Attenuation of the Apoptosis Stimulating Protein of TP53-1 (ASPP1) Is a Predictive Marker for Therapy Resistance and Overall Survival in Acute Myeloid Leukemia (AML)

Abstract ASPP1 (PPP1R13B) belongs to a family of p53-binding proteins and enhances apoptosis by stimulation of p53-transactivation of selected proapoptotic target genes. It is preferentially expressed in hematopoietic stem cells (HSC) and together with p53 preserves the genomic integrity of the HSC pool. We have previously demonstrated that ASPP1 is attenuated in AML, which is linked to methylation of the promoter region. ASPP1-interferference models suggest a functional role in leukemogenesis and therapy response. We now provide evidence that ASPP1 is highly altered in AML and attenuated expression levels associate with inferior outcomes: mRNA expression patterns of ASPP1 were assessed in an unselected patient cohort with newly diagnosed AML (n=39) - and were found to be significantly lower compared to bone marrow aspirates of 12 healthy donors (p < 0.0001, Mann-Whitney test) with a median relative expression level of 0.33 (ASPP1 AML) vs. 0.95 (ASPP1 donor). Subcohort analysis by genetic risk profile according to the European LeukemiaNet (ELN) 2017 genetic risk stratification revealed significantly lower expression levels of the intermediate/adverse risk group (n=27) compared to the favorable risk group (n=10, p=0,013) with a median relative expression level of 0.21 (ASPP1 int/adv) vs. 0.46 (ASPP1 good). In addition, analysis of patients undergoing induction chemotherapy demonstrated a significantly lower complete remission (CR) rate in patients with attenuated ASPP1 levels (p=0.0015) with a median relative expression level of 0.13 (ASPP1 nonCR) vs. 0.51 (ASPP1 CR). Of note, patients with a >10x fold decrease of ASPP2 expression were exclusively found in the patient cohort failing induction therapy. To validate these observations in an independent patient set, we next performed a database RNAseq screen on a defined, unselected AML cohort (n=142, FAB M3 were excluded). Together, ASPP1 was independently confirmed as an adverse prognostic factor: Overall survival (OS) was longer in the ASPP1 high cohort vs. the ASPP1 low group with a hazard ratio for death of 0.63. Even though this analysis closely failed significance (0.082) due to lack of power, probability to be alive after 70 months was 45% (ASPP1 high) vs. 10% (ASPP1 low) with a median survival of 19.23 months (ASPP1 low) vs. 26.4 months (ASPP1 high). Most interestingly, looking at the prognostic good risk population: attenuation of ASPP1 resulted in a dramatic decrease of survival with a hazard ratio of 0.29 and a probability to be alive after 70 months of 25% (ASPP1 low) vs. 85% (ASPP1 high). Consequently, screening for ASPP1 expression may define a patient cohort likely to fail (induction) therapy - which might be a target population for hypomethylating agents (HMA) to reconstitute ASPP1 and sensitize towards (chemo)therapy. To address this hypothesis, MOLM-14 acute leukemia cells and patient derived freshly-isolated AML samples were treated with decitabine - resulting in upregulation of ASPP1 mRNA expression levels in all assessed samples. As expected, cells were more chemo-susceptible when exposed to decitabine followed by daunorubicin and cytarabine in an ASPP1-dependent manner: MOLM-14.ASPP1i cell line strains did not reveal a significant change of proapoptotic efficacy when compared to mock strains which significantly benefitted from this approach with more than doubled proapoptotic rates (**** p < 0.0001, t-test), again confirming ASPP1 target specificity. Our results demonstrate that dysfunctional regulation of ASPP1 expression is frequently observed in AML, which associates with therapy responses and survival outcome. Prospective clinical studies are warranted to evaluate the role as a biomarker for risk stratification. Further, we provide a rationale to re-sensitize high-risk patients (defined as ASPP1 low-expressors) towards chemotherapy using HMA priming upfront - and this strategy should be followed in future trials. Disclosures Schittenhelm: BMS: Other: advisory board; Takeda: Other: advisory board; Astellas: Other: advisory board; University of Tuebingen: Patents & Royalties: patent for ASPP2k. Kampa-Schittenhelm: University of Tuebingen: Patents & Royalties: patent related to ASPP2k.

