Elderly Acute Lymphoblastic Leukemia Research Articles

Root Anomalies after Combined Chemotherapy Treatment for Acute Lymphoblastic Leukemia during Childhood: A Case Report‬‬‬‬‬‬‬‬‬‬‬‬‬

Dental abnormalities have been sporadically reported in patients treated as children for acute lymphoblastic leukemia (ALL) with multi-agent chemotherapy, with and without radiation. Reports included delayed dental development, microdontia, hypoplasia, agenesis, V-shaped roots, shortened roots, and cervical root resorption. These anomalies in pediatric patients require regular dental follow-up and sometimes highly complex treatment. We describe a well-documented case of a 19-year-old male who presented with such anomalies 12 years after treatment with several chemotherapy agents for ALL, and discuss the resultant disruption of normal tooth development that led to the ensuing abnormalities.

In-silico evaluation of fungal and bacterial L-asparaginases allergenicity

l-asparaginase (l-ASNase) is a vital therapeutic agent for acute lymphoblastic leukemia and is naturally produced by various organisms even though all the commercial L-ASNases are derived from bacteria that have allergic reactions mainly due to their prokaryotic origin. Since fungi are eukaryotes and their enzymes are expected to be similar to mammal proteins, the immune reactions against them are expected to be low. With this thought, this study aimed to provide insight that fungi are superior sources of l-ASNase for clinical use compared to bacteria. Aiming this, a total number of 120 sequences, 60 each from fungal and bacterial sources, were retrieved from the National Center for Biotechnology Information database. The datasets were grouped into three groups based on amino acid (aa) sequence length: 298–315, 340–355, and 375–390, each group consisting of 20 sequences. Subsequently, these sequences were analyzed using Bioinformatics tools. The comparative analysis of fungal and bacterial L-ASNases allergenicity using Algpred 2.0 showed that 32/60 (53.3 %) fungal L-ASNases and 56/60 (93.3 %) bacterial L-ASNases were predicted to be allergenic which was significantly different (p = 0.00). This was supported by AllerTOP 2.0 which predicted 23/60 fungal and 34/60 bacterial L-ASNases as allergenic. Surprisingly, the Allergome database has strengthened this result by indicating fungal L–ASNases are less allergenic (p = 0.003) compared to their bacterial counterparts which leads to conclude that l–ASNase allergenicity is correlated to its origin. Moreover, allergenicity of l-ASNase is also correlated with sequence length as predicted by Algpred 2.0, AllerTOP 2.0, and Allergome. In general, this study provided an outline of evidence that fungal L-ASNases have less allergenicity compared to their bacterial counterparts. Hence, fungi could be considered as potential sources of l-ASNase with reduced allergenicity.

Renal function features in pediatric acute lymphoblastic leukemia during high-dose methotrexate chemotherapy

Introduction: High-dose Methotrexate (HD-MTX), a chemotherapy agent for acute lymphoblastic leukemia, is a cytotoxic agent for some organs, including kidneys. One of the most important toxicities due to HD-MTX is acute kidney injury due to Methotrexate crystallization in renal tubules. The prevalence of Acute kidney injury due to HD-MTX administration in pediatric ALL in dr. Soetomo General Hospital was unknown. The study aims to analyze renal function features in pediatric ALL during HD-MTX chemotherapy. Methods: An analytical observational study with a prospective approach was conducted at Dr. Soetomo General Hospital Surabaya from December 2021 - July 2022. The subjects were ALL children aged 1-18 years who met inclusion and exclusion criteria. High-dose Methotrexate chemotherapy was given 3 times every 2 weeks during the consolidation phase. Laboratory examinations were performed before and after HD-MTX chemotherapy. Laboratory results were recorded to determine the GFR value. The difference test was performed using Wilcoxon signed rank test and the Friedman test with a significance value of p<0.05. Results: A total of 20 subjects, the median age was 78 months old, and boys and girls were equal. Standard Risk of ALL was in 55% of subjects, and 45% were at high risk. There was no significant difference in laboratory parameters between before and after HD-MTX chemotherapy in all cycles with a p-value >0.05. The median value of GFR in each cycle is 257.5; 243.5; 228.5. Minimal GFR was 119 ml/min/1.73 m2, and maximum GFR was 638 ml/min/1.73 m2. It showed a median value of GFR in all cycles was >175 ml/min/m2 (hyperfiltration). Conclusions: We concluded there was no AKI due to HD-MTX chemotherapy in pediatric ALL. It may be caused by providing adequate hydration, urine alkalinization, and leucovorin rescue.

