Childhood AbstractAcute Lymphoblastic Leukemia Research Articles

Abstract 5221: Identifying acute lymphoblastic leukemia risk loci in latino children via admixture mapping

Abstract Acute lymphoblastic leukemia (ALL) is the most common childhood cancer and accounts for 25% of malignancies before the age of 20. Despite decades of inquiry into ALL causes, Latino children have up to 1.4 times the rate of ALL compared to their non-Latino White counterparts. The etiology of ALL is complex and this disparity in risk has not been fully explained by social factors alone, suggesting the role of genetic variants and ancestry. Previous literature has implicated variants in IKZF1, ARID5B, GATA3, PIP42KA, and ERG with greater risk allele frequencies or population-specific risk effects in Latinos. Indigenous American ancestry has also been previously associated with ALL risk, relapse, and poor prognosis. To elucidate the genetic and ancestral etiology of ALL, we conducted the first admixture mapping analysis of 1930 childhood ALL cases and 8520 controls of self-identified Latino ethnicity. We uncovered putative admixture associations on chromosomes 2, 7, 10, and 15 (P = 3.21 x 10−5 to 7.32 x 10−8 among top associations across the four loci). Two of the four loci, on chromosomes 2 and 15, were previously not known to be associated with ALL. Following imputation with the TOPMed reference panel, we fine-mapped each putative locus by testing the ALL risk association of each variant with a minor allele frequency greater than 1%. The top associated variants at each locus showed substantial frequency differences between ancestries from the gnomAD or 1000Genomes project. For example, the top associated variant on chromosome 2 has a risk allele frequency of 43% in Native Americans from the Human Genome Diversity Panel, but a frequency of 0.10% in Non-Finnish Europeans in gnomAD. Including these variants in the admixture mapping model greatly attenuated the admixture signals at each locus, suggesting that they are likely the causal SNPs or close proxies of the causal SNPs. The top associated variants from three out of the four loci passed regional multiple testing burden and mapped to genes MGAT5, IKFZ1 and ARID5B on chromosomes 2, 7 and 10, respectively. The top two SNPs on chromosome 15, which narrowly missed the regional threshold, mapped to genes RAB11A and MEGF11. We are pursuing replication of putative novel loci on chromosomes 2 and 15 in additional Latino GWAS datasets. Our results suggest that ALL risk variants with higher frequencies in individuals with Native American ancestry may contribute to the observed increased risk of ALL in Latino children. Investigation of such risk loci can contribute to the identification of new target genes for ALL prediction and therapeutics and new insights for precision medicine, which will reduce the burden of ALL. Citation Format: Jalen Langie, Soyoung Jeon, Xiaomei Ma, Catherine Metayer, Adam J. de Smith, Joseph L. Wiemels, Charleston W. Chiang. Identifying acute lymphoblastic leukemia risk loci in latino children via admixture mapping. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5221.

A New Case of Childhood Acute Lymphoblastic B-Cell Leukemia from Pristina

Abstract Acute lymphoblastic leukemia (ALL) is a malignant disease caused by mutations in B- or T-cell precursors of bone marrow cells. Childhood acute lymphoblastic leukemia (ALL) is a subtype of pediatric cancer with a 1 in 2000 incidence. Here we present a new childhood ALL in a 3-year-old girl. As CD45/19, CD10/19, CD3, CD8, CD10, and CD19 were positive in immunohistochemically analyses of blast cells, a B-ALL was diagnosed with a causative ETV6-RUNX1 gene fusion. The patient was treated based on standard protocols BMF-ALL 2009. Interestingly, an aunt and a grandfather of the patient had experienced malignancies as well, which may be carefully interpreted as a hint on a familial cancer syndrome.

Somatic FOXC1 insertion mutation remodels the immune microenvironment and promotes the progression of childhood acute lymphoblastic leukemia

Acute lymphoblastic leukemia (ALL) is the most common malignant hematological diseases in children. An immunosuppressive microenvironment, particularly regulatory T cell (Treg) infiltration, has been documented to be highly associated with childhood ALL. This present study, based on genetic factors, was aimed at investigating the mutations potentially involved in the immunosuppressive microenvironment in childhood ALL. After whole-exome sequencing was used on DNA extracted from the T cells of ALL bone marrow samples, we found the FOXC1 H446HG induced a increased Treg while decreased cytotoxic T lymphocyte (CTL) in bone marrow. The mutation of FOXC1 in T cell promoted the proliferation of leukemia cells in vitro and in vivo. CpG islands formed by insertion mutation led to an abnormal increase in exon methylation and were associated with the suppression of FOXC1. Decreased FOXC1 attenuated the transcription of HDAC1, thus resulting in the activation of KLF10 through increasing H3K27 acetylation in the promoter region. In conclusion, the de novo insertion mutation in FOXC1 induced suppression of FOXC1, thereby promoting a Treg/CTL shift in the ALL immune microenvironment. The FOXC1 H446HG mutation might be a potential therapeutic target for ALL in the future.