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Evaluation of the Correlation between the mRNA Expression Levels of ystA and ymoA Genes in Y. enterocolitica Strains with Different Enterotoxic Properties

Pathogens ◽

10.3390/pathogens10091136 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1136

Author(s):

Agata Bancerz-Kisiel ◽

Karolina Lipczyńska-Ilczuk

Keyword(s):

Mrna Expression ◽

Clinical Symptoms ◽

Expression Level ◽

Common Source ◽

Expression Levels ◽

Relative Expression ◽

Source Of Infection ◽

Relative Expression Level ◽

Causative Agents ◽

Mrna Expression Levels

Yersinia enterocolitica is one of the main causative agents of human diarrhea. Pigs are a reservoir and the most common source of infection for humans. The aim of this study was to analyze the expression of ystA and ymoA genes in Y. enterocolitica strains with different enterotoxic properties, isolated from humans and pigs. The experiment involved two groups of Y. enterocolitica strains producing and not producing enterotoxin YstA, which were isolated from humans and pigs. All strains were ystA- and ymoA-positive. The expression of ystA and ymoA genes was analyzed by quantitative real-time PCR (qPCR). The relative expression level of the ystA gene was significantly higher than the expression level of the ymoA gene in Y. enterocolitica strains isolated from humans with clinical symptoms of yersiniosis. In other strains, a significant decrease in ystA gene transcription was observed, and the relative expression level of the ymoA gene was significantly higher than the expression level of the ystA gene. Statistically significant differences were not observed in either group of strains isolated from pigs. The results of our study revealed a correlation between mRNA expression levels of ystA and ymoA genes in Y. enterocolitica strains isolated from humans.

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The Role of TLR4 Inducing Macrophage Pyroptosis in the Pathogenesis of Severe Aplastic Anemia

Blood ◽

10.1182/blood-2021-152471 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 1114-1114

Author(s):

Rong Fu ◽

Tian Zhang ◽

Ling Deng ◽

Yang Zhao ◽

Xiaoyu Zhao ◽

...

Keyword(s):

T Cells ◽

Cd8 T Cells ◽

Culture Supernatant ◽

Expression Level ◽

Healthy Controls ◽

Expression Levels ◽

Relative Expression ◽

Caspase 1 ◽

Relative Expression Level ◽

Mrna And Protein Expression

Abstract Introduction：To investigate the expression level of macrophage pyroptosis in patients with severe aplastic anemia (SAA) and its effect on downstream effector T cells, the study explored the mechanism of Toll like receptor 4 (TLR4) inducing macrophage pyroptosis and maintaining immune homeostasis of SAA through pyruvate kinase M2 (PKM2) at cellular and molecular levels, which provided a theoretical basis for the targeted therapy of macrophages in SAA. Three groups of bone marrow macrophages of new diagnosed SAA patients, remission patients and healthy controls were induced and cultured in vitro. qRT-PCR and Western Blot were used to detect pyroptosis-related indicators IL-1β, IL-18, NLRP3, Caspase-1 and GSDMD expression level; ELISA to detect the concentration of IL-1β and IL-18 in the culture supernatant. Results: 1. The mRNA and protein levels of macrophages IL-1β, IL-18, NLRP3, Caspase-1, and GSDMD in the new diagnosed SAA group were significantly higher than those in healthy controls, and the IL-1β and IL-18 concentration in the macrophage culture supernatant increased significantly. The relative mRNA expression levels of macrophages IL-1β, IL-18, NLRP3, Caspase-1, and GSDMD in the new diagnosed group were significantly negatively correlated with WBC, RBC, Hb and PLT. 2. The relative expression level of TLR4 mRNA in macrophages in the new diagnosed group was significantly higher than that in the remission group and healthy controls, and the protein level of TLR4 was significantly increased; at the same time, the relative expression level of TLR4 mRNA was positively correlated with IL-1β, IL-18, NLRP3, Caspase-1 and GSDMD in macrophages. After knocking down TLR4 or adding TLR4 inhibitors, the mRNA and protein expression levels of IL-1β, IL-18, NLRP3, Caspase-1 and GSDMD were significantly lower than those in healthy controls, and the concentration of IL-1β and IL-18 in the culture supernatant reduced significantly. 3. The expression rate of PKM2 in macrophages of new diagnosed group by flow cytometry was significantly higher than that in the remission group and heathy controls, and the levels of PKM2 mRNA and protein were significantly increased. At the same time, the relative expression level of PKM2 mRNA was positively correlated with the expression levels of IL-1β, IL-18, NLRP3, Caspase-1, and GSDMD. After knocking down PKM2, the mRNA and protein expression levels of IL-1β, IL-18, NLRP3, Caspase-1, GSDMD were significantly lower than those healthy controls, and the concentration of IL-1β and IL-18 in the culture supernatant was also significantly reduced. After knocking down TLR4 or adding TLR4 inhibitor, PKM2 mRNA and protein expression levels in macrophages were significantly lower than those in healthy controls. 4. When CD8+ T cells were co-cultured with macrophages which were knocked down TLR4 or added with inhibitors, the expressions of perforin and granzyme B in CD8+ T cells were significantly reduced. CD8+ T cells were further co-cultured with K562 and the apoptosis level of K562 was reduced. Summary：The level of macrophage pyroptosis in SAA patients increased, and the expression levels of macrophages TLR4 and PKM2 increased and positively correlated with the level of pyroptosis. Inhibiting the expression of TLR4 and PKM2, respectively, reduced the level of macrophage pyroptosis, and at the same time inhibiting TLR4 expression reduced the expression of PKM2. Macrophages that inhibited TLR4 expression reduced the killing function of CD8+ T cells. Therefore, TLR4 may induce pyroptosis of macrophages in SAA patients through PKM2 and enhance the killing function of CD8+ T cells. Disclosures No relevant conflicts of interest to declare.