GSK3β Inhibition Prevents Macrophage Reprogramming by High-Dose Methotrexate

Methotrexate (MTX) is an antifolate drug used as a chemotherapeutic agent for acute lymphoblastic leukemia, where MTX improves patients’ prognosis. Macrophage reprogramming is being increasingly assessed as an antitumor therapeutic strategy. However, and although MTX limits the pathogenic action of macrophages in chronic inflammatory diseases, its effects on tumor-promoting macrophages have not been previously explored. We now report that MTX shapes the transcriptional and functional profile of M-CSF-dependent human macrophages, whose transcriptome is highly enriched in the gene signature that defines pathogenic tumor-associated macrophages (“large TAM”). Specifically, MTX prompted the acquisition of the gene signature of antitumoral “small TAM” and skewed macrophages toward an IL-6high IFNβ1high IL-10low phenotype upon subsequent stimulation. Mechanistically, the MTX-induced macrophage reprogramming effect correlated with a reduction of the M-CSF receptor CSF1R expression and function, as well as a diminished expression of MAF and MAFB transcription factors, primary determinants of pro-tumoral macrophages whose transcriptional activity is dependent on GSK3β. Indeed, the ability of MTX to transcriptionally reprogram macrophages toward an antitumoral phenotype was abrogated by inhibition of GSK3β. Globally, our results establish MTX as a macrophage reprogramming drug and indicate that its ability to modulate macrophage polarization may also underlie its therapeutic benefits. Since GSK3β inhibition abrogates the reprogramming action of MTX, our results suggest that the GSK3β-MAFB/MAF axis constitutes a target for the macrophage-centered antitumor strategies.

Novel bioactive hybrid Celecoxib-HDAC Inhibitor, induces apoptosis in human acute lymphoblastic leukemia cells

Acute lymphoblastic leukemia (ALL) is the most common malignancy in children. Here, we exploited the synergy between histone deacetylase inhibitors (HDACi) and cyclooxygenase 2 (COX-2) inhibitors by generating and testing a series of hybrid Celecoxib-HDAC inhibitors (selenium-containing analogues of Celecoxib) on ALL cells, of which compound 11 exhibited significant inducement to kill NALM6 cells with an average IC50 of 9.95 ± 0.44 μM compared with control Celecoxib at 28.58 ± 1.44 μM and inhibited NALM6 cells growth via the inhibition of the cell cycle in G2 phase. Furthermore, compound 11 induced apoptosis by activating PARP cleavage. Taken together, compound 11 possessed the potential to be developed further as a chemotherapeutic agent for ALL.

Panobinostat (LBH589) increase survival in adult xenografic model of acute lymphoblastic leukemia with t(4;11) but promotes antagonistic effects in combination with MTX and 6MP.

Patients diagnosed with acute lymphoblastic leukemia (ALL) bearing t(4;11)/MLL-AF4 have aggressive clinical features, poor prognosis and there is an urgent need for new therapies to improve outcomes. Panobinostat (LBH589) has been identified as a potential therapeutic agent for ALL with t(4;11) and studies suggest that the antineoplastic effects are associated with reduced MLL-AF4 fusion protein and reduced expression of HOX genes. Here, we evaluated the in vitro effects of the combination of LBH589 with methotrexate (MTX) or 6-mercaptopurine (6MP) by cell proliferation assays and Calcusyn software in ALL cell line (RS4;11); the in vivo effects of LBH589 in xenotransplanted NOD-scid IL2Rgammanull mice measuring human lymphoblasts by flow cytometry; and the expression of HOX genes by qPCR after treatment in an adult model of ALL with t(4;11). LBH589 combination with MTX or 6MP did not promote synergistic effects in RS4;11 cell line. LBH589 treatment leads to increased overall survival and reduction of blasts in xenotransplanted mice but caused no significant changes in HOXA7, HOXA9, HOXA10, and MEIS1 expression. The LBH589, alone, showed promising antineoplastic effects in vivo and may represent a potential agent for chemotherapy in ALL patients with t(4;11).

