bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Scoring System Based on Clinical, Blood Count and Immunophenotypic Data Can Predict the Risk of Disease Progression in Patients with Chronic Lymphocytic Leukemia
Authors: Alipour Sina , Leitch Heather , Vickars Linda M , Foltz Lynda M , Galbraith Paul F , Leger Chantal S , Ramadan Khaled M 
Published: April 2020

Abstract
There is limited access to advanced molecular and immunophenotypic techniques such as immunoglobulin heavy chain variable mutational status and ZAP-70 expression analysis in many parts of the world. To facilitate the development of a clinical model based on readily available clinical and well-standardized pathological data, we analyzed the well known prognostic factors in chronic lymphocytic leukemia (CLL) and proposed a scoring system. We searched the CLL database and among 465 patients (pts) we included 128 pts where the staging, blood counts, immunophenotypic and follow up data are complete. The baseline characteristics of this group and the significance of the proposed factors on the progression-free survival on univariate analysis are summarized in the table. Progression–free survival (PFS) was defined as the time from diagnosis to the time where treatment for CLL is indicated and if no treatment id indicated, the time of last follow up. In this 5 point scoring system we added one point for the presence of each one of the following factors: male gender, Rai stage 1 or more, lymphocyte count of 20 ×109/l or higher, lymphocyte doubling time less than 12 m, CD 38 expression of 20% or more. We further lumped these groups into 3 groups: no risk factors, one risk factor and tow or more risk factors. Thirty eight patients (30%) were found to have no risk factors and their 5 and 10 year PFS was 100% and 100% respectively. Forty nine pts (38%) have at least one risk factor and their 5 and 10 y PFS was 92% and 81% respectively. Forty pts (31%) have two or more risk factors and their 5 and 10 y PFS was 48% and 20% respectively. Those differences in PFS were significant (p&lt;0.00001). We conclude that readily available clinical and laboratory information in patients with CLL can be utilized to predict the risk of disease progression. Pts with no risk factors can be reliably reassured and do not require regular monitoring. Correlation of these findings with other molecular prognostic factors is needed.
Table: Risk factors for disease progression
Parameter N (%) 5-y PFS 10-y PFS p value (UVA)* *Univariate analysis Sex: Male 71 (56) 67% 43% 0.01 Female 56 (44) 91% 73% Rai stage: 0 103 (80) 87% 77% &lt;0.0001 1,2,3,4 25 (20) 49% 18% Lymphocyte count: &lt; 20×109/l 108 (84) 85% 60% &lt;0.0001 ≥20×109/l 20 (16) 40% 40% Lymphocyte doubling time ≥ 12m 111 (87) 84% 68% 0.001 &lt;12 m 17 (13) 50% 22% CD38 &lt;20% 99 (77) 83% 67% 0.006 ≥20% 29 (23) 62% 36%
Fig: Progression free survival based on the number of risk factors for disease progression (p&lt;0.00001) Fig:. Progression free survival based on the number of risk factors for disease progression (p&lt;0.00001)
Anti Hsp70 Antibodies in Untreated Patients with Chronic Lymphocytic Leukaemia
Authors: Piszcz Jaroslaw , Cichocka Edyta , Bolkun Lukasz , Galar Marzenna , Mazgajska-Barczyk Katarzyna , Milewski Marek , Kloczko Janusz 
Published: April 2020

