MV4-11) harboring Flt3-ITD mutation and one AML cell line (THP-1) without Flt3-ITD mutation as well as the primary Flt3-ITD-mutant and non-mutant AML cells from patient samples. Flow cytometry was used to detect the apoptosis of AML cell lines treated by different concentrations of 1247825-37-1.Results:Compared with the control group, 1247825-37-1 was able to significantly inhibit the proliferation of MOLM13, MV4-11 and THP-1 cells ( P<0.000 1). Besides, the cell viability of primary AML cells was also inhibited by 1247825-37-1, and a stronger inhibitory effect on non-mutant AML cells was observed. The USP7/USP47 inhibitor 1247825-37-1 could inhibit the proliferation of AML cells in a dose-dependent manner and a low dose (2 or 4 μmol/L) of 1247825-37-1 would be effective. Moreover, 1247825-37-1 was also able to efficiently induce the apoptosis of above AML cell lines in a dose-dependent manner. Conclusions:The USP7/USP47 inhibitor 1247825-37-1 significantly inhibits the proliferation of AML cells with or without Flt3-ITD mutation.

Progress of clonal evolutionary basis of FLT3-ITD-mutant acute myeloid leukemia and the effect of its different characteristics on the prognosis / 白血病·淋巴瘤

The incidence of FLT3-internal tandem duplication (ITD) mutation in acute myeloid leukemia (AML) is approximately 20%-30% and FLT3-ITD mutation generally indicates a poor prognosis. Although FLT3 inhibitors have greatly improved the efficacy of AML patients with this type of AML, relapse and drug-resistance increasingly become prominent. ITD mutations lead to dimerization and continuous self-activation of FLT3 in the absence of ligands, and cause non-ligand-dependent phosphorylation. Different characteristics of ITD, including allele ratio, length, insertion site, wild-type mutation content and co-mutated genes, could affect the prognosis of patients. The underlying mechanism of these factors has an important guiding significance for clinical prognosis, drug application and treatment strategy.

Knockout of BRCC3 aggravates acute GVHD in allogeneic hematopoietic stem cell transplant recipient mice / 中华放射医学与防护杂志

Objective:To investigate the effect and underlying mechanism of BRCC3 knockout on acute GVHD(aGVHD) of mice.Methods:A total of 12 recipient C57BL/6J mice were divided into two groups, including 6 wild type(WT) and BRCC3 -/-(KO). The recipients were exposed to 4.5 Gy + 4.5 Gy 60Co γ-rays in total body irradiation (TBI) at 30 min intervals. At 6 h post-irradiation, 1×10 7bone marrow cells and 8×10 6 splenocytes from BALB/c mice were infused into C57BL/6J mouse via tail vein to develop aGVHD mouse model. BRCC3 was specifically knocked out in aGVHD mouse model. The organ damage was examined through histopathology. The levels of serum cytokines were measured by enzyme-linked immuno sorbent assay (ELISA) and cytometric bead array (CBA), respectively. Spleen, liver and small intestine lymphocytes were isolated at 9 d post-transplantation, and the infiltration and activation of T cells in the target organs were assayed using flow cytometry. Results:The absence of BRCC3 in recipient mice significantly shortened survival ( P<0.05) with increased liver injury of aGVHD mice. In BRCC3 -/-recipient mice, the proportions of CD8+ T cells and CD8+ CD25+ T cells were significantly higher than those in the spleen( t=6.53, 5.52, P<0.05), and the proportions of CD8+ T cells and CD8+ CD25+ T cells were significantly increased in the liver ( t=3.74, 3.19, P<0.05). Similarly, the proportions of CD8+ T cells, CD8+ CD25+ T cells and CD8+ CD69+ T cells were significantly elevated in the small intestine ( t=3.52, 4.06, 3.29, P<0.05). Conclusions:BRCC3 deletion increased the proliferation and activation of donor CD8+ T cells and aggravated aGVHD, which might provide a new prevention and treatment target for aGVHD.