Could CAR-T cell therapy be promising agent in acute lymphoblastic leukemia?

Could CAR-T cell therapy be promising agent in acute lymphoblastic leukemia?

MRNA COVID-19 vaccine safety in patients with previous immediate hypersensitivity to pegaspargase

mRNA COVID-19 vaccine safety in patients with previous immediate hypersensitivity to pegaspargase

Purple sweet potato anthocyanins elicit calcium overload-induced cell death by inhibiting the calcium-binding protein S100A4 in acute lymphoblastic leukemia

This study aimed to evaluate the effects of purple sweet potato anthocyanins (PSPAs) on acute lymphoblastic leukemia (ALL) cells to elucidate the molecular mechanism underlying the anticancer properties of PSPAs. The results showed that PSPAs significantly suppressed cell proliferation by inducing obvious G2/M phase arrest. In addition, PSPAs dramatically increased the intracellular Ca2+ levels, thereby leading to calcium overload-induced cell death. Furthermore, to evaluate the relevant mechanism, bioinformatic, subcellular localization, molecular docking, and Western blot analyses were performed. PSPAs induced calcium overload by inhibiting the protein expression of S100A4. An additional assay combining S100A4 induction and PSPA treatment reconfirmed that S100A4 was the potential target of PSPAs and that PSPAs mainly exerted their antileukemic effects through the p38/c-Myc/CDK1-Cyclin B axis. Our findings indicate that PSPAs are promising therapeutic agents for ALL.

Zebrafish ALL Driven By HumanMYCand a New Technique to Biopsy Living Fish

Acute lymphoblastic leukemia (ALL) is the most common childhood cancer; B-lineage ALL (B-ALL) represents ~85% of cases. Transgenic humanMYC(hMYC), an oncogenic transcription factor in many blood cancers, is known to induce T-ALL in zebrafish. Until recently robust zebrafish B-ALL models did not exist, but we discovered fish of this transgenic line develop B-ALL also. To distinguish T-ALL from B-ALL, we use a transgenic marker,lck:GFP, where T-ALL fluoresce brightly while B-ALL fluoresce dimly. To define B- and T-ALL incidence, we monitored >600 fish by serial fluorescent microscopy and found 100% of fish developed ALL (64% T-ALL, 23% B-ALL, both 13%) by 10 months. To find transcriptional programs active in both ALL types, we performed RNA-seq on 10 T- and 14 B-ALL and compared these to T- and B-ALL induced by murineMyc(mMyc). Unexpectedly, we found B-ALL occur in distinct lineages in these related lines, withhMYCB-ALL expressingighzheavy chains, butmMycB-ALLighm. To investigate glucocorticoid responses, a key agent in ALL, we treated fishin vivowith dexamethasone (DXM).hMYCB-ALL were highly DXM-sensitive, but many recurred rapidly after treatment ceased. Studies lymphocytes or ALL cells from zebrafish typically require fish euthanization; it would be useful to obtain cells without euthanasia. Thus, we developed biopsy techniques in living fish to obtain lymphocytes from the thymus, kidney-marrow, and even scales. We found B and T cells can be obtained from thymus, with B cells abundant in scales. We can also recover ALL cells from scales of fish with B- or T-ALL. Analyses of normal lymphocytes and ALL cells from these anatomic regions prove that GFPhi cells express T cell genes, and GFPlo cells express B cell genes. These novel biopsy methods will permit studies ofex vivosamples from live fish. Overall, these new results and methods expand the utility of zebrafishhMYC-driven ALL, a model of the most important pediatric cancer. Disclosures No relevant conflicts of interest to declare.

Management of toxicities associated with novel immunotherapy agents in acute lymphoblastic leukemia.