Abstract
Background: Chronic lymphocytic leukaemia (CLL) is one of the diseases in which processes of proliferation and apoptosis are altered. It has been intensely studied in the recent years in order to understand the mechanisms of neoplastic development. Hsps or heat shock proteins are molecular chaperones involved in a number of cellular functions in stress conditions. Feng et al. (2002) have reported that heat-stressed apoptotic 12B1-D1 leukaemia cells (BCR-ABL(+)) express Hsp70 on their surface. It has also been suggested that an increased surface expression of heat shock proteins on apoptotic tumour cells results in the generation of potent antitumour T-cell responses. Furthermore, different hsps including Hsp70 have been discovered in indolent lymphoma cells. Anti Hsp70 antibodies are known to play a role in immunological and neoplastic processes. It has been well documented that this antibodies level tends to increase with age likewise an incidence of CLL. However their significance in CLL has not been clearly understood.
Aims: The aim of our study was the assessment of the anti Hsp70 antibody concentration in the patients with CLL.
Material and methods: We assessed 60 peripheral blood samples from the patients with newly diagnosed CLL. (aged 40–77), including 31 males and 29 females. A group of 20 healthy age matched subjects were used as a control group. The patients were in A-C stages of CLL according to Binet scale. Quantitative determination of anti-human Hsp70 antibodies in the serum was done using commercial test (anti Hsp70 Elisa Kits, Stressgen). The results are presented as mean ± SEM. Statistical analysis was done using Shapiro-Wilk, Mann-Whitney and Spearman’s tests.
Results: The levels of anti Hsp70 antibodies were significantly lower in the group of patients with CLL in comparison to healthy controls (105,70±16,86 ng/ml vs.243,71±60,36 ng/ml; p=0,0027). In our analysis there was no association between the levels of antibodies and the stage of the disease. There were no statistically significant correlations between the levels of anti Hsp antibodies and other parameters such as age, gender and some prognostic factors (LDH, β2-microglobulin, and lymphocyte doubling time) in the studied group.
Conclusions: The significantly lower concentrations of anti Hsp70 antibodies in CLL patients suggest that these molecules may play a role in the biology of the disease. Our study revealed lack of correlation between the level of antibodies and the disease prognostic factors. However, further studies utilizing new discovered factors are required to elucidate the role of these proteins in leukaemic biology. The encouraging results from our study suggest that it might be interesting to assess their level in CLL patient after chemotherapy.
Molecular and Functional Analysis of Stem Cell Compartment of Chronic Myelogenous Leukemia Reveals the Presence of a CD34− cell Population with Intrinsic Resistance to IMATINIB Treatment
Authors: Salvestrini Valentina , Manfredini Rossella , Fogli Miriam , Bianchi Elisa , Salati Simona , Amabile Marilina , Bertolini Francesco , Tafuri Agostino , Zini Roberta , Testoni Nicoletta , Rabascio Cristina , Rossi Lara , Martinelli Giovanni , Baccarani Michele , Ferrari Sergio , Lemoli Roberto M 
Published: April 2020

Abstract
We show the molecular and functional characterization of a novel population of lineage-negative CD34-negative (Lin−CD34−) hematopoietic stem cells from chronic myelogenous leukemia (CML) patients at diagnosis. Molecular caryotyping and quantitative analysis of BCR-ABL transcript demonstrated that about one third of CD34− cells are leukemic. CML Lin−CD34− cells showed kinetic quiescence and limited clonogenic capacity. However, stroma-dependent cultures induced CD34 expression on some cells, cell cycling, acquisition of clonogenic activity and increased expression of BCR-ABL transcript. Lin−CD34− cells showed hematopoietic cell engraftment rate in immunodeficient mice similar to Lin-CD34+ cells whereas endothelial cell engraftment was significantly higher. Gene expression profiling revealed the down-regulation of cell cycle arrest genes, genes involved in antigen presentation and processing, while the expression of genes related to tumor progression, such as angiogenic factors, was strongly up-regulated when compared to normal counterparts. Flow cytometry analysis confirmed the significant down-regulation of HLA class I and II molecules in CML Lin−CD34−cells. Imatinib mesilate did not reduce fusion transcript levels, BCR-ABL kinase activity and clonogenic efficiency of CML Lin− CD34− cells in vitro. Moreover, leukemic CD34− cells survived to BCR-ABL inhibitors in vivo. Thus, we identified a novel CD34− leukemic stem cell subset in CML with peculiar molecular and functional characteristics.
A Single Nucleotide Polymorphism in Pro-Apoptotique Protein BIM Affects Chronic Myeloid Leukaemia Evolution during Imatinib Treatment
Authors: Augis Vanessa , Belloc Francis , Airiau Kelly , Chollet Claudine , Mahon Francois-Xavier 
Published: April 2020

