Prognosis Of Acute Lymphoblastic Leukemia Research Articles

Effect of sustained measurable residue disease negativity and post-remission treatment selection on the prognosis of acute lymphoblastic leukemia in adults.

To explore the effects of monitoring measurable residual disease and post-remission treatment selection on the clinical outcomes of B-cell acute lymphoblastic leukemia (B-ALL) in adults. Between September 2010 and January 2022, adult patients with B-ALL who received combination chemotherapy, with or without allogeneic hematopoietic stem cell transplantation (allo-HSCT), were included in the retrospective study, which was approved by the Ethics Committee and the observation of Declaration of Helsinki conditions. One hundred and forty-three B-ALL patients achieved complete remission (CR) were included in the study, of whom 94 patients (65.7%) received allo-HSCT in first complete remission (CR1). Multivariate analysis showed that the most powerful factors affecting OS were transplantation (hazard ratio [HR] = 0.540, p = 0.037) and sustained measurable residue disease (MRD) negativity (HR = 0.508, p = 0.037). The subgroup analysis showed that the prognosis of the allo-HSCT group was better than that of the chemotherapy group, regardless of whether MRD was negative or positive after two courses of consolidation therapy. After consolidation therapy, the prognosis of patients with positive MRD remained significantly better in the allo-HSCT group than in the chemotherapy group. However, no significant difference was observed in the prognosis between the allo-HSCT and chemotherapy groups with negative MRD after consolidation therapy. B-ALL patients who achieve sustained MRD negativity during consolidation therapy have excellent long-term outcomes even without allo-HSCT. Allo-HSCT is associated with a significant benefit in terms of OS and DFS for patients who were with positive MRD during consolidation therapy.

Importance of genetic diagnosis for treatment and prognosis in acute lymphoblastic leukaemia (ALL) — a case report

Importance of genetic diagnosis for treatment and prognosis in acute lymphoblastic leukaemia (ALL) — a case report

T(1;4) translocation in a child with acute lymphoblastic leukemia: a case report

BackgroundAcute lymphoblastic leukemia is the most common childhood cancer, with an 80% frequency in children between 1 and 10 years old. The outcome and prognosis of acute lymphoblastic leukemia in children depends on various factors, such as age, clinical and biological features, and cytogenetic factors.Case presentationWe report the case of a pediatric patient, a 4-year-old Moroccan female who was referred to the Hematology and Oncology Department of 20 August 1953 Hospital in Casablanca and diagnosed with B-cell acute lymphoblastic leukemia associated with a rare genetic chromosomal abnormality.ConclusionTranslocation (1;4)(p21;p15) is a relatively rare chromosomal abnormality found in human leukemia and was never described isolated in pediatric B-cell acute lymphoblastic leukemia patients. It showed a good evolution by complete remission and recovery of this patient after receiving all chemotherapy and after 8 years of follow-up.

PARP1 Characterization as a Potential Biomarker for BCR::ABL1 p190+ Acute Lymphoblastic Leukemia

Detection of t(9;22), and consequent BCR::ABL1 fusion, is still a marker of worse prognosis for acute lymphoblastic leukemia (ALL), with resistance to tyrosine-kinase inhibitor therapy being a major obstacle in the clinical practice for this subset of patients. In this study, we investigated the effectiveness of targeting poly-ADP-ribose polymerase (PARP) in a model of BCR::ABL1 p190+ ALL, the most common isoform to afflict ALL patients, and demonstrated the use of experimental PARP inhibitor (PARPi), AZD2461, as a therapeutic option with cytotoxic capabilities similar to that of imatinib, the current gold standard in medical care. We characterized cytostatic profiles, induced cell death, and biomarker expression modulation utilizing cell models, also providing a comprehensive genome-wide analysis through an aCGH of the model used, and further validated PARP1 differential expression in samples of ALL p190+ patients from local healthcare institutions, as well as in larger cohorts of online and readily available datasets. Overall, we demonstrate the effectiveness of PARPi in the treatment of BCR::ABL1 p190+ ALL cell models and that PARP1 is differentially expressed in patient samples. We hope our findings help expand the characterization of molecular profiles in ALL settings and guide future investigations into novel biomarker detection and pharmacological choices in clinical practice.

Impact of XRCC3 rs861539 and XPD rs13181 gene polymorphisms on childhood acute lymphoblastic leukemia in Egyptian patients

Abstract Background There is mounting evidence that the genotypes of DNA repair proteins and susceptibility to certain malignancies are related. Few studies, however, have examined the role of the homologous repair gene X-ray repair cross-complementing group 3 (XRCC3) genotype and xeroderma pigmentosum complementation group D (XPD) in the development or prognosis of acute lymphoblastic leukaemia (ALL). Aim of the study In this study, we investigated the impact of XRCC3 rs861539 and XPD rs13181 polymorphisms on the risk of ALL. To the best of our knowledge, this is the first report of XRCC3 and XPD polymorphisms in ALL Egyptian patients. Methods Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) was used to analyse XRCC3 rs861539 and XPD rs13181 gene polymorphisms in 96 patients with ALL and in 103 disease-free controls, who were of a similar age. Results ALL risk is lower in individuals with the homozygous variant TT genotype at XRCC3 rs861539. The heterozygous variant CT genotype of XRCC3 was connected to increased disease risk of ALL in males. Additionally, C allele frequency was noticeably higher than T allele frequency in pre B ALL. In this investigation, there was no correlation between the XPD Lys751 rs13181 polymorphism and risk of ALL. Conclusion Our research reveals that genetic variation in the genes for DNA repair may influence ALL susceptibility.

