Expression of CD123 in B-acute lymphoblastic leukaemia as a predictor of Bcr/Abl rearrangement and disease relapse

Chemoimmunotherapy Combining Vincristine, Dexamethasone and Epratuzumab (hLL2) As Salvage Regimen for Older Relapsed/Refractory, CD22+ B-Acute Lymphoblastic Leukemia (B-ALL) Patients: Results of the French Non-Intensive Phase 2 Prospective Cheprall Study

目的探讨成人急性B淋巴细胞白血病（B-ALL）患者的基因突变谱，并分析其对患者预后的影响。方法收集113例2009年6月至2015年9月收治的成人B-ALL患者DNA标本，采用靶向特异的二代测序技术，针对血液恶性疾病相关的112种基因进行突变分析，通过多个数据库筛选出可能致病的基因突变，描述其发生频谱，并通过Kaplan-Meier、Cox回归模型分析突变基因对患者总生存（OS）和无复发生存（RFS）的影响。结果113例患者中103例（92.0%）发生至少一种基因突变，29例（25.6%）患者发生≥3种基因突变。所有患者中突变率较高的基因有SF1、FAT1、MPL、PTPN11、NRAS等，Ph阳性B-ALL和Ph阴性B-ALL患者中基因突变特点不尽一致。进一步分析基因突变对患者预后的影响，发现在Ph阴性B-ALL中伴JAK-STAT信号通路相关基因突变的患者（如JAK1、JAK2突变）较该信号通路未受影响的患者预后差（OS：P值分别为0.011和0.001；RFS：P值分别为0.014和<0.001），而伴PTPN11突变的B-ALL患者较不伴PTPN11突变的患者有较好的OS及RFS，但差异无统计学意义（P值均>0.05）；在Ph阳性B-ALL患者中，表观遗传学修饰相关的信号通路异常的患者预后较差（OS：P=0.038；RFS：P=0.047）。结论基因突变在成人B-ALL中存在普遍性，发生频谱因亚型而异，涉及多种信号通路，JAK家族相关基因突变常提示患者预后较差，突变基因之间的共存现象也预示着成人B-ALL患者的遗传复杂性和不稳定性。

Background: Over-activation of Wnt (derived from names of two genes; DrosophilaWingless and mouse Int-1) pathway is incriminated in leukemogenesis. Functionalloss of Wnt antagonists; DKK (Dikkopf) and SFRPs (secreted frizzled-relatedprotein), can contribute to Wnt hyper-activation. Silencing of Wnt antagonists byhypermethylation is reported in human malignancies as well as in hematopoieticmalignancies. Our aim was to estimate the frequency and the possible impact ofhypermethylation of the SFRP1 and DKK3 in acute leukemia. Methods: We evaluatedSFRP1 and DKK3 methylation status using methylation specific polymerase chainreaction (MS-PCR) in 50 acute myeloid leukemia (AML) and 30 B-acutelymphoblastic leukemia (B-ALL) patients and 20 age and sex matched controls.Results: The frequency of methylation in B-ALL patients was 40% for SFRP1, 40%for DKK-3, in AML patients; the frequency was 44% for SFRP1, 36% for DKK-3. Allthe control subjects had no aberrant methylation in either SFRP1 or DKK-3. B-ALLand AML groups showed no statistical significant difference in the frequency ofSFRP1 or DKK-3 methylation. B-ALL patients with M-SFRP1 had a significantlyhigher mean platelets count and a lower mean age compared to B-ALL patients withUM-SFRP1, no other significant clinical or hematological difference wasencountered between patients with M-SFRP1 and UM-SFRP1 or between M-DKK3and UM-DKK3 patients in the ALL or the AML group. Different B-ALL and AMLprognostic cytogenetic groups showed nearby frequency of SFRP1 and DKK3methylation. Conclusion: SFRP1 and DKK3 methylation is frequent in acuteleukemia. Treatment with demethylating agents may reverse the overactivated Wntsignaling in patients with methylated phenotype.

Bone Marrow Lymphoid Infiltrates and Aplasia after CD19-Directed CAR T Cell Therapy in Pediatric B-Lymphoblastic Leukemia

Clinical Implementation of a Testing Algorithm for the Diagnosis of Ph-like B-Cell Acute Lymphoblastic Leukemia

See related article in EMBO Molecular Medicine http://dx.doi.org/10.1002/emmm.201201703