Abstract
The tyrosin kinase inhibitor imatinib induces apoptosis in chronic myeloid leukaemia (CML) cells. RNA interference experiments have demonstrated that the pro-apoptotic protein BIM (Bcl-2 interacting mediator of cell death) was absolutely essential to this apoptosis. Thus, mutations in BIM sequence could lead to imatinib resistance independently of BCR-ABL mutations. In the current study, we performed BIM coding sequence analysis, using complementary DNA of leukocytes from CML patients mostly secondary resistant to imatinib. A total of 58 patients were analysed: secondary resistant to imatinib (n=40), or on the contrary sensitive to it (n=18). They were in chronic phase (CP, n=43), in accelerated phase (AP, n= 12), or in blastic phase (BP, n=3). Four patients were mutated in the tyrosin kinase domain of BCR-ABL. We did not find any mutation with amino acid change on the coding sequence of BIM. However, we observed a single nucleotide polymorphism (SNP) located in the fifth exon which encode for the BH3 alpha helix part of the protein. The impact of this previously described silent polymorphism C465T (rs724710) has never been studied before. We found the 3 following genotypes : C/C homozygous, T/T homozygous and T/C heterozygous. T/T genotype was the less frequent (It represents about 5% in the European population described in the international Hapmap project). Although not significant (10%, p=0.1468), we observed an increased T/T genotype frequency in the resistant population. In this resistant group, the T/T genotype frequency was increased in advanced phases (AP+BP, 20%) as compared to CP (4%, p=0.0016) or to sensitive cohort (5.6%, p=0.0153) (Fig.1). T/T genotype could affect RNA stability, translation speed and protein folding as described by Kimchi-Sarfaty et al. for the MDR1 gene (Science2007;315:525–8). This genotype can possibly be in linkage disequilibrium with others SNPs or abnormalities (i.e. in the promoter, in splicing sites) in the non coding sequence. The analysis of this polymorphism could be useful to predict the evolution of the disease during imatinib treatment and discriminate resistant patients particularly in advanced phase of the disease. Fig 1 : Genotypes distribution of the C465T polymorphism.
Fig 1. Genotypes distribution of the C465T polymorphism. Fig 1. Genotypes distribution of the C465T polymorphism.
Preliminary Functional Studies of hnRNPK in Progression of CML
Authors: Xiaoli Liu , Zhu Hongqian , Zhang Song , Du Qingfeng , Gong Junmei , Liu Zhi , Ouyang Zhao , Ouyang Lingyun 
Published: April 2020

Abstract
The blast crisis(BC) is terminal phase of chronic myeloid leukemia (CML), which is accompanied by an increase in both BCR/ABL mRNA and protein level and imatinib(IM) resistance. The BCR/ABL oncoprotein is responsible for inducing and sustaining the leukemic phenotype through its deregulated tyrosine kinase activity which is essential for recruitment and induction of signaling pathways leading to cytokine-independent proliferation, resistance to apoptosis, and impaired differentiation of BCR/ABL-expressing myeloid and lymphoid progenitors. However, the molecular mechanisms responsible for blastic transformation remain poorly understood, although a reasonable assumption is that the unrestrained activity of BCR/ABL in hematopoietic stem/progenitor cells is the primary determinant of disease progression. We had analyzed the changes of protein expression between the bone marrow mononuclear cells in CML-CP and that in CML-BC by the technique of proteome. Compared to CML-CP, the protein expression of heterogeneous nuclear ribonucleoprotein K(hnRNPK) increases in CML-BC. In present study, the over-expression of hnRNPK in CML-BC was verified by western blotting. Furthermore, the expression of hnRNPK increases in a new imatinib-resistant BCR/ABL-positive cell line, K562-R, than in the wild-type K562. After K562 and K562-R were treated by different inhibitors such as PD98059, LY294002, AG490 and imatinib, the protein expression and mRNA level of hnRNPK were detected by western blotting and QRT-PCR. Inhibition of BCR-ABL by imatinib significantly decreased hnRNPK expression in K562 and inhibition of MEK by PD98059, downstream of the BCR-ABL signaling cascade, decreased hnRNPK expression in both K562 and K562-R cells. In addition, down-regulation of hnRNPK by small double-stranded RNA specifically complementary to hnRNPK (siRNA-hnRNPK) can inhibite cell growth and induce maximal G2/M block at 48 hours in both K562 and K562-R cells, whereas cell appototis was observed only at 72 hours by AnnexinV-PI assay. The combination of siRNA-hnRNPK and imatinib induce more cell death in K562-R cells than either treatment alone. Our data therefore suggest that hnRNPK is regulated by the BCR-ABL/MAPK cascade in Ph+ CML. The results that down-regulating hnRNPK expression induced cell-growth arrest and subsequent cell death suggest the potential therapeutic utility of this strategy in patients with CML.
Systems Understanding of Synergism Between As4S4 and Imatinib in Treating BCR/ABL Leukemia Model and in Attenuating BCR/ABL Oncoprotein as Well as Related Regulatory Networks
Authors: Zhang Qun-Ye , Mao Jian-Hua , Liu Ping , Huang Qiu-Hua , Lu Jing , Zhang Yan , Weng Lin , Chen Quan , Chen Sai-Juan , Chen Zhu 
Published: April 2020