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Topology estimation of gene regulatory networks with relative expression level variations

2012 American Control Conference (ACC) ◽

10.1109/acc.2012.6314870 ◽

2012 ◽

Author(s):

Yali Wang ◽

Tong Zhou

Keyword(s):

Gene Regulatory Networks ◽

Regulatory Networks ◽

Expression Level ◽

Relative Expression ◽

Relative Expression Level ◽

Gene Regulatory ◽

Topology Estimation

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Ph-Negative Chronic Myeloproliferative Neoplasms – Population Analysis, a Single Center 10-years’ Experience

Blood ◽

10.1182/blood.v124.21.5556.5556 ◽

2014 ◽

Vol 124 (21) ◽

pp. 5556-5556

Author(s):

Vasily Shuvaev ◽

Irina Martynkevich ◽

Alla Abdulkadyrova ◽

Vera Udaleva ◽

Tatyana Zamotina ◽

...

Keyword(s):

Overall Survival ◽

Risk Stratification ◽

Molecular Mechanisms ◽

Conflicts Of Interest ◽

Risk Group ◽

Myeloproliferative Neoplasms ◽

Population Analysis ◽

Myelogenous Leukemia ◽

Rank Test ◽

Chronic Myeloproliferative Neoplasms

Abstract Objectives and background. Nowadays chronic myeloproliferative neoplasms (MPN) other than chronic myelogenous leukemia undergo renaissance of interest. It results from advances in decryption of molecular mechanisms of pathogenesis and invention of target drugs. Epidemiological information is needed to assess potential effect and additional costs of new diagnostic and therapeutic techniques. The objective of our study was to review experience of MPN diagnostic and treatment in our center for past ten years. Methods. Our institution serves as primary hematological outpatient department for a half of Saint-Petersburg city with about 2 million inhabitants. We reviewed patients' charts to obtain information about incidence, symptoms, diagnostic test results, treatment options and relationship to prognostic factors. Statistical methods included descriptive statistics, nonparametric ANOVA for frequencies comparisons and Kaplan-Meyer method with log-rank test for survival comparisons in Statistica 7.0 package. Results. Since 2004 to 2013 there were 570 newly diagnosed MPN patients (pts) in our center. This group consisted of primary myelofibrosis (PMF) (203 pts; 126 female, 77 male; median age 63 years, range 16-83 years), essential thrombocythemia (ET) (201 pts; 146 female, 55 male; median age 58 years, range 23-78 years), polycythemia vera (PV) (166 pts; 96 female, 70 male; median age 57 years, range 20-85 years). The incidence rates were stable during study period: PMF incidence varied from 0.65 to 1.35 with mean of 1.01 new patient per 100 000 inhabitants per year; ET had incidence from 0.60 to 2.1 with mean of 1.00 and PV had incidence from 0.5 to 1.15 with mean of 0.83. The most prevalent symptoms of disease were: splenomegaly (65.5%), constitutional symptoms (fever, night sweats, weight loss) (31.0%), anemia (36.3%) thrombosis (24.1%) for PMF; fatigue (33.2%), headache and dizziness (25.6%), arthralgia (21.8%), erythromelalgia (15.8%) for ET; plethora (82.5%), headache and dizziness (52.4%), fatigue (31.3%) for PV. JAK2V617F was detected in 49.7% of PMF pts, 57.8% of ET pts and in 97.7% of PV pts. Thrombosis rates according WHO IPSET-thrombosis system risks` groups of ET and PV pts were: low-risk group 3.33% (3/90), intermediate-risk group 11.1% (13/117) and 39.4% (63/160) in high-risk group with highly significant (p<0.0001) differences between risks' groups. There were 169 lethal outcomes in the analysed group (102 PMF; 31 ET; 36 PV). Ten-years overall survival rates were 49.8% in PMF pts, 84.6% in ET pts and 78.3% in PV pts. (fig.1). Overall survival in PMF was significantly influenced by risk stratification as IPSS, DIPSS and DIPSS+. Survival curves according DIPSS+ groups are presented in fig.1. Conclusions. Patients with MPN are presented in substantial number; therefore need much finance for novel therapy introduction. Risk stratification systems has high predictive value. Innovative drugs treatment results should be evaluated in comparison with historical control. Figure1 Overall survival in PMF patients according to DIPPS+ stratification groups. Figure1. Overall survival in PMF patients according to DIPPS+ stratification groups. Disclosures No relevant conflicts of interest to declare.