The advent of novel immunotherapies, such as blinatumomab, inotuzumab, and chimeric antigen receptor (CAR) T cell therapy, has revolutionized the therapeutic landscape in the treatment of relapsed/refractory B cell acute lymphoblastic leukemia, but can be associated with specific toxicities. We review unique toxicities of each of these in this article. Blinatumomab, a bispecific T cell engager, has been associated with cytokine release syndrome (CRS) and neurological toxicities, both of which can be prevented and managed with corticosteroids. Inotuzumab is a calicheamicin-conjugated CD22 targeting antibody. The calicheamicin component of the drug is likely associated with the hepatotoxicity seen with inotuzumab, especially sinusoidal obstruction syndrome, which can happen both in the context of the drug alone, and also with allogeneic stem cell transplantation. QT prolongation has also been noted with inotuzumab. CAR T therapy uses genetically modified autologous T cells directed against CD19, a known target on B cells. CRS and neurological symptoms, formally termed as immune-effector-cell-associated neurological syndrome, have been described along with hypogammaglobulinemia, cytopenias, and infections.

Using Pharmacogenomics Testing to Optimize Care in Children with Acute Hematologic Malignancies

Background Inter-patient variation in the pharmacokinetics of drugs can result in unpredictable variability in occurrence of adverse events and toxicity as well as variation in therapeutic efficacy. Although a multitude of factors contribute to this observed variation, inherited variation in genes involved in drug metabolism and transport has been shown to play a significant role. Genetic polymorphisms can influence the gene expression and/or activity, thereby impacting drug pharmacokinetics and causing inter-individual variation in drug levels which can alter risk of toxicity or therapeutic efficacy. The goal of this study is to identify pharmacogenomic biomarkers predicting drug response and toxicity in children receiving chemotherapy for acute hematologic malignancies. Identification of such biomarkers will establish relevant biomarkers for allowing for personalizing therapy to achieve maximum benefit and minimal side effects in future studies. Study Design and Methods: Study Population Male and female participants are included with no restrictions based on racial and/or ethnic origin. All patients aged 26 years of younger who have received at least one year of treatment for acute lymphoblastic leukemia (ALL) or acute myeloid leukemia (AML) and have experienced organ toxicity CTCAE v5.0 Grade 3 or higher at the University of Florida are offered participation. Subjects meeting eligibility criteria are excluded if they cannot provide a blood specimen. Study Design This pilot study proposes to identify and characterize pharmacogenomic variations that lead to different systemic toxicity profiles in pediatric patients receiving commonly used chemotherapy agents for ALL and AML. Agents of interest include drugs used during induction chemotherapy cycles which include Vincristine, Methotrexate, Cytarabine, Daunorubicin, Prednisone, Dexamethasone, 6-thioguanine, Cyclophosphamide, 6-mercaptopurine, and Etoposide. A single-nucleotide polymorphism (SNP) panel to evaluate SNPs in pharmacokinetic pathway genes of the selected drugs has been created. Genotyping is performed using Taqman genotyping assays and the Quant studio real time PCR system. We are investigating genes involved in the pharmacokinetics of the selected chemotherapy agents such as ABCB1, GSTM1, GSTT1, BCL2, MCL1, BCL2L11, MTHFR, SLC29A1, FMO3, DYNC2H1, TPMT, NUDT15, GSTP1, SOD2, and TP53. Toxicities that are correlated to SNP genotypes include the following: renal injury defined as creatinine clearance 29-15 ml/min/1.73 m2 or less; hepatic injury defined by increase in serum total bilirubin > 3.0 times the upper normal limit; hematological impairment including severe neutropenia, thrombocytopenia, and anemia leading to treatment delays; number of neutropenic fever hospitalizations; thromboembolic events; pancreatitis; cardiotoxicity defined by decline in ejection fraction by > 20% of baseline; neurotoxicity defined by examination and alterations in function; and insulin-dependent glucose resistance. The relationship and prevalence of SNP variations to the defined toxicities is analyzed. Statistical Considerations and Study Endpoint Complete case analysis will be performed. Toxicities per CTCAE v5.0 are graded by ordinal statistics. We aim to describe rates of pharmacogenomic SNP variants and the prevalence of specific toxicities related to these variants. The sample size of 100 patients was selected because this number should allow us to be relatively precise in our conclusions while still allowing for estimates of variability. Factors such as the rarity of pediatric cancer and the numbers of patients treated at our institution were considered in selecting this sample size. A chi-square test is used to evaluate for an association between SNP variants and toxicity grading. To quantify identified associations, multinomial logistic regression analysis will be performed. A P-value <0.05 is considered statistically significant. Study Endpoints: The study endpoint is to identify pharmacogenomic SNP variants of children undergoing treatment for acute hematologic malignancies and to provide an analysis of the relationship between these variants and clinically graded toxicities. Identification of these biomarkers would help guide subsequent validation in a larger cohort and establish clinically relevant biomarkers for personalization of acute leukemia therapy. Disclosures No relevant conflicts of interest to declare.