Elevated expression of CDT1 in childhood acute lymphoblastic leukemia promotes cell proliferation, invasion and migration through activation of EMT.

Acute lymphoblastic leukemia (ALL) is a malignant disease of the hematopoietic system. At present, the mechanism and pathogenesis of ALL have not been fully clarified. This study aimed to illustrate the roles of Cdc10 protein-dependent transcript 1 (CDT1) in ALL. Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR) was performed to examine serum levels of CDT1 in childhood ALL patients and healthy volunteers. The interaction between CDT1 expression and prognosis of childhood ALL was analyzed. Meanwhile, expressions of CDT1 in ALL cell lines were determined. Furthermore, CDT1 knockdown model was constructed in ALL cells, and Cell Counting Kit-8 (CCK-8), and Transwell assays were conducted to analyze the effect of CDT1 on the biological functions of ALL cells. Potential mechanism was further explored through detecting the expressions of Epithelial-to-mesenchymal transition (EMT)-related genes. RT-qPCR results indicated that serum level of CDT1 in childhood ALL patients was remarkably higher than that of healthy volunteers. Childhood ALL patients with high expression of CDT1 had lower overall survival rate compared with those expressing low expression of CDT1. CDT1 knockdown remarkably decreased the proliferation and metastasis abilities of pediatric ALL cells. Results of western blot showed that CDT1 might contribute to the malignant progression of childhood ALL via activating EMT. The findings showed that elevated CDT1 facilitated ALL metastasis by promoting EMT, suggesting that CDT1 played a pivotal role in ALL metastasis and may serve as a novel prognostic biomarker and therapeutic target.

Long-term ovarian reserve and fertility outcomes in female survivors of childhood acute lymphoblastic leukemia

Acute lymphoblastic leukemia (ALL) is the most common childhood malignancy, yet data regarding long-term ovarian reserve of female survivors are limited. The aim of this study was to investigate whether there is a differential pattern of anti-Mullerian hormone (AMH) levels in female childhood ALL survivors compared with the normal age-matched population. In a cohort of 56 female childhood ALL survivors (median age 29 years; median follow-up 20.6 years), a negative correlation was found between age at leukemia diagnosis and age-adjusted anti-Mullerian hormone (AMH) levels (r = −0.334, p = .031). Despite alkylating agent therapy, AMH levels did not differ significantly from age-related nomograms (age < 30, p = .17; age ≥ 30, p = .94). The mean number of children per fertile woman adjusted for maternal age was similar to the national average (2.76 versus 3.11, p = .19). Our results imply that reproductive outcomes are not significantly hampered in female pediatric ALL survivors. Long-term surveillance of ovarian reserve may enable personalized survivorship counseling.

Altered proteome of high-density lipoproteins from paediatric acute lymphoblastic leukemia survivors

Acute lymphoblastic leukemia (ALL) is the most frequent malignancy in children. With the use of more modern, efficient treatments, 5-year survival has reached more than 90% in this population. However, this achievement comes with many secondary and long-term effects since more than 65% of the survivors experience at least one severe complication, including the metabolic syndrome and cardiovascular diseases. The main objective of the present work was to characterize the composition of HDL particles isolated from pediatric ALL survivors. HDLs from 8 metabolically healthy ALL survivors, 8 metabolically unhealthy ALL survivors and 8 age- and gender-matched controls were analyzed. The HDL fraction from the survivors contained less cholesterol than the controls. In addition, proteomic analyses revealed an enrichment of pro-thrombotic (e.g., fibrinogen) and pro-inflammatory (e.g., amyloid A) proteins in the HDLs deriving from metabolically unhealthy survivors. These results indicate an alteration in the composition of lipid and protein content of HDL from childhood ALL survivors with metabolic disorders. Although more work is needed to validate the functionality of these HDLs, the data seem relevant for survivor health given the detection of potential biomarkers related to HDL metabolism and functionality in cancer.

Interactions Between Pseudomonas Immunotoxins and the Plasma Membrane: Implications for CAT-8015 Immunotoxin Therapy.