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Ethnic Differences in MicroRNA-375 Expression Level and DNA Methylation Status in Type 2 Diabetes of Han and Kazak Populations

Journal of Diabetes Research ◽

10.1155/2014/761938 ◽

2014 ◽

Vol 2014 ◽

pp. 1-7 ◽

Cited By ~ 14

Author(s):

Xiangyun Chang ◽

Siyuan Li ◽

Jun Li ◽

Liang Yin ◽

Ting Zhou ◽

...

Keyword(s):

Type 2 Diabetes ◽

Methylation Status ◽

Cpg Methylation ◽

Expression Level ◽

Related Factor ◽

Expression Levels ◽

Relative Expression ◽

Cpg Sites ◽

Significant Difference

Han population is six times as likely as Kazak population to present with type 2 diabetes mellitus (T2DM) in China. We hypothesize that differential expression and CpG methylation of miR-375 may be an ethnic-related factor that influences the incidence of T2DM. The expression level of miR-375 was examined using real-time PCR on Kazak and Han T2DM plasma samples. Furthermore, the methylation levels of CpG sites of miR-375 promoter were determined by MassARRAY Spectrometry in these samples. The relative expression levels of plasma miR-375 in Kazak T2DM samples are 1, and the relative expression levels of plasma miR-375 in Han T2DM samples are 3. The mean level of miR-375 methylation, calculated from the methylation levels of the CpG sites, was 8.47% for the Kazak T2DM group and 10.38% for the Han T2DM group. Further, five CpG units showed a statistically significant difference between Kazak and Han T2DM samples, and, among them, four were hypomethylated and only one CpG unit showed hypermethylation in Kazak T2DM samples. These findings indicate that the expression levels of plasma miR-375 and its CpG methylation in the promoter region are ethnically different, which may contribute to the different incidence of diabetes observed in Kazak and Han populations.

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Hydrogen-Rich Water Ameliorates Murine Chronic Graft-versus-Host Disease through Antioxidation

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/1165928 ◽

2021 ◽

Vol 2021 ◽

pp. 1-8

Author(s):

Liren Qian ◽

Jiaxin Liu ◽

Weina Ma ◽

Yu Liu ◽

Xiaona Wang ◽

...

Keyword(s):