A Comprehensive Study of the Chemosensitizing Effects of AZD‐1775, WEE‐1 Inhibitor in Combination with DNA Damaging Treatments in the Jurkat Leukemia Cell Model

Current treatment for leukemia focuses on DNA damaging treatments (DNAdt), including chemo (cytarabine, vincristine, doxorubicin, etc.) and/or radiotherapy. However, Acute Lymphoblastic Leukemia (ALL) mechanisms of resistance limit the value of these treatments. Recent studies evaluated the G2‐M gatekeeper Wee1 inhibition as a possible alternative treatment for other types of cancer, such as: breast, prostate, pancreatic, lung, leukemia, glioblastoma, among others. AZD‐1775, a well‐documented Wee1 (G2‐M‐Gatekeeper) inhibitor, has shown chemosensitizing, synergistic effects, and currently is undergoing clinical trials. Therefore, our working hypothesis is that: AZD‐1775 sensitizes Leukemia cell lines to DNA‐damaging drugs, via a mitotic catastrophe process. This study evaluates the cytotoxic and chemo‐sensitization effect of AZD‐1775 on an ALL T‐cell model (Jurkat). Even more, this study innovates the understanding of AZD‐1775 by carefully crafting a panel of drugs with different mechanisms that induce DNA damage. Cell viability assays (resazurin‐based; high throughput 384 well plates) were done to determine the inhibitory effects of AZD‐1775 and a series of DNAdt drugs alone and in combinations. The corresponding IC50 value for all curves obtained were calculated using GraphPAD Prism (Vers.7). The nature of the interactions (synergy, additivity, antagonism) between AZD‐1775 and the different DNAdt drugs was determined with Combenefit and the CompuSyn Software. The Combenefit freeware permitted fitting and analysis of the data to various synergy models (Loewe‐additivity, Bliss‐independence and Highest‐single Agent); while, GraphPad Prism and CompuSyn allowed estimates of drug combination indexes (CI) and dose‐reduction indexes (DRI). The individual agents exhibited the following rank order of inhibitory potencies (IC50±SEM): cytarabine (160nM±6nM SEM) ≥ AZD‐1775 (370nM±9nM SEM) >> etoposide (2μM±60nM SEM) = cisplatin (3μM±127nM SEM) > 5′‐fluorouracil (34μM±2.7μM SEM). The results demonstrate a significant synergistic effect of AZD‐1775 with cytarabine, 5FU, cisplatin and etoposide. Conversely, most of these drugs were also capable of potentiating the effect of AZD‐1775 in a synergistic manner. In all models of synergy analysis, statistically significant synergistic interactions were confined to specific concentration ranges of drug‐combinations. For example, the most significant synergistic interactions were obtained with AZD‐1775 concentrations of 58nM to 270nM, and cytarabine concentrations of 8nM to 125nM, with the Bliss‐independence model. In conclusion, our findings strongly suggest that AZD‐1775 can be used as a chemosensitizing agent in ALL, capable of potentiating the effect and susceptibility to DNAdt; whilst, allowing the patient to require lower doses of these chemotherapies to decrease adverse effects.Support or Funding InformationPartially supported by NIGMS/INBRE award P20GM103475. The content is solely the responsibility of the author and does not necessarily represent the official views of any of the supporting agencies.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