Acute Lymphoblastic Leukemia (ALL) remains the most frequent cause of cancer-related mortality in children and novel therapies are needed for the treatment of relapsed/refractory childhood ALL. One approach is the targeting of ALL blasts with the Pseudomonas immunotoxin CAT-8015. Although CAT-8015 has potent anti-leukemia activity, with a 32% objective response rate in a phase 1 study of childhood ALL, haemolytic-uremic syndrome (HUS) and vascular leak syndrome (VLS), major dose-limiting toxicities, have limited the use of this therapeutic approach in children. Investigations into the pathogenesis of CAT-8015-induced HUS/VLS are hindered by the lack of an adequate model system that replicates clinical manifestations, but damage to vascular endothelial cells (ECs) and blood cells are believed to be major initiating factors in both syndromes. Since there is little evidence that murine models replicate human HUS/VLS, and CAT-8015-induced HUS/VLS predominantly affects children, we developed human models and used novel methodologies to investigate CAT-8015 interactions with red blood cells (RBCs) from pediatric ALL patients and ECs of excised human mesenteric arteries. We provide evidence that CAT-8015 directly interacts with RBCs, mediated by Pseudomonas toxin. We also show correlation between the electrical properties of the RBC membrane and RBC susceptibility to CAT-8015-induced lysis, which may have clinical implication. Finally, we provide evidence that CAT-8015 is directly cytototoxic to ECs of excised human mesenteric arteries. In conclusion, the human models we developed constitutes the first, and very important, step in understanding the origins of HUS/VLS in immunotoxin therapy and will allow further investigations of HUS/VLS pathogenesis.

Significance of 5,10-methylenetetrahydrofolate reductase gene variants in acute lymphoblastic leukemia in Indian population: an experimental, computational and meta-analysis

Acute lymphoblastic leukemia (ALL) arises due to several genetic alterations in progenitor cells, and methotrexate is frequently used as part of the treatment regimen. Although there is evidence for an effect of 5,10-methylenetetrahydrofolate reductase gene (MTHFR) C677T and A1298C variations on drug response in ALL, its risk association for ALL is still unresolved. In a case–control study of 203 patients with ALL and 246 controls and meta-analysis in the Indian population, we showed an insignificant association of MTHFR C677T and A1298C genotypes with childhood and adult ALL. Comprehensive in silico characterization of non-synonymous single nucleotide polymorphisms (nsSNPs) and SNPs of the 3′ untranslated region (UTR) revealed nine nsSNPs as deleterious, and three SNPs in the 3′UTR could possibly alter the binding of miRNAs. The study revealed that several overlooked SNPs may contribute to the risk of ALL susceptibility and further studies of these SNPs with functional characterization in a large sample size are required to understand the significant role of MTHFR in ALL development.

10p12.31-12.2 variants and risk of childhood acute lymphoblastic leukemia in a Chinese population

Acute lymphoblastic leukemia (ALL) is a clonal hematologic disease, and is the most common cause of childhood malignancy. Recently, a new locus was identified at 10p12.31-12.2 through a genome-wide association study (GWAS) that included racially and ethnically diverse populations. We conducted a replication study with 570 cases of ALL and 673 cancer-free controls to validated the association of this locus with ALL susceptibility in a Chinese population. The results of our study confirmed that the 10p12.31-12.2 locus was linked to childhood ALL susceptibility in the Chinese population. Interestingly, we also found that the single nucleotide polymorphisms (SNPs) in this locus had a larger effect on susceptibility to high-risk ALL than on susceptibility to low-risk ALL.

Polymorphism in IKZF1 gene affects age at onset of childhood acute lymphoblastic leukemia

Acute lymphoblastic leukemia (ALL) is the most common childhood cancer, characterized by a peak of incidence between 2 and 5 years. Since recently conducted genome-wide association (GWA) studies revealed that the common low-penetrance susceptibility allele at 7p12.2 (IKZF1 gene) confers an increased risk of pediatric ALL, we investigated whether the risk allele at rs4132601 also coexists with well-established prognostic factors, among 508 Polish pediatric patients with newly diagnosed ALL. Additionally, to verify whether the risk allele is favored by somatic tumor evolution, we examined the incidence of IKZF1 deletions in leukemic clones derived from 153 previously genotyped cases of pediatric ALL. Results of the analysis provide statistically significant support for an association between the rs4132601 polymorphic site and age at diagnosis of childhood ALL (p = 0.04). No association between allele variant and occurrence of IKZF1 deletions was found. These data provide further evidence of a biological role of gene variants in the development of ALL.