Inherited polymorphic sequence variations in drug transport genes like ABCB1 impact a portion of patients with hematologic malignancies that show intrinsic or acquire resistance to treatment. Keeping in view inter-individual sensitivities for such drugs, we through this case-control study tested whether ABCB1 C3435T and G2677T polymorphisms have any influence on the risk and treatment response in patients with chronic myeloid leukemia (CML) and B-acute lymphoblastic leukemia (B-ALL). Genotyping for ABCB1 polymorphisms was performed by polymerase chain reaction-restriction fragment length polymorphism in 100 CML and 80 B-ALL patients along with 100 age and gender matched healthy controls. ABCB1 C3435T and G2677T polymorphism showed no association with CML. Genotype distribution revealed significant higher frequency of TT genotype for both SNPs in B-ALL cases and associated with increased B-ALL risk (OR 2.5, p = 0.04 for 3435TT; OR 2.4, p = 0.04 for 2677TT). There was no significant difference in genotype frequency of 3435C > T and 2677G > T among resistant and responsive groups for the two leukemia types. Kaplan-Meier survival plots revealed significantly lower event free survival in CML and B-ALL patients that were carriers of 3435TT genotype (p < 0.05). Multivariate analysis considered 3435TT genotype as independent risk factor for imatinib resistance in CML cases (HR 6.24, p = 0.002) and increased relapse risk in B-ALL patients (HR 4.51, p = 0.03). The current study provides preliminary evidence of a significant association between variant TT genotype and increased B-ALL risk. Also, results suggest that ABCB1 3435TT genotype increases imatinib resistance in CML and influence therapeutic outcome in B-ALL.

Development of CAR T Cells Targeting U5 snRNP200 for the Treatment of Acute Myeloid and B-Lymphoid Leukemias

The aim of this study was to clarify the clinical significance of CD37 expression in B cells from B acute lymphoblastic leukemia (B-ALL) and B cell non-Hodgkin's lymphoma (B-NHL). The expression level of CD37 on B cells from bone marrow samples of normal controls (n = 19), B-ALL patients [including untreated cases (n = 5) and cases with minimal residual disease (MRD, n = 15)] and B-NHL patients (n = 25) whose bone marrow was involved by lymphoma cells, was detected by multiple parameter flow cytometry. The results indicated that the B cells from both untreated cases and cases with MRD lowly expressed CD37 (1.04 ± 0.24 and 1.50 ± 0.89), the normal precursor B cells (control cases) also lowly expressed CD37 (1.64 ± 0.52). There was no difference of CD37 expression level between 3 groups of cases(P > 0.05). Meanwhile the normal mature B cells and B-NHL cells highly expressed CD37 (14.23 ± 7.84 and 14.53 ± 10.93), but there was no difference of CD37 expression between them (P > 0.05). The comparison of CD37 expression level in normal B cells of development stages showed that the progenitor B cells lowly expressed CD37 (0.88 ± 0.17), the CD37 expression of precursor B cells was enhanced (2.44 ± 0.69), while the CD37 expression level of mature B cells was highest. It is concluded that the low expression of CD37 is not the characteristic of B- ALL cells. The expression level of CD37 increases gradually during the mature process of B cells, i.e, the expression level of CD37 does not associate with benignity or malignancy of B cells.

Background:This study was conducted to evaluate CD30 expression in minimum residual disease after chemotherapy in B-acute lymphoblastic leukemia (B-ALL).Materials and Methods:This was a cross-sectional study on 30 new cases of B-ALL between 2018 and 2019. We checked CD30 expressions in fresh bone marrow aspirates by flow cytometry. After 28 days of routine chemotherapy, we calculated minimal residual disease in CD30 positive and negative patients and compare them by Kolmogorov–Smirnov test.Results:Thirty patients with B-ALL with a mean age of 15.62 ± 20.488 were included in the study. CD30 marker was positive in about 10 patients and was negative in about 20 participants. Mean blast count in baseline in CD30 positive group was 77 ± 7.88%, in negative group was 76.3 ± 17.78 % (P = 0.292). After 28 days of chemotherapy mean minimal residual disease (MRD) was 1.07 ± 3.754 in the negative group, 0.12 ± 0.034 in the positive group (P = 0.025).Conclusion:Lower MRD on day 28 after chemotherapy was seen in B-ALL patients with baseline CD30 expression.