A Single Mouse Trial Platform for Evaluation of Novel Agents in Acute Lymphoblastic Leukemia By the Pediatric Preclinical Testing Consortium

Introduction: While children diagnosed with the most common pediatric malignancy, acute lymphoblastic leukemia (ALL), now experience close to a 90% likelihood of cure, the outcome for several high-risk subtypes remains poor. Furthermore, since standard-of-care drugs are extremely effective in this disease, and there are relatively few patients eligible for early phase clinical trials, only the most promising new agents are advanced for clinical evaluation following rigorous preclinical testing. However, conventional preclinical testing of novel agents is not sufficiently resourced to be able to encompass the vast heterogeneity of pediatric ALL, and new approaches to preclinical testing are required in this disease. The purpose of this study was to evaluate the utility of a single mouse trial (SMT) platform for preclinical assessment of novel agents on an almost clinical trial scale, to encompass the broad heterogeneity of pediatric ALL in a single experiment, and to identify molecular biomarkers associated with in vivo drug responses when carried out using molecularly-annotated patient-derived xenografts (PDXs).Methods: Eighty pediatric ALL PDXs broadly representative of all pediatric ALL subtypes were characterized in terms of engraftment kinetics in immune-deficient NSG mice, and molecularly annotated by RNA-seq, exome-seq and DNA copy number analysis. Between 2-5 million cells from each PDX were inoculated via the tail vein into 2 NSG mice/PDX. Starting at 2 weeks post inoculation engraftment was monitored by flow cytometric enumeration of the proportion of human CD45+cells in the murine peripheral blood (%huCD45+). When the %huCD45+ for each PDX reached >1% one mouse/PDX was treated with the established drug and topoisomerase I inhibitor topotecan (Tpt, 0.6 mg/kg IP daily x 5 x 2 weeks, repeated at 21 days) and the other mouse was treated with the experimental drug and second mitochondria-derived activator of caspases (SMAC)-mimetic birinapant (Bpt, 15 mg/kg IP every 3 days x 5). Treatment response was monitored using stringent objective response criteria modeled after the clinical setting, by mouse event-free survival (EFS where an event was defined as 25% huCD45+), and by waterfall plots comparing the maximum decrease in %huCD45+at any point post treatment initiation. The authenticity of each PDX was verified using a 60-allele SNP array both at the time of inoculation and at relapse post drug treatment for all mice.Results: Retrospective analysis of 1,000 samples of “single mouse” data previously showed that the single mouse results predicted the overall group response from conventional testing 75.3% of the time, which increased to 94.3% if a deviation of ± one response category was allowed. SMT results were achieved for 71 (88.8%) and 73 (91.3%) of the intended 80 mice for Tpt and Bpt, respectively. Waterfall plots revealed that 60/71 (84.5%) and 30/73 (41.1%) of PDXs achieved regressions in response to Tpt and Bpt treatment, respectively. When compared with historical objective response measures from conventional drug testing carried out by the Pediatric Preclinical Testing Consortium (PPTC) the SMT results showed high concordance for both Tpt (R=0.904; P=0.014; n=7 PDXs) and Bpt (R=0.804; P<0.0001; n=20 PDXs). Moreover, the SMT confirmed the preferential in vivo efficacy of Bpt against B-ALL compared with T-ALL, early T-cell precursor ALL (ETP-ALL) and ALL with mixed linage leukemia gene rearrangements (MLLr-ALL) that was previously revealed by conventional testing carried out by the PPTC. Analysis of divergent responses observed within the MLLr-ALL subpanel (n=9 PDXs) to Tpt revealed a potential 40 gene signature model, with up-regulated genes in Responders being associated with transcription regulation, cellular proliferation and differentiation.Conclusions: This study has shown that SMTs provide an accurate and cost-effective platform for preclinical drug testing in pediatric ALL on an almost clinical trial scale. Moreover, SMTs can almost encompass the heterogeneity of pediatric ALL in a single experiment, and are likely to be useful for large-scale correlations with in vitro drug sensitivity data. Finally, SMTs have the power to identify molecular biomarkers of in vivo response to established and novel drugs in pediatric ALL when combined with exome-seq, RNA-seq and DNA copy number analysis of molecularly-annotated PDXs. Supported by U01CA199000 from the NCI. DisclosuresNo relevant conflicts of interest to declare.

