bluuebunny/crossref_metadata_2025_split huggingface dataset:
CDKN2 Gene Deletion As Poor Prognosis Predictor Involving in the Progression of Adult B-Lineage Acute Lymphoblastic Leukemia Patients
Authors: Li Yuling , Xu Na , Zhou Xuan , Huang Jixian , Li Lin , Lu Qisi , Lu Ziyuan , He Bolin , Pan Chengyun , Liu Qifa , Xiaoli Liu 
Published: October 2019

Abstract
Background : Genes CDKN2A (MTS1) and CDKN2B (MTS2), which encode for p16INK4a/p14ARF and p15INK4b respectively from the 9p21.3 locus, are thought to be an important growth suppressor gene. INK4a/ARF regulates p53 tumor suppressor function by interacting with MDM2. P15/INK4b works as a CDK inhibitor, which inhibits phosphorylation of Rb by inactivating cyclin/ CDK-4/6 complexes and thereby controls cell-cycle G1 progression. Deletion of CDKN2A/B was well known in many hematologic malignancies, but only few report investigated this deletion effect on clinical prognosis.
Methods: This study performed analysis of the CDKN2 deletion in 215 adult B-ALL patients, and relation with cytogenetic prognostic factors (BCR/ABL; E2A/PBXl; TEL/AML1; MLL rearrangement; MYC, IGH translocation).
Results: The prevalence of CDKN2 deletions in all study population was 28.4%. There is no difference between patients with CDKN2 deletion and wild-type patients in sex, age, white blood cells(WBC) count, BM blast percentage,extra infiltration and induction complete remission(CR) rate. Analysis at relapse patients revealed increased rate of CDKN2 loss (44.6%) compared with the rate in all patients (28.4%) (p=0.006). Deletions of CDKN2 were significantly associated with poor outcomes including decreased overall survival (OS) (p<0.001), lower disease free-survival (DFS) (p<0.001), and increased cumulative incidence of relapse (p=0.002); Also, CDKN2 deletions were strongly associated with IGH translocation (p=0.021); and had an adverse effect on patients who with BCR-ABL fusion gene or MLL rearranged. Among of patients with the CDKN2 deletions , the subgroup of received Allo-HSCT treatment had a better OS and DFS compared with the subgroup received chemotherapy only (OS: p<0.000; DFS: p<0.000).
Conclusion: Patients with CDKN2 gene deletion were benefited from allogeneic hematopoietic stem cell transplantation (Allo-HSCT). Deletion of CDKN2 gene was commonly observed through leukemia progression and was a possible poor prognostic marker in long-term outcomes.

Disclosures
No relevant conflicts of interest to declare.
Tumor Vs Normal Gene Expression Profiles Identify Deregulated Pathways in ETV6/RUNX1 Positive Acute Lymphoblastic Leukemia: A Microarray-Based Meta-Analysis
Authors: Heiden Thomas , Nickel Janette , Preissner Robert , Seeger Karl 
Published: October 2019

Abstract
Acute lymphoblastic leukemia (ALL), a clinically and biologically heterogeneous disease, is the most common type of childhood cancer. Approximately 25% of B-cell precursor ALL (BCP-ALL) carry the cryptic chromosomal translocation t(12;21)(p13;q22), the most frequent genetic aberration in pediatric ALL. This translocation combines two transcription factors and essential regulators of normal hematopoiesis, ETV6 and RUNX1, generating the fusion oncogene ETV6/RUNX1 (synonym TEL/AML1). Recent studies in various animal models have strengthened the view that ETV6/RUNX1 positive cells give rise to preleukemic clones with a differentiation block in the pro/pre-B stage of B cell development that, after acquisition of additional mutations, may transform into full malignancy. However, the effects triggered by the expression of ETV6/RUNX1 and the associated additional mutated genes are not well understood.
We compared global gene expression data from ETV6/RUNX1 positive ALL and normal hematopoietic progenitor cells to identify expression signatures underlying the malignant phenotype. Our meta-analysis comprised Affymetrix HG-U133A und HG-U133 Plus 2.0 data from a total of 321 patients with ETV6/RUNX1 positive ALL, different ETV6/RUNX1 positive cell models, leukemias with 11q23 rearrangements and hyperdiploid ALL, as well as 251 samples from various normal controls including normal bone marrow, hematopoietic stem cells (HSCs) and normal B-(progenitor) cells (multipotent progenitor, pro-, pre-, immature-, naive-, centrocyte-, centroblast-, memory-, plasmablast-B cells). To eliminate potential lab batch effects, we performed RMA normalization of all microarray CEL files and next applied the batch effect removal algorithm ComBat. For determination of differentially expressed genes and pathway analyses, the R/Bioconductor packages limma and SPIA were used, respectively.
We generated a gene expression landscape of the normal B-cell development and were able to show that pre-B cells are the closest normal counterpart of ETV6/RUNX1 positive ALL. Furthermore, in comparisons with normal HSCs the p53-, mTOR-, PI3K-AKT-, Apoptosis-, and JAK-STAT signaling pathways were found to be deregulated in those ALL. The combination of multiple global gene expression data sets from ETV6/RUNX1 positive ALL offers the possibility of an integrated large-scale meta-analysis and reveals novel insights into deregulated gene networks in this type of leukemia.