Characteristics of Molecular Genetic Mutations and Their Correlation with Prognosis in Adolescent and Adult Patients with Acute Lymphoblastic Leukemia

Introduction: The prognosis of acute lymphoblastic leukemia (ALL) in adolescents and adults is poor, and recurrence is an important cause of their death. Changes of genetic information play a vital role in the pathogenesis and recurrence of ALL; however, the impact of molecular genetic mutations on disease diagnosis and prognosis remains unexplored. This study aimed to explore the frequency spectrum of gene mutations and their prognostic significance, along with the minimal residual disease (MRD) level and hematopoietic stem cell transplantation (HSCT), in adolescent and adult patients aged ≥15 years with ALL. Methods: The basic characteristics, cytogenetics, molecular genetics, MRD level, treatment regimen, and survival outcome of patients with untreated ALL (≥15 years) were collected, and the correlation and survival analysis were performed using the SPSS 25.0 and R software. Results: This study included 404 patients, of which 147 were selected for next-generation sequencing (NGS). NGS results revealed that 91.2% of the patients had at least one mutation, and 67.35% had multiple (≥2) mutations. NOTCH1, PHF6, RUNX1, PTEN, JAK3, TET2, and JAK1 were the most common mutations in T-ALL, whereas FAT1, TET2, NARS, KMT2D, FLT3, and RELN were the most common mutations in B-ALL. Correlation analysis revealed the mutation patterns, which were significantly different between T-ALL and B-ALL. In the prognostic analysis of 107 patients with B-ALL, multivariate analysis showed that the number of mutations ≥5 was an independent risk factor for overall survival and the RELN mutation was an independent poor prognostic factor for event-free survival. Discussion: The distribution of gene mutations and the co-occurrence and repulsion of mutant genes in patients with ALL were closely related to the immunophenotype of the patients. The number of mutations ≥5 and the RELN mutation were significantly associated with poor prognosis in adolescent and adult patients with ALL.

Association of CDKN2A/B mutations, PD-1, and PD-L1 with the risk of acute lymphoblastic leukemia in children

PurposeCurrently, the significance of CDKN2A/B mutations in the pathogenesis and prognosis of acute lymphoblastic leukemia (ALL) is inconclusive. In this study, we analyzed the genetic and clinical features of children with CDKN2A/B mutations in ALL. In addition, we evaluated the expression and significance of programmed cell death protein 1 (PD-1) and programmed cell death ligand 1 (PD-L1) in serum and explored their role in the susceptibility of childhood ALL.MethodsWe sequenced CDKN2A/B in the peripheral blood of 120 children with ALL and 100 healthy children with physical examination. The levels of CD4+ T, CD8+ T, and NK cells were measured by flow cytometry (FCM). Furthermore, the expression of PD-1 and PD-L1 was detected by ELISA.ResultsWe found 32 cases of CDKN2A rs3088440 and 11 of CDKN2B rs2069426 in 120 ALL children. Children with ALL in the CDKN2A rs3088440 were more likely to have hepatosplenomegaly (P = 0.019) and high risk (P = 0.014) than the wild group. In contrast, CDKN2B rs2069426 was more likely to develop lymph node metastasis (P = 0.017). The level of PD-L1 in the serum of ALL children was significantly higher than that of the control group, and there was no significant difference in PD-1 (P < 0.001). Additionally, children with CDKN2A rs3088440 had reduced CD8+ T cell counts than the wild group (P = 0.039).ConclusionCDKN2A rs3088440 and CDKN2B rs2069426 may be related to the occurrence and development of ALL in Chinese children. Additionally, PD-1/PD-L1 may be involved in the immune escape process of ALL, which is expected to become a new target for the treatment of the disease.

Effect of XRCC1 Arg399Gln Gene Polymorphism on Survival in Lymphoblastic Leukemia.

The relevance of the research of the article is conditioned upon the problem of the development of molecular genetic diagnostics to determine the effectiveness of treatment for acute lymphoblastic leukemia in children. The purpose of the article is to identify the polymorphism parameters of the P53 Arg72Pro and XRCC1 Arg399Gln genes in acute lymphoblastic leukemia with criteria for determining the survival rates of sick children. Methods for the study of the identified problem are the study of the medical histories of children with acute leukemia, which allowed selection of the necessary contingent of patients for further genetic study of their frozen blood, where the genomic part of deoxyribonucleic acid was isolated from the frozen blood in a standard way using molecular biological research when performing a polymerase chain reaction. The article presents the results of a study that shows that in children with acute lymphoblastic leukemia, the frequency of genotypes of the XRCC1 Arg399Gln gene is variable. The most common genotypes are Arg/Gln and Arg/Arg, approximately 48% each. The Gln/Gln genotype is less common. Relapse-free survival of children with the Arg/Gln and Gln/Gln genotypes was the highest, slightly lower rates were noted with the Arg/Arg genotype. It was identified that the frequency of genotypes of the XRCC1 Arg399Gln gene can be a predictor of prognosis in acute lymphocytic leukemia in children, which can be considered when choosing treatment tactics, and this has practical significance for the field of medicine.

Mutation of the TP53 gene in acute lymphoblastic leukemia does not affect survival outcomes after haploidentical hematopoietic stem cell transplantation.