Background: The study focuses on B-acute lymphoblastic leukemia (B-ALL), a common childhood blood cancer marked by an overproduction of B lymphocytes. It explores the role of cytokine expression changes, particularly Interleukin-33 (IL-33), which has both anti- and pro-tumor effects. The research assesses the expression of IL-33 and its receptor, suppression of tumorigenicity 2 (ST2), in B-ALL patients before and after chemotherapy treatment. Materials and Methods: In this quasi-experimental investigation, peripheral blood specimens were taken from 33 newly-diagnosed/untreated ALL patients. An induction chemotherapy course was administered to ALL patients for 30 days following diagnosis. Blood samples were taken again after the completion of induction chemotherapy. 30 healthy individuals with matched age and gender were also recruited as a control group. Serum IL-33 quantities were assessed using the ELISA method. Total RNA was extracted from peripheral blood samples, and the gene expression of IL33 and ST2 was measured using real-time PCR. Results: In B-ALL patients, the pre-treatment serum IL-33 concentrations (33.89 ± 4.38 Pg/mL) were substantially greater than the healthy group (20.66 ± 2.20 Pg/mL, P&lt;0.02). Pre-treatment expression of IL-33 and ST2 mRNA (1.79 ± 0.18 and 1.42 ± 0.18) was also significantly higher than those in the healthy group (1.00 ± 0.10 and 1.00 ± 0.09; P&lt;0.001 and P&lt;0.05, respectively). After chemotherapy, serum IL-33 concentrations (22.40 ± 2.60 Pg/mL) and mRNA expression of IL-33 (0.91 ± 0.15) were significantly reduced compared to pre-treatment levels (P&lt;0.04 and P&lt;0.001, respectively). There were no notable differences in serum IL-33 levels or mRNA expression of IL-33 and ST2 between controls and treated patients (P&lt;0.61, P&lt;0.63, and P&lt;0.48, respectively). Conclusion: Greater expression of IL-33 and ST2 was observed in B-ALL patients, which were decreased at the end of a therapy course.

Background:CD34 antigen is expressed by early hematopoietic progenitor cells and acute leukemia cells. Its expression is associated with good prognosis in acute myeloid leukemia. Literature is sparse on its prognostic significance in B- acute lymphoblastic leukemia (B-ALL) especially from India. Hence the present study was undertaken to analyse the frequency of CD34 expression in B-ALL in Indian patients and determine its prognostic significance by associating with other prognostic markers and aberrant antigen expression. Methods:Seventy-five B-ALL patients diagnosed by flow cytometry over a period of 3½ year were studied. Correlation of CD34 expression was studied with gender, age, total leucocyte count (TLC), French-American-British (FAB) morphological type, immuno-phenotypic markers, cytogenetics and minimal residual disease. Differences between groups were evaluated using Student’s T-test for quantitative data and Chi-square test/Fishers exact T-test for qualitative variables. P value <0.05 was considered significant. Results:CD34 was positive in 66.66% (50/75) cases while it was negative in rest (33.33%; 25/75) cases. CD13, CD33, CD5, CD7 and CD11b were more frequently expressed in CD34 negative B-ALL (p=0.025). No association of CD34 expression was found with gender, age, TLC, FAB morphological type, other immune-phenotypic markers, MRD and cytogenetics studied.Conclusions:The expression of CD34 does not associate with known prognostic markers in B-ALL. However, absence of CD34 is associated with aberrant immunophenotypic expression (myeloid + T-cell antigens) in these patients. Larger studies with larger sample size and more extensive immunophenotypic panel needs to be done in Indian setup to confirm these findings.

Background: B-acute lymphoblastic leukemia (B-ALL) is a hematological neoplasm of the stem lymphoid cell of the B lineage, characterized by the presence of genetic alterations closely related to the course of the disease. The number of alterations identified in these patients grows as studies of the disease progress, but in clinical practice, the conventional techniques frequently used are only capable of detecting the most common alterations. However, techniques, such as next-generation sequencing (NGS), are being implemented to detect a wide spectrum of new alterations that also include point mutations. Methods: In this study, we designed and validated a comprehensive custom NGS panel to detect the main genetic alterations present in the disease in a single step. For this purpose, 75 B-ALL diagnosis samples from patients previously characterized by standard-of-care diagnostic techniques were sequenced. Results: The use of the custom NGS panel allowed the correct detection of the main genetic alterations present in B-ALL patients, including the presence of an aneuploid clone in 14 of the samples and some of the recurrent fusion genes in 35 of the samples. The panel was also able to successfully detect a number of secondary alterations, such as single nucleotide variants (SNVs) and copy number variations (CNVs) in 66 and 46 of the samples analyzed, respectively, allowing for further refinement of the stratification of patients. The custom NGS panel could also detect alterations with a high level of sensitivity and reproducibility when the findings obtained by NGS were compared with those obtained from other conventional techniques. Conclusions: The use of this custom NGS panel allows us to quickly and efficiently detect the main genetic alterations present in B-ALL patients in a single assay (SNVs and insertions/deletions (INDELs), recurrent fusion genes, CNVs, aneuploidies, and single nucleotide polymorphisms (SNPs) associated with pharmacogenetics). The application of this panel would thus allow us to speed up and simplify the molecular diagnosis of patients, helping patient stratification and management.

Inotuzumab Use: A Single-Center Experience in B-Cell Acute Lymphoblastic Leukemia

Single-Cell Analyses Identify Transcriptional Characterizations of Subsets Associated with Efficacy and Toxicity for CAR-T Immunotherapy in B-ALL Patients