Disclosures
No relevant conflicts of interest to declare.
Targeting Heme Oxygenase-1 Improves Poor Therapeutical Outcomes of BCR-ABL-Positive Adult B-Allwithikaros Isoform 6
Authors: Wang Jishi , Wang Ziming , Lin Xiaojing , Fang Qin , Chen Shuya , Huang Jun , Zhe Nana , Yu Meisheng , Zhang Yaming , Zhou Bingfeng 
Published: October 2019

Abstract
Genomic profiling studies have demonstrated that the dominant-negative Ikaros isoform 6 (IK6) is overexpressed in BCR-ABL1-positive B-lineage acute lymphoblastic leukemia (BCR-ABL1-positive B-ALL), and it is strongly associated with a poor outcome. In addition, IK6 expression leads to BCR-ABL-positive B-ALL patients insensitive to tyrosine kinase inhibitors (TKIs). Moreover, a study has revealed that Heme oxygenase-1 (HO-1), an essential survival factor, could be a potential target in treating ALL. Therefore, we were wondering whether targeting HO-1 could improve poor therapeutical outcomes of BCR-ABL-positive adult B-ALL with IK6. Firstly, we detected that IK6 existed in 20 of 42 (47.6%) adult BCR-ABL1-positive B-ALL by using reverse transcribed polymerase chain reaction (PCR) and nucleotide sequencing. We also found that early clinical response was poor in BCR-ABL1-positive B-ALL patients with IK6, and they were usually high WBC counts at diagnosis and a high relapse rate. Compared to healthy donors' CD34+ cells, the levels of HO-1 expression were higher in primary CD34+ cells derived from adult BCR-ABL1-positive B-ALL patients with IK6 by qRT-PCR and western blot studies. And there was a strong correlation between the expression of IK6 and HO-1. In primary CD34+ leukemic cells derived from our IK6-positive patients' pool, silencing HO-1 resulted in the reduce of proliferation and a substantial proapoptotic effect, including a significantly enriched subG1 population, an increase of Annexin V-positive cells and of caspase3 cleavage. Furthermore, silencing HO-1 also increased their sensitivity to TKIs. This phenomenon was also supported by the observation that ZnPP IX substantially augmented the growth-inhibitory effects of imatinib on primary leukemic cells. Finally, we found that IK6 led to STAT5 activiation, and HO-1 was one of the downstream target genes of STAT5. In conclusion, the existence of extra IK6 seems to exacerbate poor prognosis and risks of relapse in adult BCR-ABL1-positive B-ALL. HO-1 plays an important role as a survival factor in BCR-ABL1-positive B-ALL with IK6, and targeting HO-1 can probably improve the clinical outcomes.

Disclosures
No relevant conflicts of interest to declare.
Dynamic Telomere Shortening and Chromosomal Instability in Irradiated CD34+ Cells Transduced with TP53 Hotspot Mutations R175H, R248W and R249S
Authors: Salari Azam , Thomay Kathrin , Schienke Andrea , Hagedorn Maike , Ebersold Juliane , Krueger Andreas , Schambach Axel , Schlegelberger Brigitte , Göhring Gudrun 
Published: October 2019