Elderly acute lymphoblastic leukemia: a Mayo Clinic study of 124 patients

Poor outcomes in elderly acute lymphoblastic leukemia (ALL) are well recognized, but the contributors are ill-defined. We characterized 124 patients ≥60 years old at our institution. The majority (n = 102, 82%) were treated with intensive chemotherapy. Of these, 8/102 (8%) died within the first 100 days; 92/102 (90%) achieved complete remission (CR/CRi). Only 31/124 (25%) patients underwent allogeneic hematopoietic stem cell transplantation. The median overall survival (OS) for the entire cohort was 19.8 months. In a multivariate analysis, ECOG performance status ≥2, high white blood cell count, and high lactate dehydrogenase (at time of diagnosis) negatively influenced OS (p<.01). In a subgroup analysis of the intensive treatment group, BCR-ABL1+ patients had markedly better OS (hazard ratio 0.3, 95% CI 0.1–0.7; p<.01). In summary, despite few early deaths and a high CR/CRi rate, elderly ALL continues to have a poor prognosis, underscoring the need for more effective therapies.

Personalized nanomedicine: a rapid, sensitive, and selective UV-vis spectrophotometry method for the quantification of nanostructured PEG-asparaginase activity in children's plasma.

PurposePEGylated asparaginase (PEG-ASNase), which hydrolyzes asparagine to ammonia and aspartic acid, is an effective nanostructured antitumor agent for acute lymphoblastic leukemia (ALL). In order to monitor the activity of PEG-ASNase in plasma and design an individualization project, a rapid and sensitive method to determine PEG-ASNase activity in plasma using ultraviolet–visible spectrophotometry was established.MethodsPEG-ASNase is commonly used in acute lymphoblastic leukemia. With Nessler’s reagent as the chromogenic reagent of ammonia, a stable yellow complex was produced. The units of enzyme activity were defined as micromoles of ammonia released per minute.ResultsCalibration curves fitted by plotting the OD at 450 nm of the Nessler product vs concentration were linear in the range of 27.8–1,111.0 IU/L with r2=0.999. The lower limit of quantification for PEG-ASNase activity in human plasma was 20 IU/L with good accuracy and precision. The intra- and interday precision (relative standard deviation) values were below 10% and accuracy ranged from 90% to 110% at all quality control levels. Analytical recoveries were determined between 90% and 110% for all quality control samples.ConclusionThis study proved that the Nessler method is well validated and can be successfully applied in the determination of plasma samples in the clinical setting for patients with ALL. It takes personalized nanomedicine to an entirely new level.

Recent Developments in Adolescent and Young Adult (AYA) Acute Lymphoblastic Leukemia.

Adolescent and Young Adult (AYA) Oncology is a relatively new field encompassing research in the unique pathophysiology, clinical care, and psychosocial issues facing patients between the ages of 15 and 40 with cancer. About 100,000 of the approximately 1.5 million people diagnosed annually with cancer in the USA are in this age range. This chapter will review notable new developments in the care of adolescents and young adults with acute lymphoblastic leukemia (ALL) within the last 3years. The preponderance of data favors the treatment of AYA ALL patients with pediatric-inspired treatment regimens due to better relapse-free and overall survival. Minimal residual disease (MRD) measurement is emerging as an important prognostic factor and can serve as a new measure of efficacy of the addition of novel therapies to the treatment of patients with new diagnoses. There have been several treatment advances ranging from new cytotoxic agents for ALL to new antibody-based therapy to novel immune therapies such as CAR-T cells. The care of AYA ALL patients is improving as the unique issues for this patient population are addressed.

ALL in Older Adults

ALL in Older Adults

Regulation of p53 and survivin by prodigiosin compound derived from Serratia marcescens contribute to caspase-3-dependent apoptosis in acute lymphoblastic leukemia cells.