Abstract 4783: Proteomic analysis of acute lymphoblastic leukemia: de novo versus relapse

Abstract Acute lymphoblastic leukemia (ALL) is a heterogeneous disease with distinct biological and prognostic grouping. The initial response to induction chemotherapy is the most important prognostic factor in patients with acute lymphoblastic leukemia. Patients with rapid early response (RER &amp;lt; 5% of blast in bone marrow at day 7 after induction chemotherapy) have better event-free survival compared to slow early responders (SER ≤ 25% of blast). However, no predictive marker for relapse or treatment failure is currently available. This study aimed to find plausible biomarkers of drug sensitivity in bone marrow mononuclear cells (BMMNCs) of childhood ALL cases using 2-dimensional gel electrophoresis and subsequent MALDI-TOF/mass spectrometry. The 714 spots in RER and 555 spots in SER were visualized in gels. Of these 28 up-regulated and 25 down-regulated proteins in SER_as compared to RER_were identified. The detected proteins were 60-kDa heat shock protein, actin, lamin-B1, heterogeneous nuclear ribonucleoprotein (hnRNP) K, and so on. HnRNPK, down-regulated in SER was further verified by Western blot analysis. In future studies, the roles of HnRNPK in drug sensitivity/resistance are undergoing to elucidate as a potential biomarker for ALL therapy. [Funding source: NRF grant (R13-2005-012-01003-0)]. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4783. doi:1538-7445.AM2012-4783

Extra Copies of der(21)t(12;21) plus Deletion ofETV6Gene due to dic(12;18) in B-Cell Precursor ALL with Poor Outcome

Acute lymphoblastic leukemia (ALL), CD10+ B-cell precursor, represents the most frequent type of childhood ALL from 3 to 6 years of age. The t(12;21)(p13;q22) occurs in 25% of cases of B-cell precursor ALL, it is rare in children less than 24 months and have been related to good prognosis. Some relapse cases and unfavorable prognosis in ALL CD10+ are associated with t(12;21) bearing additional aberrations as extra copies of chromosome 21 and ETV6 gene loss. This report describes the case of a 15 month-year old girl, who displayed a karyotype with addition on chromosome 12p plus trisomy 10 and tetrasomy of chromosome 21. Molecular cytogenetic studies revealed two extra copies of the der(21) t(12;21), trisomy 10 and deletion of the second ETV6 gene due to the dic(12;18). These findings show the great importance of molecular cytogenetic studies to clarify complex karyotypes, to define prognostic, to carry out risk group stratification and to support correctly disease treatment in childhood acute lymphoblastic leukemia.

The clinical significance of methylenetetrahydrofolate reductase (MTHFR) polymorphisms in acute lymphoblastic leukemia

Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy. Genetic polymorphisms in the folate metabolic pathway may contribute to the susceptibility to childhood ALL because they affect the DNA synthesis, methylation, and repair. The most common polymorphisms are methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C. The current study aimed at detecting the frequency of these two genetic polymorphisms in de novo ALL patients, and to clarify their impact on the response to induction chemotherapy, as well as treatment toxicity. MTHFR C677T and A1298C polymorphisms were tested in 30 de novo ALL patients by restriction fragment length polymerase chain reaction technique. Thirty normal age- and sex-matched subjects were subjected to the same analysis as a control group. The frequency of MTHFR A1298C gene polymorphism was significantly lower in ALL patients than the controls thus showing a protective effect. The two polymorphisms had no effect on the response to induction chemotherapy. As regards the treatment toxicity, MTHFR C677T polymorphism was associated with marked thrombocytopenia, while A1298C polymorphism was associated with hepatic toxicity. Identifying predictors of methotrexate sensitivity may lead to the development of individualized treatment strategies with improved efficacy and reduced toxicity as well as adjusting the initial methotrexate dose.

New Decision Support Tool for Treatment Intensity Choice in Childhood Acute Lymphoblastic Leukemia

Acute lymphoblastic leukemia (ALL), the most common cancer in childhood, has its treatment modulated by the risk of relapse. An appropriate estimation of this risk is the most important factor for the definition of treatment strategy. In this paper, we build up a new decision support tool to improve treatment intensity choice in childhood ALL. Our procedure was applied to a significant cohort of Brazilian children with ALL, the majority of the cases treated in the last decade in the two main University Hospitals of Rio de Janeiro. Some intrinsically difficulties of this dataset introduce an assortment of challenges, among those the need of a proper selection of features, clinical and laboratorial data. We apply a mutual information-based methodology for this purpose and a Neural Network to estimate the risk. Among the relapsed patients, 98.2% would have been identified as high-risk by the proposed methodology. The proposed procedure showed significantly better results when compared to the BFM95, a widely used classification protocol.