Abstract
The combination of Imatinib and arsenic sulfide (As4S4) exerts more profound in vivo therapeutic effects on chronic myeloid leukemia (CML) at both organism and cellular levels, as evident for the first time in this paper by significantly prolonged lifespan in mouse leukemia model bearing BCR/ABL and induction of apoptosis of BCR/ABL expressing cells. To address mechanisms underlying this synergy, we performed systematic analysis of the dynamic change of proteome, phosphoproteome and transcriptome in K562 cells after As4S4 and/or Imatinib treatment with the support of principal component analysis (PCA) and self-organization maps (SOM). Moreover, protein biochemistry experiments were performed to confirm the important conclusions from multiomics study. The integrated information indicated that As4S4 promoted the unfolding protein reaction (UPR) and activity of ubiquitination pathway, which can be considered as a major biochemical basis of the pharmacological effects of this ancient medicine. In this context, As4S4 was shown to target BCR/ABL through ubiquitination of key lysine residues and led subsequently to its degradation by proteasome. Meanwhile, Imatinib inhibited the PI3K/AKT/mTOR pathway with synergism of As4S4 and arrested cell cycle. Combination of the two agents synergistically decreased activity and quantity of BCR/ABL and activated intrinsic and extrinsic apoptosis pathways. These complex multi-molecular target and multi-pathway modifications at protein level, together with those at transcriptional regulation level, ultimately resulted in the synergistic attenuation of BCR/ABL oncoprotein and the therapeutic effects on CML model.
FLT3, CD32, PU.1, ERG, uPAR, and TAP2 Are Strongly Associated with the Progression of Chronic Myeloid Leukemia and Combination of Small Interference RNA of FLT3 and STI571 Synergistically Induced Apoptosis of K562 Cells
Authors: Kim Kyung Im , Ahn Kwang-Sung , Park Juwon , Lee Chansu , Kim Byoung Kook , Lee Young-Yiul , Oh Jung Mi , Yoon Sung-Soo 
Published: April 2020

Abstract
To characterize molecular mechanisms by which transition from chronic phase to blast crisis in chronic myeloid leukemia (CML) for developing novel therapeutic targets, we analyzed gene-expression profiles of leukemic cells from 12 patients in chronic phase and 9 patients in blast crisis using a 8.7K cDNA chip. We identified 89 genes that were up-regulated as well as 54 genes that were down-regulated in blast crisis of CML. The expression profile included oncogenes, tumor suppression genes, and human genes encoding proteins involved in transcription, signal transduction, metabolism, cell growth, differentiation, apoptosis and immune functions. 18 genes were selected among the up-regulated group for analysis using real-time PCR. Real-time PCR data indicated that the expression of FLT3 (p &lt; 0.001), CD32 (p &lt; 0.001), ERG (p &lt; 0.001), uPAR (p &lt; 0.001), MAD (p &lt; 0.001) and TAP2 (p &lt; 0.001) showed statistically significant difference between chronic phase and blast crisis. For further analysis, we utilized small hairpin RNAs (shRNAs), also referred to as small interfering RNAs, to target human FLT3. These small interfering RNA constructs significantly inhibited FLT3 expression at mRNA and protein levels in K562 cells. After treating both the FLT3 knockdown cells and control cells (FLT3 wild type) with STI571, MTT assay and the expression patterns of apoptosis related genes (PARP, caspase-3, Bax) were examined. MTT assay and caspase-3 activity assay showed that silencing of the gene for FLT3 significantly reduced cell viability and ultimately facilitated the induction of apoptotic cell death by STI571. These findings uncovered evidence of a complex signaling network operating down-stream of FLT3 that actively contributes to tumor progression. Thus, RNA interference-directed targeting of FLT3 can be a potential candidate anticancer agent in association with STI571 against chronic myeloid leukemia.
HHGV678, a Novel Tyrosine Kinase Inhibitor, Inhibits the Cell Growth of Imatinib-Sensitive and Resistant BCR-ABL Positive Cells
Authors: Qiu Lin , Wang Xiao-dan , Ge Fang , Wang Xiu-li , Zhang Bo-long , Ma Jun 
Published: April 2020