Tumor suppressor p53 and proto-oncogene survivin are challenging targets for anticancer drugs in acute lymphoblastic leukemia (ALL) which are associated with chemoresistance. Yet, no p53 and survivin-modulating drug with low toxicity and high efficacy has been approved for clinical application in ALL. Consequently, the search for novel compounds which target p53 or survivin is needed to further advance ALL treatment. Prodigiosin, a secondary metabolite of Serratia marcescens induces apoptosis in cancer cells with no toxicity on normal cells. However, the possible potential of prodigiosin as p53- and survivin-modulating agent in ALL cells has not been investigated. Wt-p53 Molt-4 cells were treated with 100 to 600 nM prodigiosin, after which, viability, cell proliferation rates, survivin and p53 protein levels, caspase-3 activation, and apoptosis were evaluated. After 24-, 48-, and 72-h treatments with 100 to 600 nM prodigiosin, cell proliferation rates were measured to be 93.7-77.3%, 75.5-58.3%, and 55-23.3%, respectively. Treatment for 48 hours with 100 to 600 nM prodigiosin resulted in 41-19% decrease in survivin protein levels followed by 450-950% increases in caspase-3 activation levels. Prodigiosin induced remarkably p53 accumulation and increased p53/survivin and caspase-3/survivin ratios by 6.1 to 11.3 and 10.3 to 47.5-fold at 100 to 600 nM, respectively. Survivin protein levels were inversely proportional to p53 accumulation levels. Low survivin protein levels combined with high levels of p53 accumulation were correlated to higher apoptotic rates. P53 and survivin as molecular targets of prodigiosin contribute to caspase-3-dependent apoptosis in ALL cells and this compound represents an attractive p53- and survivin-modulating agent in ALL.

Abstract 765: Prognostic impact of Philadelphia chromosome in elderly acute lymphoblastic leukemia patients ineligible for hematopoietic stem cell transplantation

Abstract Elderly acute lymphoblastic leukemia (ALL) patients are often unfit for neither dose-intensive chemotherapy nor stem cell transplantation, thus their clinical outcomes have been dismal. The increased incidence of BCR-ABL translocation with increasing age also contributed to poor prognosis but, it was our conjecture that the introduction of potent tyrosine kinase inhibitors (TKI) targeting ABL, BCR-ABL translocation might not deliver negative prognostic impact in elderly ALL. Considering that many elderly Ph- ALL patients do not tolerate cytotoxic chemotherapy required for effective tumor control, we supposed that efficacy of ABL TKIs and their combination strategy with low dose cytotoxic chemotherapy would result in better survival for elderly Ph+ ALL compared to their Ph- counterparts. This study was a multicenter longitudinal cohort study of elderly de novo ALL (≥ 60 years) recruited from 11 hospitals in Korea from January 2005 to December 2015. We considered Ph+ if it was positive in at least one of three techniques: conventional cytogenetics, fluorescence in situ hybridization, or quantitative real-time polymerase chain reaction (qRT-PCR). Among the 96 patients enrolled, there were 50 Ph- ALL patients (52.1%) and 46 Ph+ ALL patients (47.9%). There were no differences between the 2 groups regarding age, sex distribution, performance status, and induction regimens. For treatment outcomes evaluation, the 5 Ph+ ALL patients not receiving TKI were eliminated and 41 Ph+ ALL patients treated with TKI were compared with 50 Ph- ALL patients (Table). Complete remission (CR) was attained in 27 (54.0%) of patients in the Ph- group versus 35 (85.4%) in the Ph+ group (P=0.001). The median RFS was significantly longer in the Ph+ group (3.1 months for Ph- vs. 8.9 months for Ph+, P=0.017). Induction death and induction failure occurred significantly more often in the Ph- group. The median OS was 6.3 months for Ph- group versus 10.3 months for Ph+ group (P=0.033). In conclusion, using real world data, this study reports the possible favorable impact of Ph+ in TKI-era for elderly patients. Citation Format: Ja Min Byun. Prognostic impact of Philadelphia chromosome in elderly acute lymphoblastic leukemia patients ineligible for hematopoietic stem cell transplantation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 765. doi:10.1158/1538-7445.AM2017-765