Treatment of pediatric acute lymphoblastic leukemia: Progress achieved and challenges remaining

Acute lymphoblastic leukemia (ALL) is the most common cancer in children. Careful building upon past clinical trials and thoughtful application of our limited knowledge of pharmacology have provided steady improvement in outcome for newly diagnosed patients. Precise identification of the many patients who are unlikely to relapse with current effective regimens is required to avoid the morbidity of further intensification of therapy. Progress is sorely lacking for relapsed patients. Most patients who relapse die. Gene expression arrays and comparative genomic hybridization have further extended our appreciation of the known immunophenotypic and genetic diversity of childhood ALL. Insight into the molecular mechanisms of treatment failure may provide guidance for future efforts.

Gene – gene interactions in the folate metabolic pathway influence the risk for acute lymphoblastic leukemia in children

Acute lymphoblastic leukemia (ALL) is the most common pediatric cancer. Genetic polymorphisms in the folate metabolic pathway may contribute to the susceptibility to childhood ALL because they affect the DNA synthesis, methylation and repair. We analysed common genetic polymorphisms of 5,10-methylenetetrahydrofolate reductase (MTHFR), thymidylate synthase (TS), methionine synthase (MS) and methionine synthase reductase (MTRR) in 68 children with ALL and 258 healthy controls to investigate their influence on the risk for ALL. No significant differences in frequencies of separate polymorphisms were observed between both groups. Combined MTHFR 677CT/TT and MS 2756AG/GG genotypes showed a nonsignificant tendency to reduce the risk for ALL 2.24-fold (CI: 0.191 – 1.037, P: 0.061). The risk was significantly reduced in carriers of combined MTHFR 677CT/TT, MS 2756AG/GG and MTRR 66AG/GG genotypes (OR: 0.312; CI: 0.107 – 0.907; P: 0.032). Our results suggest that gene – gene interactions that may decrease the methylation capacity might have a protective effect on the risk for childhood ALL.

Acute Lymphoblastic Leukemia: Current Treatment

Acute lymphoblastic leukemia (ALL) has served as a model for cancer research in children as well as adults. Because of the major advances in diagnosis, development of rational use of combination chemotherapy, specific central nervous system (CNS) preventive therapy and improved supportive care, the cure rate in childhood ALL has increased tremendously and at present approximately 70–80% of children with ALL are potentially cured. This manuscript provides an overview of current concepts in classification, prognosis and treatment of ALL.

The Investigation of Karyotypic Instability in the High-Hyperdiploidy Subgroup of Acute Lymphoblastic Leukemia

Acute lymphoblastic leukemia (ALL) of childhood has been cytogenetically well characterized, and approximately 25% of cases will have a high-hyperdiploid (51–68) chromosome complement. In a 5 year period a consecutive series of 152 presentation ALL's were karyotyped. In all cases a result was obtained and 138 (91%) had a detectable abnormal clone of which 44 (29%) were high-hyperdiploid. Within the high-hyperdiploidy group karyotypic cell to cell variation was observed in many cases. To provide further evidence of this phenomenon a dual-color fluorescence in-situ hybridization (FISH) experiment was performed on stored fixed suspension from 14 ALL's with such a karyotype. In each case 4–6 probes were investigated, employing probes to centromeres of chromosomes X, 4, 6, 8, and 10 and a locus specific probe to chromosome 21q22. It was found that the FISH produced results that were generally in good agreement with the G-banding findings and supported the notion of karyotypic cell to cell variation. FISH further showed that most of cases would have two extra copies of chromosome 21 in the majority of leukemic cells and a single extra copy in the minority. A further finding was that fewer cells contained extra copies of chromosomes 6, 8 and 10 than was expected based on the comparison of the signal number of the other probes investigated. In contrast chromosomes X, 4, and 21 seldom displayed this feature. We have demonstrated that karyotypic instability as defined by karyotypic cell to cell variation is a feature of the high-hyperdiploid subgroup in childhood ALL. It is questioned whether the underlying defect resulting in the observed karyotypic instability of this subgroup is one of the primary causative events in the formation of the leukemia.