Granulocyte colony-stimulating factor-primed bone marrow: an excellent stem-cell source for transplantation in acute myelocytic leukemia and chronic myelocytic leukemia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Steady-state bone marrow (SS-BM) and granulocyte colony-stimulating growth factor-primed BM/peripheral blood stem-cell (G-BM/G-PBSC) are the main stem-cell sources used in allogeneic hematopoietic stem-cell transplantation. Here, we evaluated the treatment effects of SS-BM and G-BM/G-PBSC in human leucocyte antigen (HLA)-identical sibling transplantation.</p><p><b>METHODS</b>A total of 226 patients (acute myelogenous leukemia-complete remission 1, chronic myelogenous leukemia-chronic phase 1) received SS-BM, G-BM, or G-PBSC from an HLA-identical sibling. Clinical outcomes (graft-versus-host disease [GVHD], overall survival, transplant-related mortality [TRM], and leukemia-free survival [LFS]) were analyzed.</p><p><b>RESULTS</b>When compared to SS-BM, G-BM gave faster recovery time to neutrophil or platelet (P < 0.05). Incidence of grade III-IV acute GVHD and extensive chronic GVHD (cGVHD) was lower than seen with SS-BM (P < 0.05) and similar to G-PBSC. Although the incidence of cGVHD in the G-BM group was similar to SS-BM, both were lower than G-PBSC (P < 0.05). G-BM and G-PBSC exhibited similar survival, LFS, and TRM, but were significantly different from SS-BM (P < 0.05). There were no significant differences in leukemia relapse rates among the groups (P > 0.05).</p><p><b>CONCLUSIONS</b>G-CSF-primed bone marrow shared the advantages of G-PBSC and SS-BM. We conclude that G-BM is an excellent stem-cell source that may be preferable to G-PBSC or SS-BM in patients receiving HLA-identical sibling hematopoietic stem-cell transplantation.</p>

Therapeutic effect of CY-fTBI and BMM conditioning regimen in the process of allo-HSCT treating Ⅲ,Ⅳ non-Hodgkin lymphoma: 15 years analysis of single-center / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy of allogeneic hematopoietic stem cell transplantation（allo-HSCT）in the treatment of patients with Ⅲ,Ⅳ non-Hodgkin lymphoma（NHL）, and compared the efficacy between Cy- fractionated to talbody irradiation（fTBI）based conditioning regimen and Maryland, horse flange and mitoxantrone（BMM）.</p><p><b>METHODS</b>The clinical data of 47 patients with Ⅲ, Ⅳ NHL after allo- HSCT from November 1998 to May 2014 were collected and retrospectively analyzed. To observe the hematopoietic reconstruction recovery after transplantation, cumulative incidence of acute graft- versus- host- disease （aGVHD） and chronic graft- versus- host- disease （cGVHD）, transplantation related mortality （TRM）, recurrence rate （RR）, disease- free survival （DFS）, overall survival（OS）. Compare the efficacy of fTBI and BMM conditioning regimen at the same time.</p><p><b>RESULTS</b>Neutrophils achieving 0.5×10⁹/L and platelets achieving 50×10⁹/L on day 17 （range, 10- 72） post transplantation. Acute GVHD occurred in 53.19%, among them, grade Ⅰ-Ⅱ occurred in 42.55%, grade Ⅲ-Ⅳ occurred in 10.65%, and cGVHD occurred in 21.28%. 21 patients were alive with a median follow up of 9.7 months（0.2-149.1 months）. Overall survival（OS）was 73.5%, 49.3%, 40.1% respectively in the first, third and fifth year in Cy-fTBI group; in BMM group it was 67.8%, 32.9% and 31.4% respectively, and disease-free survival（DFS）was 65.3%, 45.6%, 30.2% respectively in the first, third and fifth year. In Cy-fTBI group, the recurrence rate（RR）and transplantation related mortality（TRM）in the first year were 18.9%, 23.0% respectively, the third year were 19.5%, 38.3% and the fifth year were 35.2%, 39.2%. In BMM group, RR and TRM in the first year were 27.4%, 24.5% respectively, the third year were 38.9%, 46.4% and the fifth year were 39.2%, 48.2%. However, there was no significant difference in the indicator of OS, DFS, RR, TRM in the two groups.</p><p><b>CONCLUSION</b>Allo-HSCT could make some Ⅲ,Ⅳ NHL patients achieve long-term disease- free survival, but the TRM was still high relatively. Moreover, compared with the program of BMM conditioning regimen, Cy-fTBI might reduce the TRM and RR, meanwhile, increase the DFS and OS. However, due to the small number cases of two groups, there was no statistical significant difference.</p>