Abstract
Patients with MDS and a complex karyotype have a very short median survival and a high risk of transformation into AML. We showed earlier that TP53 mutations are associated with complex karyotype and disease progression. However, it is poorly understood how TP53 mutations contribute to the induction of chromosomal instability in hematopoietic stem and progenitor cells.
We therefore established a long-term cell culture (LTC) model and investigated the role of TP53 in human CD34+ hematopoietic stem and progenitor cells (HSCs) isolated from cord blood and in HT1080 cells over 6 weeks. We chose 3 different modifications: 1) TP53-deficient HSCs via shRNA knockdown, 2) HSCs with different lentivirally introduced TP53 hotspot mutations (R248W, R175H, R273H, R249S) and 3) HT1080 cells with different lentivirally introduced TP53 hotspot mutations (R248W, R175H, R273H, R249S). We performed each LTC at least three times. In order to stress the cells and induce chromosomal instability, we irradiated half of the cells. Besides functional assays in the first week, we performed detailed cytogenetic analysis including telomere length measurement at weeks 1, 3 and 6.
TP53 mutations and the downregulation of TP53 led to impaired hematopoiesis with decreased erythroid differentiation, increased apoptosis and decreased proliferation. None of the modifications induced chromosomal instability in cells without irradiation. In the irradiated cells, all cells carrying a TP53 mutation or a TP53 downregulation developed a chromosomal instability in comparison to the cells transduced with control vectors. However, no stable complex clones developed. Telomeres shortened during follow-up in HSCs carrying the mutations R175H, R248W and R249S. No other cells showed a dynamic response in telomere length. In order to analyze the DNA repair capacity, we performed yH2AX foci assay. Surprisingly, the same cells which showed a telomere shortening showed a lower amount of foci. This could be due to a lower amount of double-strand breaks or to a lower ability to form foci.
In summary, TP53 mutations and the downregulation of TP53 led to an increased chromosomal instability in irradiated cells only. Modifications alone did not lead to the development of complex karyotypes. Furthermore, only the mutations R175H, R248W and R249S led to a telomere shortening. In conclusion, a TP53 mutation seems to require additional passenger mutations in order to lead to MDS or AML with complex karyotype.

Disclosures
No relevant conflicts of interest to declare.
Peg-PLGA-PLL Nanoparticles in Combination with Gambogic Acid for Reversing Multidrug Resistance of K562/A02 Cells to Daunorubicin
Authors: Xu Peipei , Wang Ruju , Ouyang Jian , Chen Bing 
Published: October 2019

Abstract
Purpose: The present chemotherapy in malignancy treatment, including leukaemia, is plagued by one main problem: multidrug resistance (MDR) which is often related to excessive expression of adenosine triphosphate-dependent efflux pump. In recent years, attempts have been focused on searching competitive inhibitors. However, high toxicity and severe side effects were the main impediments for the application of inhibitors.
Methods: this study investigated a new strategy to utilise polyethylene glycol-polylactic-co-glycolic acid-poly-L-lysine (PEG-PLGA-PLL) as a delivery system to combine with gambogic acid (GA) with the purpose of delivering daunorubicin (DNR) to the target location to reserve MDR with maximised therapeutic efficacy and minimised side effects. The anti-tumour ability of DNR in the presence of GA-PEG-PLGA-PLL was evaluated by methyl thiazolyl tetrazolium (MTT) assay, flow cytometry and 4',6-diamidino-2-phenylindole (DAPI) staining. The possible signalling pathway was demonstrated by quantitative real-time polymerase chain reaction (QPCR) and Western blot assay.
Results: The intracellular concentration of DNR can evidently increase with the presence of GA-PEG-PLGA-PLL, and the resistance of K562/A02 cells to DNR can be reversed when combined with GA-PEG-PLGA-PLL, which acted as a delivery system. The anti-tumour efficiency was promoted.
Conclusion: GA-PEG-PLGA-PLL can be a promising agent in reversing MDR of K562/A02 to DNR.