Survival Rate ◽

Treatment Option ◽

Graft Versus Host Disease ◽

Caspase 3 ◽

Expression Level ◽

Therapeutic Effects ◽

Expression Levels ◽

Relative Expression ◽

Graft Versus Host ◽

Staining Score

Background. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an important treatment option for various hematopoietic diseases and certain hereditary diseases. Chronic graft-versus-host disease (cGVHD) has become the main life-threatening complication and cause of death in later stage postallo-HSCT. Current treatment options for cGVHD are limited. Hydrogen gas (H2) has been demonstrated that has antioxidative, anti-inflammatory, and antifibrosis effects. The aim of this study was to confirm whether oral administration hydrogen-rich water exerted therapeutic effects on a scleroderma cGVHD mouse model and tried to explain the mechanism underly it. Methods. A mouse cGVHD model was established by haploidentical bone marrow transplantation. To evaluate therapeutic effects of H2 on cGVHD, survival rate, changes in clinical scores, and skin pathologic characteristics of cGVHD mice were observed. To evaluate its therapeutic mechanism, we detected the expression levels of antioxidative enzymes heme oxygenase-1(HO-1) and NAD (P)H: quinone acceptor oxidoreductase 1(NQO1) in skin homogenates. We also detected the expression level of the apoptotic protein caspase-3 in skin homogenates. Results. 1-month survival rate of cGVHD mice in the hydrogen group reached 93.3%, significantly higher than 66.7% in the nonhydrogen group ( p < 0.05 ). Clinical score of cGVHD mice was improved by hydrogen-rich water at 96 days posttransplantation (2.2 versus 4.5, p < 0.05 ). The skin pathological condition of cGVHD mice was significantly improved by hydrogen-rich water. At 96 days posttransplantation, average skin pathological hematoxylin and eosin (HE) staining score in the hydrogen group was 1.05, which was significantly lower than 3.2 in the nonhydrogen group ( p < 0.01 ). Average Masson staining score was 0.6 point in the hydrogen group, lower than 0.9 point in the nonhydrogen group ( p < 0.05 ). Both the relative expression levels of HO-1 and NQO1 proteins in skin specimens of cGVHD mice in the hydrogen group were lower than that in the nonhydrogen group (2.47 versus 6.21 and 1.83 versus 3.59, p < 0.05 ). The relative expression level of caspase-3 protein in skin specimens of cGVHD mice increased to 7.17 on the 96th day after transplantation, significantly higher than 4.36 in the hydrogen group. Conclusion. In this study, we found that oral hydrogen-rich water improved the survival rate and clinical symptoms of cGVHD mice by antioxidant and antiapoptosis. This study would pave the way for further clinical study, which may provide a new treatment option for cGVHD.

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Correlation of Vitamin D3 with the Expression of RORγt and Foxp3 mRNAs in the Peripheral Blood of Myasthenia Gravis Patients

NeuroImmunoModulation ◽

10.1159/000510861 ◽

2020 ◽

pp. 1-7

Author(s):

Pan Huang ◽

Xiao-ying He ◽

Min Xu

Keyword(s):

Myasthenia Gravis ◽

Peripheral Blood ◽

Normal Subjects ◽

Orphan Receptor ◽

Expression Level ◽

Control Group ◽

Foxp3 Mrna ◽

Expression Levels ◽

Relative Expression ◽

The Relationship

Objectives: to investigate the expression levels of 1,25(OH)2D3 in the peripheral blood from patients with myasthenia gravis (MG) and to correlate levels with retinoid-related orphan receptor γt (RORγt) and forkhead or winged-helix transcription factor 3 (Foxp3) mRNA expression. Methods: Sixty-seven patients with MG were enrolled in the experimental group, and 50 normal subjects were selected as the control group. The expression levels of 1,25(OH)2D3 and RORγt and Foxp3 mRNAs were measured in the serum of the 2 patient groups and the relationship between factors were correlated with the severity score of MG. The relationship between the levels of 1,25(OH)2D3 and the relative expressions of RORγt and Foxp3 mRNAs was determined. Results: There were no differences between groups regarding patient’s baseline data. 1,25(OH)2D3 and RORγt and Foxp3 mRNAs are differentially expressed in the MG group and the control group (p < 0.05). QMG score is negatively correlated with the expression level of peripheral blood 1,25(OH)2D3 and Foxp3 mRNA (r = −0.797, −0.543; p < 0.01) and positively correlated with the relative expression level of RORγt mRNA (r = 0.539; p < 0.01). 1,25(OH)2D3 expression level was negatively correlated with the relative expression of RORγt mRNA (r = −0.559; p < 0.01) and positively correlated with the relative expression of Foxp3 mRNA (r = 0.390; p < 0.01). Conclusions: The levels of 1,25(OH)2D3 were shown to be lower in patients with MG compared to normal controls. The observed low levels of 1,25(OH)2D3 may lead to changes in the expression of RORγt and Foxp3 mRNAs involved in MG.

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Heterogeneity of MYC Abnormalities in Multiple Myeloma

Blood ◽

10.1182/blood-2020-140844 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 2-3

Author(s):

Neeraj Sharma ◽

James B Smadbeck ◽

Nadine Abdallah ◽

Kathryn E. Pearce ◽

Yan Asmann ◽

...