Previous studies have demonstrated that TP53 mutation is correlated with insufficient therapy response and unfavorable prognosis in acute lymphoblastic leukemia (ALL). Few studies have investigated the impact of TP53 mutation in ALL patients after haploidentical hematopoietic stem cell transplantation (haplo-HSCT). We completed a retrospective study of 65 ALL patients with available TP53 status who underwent haplo-HSCT. They were divided into a TP53 mutation group (TP53mut ) and a TP53 wild-type (TP53wt ) group. TP53mut showed comparable 2-year cumulative incidence of relapse (CIR) rates (13.1% vs 12.5%, P=.96) and 2-year leukemia-free survival (LFS) (74.2% vs 77.4%, P=.80) with TP53wt . No significant differences in 2-year overall survival (OS) rates (82.9% vs 87.3%, P=.61) or 2-year NRM rates (12.7% vs 10.2%, P=.69) were observed in TP53mut and TP53wt patients. Multivariate analysis suggested that white blood cell (WBC) count at initial diagnosis (>50 × 109 /L: hazard ratio [HR]=3.860, P=.016) and age (>40 years old: HR=4.120, P=.012) are independent risk factors for 2-year LFS. Our study showed that TP53 mutations may not be related to the unfavorable impact on survival in ALL patients after treatment with haplo-HSCT. The present results suggested that haplo-HSCT may eliminate the poor prognosis effect of TP53 mutation in ALL.

Impact of IKZF1 Deletions in the Prognosis of Childhood Acute Lymphoblastic Leukemia in Argentina.

Simple SummaryAn association of deletions in the IKZF1 gene (IKZF1del) with poor prognosis in acute lymphoblastic leukemia (ALL) has been demonstrated. However, the co-occurrence of IKZF1del with deletions in other genes (IKZF1plus) seems to better define prognosis. Our aim was to analyze the influence of IKZF1del and/or IKZF1plus profiles on the survival of children with ALL. We analyzed 1023 eligible cases who were classified into three subsets: IKZF1not-del: IKZF1-not-deleted (n = 835), IKZF1del: IKZF1deleted (n = 94) and IKZF1plus: IKZF1del plus deletion of other genes (n = 94). Leukemia-free-survival probability (standard deviation) (LFSp(SE)) was 75 (2)% for IKZF1not-del, 51 (6)% for IKZF1del and 48 (6)% for IKZF1plus (p-value < 0.00001). The LFSp(SE) according to ALL-IC risk-group stratification showed a statistically significant difference within Intermediate-Risk patients, LFSp (SE) being 77 (2)% for IKZF1not-del, 61 (10)% for IKZF1del and 54 (7)% for IKZF1plus (p-value = 0.0005). The LFSp(SE) of the high-risk group was: 61 (4)% IKZF1not-del, 38 (8)% IKZF1del and 35 (9)% IKZF1plus (p-value = 0.0102). Thus, the IKZF1 status was shown to define a population of patients with a poor outcome, mainly in those with intermediate risk. However, not-significant differences were observed between the LFSp of IKZF1del versus IKZF1plus groups. The study of the IKZF1 status should be incorporated into the risk-group stratification of pediatric ALL.An association of deletions in the IKZF1 gene (IKZF1del) with poor prognosis in acute lymphoblastic leukemia (ALL) has been demonstrated. Additional deletions in other genes (IKZF1plus) define different IKZF1del subsets. We analyzed the influence of IKZF1del and/or IKZF1plus in the survival of children with ALL. From October 2009 to July 2021, 1055 bone marrow samples from patients with ALL were processed by Multiplex ligation-dependent probe amplification (MLPA). Of them, 28 patients died during induction and 4 were lost-in-follow-up, resulting in an eligible 1023 cases. All patients were treated according to ALLIC-BFM-2009-protocol. Patients were classified into three subsets: IKZF1not-deleted (IKZFF1not-del), IKZF1deleted (IKZF1del) and IKZF1del plus deletion of PAX5, CDKN2A, CDKN2B and/or alterations in CRLF2 with ERG-not-deleted (IKZF1plus). The LFSp and SE were calculated with the Kaplan–Meier calculation and compared with a log-rank test. From the 1023 eligible patients, 835 (81.6%) were defined as IKZF1not-del, 94 (9.2%) as IKZF1del and 94 (9.2%) as IKZF1plus. Of them, 100 (9.8%) corresponded to Standard-Risk (SRG), 629 (61.5%) to Intermediate-Risk (IRG) and 294 (28.7%) to High-Risk (HRG) groups. LFSp(SE) was 7 5(2)% for IKZF1not-del, 51 (6)% for IKZF1del and 48 (6)% for IKZF1plus (p-value < 0.00001). LFSp(SE) according to the risk groups was: in SRG, 91 (4)% for IKZF1not-del, 50 (35)% IKZF1del and 100% IKZF1plus (p-value = ns); in IRG, 77 (2)% IKZF1not-del, 61 (10)% IKZF1del and 54 (7)% IKZF1plus (p-value = 0.0005) and in HRG, 61 (4)% IKZF1not-del, 38 (8)% IKZF1del and 35 (9)% IKZF1plus (p-value = 0.0102). The IKZF1 status defines a population of patients with a poor outcome, mainly in IRG. No differences were observed between IKZF1del versus IKZF1plus. MLPA studies should be incorporated into the risk-group stratification of pediatric ALL.