Abstract
Imatinib (IM) is a higherly effective targeted drug for CML. However, some CML patients, especially accelerated and blast crisis phase, often relapse due to drug resistance resulting from the emergence of IM-resistant point mutations within the BCR-ABL tyrosine kinase domain. This stimulates the development of new kinase inhibitors that are able to override resistance to IM. HHGV678 is a novel tyrosine kinase inhibitor and we employed IM-sensitive (K562 and 32Dp210) and resistant (K562R and fifteen 32Dp210 mutants) BCR-ABL+ cell lines to compare HHGV678 with IM on growth inhibition. In addition, synergistic effect of HHGV678 with IM was observed in 32Dp210 and 5 BCR-ABL mutants frequently observed in CML patients. MTT assay results showed that the estimated IC50 value of HHGV678 for K562 and 32Dp210 were 15.5 and 28-fold, for K562R and 15 BCR-ABL mutants, were 1.4–124.0-fold lower than that of IM, indicating that HHGV678 was a more effective than IM against cell growth of IM-sensitive and resistant cells. Using combination index analysis, HHGV678 displayed synergistic growth inhibition when used with IM in BCR-ABL mutants (M244V, Q252H, Y253H, E255K and T315I). HHGV678, combined with IM at their IC50 concentration induced apoptosis 2–5 fold higher than that of HHGV678 alone in BCR-ABL mutants respectively, by annexin-V staining. At the same condition, HHGV6787 resulted in remarkable decrease in CrKL phosphorylation as determined by western blot. We conclude that HHGV678 have significant activity against IM-sensitive and resistant BCR-ABL+ cell, especially when it combined with IM that warrant further investigation in clinical trials.
Preliminary Studies on the Role of a Small GTPase, RhoA and Its Regulation in Imatinib Resistance
Authors: Zhang Song , Du Qingfeng , Zhu Hongqian , Li Rong , Liu Zhi , Yang Jun , Wang Yu , Ouyang Zhao , Xiaoli Liu 
Published: April 2020

Abstract
Imatinib has impressive response rates and good tolerability quickly led to its adoption as frontline therapy for all patients with chronic-phase CML (chronic myeloid leukemia), but the therapeutic effect of imatinib is poor in the blast crisis, and imatinib resistance has become a major problem in CML. The possible mechanisms of imatinib resistance include the amplification of BCR-ABL fusion gene and its expression increase, the point mutant of BCR-ABL kinase domain and the effects of other tyrosine kinases such as Src, Hck and Lyn and so on. However, the second-generation tyrosine kinase inhibitors (such as nilotinib and dasatinib), which were developed to overcome imatinib resistance resulting from the point mutant or the activation of other tyrosine kinases, even can not prevent all patients with CML progression to drug resistance. So there would be the other potential factor in imatinib resistance. Our previous studies generated a new imatinib-resistant BCR/ABL-positive cell line, K562-R. The 50% inhibitory concentration of imatinib was 15-fold higher in K562-R than in the wild-type K562. The expression of RhoA gene is up-regulated in K562-R by microarray analyses. RhoA, a small GTPase (24KD), has been found overexpression in breast, colon, head and neck squamous cell carcinoma, bladderand testicular cancer, lung and gastric cancer. It plays an important role in the initiation as well as the progression of human cancer, but the potential role of RhoA related to imatinib resistance has yet been unknown. In this study, we firstly detect the biologic characteristic of K562-R cells with RhoA down-expression by RNA interference. When K562-R cells were transfected with 150nM siRNA-RhoA for 48 hours, the percentage of apoptotic K562-R cells is respectively 12.82% by AnnexinV-PI assay and 9.0% by Hoechst 33258 staining and both have significant increase, cell cycle analysis found significant G0/G1 arrest, the expression of CD29 increase and that of CD71 and GPA have no difference. Secondly, The K562-R cells were treated with three selective inhibitors, including PD98059 (Ras/MAPK inhibitor), LY294002 (PI3K/AKT inhibitor) and AG490(JAK/STAT inhibitor) for 2,4 and 8 hours and the expression of RhoA were analyzed by Western-Blotting. The expression of RhoA is arrested in the K562-R cells treated with PD98059 and AG490 and no different with LY294002. These results indicate that RhoA would be an important target in the down-stream of multi-signal pathways related to imatinib resistance and the potential function of RhoA in imatinib resistance involve in increasing of cell proliferation, resistance to cell apoptosis and changes of cell adhesion.