Keyword(s):

Gene Expression ◽

Multiple Myeloma ◽

Overall Survival ◽

Clinical Trials ◽

Genome Sequencing ◽

Mayo Clinic ◽

Research Funding ◽

Advisory Board ◽

Expression Levels ◽

Myc Gene

Background: Multiple myeloma (MM) is an incurable plasma cell malignancy and genetic abnormalities contribute to disease heterogeneity and outcome. Primary abnormalities, namely recurrent immunoglobulin (Ig) heavy chain translocations and hyperdiploidy, occur early in disease course. Secondary events, such as MYC abnormalities occur upon progression. Earlier studies showed MYC abnormalities detected by FISH or by capture sequencing were independently associated with poor outcome (Walker, et al., BCJ, 2014), while recent studies using WGS did not support this finding (excluding MYC/IGL) (Mikulasova, et al. Haematologica, 2020; Misund, et al. Leukemia, 2020). We hypothesize these discrepancies are due to differences in methods and sensitivities of detection of MYC abnormalities by FISH vs. WGS. Given that MYC abnormalities often display remarkable genomic heterogeneity with numerous gene partners, reduced detection of MYC abnormalities by FISH is not unexpected. This hypothesis is supported by lower frequencies of MYC abnormalities found by FISH (15%) vs. NGS (30-35%) consistent with ~50% false-negative rate of the MYC FISH probe (Smadbeck, et al. BCJ, 2019). To better understand the role of MYC in myeloma disease outcome, we compared the MYC abnormality subtype identified by FISH or NGS vs. MYC gene expression levels and overall survival. Methods: We performed a retrospective study of newly diagnosed MM patients seen at Mayo Clinic or enrolled in the MMRF CoMMpass trial. For Mayo cases, MYC FISH results (breakapart probe, Abbott) were obtained from the Mayo Clinic Genomics database (N=1342) and mate pair sequencing (MPseq) was performed on 140 cases. For CoMMpass cases, we obtained tumor long-insert whole genome sequencing (WGS), RNA sequencing (RNAseq) for gene expression and clinical outcome data. Overall survival (OS) was defined as time from diagnosis to death from any cause or to last follow up. Survival curves were estimated using Kaplan Meier and compared using the Log-Rank test. Statistical analyses performed using SPSS and JMP with significance determined when P <0.05. Results: We first evaluated the impact of MYC abnormalities on OS when detected by FISH or NGS. In Mayo cases, OS was significantly shorter in patients with MYC abnormalities compared to patients without MYC abnormalities using FISH (5.3 vs. 8.0 years, P<0.001, N=1342). In contrast, there was no significant difference in OS between patients with or without MYC or abnormalities using MPseq or WGS in both the Mayo and CoMMpass cohorts (Mayo: 6.4 vs. and 6.9 years P=0.78, N=140; CoMMpass: 4.9 vs. and 5.1 years P=0.74, N=546). Since FISH-detected MYC abnormalities were associated with poor outcome, we evaluated differences in the types of MYC abnormalities identified FISH and genome sequencing; 270 of 658 CoMMpass cases had a MYC abnormality and 12 abnormality subgroups were identified. In the Mayo cases, FISH preferentially detected translocations and complex abnormalities and missed insertions with flanking duplicating sequences or terminal tandem duplications (TTD) that occur telomeric to MYC. Since the level of MYC expression should be a consequence of the various genomic abnormalities altering the MYC gene region, we compared MYC expression levels in relation to MYC abnormality subgroups. Highest expression was seen with MYC amplification, followed by Ig abnormalities, non-Ig abnormalities, complex deletion/duplications, proximal deletions, non-Ig insertions, terminal deletions, TTD, trisomy 8, no MYC structural variation, monosomy 8 and cases with MAX mutations had the lowest expression. Abnormalities identified by FISH had higher MYC expression (83.5 TPM) compared to cases predicted to be missed by FISH (63.2 TPM). We tested if high MYC expression, irrespective of MYC structural abnormality, was associated with differences in OS. Boxplot analysis was used to categorize MYC expression in 631 CoMMpass patients as top quartile/high MYC expression (Q4≥ 75 TPM, n=159) and bottom quartile/low MYC expression (Q1≤ 16.5 TPM, n= 158) (see Figure). OS was significantly shorter in patients with high MYC expression compared to patients with low MYC expression (4.6 vs. 5.3 years, P <0.038). Conclusion: We show that FISH detects only a subset of the MYC abnormalities detected by genome sequencing, and that FISH-detected MYC abnormalities are associated with higher MYC gene expression and decreased survival. Figure 1 Disclosures Kumar: Kite Pharma: Consultancy, Research Funding; Janssen Oncology: Other: Research funding for clinical trials to the institution, Consulting/Advisory Board participation with no personal payments; AbbVie: Other: Research funding for clinical trials to the institution, Consulting/Advisory Board participation with no personal payments; Oncopeptides: Consultancy, Other: Independent Review Committee; IRC member; Dr. Reddy's Laboratories: Honoraria; Cellectar: Other; Takeda: Other: Research funding for clinical trials to the institution, Consulting/Advisory Board participation with no personal payments; Novartis: Research Funding; Tenebio: Other, Research Funding; Carsgen: Other, Research Funding; Amgen: Consultancy, Other: Research funding for clinical trials to the institution, Consulting/Advisory Board participation with no personal payments, Research Funding; Merck: Consultancy, Research Funding; Genecentrix: Consultancy; BMS: Consultancy, Research Funding; Karyopharm: Consultancy; Celgene/BMS: Other: Research funding for clinical trials to the institution, Consulting/Advisory Board participation with no personal payments; Genentech/Roche: Other: Research funding for clinical trials to the institution, Consulting/Advisory Board participation with no personal payments; Sanofi: Research Funding; MedImmune: Research Funding; Adaptive Biotechnologies: Consultancy.