Clinical features and prognosis of acute lymphoblastic leukemia in children with Epstein-Barr virus infection

BackgroundAcute lymphoblastic leukemia (ALL) is one of the most common malignant diseases of the hematopoietic system in children. Although the etiology of ALL is unknown, it has been reported that it may be associated with Epstein-Barr virus (EBV) infection. The aim of this study was to analyze the impact of EBV infection on the clinical features and prognosis of childhood ALL.MethodsA total of 162 children with ALL admitted to Heilongjiang Provincial Hospital from January 2018 to December 2020 were selected for this stud, and were divided into 2 groups, infected group and non-infected group, according to whether they had EBV infection. Differences in clinical characteristics between the 2 groups were analyzed by χ2 or t-test. The impact of EBV infection on the prognosis of children was analyzed by Kaplan-Meier survival and Cox regression analysis.ResultsThe 2 groups were statistically significantly different (P<0.05) according to comparison of characteristics such as first symptoms, karyotype, immunophenotyping, clinical risk, whether secondary infection occurred during chemotherapy, and lymphocyte subsets. Logistic regression results suggested that first symptoms, karyotype, immunophenotyping, clinical risk, the presence of secondary infection during chemotherapy, and lymphocyte subsets were independently associated with EBV infection in children with ALL (P<0.05). The complete remission rate at 46 days after chemotherapy, event-free survival (EFS), overall survival (OS), and survival rate were lower in the infected group than non-infected group, and the complete remission recurrence rate was higher than non-infected group (P<0.05). The EBV DNA levels were statistically lower in the good prognosis group (1.07±0.25×103 copies/L) than poor prognosis group (8.86±1.14 ×103 copies/L) (P<0.01). The area under the curve (AUC) for EBV to predict prognosis in children with ALL was 0.921, sensitivity and sensitivity were 86.57%, 80.16%.ConclusionsInfection with EBV is associated with first symptoms, karyotype, immunophenotyping, clinical risk, secondary infection during chemotherapy, and lymphocyte subpopulation index levels in children with ALL, and children with EBV infection have a reduced clinical remission rate and poor prognosis. Therefore, the detection of EBV DNA is clinically important for assessing the prognosis of their disease.

Genomic heterogeneity contributed to different prognosis between adult and pediatric acute lymphoblastic.

The prognosis of acute lymphoblastic leukemia (ALL) in adults is inferior to that in children. Hence, ALL remains challenging to cure in the adult population. Aberrant genetic alterations have been observed in ALL, although the patterns of differential gene alterations in adult and pediatric ALL have not been comprehensively determined on a genome-wide scale. We investigated the biologic differences in genomic profiles between adults (n=64) and children (n=54) with ALL and relationship between genomic heterogeneity and prognosis. The 2 populations showed similar common mutation types but an increased prevalence of genetic alterations in adult ALL. The median numbers of gene mutations were 17 (range: 1-53) and 4.5 (range: 1-19) per sample in adult and pediatric ALL, respectively (p<0.001). An increased number of gene mutations and age were significantly correlated (R2 =0.5853, p<0.001). We identified 122 and 53 driver genes in adult and pediatric ALL samples, respectively. IKZF1, IDH1, and TTN mutations were significantly enriched in adult patients with ALL. KRAS, ARID1A, and CREBBP mutations were significantly enriched in pediatric patients with ALL (p<0.05). The incidence of relapse was 40.0% and 9.6% in adult and pediatric patients with ALL, respectively (p=0.003). The overall survival and relapse-free survival of adult patients with ALL were poorer than those of pediatric patients with ALL (p=0.002 and p<0.001, respectively). This genomic landscape enhances the understanding of the biologic differences in ALL between the 2 populations and provides insight for developing therapeutic approaches.

Impact of cytogenetic abnormalities on outcomes of adult Philadelphia-negative acute lymphoblastic leukemia after allogeneic hematopoietic stem cell transplantation: a study by the Acute Leukemia Working Committee of the Center for International Blood and Marrow Transplant Research

Cytogenetic risk stratification at diagnosis has long been one of the most useful tools to assess prognosis in acute lymphoblastic leukemia (ALL). To examine the prognostic impact of cytogenetic abnormalities on outcomes after allogeneic hematopoietic cell transplantation, we studied 1731 adults with Philadelphia-negative ALL in complete remission who underwent myeloablative or reduced intensity/non-myeloablative conditioning transplant from unrelated or matched sibling donors reported to the Center for International Blood and Marrow Transplant Research. A total of 632 patients had abnormal conventional metaphase cytogenetics. The leukemia-free survival and overall survival rates at 5 years after transplantation in patients with abnormal cytogenetics were 40% and 42%, respectively, which were similar to those in patients with a normal karyotype. Of the previously established cytogenetic risk classifications, modified Medical Research Council-Eastern Cooperative Oncology Group score was the only independent prognosticator of leukemia-free survival (P=0.03). In the multivariable analysis, monosomy 7 predicted post-transplant relapse [hazard ratio (HR)=2.11; 95% confidence interval (95% CI): 1.04-4.27] and treatment failure (HR=1.97; 95% CI: 1.20-3.24). Complex karyotype was prognostic for relapse (HR=1.69; 95% CI: 1.06-2.69), whereas t(8;14) predicted treatment failure (HR=2.85; 95% CI: 1.35-6.02) and overall mortality (HR=3.03; 95% CI: 1.44-6.41). This large study suggested a novel transplant-specific cytogenetic scheme with adverse [monosomy 7, complex karyotype, del(7q), t(8;14), t(11;19), del(11q), tetraploidy/near triploidy], intermediate (normal karyotype and all other abnormalities), and favorable (high hyperdiploidy) risks to prognosticate leukemia-free survival (P=0.02). Although some previously established high-risk Philadelphia-negative cytogenetic abnormalities in ALL can be overcome by transplantation, monosomy 7, complex karyotype, and t(8;14) continue to pose significant risks and yield inferior outcomes.