Disclosures
No relevant conflicts of interest to declare.
Drug Induced MLL Fusion Degradation Via Hsp90 Inhibition
Authors: Cantilena Sandra , Goulden Nicholas , Williams Owen , de Boer Jasper 
Published: October 2019

Abstract
The survival rate for infants is less than 50%. The fast majority of infant acute leukemias are characterized cytogenetically by balanced chromosomal translocations involving the mixed lineage leukemia (MLL) gene. Leukemic therapies that degrade the driver oncogene are associated with loss of cancer cell self-renewal and excellent cure rates. Therefore, therapy that degrades the MLL fusion gene would offer new hope to these patients.
Recently it was shown that the Drosophila Trithorax gene, an analogue of the human MLL, is degraded by Radicicol. Radicicol is a natural compound and a well-known Hsp90 inhibitor.
Here, we show that Radicicol is able to induce a dose dependent degradation of the MLL-fusion protein in a panel of human MLL rearranged cell lines and in human cord blood-derived MLL-AF9 immortalised myeloid cells. This drug induced degradation of the MLL-fusion gene results in down-regulation of the expression of MLL target genes, including HOXA9, MEIS1 and c-MYB. Functionally, this results in a loss of self-renewal of the leukemic stem cells, as shown by methylcellulose colony forming assays.
Radicicol proved ineffective and too toxic for in vivo use. One of the best tolerated Hsp90 inhibitors is Ganetespib. It is currently in phase II/III clinical trials.
We extended our Radicicol data to Ganetespib. Like Radicicol, Ganetespib induces MLL-fusion protein degradation and downregulation of MLL target genes. Treatment of MLL rearranged leukemia with Ganetespib results in a loss of leukemic stem cell activity.
In conclusion, we show how the inactivation of the MLL-fusion and down-regulation of MLL target genes results in a block of leukemic stem cell self- renewal. We will validate these findings in a pre-clinical in vivo model in the near future.

Disclosures
No relevant conflicts of interest to declare.
Tissue Transglutaminase Regulates Steroid Resistance in Acute Lymphoblastic Leukemia Via Nuclear Factor-Kappa B Related Signaling Pathways
Authors: Jung Hyun Joo , Han Eunhee , Park Jun Eun 
Published: October 2019

Abstract
Steroid resistance is lethal to the patients with acute lymphoblastic leukemia (ALL). To improve survival for steroid-resistant ALL patients, it is necessary to develop innovative therapeutic strategies to overcome steroid resistance. Tissue transglutaminase (TG2) is an intracellular calcium-dependent protein cross-linking enzyme reported to be over-expressed in various metastatic or chemotherapy-resistant cancer cells including lymphoma cells. Chronic expression of TG2 constitutively activates nuclear factor kappa B (NF-kappaB) and it is directed linked to NF-kappaB-related signaling networks including hypoxia-inducible factor-1 (HIF-1), Zeb, Snail, or Twist that influence drug resistance or metastasis in inflammatory or cancer cells. In the present study, we used biochemical and molecular methodologies to demonstrate that steroid resistance was associated with TG2-regulated signaling pathways including NF-kappaB and its downstream transcriptional factors in steroid-resistant ALL. We generated steroid-adapted subclones of ALL cell lines that were extremely more resistant to steroid than the parent cells. We found that steroid-resistant ALL cells expressed elevated levels of TG2 and the modification of TG2 activities altered NF-kappaB expression in this cells. TG2-related NF-kappaB activation modulated the expression of HIF-1 and NF-kappaB-targeted genes such as Zeb, Snail, and Twist in steroid-resistant ALL cells. Also, TG2 inhibition improved the cytotoxicity of steroid in steroid-resistant ALL cells. This is the first study to show the link between TG2, NF-kappaB, its downstream signaling networks, and steroid resistance in ALL. TG2 inhibition could be used as an alternative target to overcome steroid resistance in ALL. Furthermore, TG2 and HIF-1 expression could serve as new biomarkers of steroid resistance evolution or cancer recurrence in ALL.

Disclosures
No relevant conflicts of interest to declare.
FLAG-IDA As First Line Induction Treatment for Children De Novo AML
Authors: Feng Xiaoqin , Li Chunfu , He Lan 
Published: October 2019