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Low Expression Of BCL11B Predicts Poor Overall Survival In Adult Standard Risk T-ALL

Blood ◽

10.1182/blood.v122.21.1362.1362 ◽

2013 ◽

Vol 122 (21) ◽

pp. 1362-1362

Author(s):

Isabelle Bartram ◽

Nicola Gökbuget ◽

Cornelia Schlee ◽

Sandra Heesch ◽

Lars Fransecky ◽

...

Keyword(s):

Overall Survival ◽

T Cell ◽

Poor Prognosis ◽

Clinical Characteristics ◽

Risk Group ◽

High Expression ◽

Prognostic Impact ◽

Expression Levels ◽

Standard Risk ◽

Low Expression

Abstract Introduction Although risk stratification, detection of minimal residual disease (MRD) and implementation of novel therapeutic agents have improved outcome in acute lymphoblastic leukemia (ALL), survival in adult T-ALL patients still remains unsatisfactory. Therefore, new prognostic markers and potential therapeutic targets are warranted. BCL11B, a key player in normal T-cell development, has recently gained interest due to its high mutation rate (9-16%) in T-ALL. We investigated the frequency of BCL11B mutations, expression levels and the prognostic value in a large uniformly treated cohort of adult T-ALL. Patients and methods We analyzed bone marrow (BM) samples of 201 adult T-ALL patients sent to the reference laboratory of the German Multicenter Study Group for Adult ALL (GMALL). BCL11B expression was determined in 195 patients by qRT-PCR, BCL11B mutations were assessed in 178 patients by Sanger sequencing of exon 4 (including all 6 zink finger [ZF] domains). Low expression of BCL11B was defined by expression levels in the lowest quartile (BCL11Blow), high expression by levels in the three remaining quartiles (BCL11Bhigh). Samples had previously been characterized for expression of BAALC, IGFBP7, MN1, WT1, GATA3, ERG as well as for the mutations status of NOTCH1, WT1, and TCR rearrangements. Clinical data were available for 169 patients enrolled on the GMALL trials. We generated BCL11B associated gene expression profiles (GEP) derived from an independent set of 86 T-ALL patients enrolled in the Microarrays Innovations in LEukemia multicenter study. Results BCL11B was aberrantly expressed in adult T-ALL with significantly higher expression levels in thymic compared to early T-ALL (0.6 vs. 0.3, P=0.01). Expression of genes associated with a prognostic impact (BAALC, IGFBP7, ERG) or/and T-cell stage dependent expression profile (GATA3, IGFBP7) showed that BCL11Blow (n=49) had higher MN1 (1.6 vs, 0.3, P=0.01), IGFBP7 (1.3 vs. 0.5, P=0.02), and lower GATA3 expression (2.1 vs. 5.7, P<0.01) compared to BCL11Bhigh patients (n=146). This maturation stage specific expression of BCL11B was stressed by a higher rate of TCR rearrangement in BCL11Bhigh patients (73% vs. 27%, P=0.005) and further underlined by the BCL11B derived GEP linking low BCL11B to an immature molecular signature characterized by high expression of BAALC, IGFBP7, FLT3, CD34. Regarding clinical characteristics, low BCL11B expression was associated with a poor prognosis (5-year overall survival (OS): low 35% (n=40) vs. high 53% (n=129), P=0.02) in the overall T-ALL cohort. In the standard risk group of thymic T-ALL (n=102), BCL11Blow identified patients with an unexpected poor outcome (5-year OS: 20%, n=18) compared to BCL11Bhigh (62%, n=84, P<0.001). In addition, BCL11Blow thymic T-ALL patients showed a lower remission rate (5 years: 38% vs. 72%, P=0.02). BCL11B mutations were found in 24 of the 178 (13.5 %) T-ALL patients. In 9 patients, mutations resulted in frame shifts, whereas the remaining, except for a single one, missense mutations were located in the ZF domains: three resulting in introduction of a stop codon. BCL11B mutations were enriched in the mature immunophenotype (thymic: 20%, mature: 8%, early: 3%, P=0.03). No differences were observed in mRNA levels for BAALC, IGFBP7, MN1, WT1, GATA3, ERG, but patients with BCL11B mutations were less frequently assigned to the BCL11Blow group compared to those with high expression (5% vs. 95%, P=0.02). Regarding the clinical characteristics, BCL11B mutations had no prognostic impact regarding OS, neither in the total T-ALL cohort (5-year OS: 56% vs. 48%, n.s.) nor in the thymic T-ALL subgroup (5-year OS: 65% vs. 51%, n.s). Conclusion Our data confirmingly show a high frequency of BCL11B mutations (13.5%) in the so far largest cohort of adult T-ALL patients. As loss of function mutations were restricted to functional ZF domains and recurrently occurred in thymic T-ALL, these data stress a potential pathogenetic role of BCL11B as T-cell specific transcription factor. Importantly, low expression was associated with poor prognosis; in particular in the standard risk group of thymic T-ALL, BCL11Blow is a novel marker that identifies patients with an unacceptable poor prognosis. These findings might help to improve risk stratification in a significant proportion of adult T-ALL patients, which fail to standard therapy despite the favourable immunophenotypic characteristics. Disclosures: No relevant conflicts of interest to declare.

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Litchi Flowering is Regulated by Expression of Short Vegetative Phase Genes

Journal of the American Society for Horticultural Science ◽

10.21273/jashs04316-17 ◽

2018 ◽

Vol 143 (2) ◽

pp. 101-109

Author(s):

Jiaqi Hu ◽

Hye-Ji Kim ◽

Houbin Chen ◽

Biyan Zhou

Keyword(s):

Low Temperature ◽

Expression Patterns ◽

Bioinformatic Analysis ◽

Molecular Characteristics ◽

Vegetative Phase ◽

Expression Levels ◽

Relative Expression ◽

Short Vegetative Phase ◽

Relative Expression Level

Short vegetative phase (SVP), a MADS-domain transcription factor, was shown to act as a repressor of flowering in arabidopsis (Arabidopsis thaliana). Although the role of SVPs in flowering is well characterized in the model plant arabidopsis, little is known in evergreen woody litchi (Litchi chinensis). In this study, three litchi SVP homologs (LcSVP1, LcSVP2, and LcSVP3) were cloned, and the bioinformatic analysis of the LcSVPs was carried out to identify their molecular characteristics. Their expression patterns in the apical meristem (AM) during the transition from vegetative to reproductive phase were studied under natural flowering inductive conditions. Also, brassinosteroid (BR) treatment under low temperature conditions was performed to elucidate the role of LcSVPs in the BR-regulated flowering. The results showed that LcSVPs belonged to the MADS superfamily. LcSVP relative expression levels in AMs of the early- and late-flowering cultivars showed decreasing trends with the transition from vegetative to reproductive growth. Under low temperature condition, relative expression levels of LcSVP1, LcSVP2, and LcSVP3 in AMs or panicle primordia showed decreasing trends, whereas those in the AMs of the BR-treated trees remained at relatively high levels. Relative expression analysis of the litchi homolog, flowering locus t 1 (LcFT1), showed that the BR-treated leaves had lower relative expression level than nontreated control leaves. The findings suggest that LcSVPs act as repressors involved in flowering in natural conditions and the BR-regulated flowering.

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