The Landscape of Circular RNAs in Drug Resistant TCF3-Rearranged Childhood Leukemia

Treatment resistance is associated with poor prognosis in acute lymphoblastic leukemia (ALL). The underlying causes are largely unknown and specific risk markers and treatment options are consequently lacking. Efforts need to be made to identify new therapeutic targets in drug resistant patients who are left without other treatment options. This applies in particular to ALL patients bearing the t(17;19)(q22;p13) translocation that leads to the expression of the oncogenic chimeric transcription factor TCF3-HLF . Due to treatment resistance these patients typically suffer from relapse and die within two years from diagnosis.Circular RNAs may present such novel targets. Similar to linear RNAs, circRNAs are derived from parental genes. They are circularized by backsplicing and are remarkably stable. CircRNAs may regulate the expression of their parental gene, e.g. by sequestration of miRNAs or RNA binding proteins. CircRNAs have been shown to play a role in cancer by regulating microRNAs with tumor suppressive or oncogenic function (including let-7 and miR-7) and dysregulated circRNA expression has been observed in a variety of human tumors.Therefore we analyzed the landscape of circular RNAs in the drug resistant TCF3-HLF + cell line HAL01 and compared our findings to RNA expression data of a cohort of TCF3-HLF + patients (n=5) at diagnosis and remission. As controls, we used a cell line (697) and data derived from matched diagnosis/remission samples of patients (n=5) with the drug sensitive ALL subtype TCF3-PBX1 .Cell lines underwent whole-exome, -transcriptome and -circRNome sequencing. Single nucleotide variation, indel and copy number analysis revealed only few and not previously in TCF3-HLF+ ALL reported somatic mutations (MSH6, EP300) in HAL01. Transcriptome sequencing confirmed the expression of a TCF3-HLF fusion transcript joining exon 16 of TCF3 to exon 4 of HLF. 54 inserted nontemplate nucleotides indicated terminal deoxynucleotide transferase activity characteristic of an early B cell stage. Similar to leukemic TCF3-HLF+ patient samples, expression of the B cell transcription factor PAX5 was halved, which may account for the block in B cell development. Gene expression analysis restricted to genes specifically expressed in HAL01 and not expressed in 697 revealed a functional cluster comprised of GO terms implicated in drug resistance as most highly enriched.We used a bioinformatic approach to identify circRNAs in RNAseq data of TCF3-HLF+ and TCF3-PBX1+ patient samples. We detected 556 putative circRNAs. 249 genes expressed specific circRNAs only in leukemic and not in remission samples. Of those, 109 circRNAs were exclusively expressed in drug resistant TCF3-HLF -positive leukemia and 9 derived from parental genes such as the lysine demethylase KDM1A involved in p53 mediated DNA damage signaling were recurrently detected.Enrichment of circRNAs by RNase R digest and subsequent sequencing led to detection of 14,311 circular RNAs in HAL01 and 697. 7,918 of those were described in the circBase database, 6,393 were novel. The circRNAs were derived from 4,652 parental genes. 1 to 39 circRNAs were transcribed per gene. The most highly, but ubiquitiously expressed circRNA was derived from the gene NRIP1, a corepressor that modulates a variety of transcription factors. Several circRNAs derived from genes known to be involved in ALL pathogenesis (PAX5, IKZF1 and RUNX1) were detected. CircRNAs derived from the drug transporter gene Multidrug resistance-associated protein 1 (MRP1) were expressed in drug resistant but not in sensitive cells. Two of those were also detected in a leukemic TCF3-HLF+ patient sample. MRP1 detoxifies chemotherapeutic drugs by export of these substances out of the cells. Clinically relevant substrates of MRP1 are anticancer drugs, including vincristine, etoposide, topotecan and methotrexate. Drug screening of TCF3-HLF+ patient derived xenografts previously demonstrated explicit resistance against these drugs. Overexpression of MRP1 and similar transporters is considered a major cause of multidrug resistant tumor phenotypes and may be due to promoter hypomethylation or gene amplification. Upregulation of these transporters up to 10 fold has been observed in TCF3-HLF+ leukemia. Our results may add another layer of regulation and may indicate circular RNAs as novel biomarkers or therapeutic targets for patients with therapy resistant, fatal disease. DisclosuresNo relevant conflicts of interest to declare.

Nucleolin As a Survival Predictor in Acute Lymphoblastic Leukemia and Its Promotion to Drug Resistance By ERK Pathway