Abstract
Objective: The complete remission after induction therapy is very important for the prognosis of AML. Fludarabine, cytarabine, granulocyte colony-stimulating factor (G-CSF), and idarubicin (FLAG-IDA) regimen has been proven to be an effective chemotherapy for relapsed or refractory acute myeloid leukemia. The aim of this study was to evaluate complete remission rate and toxicity in children with de novo acute myeloid leukemia (none APL) who received the FLAG-IDA regimen as induction therapy.
Method: Between March 2014 and July 20015, 14 children with de novo acute myeloid leukemia (none APL) were received FLAG-IDA regimen as first line induction treatment in our center. The regimen including: Fludarabine 30 mg/(m2.d), PI 0.5hr, qd, d2-6;
Ara-C 2g/(m2.d), PI 3hr, d2-6; Ida (Idarubicin) mg/(m2.d), PI 1hr, qd, d4-6; G-CSF 5μg/(Kg.d), s.c./i.v. qd,d1-7.
Of the 14 children, age 1-13 years old (median age 7 years old), including 2 (14.3%) children with favorable gene, 4 (28.6%) children with high risk gene. 3(21.4%) children received 1 cycle, 11 (78.6%) received 2 cycles of FLAG-IDA regimen. The CR rate and toxicity in total 25 cycles were evaluated.
Result: After 25 cycles of FLAG-IDA regimen, only 1 child (7.1%) did not get CR, 13/14 (92.9%) got CR after 1 or 2 cycles of FLAG-IDA induction treatment. In 25 FLAG-IDA cycles, the neutropenic time was from 13-43 days (mean 24.8 days). In the induction period, 2(14.3%) children suffered sepsis, 3 children (21.4%) had proven invasive fungal infection. Of 14 children, 2 children(14.3%) combined with cutaneous anaphylaxis, 2 children combined with transient fever after venous transfusion of chemicals. None obvious cardiac toxicity (arrhythmia or heart failure) was proved. One child died of ARDS in the neutropenic period after 2nd FLAG-IDA cycle. Of 14 children, 12 children (85.7%) got continues CR with the follow up 1-19 months (median 8 months).
Discussion: Our primary study showed quite high CR rate combined with quite toxicity but can be tolerated using the FLAG-IDA regimen as first line induction treatment. Comprehensive supportive care should be given during the induction therapy. The LFS and OS need to be feedback after long time follow up.

Disclosures
No relevant conflicts of interest to declare.
Arsenic Trioxide As a Front-Line Therapy for Newly Diagnosed Acute Promyelocytic Leukemia
Authors: Chen Weihong , Du Xin , Zhuo Jiacai 
Published: October 2019

Abstract
PURPOSE:
Arsenic trioxide (ATO) is a very effective drug for acute promyelocytic leukemia (APL). However, little has been reported concerning its use as front-line therapy for complete remission of newly diagnosed APL with t(15;17) chromosomal abnormality.
METHODS:
The adult patients newly diagnosed with APL with t(15;17) chromosomal abnormality (which would invariably lead to a positive PML/RAR alpha fusion gene) were received 4-5 weeks of ATO administered intravenous drip at a daily dose of 10mg(concentration 2%)/day as induced chemotherapy. When a complete remission was achieved by the ATO induction, a three weeks interval would be observed, after which 5 + 3 days of maintenance (with cytarabine and anthracycline, respectively) would be carried out. A further interval of 2 weeks would then take place, after which all-trans retinoic (ATRA) would be administered oral for another 2 weeks. A final interval of 2 weeks after the administration of ATRA would take place before a recurrence of the chemotherapy regimen, which would restart again with ATO induction. After PML/RAR alpha fusion gene Q-PCR tested negative, the regimen would continue recurring, ensuring negativity of the gene for 2 years. After 2 years the regimen would end, and PML/RAR alpha fusion gene of the patients were to be observed/monitored once every 3-6 months for 10 years. Central nervous system leukemia (CNSL), which might be caused by APL, was to be prevented by lumbar punctures chemotherapy once every 6-8 weeks after complete remission of the patients. Six times were taken.
RESULTS:
Out of 496 for newly diagnosed APL from 2005 to 2015, we identified 113 patients as newly diagnosed APL. All patients achieved complete response after treatment with ATO induced chemotherapy. Sixty-two out of 65 (95.38%) patients with APL were cured with ATO induction and subsequent 5 + 3 maintenance (cytarabine and anthracycline) therapy from 2005 to 2010. The other three patients were relapsed or had refractory disease and then died. Forty-six out of 48 patients (95.83%) achieved complete cytogenetic remission from 2011 to 2015. The other two patients were relapsed or had refractory disease and then died. All patients had no serious adverse events. Five patients had dyspnea, fever, weight gain, peripheral edema and were successfully treated with dexamethason and furosemide.
CONCLUSIONS:
Arsenic trioxide is a very effective drug as the front-line therapy for newly diagnosed acute promyelocytic leukemia.

Disclosures
No relevant conflicts of interest to declare.