Nucleolin (NCL) is an important nucleocytoplasmic multifunctional protein.It plays multiple functions in cell proliferation, apoptosis and angiogenesis. NCL displays oncogenic roles in a large variety of cancer cells, which was reported as high expression in most cancers. In these cancer cells, NCL overexpression influences cell survival, proliferation and invasion through its action on different cellular pathways. However, less research on the roles of NCL in hematologic malignancies was noted. Our previous studies showed that overexpression of NCL is associated with worse prognosis in the patients with acute leukemia and NCL expressed higher in resistant HL-60/ADR than in sensitive HL-60 cells. The potential mechanisms of NCL in chemo-resistance have yet to be investigated. Here we presented the association of NCL with clinical prognosis of acute lymphoblastic leukemia (ALL) and the role of NCL in drug resistance in T-ALL lines. Clinically, the results showed that overexpressed NCL at both mRNA and protein level was associated with a poorer overall survival (OS) and relapse free survival (RFS), as an independent prognostic marker in ALL. mRNA level of NCL in de novo ALL was quantitatively higher than in complete remission(CR) status, and refractory/relapse ALL had the highest level, which relative levels using 2-∆∆Ct method were 1.386, 0.682, and 2.159 respectively (Median(QR), P <0.05) . Expression of NCL protein was observed in 58.3% (35/60) de novo samples, 8.0% (7/87) CR samples and 27.7% (13/47) refractory/relapse samples (P <0.05). Two-year OS rates (Figure 1 ) were respectively 33.3%±8.6% vs 4.0%±1.0% for patients with NCL- negativity and NCL-positivity (P <0.05). Two-year RFS rates were respectively 16.3%±7.2% vs 3.0%±1.0% for patients with NCL-negativity and NCL-positivity (P <0.05). Upon above clinical data, we farther investigated the mechanism(s) by which NCL regulates cellular drug resistance in T-ALL Jurkat cell line. We established cell models with NCL different expression which was transfectedby lentivirus-mediated NCL overexpression vector or siRNA vector, in order to produce NCL overexpression and NCL knockdown models for comparison drug sensitivity. The IC50 of adriamycin on Jurkat cells with NCL-upregulation and NCL- downregulation were 1.362±0.271μg/ml and 0.077±0.010μg/ml respectively (P <0.05). The results of intracellular drug accumulation and effluxion analysis further strengthened the unambiguous promotion impact of NCL on insusceptibility to anticancer agent. Furthermore, we found that Ras/Raf/MEK/ERK signaling and its downstream cell apoptosis factors including c-Myc, Bcl-2 and drug resistance-associated molecules BCRP and MRP1 were all significantly up-regulated after NCL overexpression, conversely, down-regulated after NCL knockdown. Overexpressed NCL up-regulated c-Myc, Bcl-2, BCRP, LRP, MRP1 at mRNA levels, with 2-∆∆Ct values were 7.40±0.40, 1.43±0.11, 2.88±0.07, 2.13±0.02, and 3.35±15, respectively. On the contrary, knockdown of NCL led to reversed results, 2-∆∆Ct values of them as following: 0.24±0.03, 0.32±0.09, 0.03±0.01, 0.69±0.04, 0.55±0.08. Protein levels were in accordance with mRNA level. Notably, co-IP assays confirmed the NCL-Ras, NCL-ERK, NCL-BCRP and NCL-MRP1 interaction. When Ras/EKR signaling was inhibited, levels of BCRP, MRP1 were both decreased, but did not affect to NCL expression. In summary, our findings revealed the novel understanding role of NCL in ALL treatment unresponsiveness and the emerging marker of NCL as a survival predictor and treatment target in ALL. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

TP53 Germline Variations Influence the Predisposition and Prognosis of Acute Lymphoblastic Leukemia in Children

Germline TP53 mutations is the genetic basis of Li-Fraumeni syndrome (LFS), a highly penetrant cancer predisposition condition. Recent reports of germline TP53 variants in children with hypodiploid acute lymphoblastic leukemia (ALL) suggest that childhood leukemia is another manifestation of LFS. However, the pattern, prevalence, and clinical relevance of pathogenic variants in TP53 in childhood ALL remain unknown.To comprehensively characterize genetic variants in TP53 in childhood ALL, we performed targeted sequencing of TP53 coding regions in 3858 children from the Children's Oncology Group front-line ALL clinical trials (COG AALL0232 and P9900). In total, 49 different non-silent coding rare TP53 variants were identifiedin 77 (2.0%) patients sequenced, including 40 missense, 1 nonsense, 6 frameshift, and 2 inframe deletion variants. To determine which TP53 variants are potentially pathogenic and related to ALL risk, we examined their experimentally-validated p53 transcriptional activity, bioinformatically-predicted damaging effects on TP53 function, and also the variant frequency in non-ALL populations. Twenty-two TP53 variants were classified as ALL risk-related (VAR), including 7 protein-truncating and 15 missense variants, all of which were either absent or exceedingly rare in non-ALL populations (i.e., allele frequency < 0.006% in the ExAC dataset). Fourteen missense VAR reside in the p53 core DNA-binding domain, with two affecting residues essential for DNA contact and seven located at residues that are important for the overall architecture of the DNA-binding surface. The 27 remaining TP53 variants were classified as “variants of unknown significance” (VUS). Together, of 3858 children in this ALL cohort, 26 (0.67%) patients had a predicted VAR and 51 (1.3%) had a VUS in the TP53 gene.We next evaluated the association of germline TP53 variants with patient demographics and ALL tumor genetic characteristics. Children with a TP53 VAR were significantly older at diagnosis (P < 0.0001), and had significantly lower leukocyte count at presentation than those with a VUS or wild-type (WT) genotype (P= 0.006). Of 26 patients carrying a germline TP53 VAR, 17 (65.4%) exhibited hypodiploidy in ALL blasts (mostly low hypoploidy or masked hypoploidy). In contrast, hypodiploid ALL was only present in 3.9% and 1.2% of children with TP53 VUS or WT genotype, respectively. The prevalence of TP53 VAR did not differ by genetic ancestry (P= 0.9). We also examined the relationship between germline TP53 variants and treatment outcomes of ALL therapy. In the COG AALL0232 cohort, the presence of a TP53 VAR was associated with a significantly lower event free survival (EFS) and overall survival (OS) compared to patients without a risk variant (hazard ratio [HR] of 2.84, P= 0.0007 for EFS; and HR of 3.13, P= 0.003 for OS). In multivariate analyses, TP53 VAR remained prognostic after adjusting for ancestry, age, leukocyte count at diagnosis, and minimal residual disease at the end of remission induction therapy (HR of 3.36, P= 0.0002 for EFS; HR of 2.88, P= 0.007 for OS). Importantly, there was a markedly higher risk of second cancers among patients with TP53 VARs compared to those with TP53 VUS or WT TP53: 5-year cumulative incidence of 25.1% vs. 0.7%, P= 5.4 × 10-11. In the COG P9900 cohort, only 4 patients harbored TP53 VAR, of whom 3 cases experienced adverse events (one patient each with ALL relapse, second cancer, or death in remission), and both EFS and OS were significantly worse than patients with TP53 VUS or WT genotype (HR of 7.14, P= 0.0008 for EFS; HR of 14.19, P= 1.1 x 10-5 for OS). Taken together, 14 of 26 patients with a TP53 VAR experienced adverse events, with 5 ALL relapses (36% of all events) and 5 second cancers (36%). This pattern of adverse events was dramatically different from those in patients with WT TP53 or VUS, for whom ALL relapse accounted for 75% of all events with only 4% as second cancers (P= 1.2 × 10-7). In fact, even within hypodiploid ALL patients who experienced adverse events, the frequency of second cancer was significantly higher in those with a TP53 VAR than those without (50% vs 5%, P= 0.01), further suggesting that germline TP53 variation was the underlying cause of second cancers in these cases.In conclusion, our findings identified germline TP53 variants as an important determinant of ALL predisposition and adverse treatment outcomes, particularly the risk of second malignant neoplasms. DisclosuresBorowitz:Beckman Coulter: Honoraria; Becton-Dickinson Biosciences: Research Funding; HTG Molecular: Honoraria. Mullighan:Loxo Oncology: Research Funding; Amgen: Consultancy. Hunger:Jazz Pharmaceuticals: Honoraria; Novartis: Consultancy; Erytech Pharmaceuticals: Consultancy; Amgen: Consultancy, Equity Ownership.

IKZF1 Deletions at Diagnose and Relapse of Childhood B-ALL

The transcription factor Ikaros is an essential regulator of lymphopoiesis and IKZF1 gene deletions have been associated with poor prognosis in acute lymphoblastic leukemia (ALL). Here we aim to assess the frequency and the prognostic impact of IKZF1 deletions in Turkish childhood B-ALL by using both deep sequencing and Multiplex Ligation-dependent Probe Amplification (MLPA).The patients cohort was comprised of 78 diagnose and 44 relapse samples of childhood B-ALL (17 paired diagnostic/relapse samples). The median age was 5 years (range, 0.4-17 years), and the median blast percentage at the time of diagnosis was 95% (range, 30-100). All patients were treated according to Berlin-Frankfurt-Munster (BFM) regimen. Patients were sequenced for the most frequent deletions of IKZF1 Δ 2-7, Δ4-7 and Δ4-8 by targeted deep sequencing (454 GS Junior platform, Roche) and also screened by MLPA (Multiplex ligation dependent probe amplification, SALSA kit, MRC Holland) for all exons of IKZF1 . IKZF1 deep sequencing plates were designed by IRONII study consortium. We used Coffalyser v9.4 software for the MLPA analyses, AVA and GENOMIZE programs for the deep sequencing results.IKZF1 deletions were detected in 13% of the diagnose samples and 19% of the first relapsed materials. The detected Δ IKZF1 were listed in Table 1. Δ4-7 was the most common deletion both in diagnose and relapse that results in the expression of a dominant-negative IKAROS variant. The P#15,16,17 were wild type at the time of diagnosis however, gained Δ4-7 in the relapse. Furthermore, the P#3 displayed Δ4-7 both in diagnose and relapse time. We detected Δ2-7 in two first relapsed samples. The P#12 gained Δ4-8 deletion at second relapse in addition to Δ2-7 deletion. Deletions were both confirmed by deep sequencing and MLPA. In addition to NGS results, Δ1-6, Δ2-8, Δ2-3, exon1 and exon3 deletions were determined in five independent cases by MLPA. None of the patients who contained IKZF1 deletions showed somatic point mutations for the screened genomic regions. In the survival analysis, IKZF1 deletions were not found associated with lower overall survival rate (P=0,169).According to these results, IKZF1 deletions occur more often in relapse than diagnosis, although based on limited patient numbers the event is showing a continuous and/or accumulated pattern. I KZF1 deletions is associated with increased risk of relapse and poor outcome in the literature. Here, we could not observe such effect. However, we detected more frequently IKZF1 deletions in the relapsed samples, which may contribute to development of therapy resistance in B-ALL.AcknowledgmentsThis work was supported by TUBITAK (Project Number: 114S038) and and Scientific Research Coordination Unit of Istanbul University (Project no. 35624 and 20520). IKZF1 deep sequencing plates were designed by IRONII study consortium. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Akut Lenfoblastik Lösemide Flow Sitometri ile İmmünofenotiplemenin Prognoz Üzerine Etkileri

Background: The identification of immunophenotype subgroups is very important for the diagnosis and prognosis of acute lymphoblastic leukemia(ALL). Material and Methods: The study included 105 children with ALL(65 males, 40 females; mean age 5.9±3.8 years) who were treated TR-ALL 2000(modified) BFM treatment protocol. Results: The distributions of EGIL classification were pro-B ALL(n=1), common B ALL(n=46), pre-B ALL(n=40), pre-T ALL(n=8), cortical T ALL(n=6), and mature T ALL(n=4). Leukocyte≥100,000/mm³, lymphadenopathy≥2 cm, mediastinal involvement were commonly identified in T ALL group. T ALL had a poor response to chemotherapy according to 8th-day peripheral circulation blast counts and 15th-day bone marrow aspiration(BMA) blast counts. The recurrence, mortality, and death rate in the induction period of treatment were frequently detected in T ALL group. The variables that had prognostic potential, as indicated by univariate analyses, were leukocyte count, hepatomegaly, splenomegaly, and lymphadenopathy at the time of diagnosis, 8th-day steroid response, 15th-day BMA response, risk group, recurrence, and immunophenotyping. Multivariate Cox regression analysis demonstrated that only the leukocyte count(HR 2.51, p &amp;lt; 0.001) was a predictor of prognosis. Conclusion: Immunophenotyping may be effective in the diagnosis and prognosis of ALL, identification of risk groups, and in risk-based treatment planning. T ALL had a poor prognosis.

Comparison of Five-Year Survival Rate Between Black and White Children With Acute Lymphoblastic Leukemia.

IntroductionDespite improvements in the prognosis of acute lymphoblastic leukemia (ALL), it is still the most common childhood cancer. The goal of this study was to investigate if there was a significant difference in the five-year survival between Black and White children with ALL, specifically up to the year 2016 which has not been researched.MethodsA retrospective cohort study of Black and White children diagnosed with ALL between 1975 and 2016 was carried out using the Surveillance, Epidemiology, and End Results (SEER) Program database. Children aged 0-19 were separated into Black or White, and then survival analysis was used to compare five-year survival. A multivariate cox regression analysis was carried out to determine the association between race and five-year survival with ALL.ResultsOur sample included 17,663 cases consisting of 16,238 White children and 1,425 Black children. White children had a significantly increased five-year mortality survival when compared to Black children. Upon using multivariate cox regression analysis, both unadjusted and adjusted models showed a significantly higher risk of death in Black children when compared to White children.ConclusionsOur study found that there is a significant difference in the five-year survival between Black and White children diagnosed with ALL. The difference in survival persists even when controlling for sex, age at diagnosis, year of diagnosis, and histology. Future studies should be carried out to control for more confounders that the SEER database is unable to control for.

Genomic Heterogeneity Contributed to the Different Prognosis of Adult and Pediatric Acute Lymphoblastic Leukemia

Prognosis of acute lymphoblastic leukemia (ALL) in adults is inferior than children. Hence, ALL is still a challenging disease to be cured in adults population. Aberrant genetic alterations has been observed previously in acute lymphoblastic leukemia (ALL), while the patterns of differential gene alteration have not much been comprehensively determined in the adult and pediatric ALL on a genome-wide scale. This study attempted to investigate the biological differences in genomic profiling between the adults and children with ALL, and the correlation between the genomic heterogeneity and prognosis. We collected the survival informations and surveyed the gene mutations profile using whole-exome sequencing (WES) in samples from 39 adults and 54 children of de novo ALL in our institution.The sequencing revealed 663 high-confidence somatic single-nucleotide variants (SNVs), 25 stopgain and 17 frameshift insertion in 690 genes in the 39 adult ALL samples. While in the 54 pediatric ALL samples, 232 SNVs, 12 stopgain and 10 frameshift insertion in 284 genes were identified. The results showed the similar common mutation types in adult and pediatric ALL. However, the median number of detected gene mutations was 19 (range: 1-53) per sample in adult ALL and 4.5(range: 1-19) in pediatric ALL(P&amp;lt;0.001), indicating the reduced prevalence of genetic alterations in pediatric ALL. A significant correlation between the increased of number of gene mutations and age was found (R2 = 0.3096, P&amp;lt;0.001,FigureC). The most frequently mutated genes were NOTCH1 in 6 samples (15.38%), TTN in 5 samples(12.8%), IKZF1 in 4 samples(10.2%) and NRAS in 4 samples(10.2%) in adult ALL. In pediatric ALL, the most frequently mutated genes were KRAS in 15 samples(27.7%), NOTCH1 in 13 samples (24.1%), NRAS in 9 samples(16.7%) and CREBBP in 6 samples(11.1%). The SETD2 and TTN mutations were significantly enriched in adult ALL. While the KRAS, ARID1A and CREBBP mutations were significantly enriched in pediatric ALL. (P&amp;lt;0.05,FigureA, B). Moreover, TP53 mutation was found in 1(0.03%) adult and 2(0.03%) pediatric ALL samples. 67 and 53 driver genes were identified in adult and pediatric ALL samples, respectively. Analysis of transcriptome sequencing data identified transcripts derived from 41 gene rearrangements in 25 (60.9%) adult ALL samples and 20 gene rearrangements in 15 (27.7%) pediatric ALL samples. The most frequent gene rearrangements were BCR/ABL1 fusions (13 samples), fusions of PAX5/ZCCH7 (4 samples) in adult ALL. The most common gene rearrangements in pediatric ALL were MLL fusions(7 samples), TEL-AML1 fusions (6 samples). Using integrated genomic analysis, we identified 3 functional pathways recurrently mutated in adult ALL: transcriptional regulation, NOTCH1 signaling, Ras signaling, and 5 in pediatric ALL: PI3K-AKT-mTOR signaling, JAK-STAT signaling, NOTCH1 signaling, Ras signaling, microRNA processing. The incidence of relapse was 33.3% and 7.7% in the adult and pediatric ALL, respectively(P=0.003). The overall survival(OS) and relapse free survival (RFS) of adult ALL were poorer than pediatric ALL(P=0.003, P&amp;lt;0.001, respectively,FigureD, E), indicating an unfavorable prognosis. Moreover, the number of gene mutations seems to be related with the decreased of times to relapse(R2 = 0.0571, P=0.39). Patients with different genetic subtypes were assigned to the different subgroups. The signatures may related to the inferior outcome of adults compared to children were identified. Adult ALL patients had more enrichment for alterations of amino acid degradation and transcription misregulation, which may explain in part the disparity in the different responses to treatment of the two populations. The study in genomic profiling across the age spectrum elucidated the genomic heterogeneity between adult and pediatric ALL, including the different in the counts of gene mutation, the frequently mutated genes and the fusion genes, which may be the contributing factors that influence prognosis. This genomic landscape enhanced the understanding of the biological differences of disease between the two populations and provided a clue for novel therapeutic approaches. Disclosures No relevant conflicts of interest to declare.