Rhoa Mutation Is a Potential Biomarker Associated with Adverse Prognosis and High- Tumor Burden in Patients with Nodal Peripheral Lymphomas with T-Helper Follicular Phenotype (nPTCL-Thf): Data from a Brazilian Retrospective Cohort of Nodal PTCL

Abstract Introduction: Nodal PTCL constitute a rare group of aggressive malignancies with heterogeneous clinical-biological presentation and outcomes. In the last decade, its pathophysiological knowledge has been improved, with descriptions of gene mutations associated with epigenetic phenomena (IDH2, TET2 and DNMT3A) and of the RHOA G17V mutation playing a fundamental role in the lymphomagenesis. However, the prognostic impact of these alterations is still controversial, and particularly, in the case of the RHOA mutation, it has never been previously accessed in the literature. Our group has described and validated biomarkers with potential prognostic impact in nPTCL patients, including overexpression of the genes CCNA2, GATA3 and monocytosis in peripheral blood [1,2,3]. The identification of new potential molecular biomarkers can refine the prognostic stratification of these tumors and allow identification of targets for future specific therapies. The aim of this study was to evaluate the frequency and prognostic impact of mutations in the IDH2, TET2, DNMT3 and RHOA genes in Brazilian patients with nPTCL. Methods: In this observational, retrospective and unicentric study, we analyzed the clinical-epidemiological characteristics, outcomes and mutational profile of 59 Brazilian patients with nPTCL treated at the HC-FMUSP, from January 2000 to December 2017. All cases were submitted to centralized histopathological review and were classified according to the criteria proposed by WHO-2008. Cases initially categorized as PTCL/NOS were later reclassified according to WHO-2016 criteria. FFPE-tumor samples from patients with PTCL/NOS, AITL, ALK+/ALCL and ALK-/ALCL were submitted to DNA extraction using QIAmp DNA FFPE kit. For amplification of specific products of target-genes primers flanking the hot spots regions were designed. After this step, the PCR products were submitted to first generation sequencing in 3500 Genetic Analyzer. Absolute and relative frequencies of mutations were accessed for the total cohort and its pathological subtypes. OS and PFS curves were constructed using the Kaplan-Meier method. Log-rank test was used to estimate the prognostic impact of mutations. Results: The clinical-epidemiological characteristics of the 59 Brazilian-patients with nPTCL are summarized in Table 1. With a median follow-up of 3.70 years (0.90-12.4 years), the estimated 2-years OS and PFS were 59.1% and 47.2%, respectively. ORR was 55.9% (33/59), with early relapse rate (< 12 months) of 14.3% (5/59) and global death rate of 52.5% (31/59). In the total cohort, we found a mutation frequency of 3.4% (2/59) for the IDH2 gene, 62.7% (37/59) for DNMT3A, 23.7% (14/59) for RHOA and 50.8% (30/59) for TET2. There was no statistically significant difference in the frequency distribution of IDH2, DNMT3A and TET2 mutations between the different histological subtypes of nPTCL. However, there was a statistical trend towards a higher occurrence of the RHOA mutation in the AITL and PTCL/NOS subtypes (3/9-33.3% and 7/16-43.7%, respectively; p=0.07). Among 7 cases with RHOA mutation classified as PTCL/NOS according WHO-2008 criteria, 6/7 (85.7%) expressed the PD-1 marker in immunohistochemistry, being reclassified as nPTCL with THf phenotype according to WHO-2016 criteria. So, the mutation RHOA was predominantly found in THf cell-derived neoplasms in our cohort. The mutational status of DNMT3A, RHOA and TET2 genes had no prognostic impact on OS, with p=0.85, p=0.13 and p=0.95, respectively. The same was observed in relation to PFS for the DNMT3A (p=0.70) and TET2 (p=0.52) mutations. However, the presence of the RHOA mutation was associated with the unfavorable PFS in our cohort (HR:1.98, p=0.05). We observed 2-year PFS of 28.6% (95% CI: 8.8-52.4%) for mutated-RHOA cases versus 52.9% (95% CI: 37.3-66.3%) for wild-type-RHOA patients (p=0.05) [Figure 1]. We also demonstrated that RHOA mutation was a predictor of lower ORR to first-line therapy (p=0.01) and was associated with high tumor burden (p=0.03) [Figure 2]. Conclusion: In this study, for the first time was demonstrated the unfavorable prognostic impact of the RHOA mutation in patients with nPTCL-Thf (AITL and nPTCL-THf), making it a potential molecular biomarker predictor of poor-PFS, associated with resistance to primary therapy and with high tumor burden. Such results are preliminary and will need to be validated in series with a larger number of cases. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

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Impact of pretreatment anemia on upfront abiraterone acetate therapy for metastatic hormone-sensitive prostate cancer: a multicenter retrospective study

BMC Cancer ◽

10.1186/s12885-021-08206-8 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Teppei Okamoto ◽

Daisuke Noro ◽

Shingo Hatakeyama ◽

Shintaro Narita ◽

Koji Mitsuzuka ◽

...

Keyword(s):

Prostate Cancer ◽

Prognostic Factor ◽

Abiraterone Acetate ◽

Tumor Burden ◽

Hazard Ratio ◽

Historical Cohort ◽

High Tumor Burden ◽

Significant Difference ◽

Hormone Sensitive ◽

The Impact

Abstract Background Anemia has been a known prognostic factor in metastatic hormone-sensitive prostate cancer (mHSPC). We therefore examined the effect of anemia on the efficacy of upfront abiraterone acetate (ABI) in patients with mHSPC. Methods We retrospectively evaluated 66 mHSPC patients with high tumor burden who received upfront ABI between 2018 and 2020 (upfront ABI group). We divided these patients into two groups: the anemia-ABI group (hemoglobin < 13.0 g/dL, n = 20) and the non-anemia-ABI group (n = 46). The primary objective was to examine the impact of anemia on the progression-free survival (PFS; clinical progression or PC death before development of castration resistant PC) of patients in the upfront ABI group. Secondary objectives included an evaluation of the prognostic significance of upfront ABI and a comparison with a historical cohort (131 mHSPC patients with high tumor burden who received androgen deprivation therapy (ADT/complete androgen blockade [CAB] group) between 2014 and 2019). Results We found that the anemia-ABI group had a significantly shorter PFS than the non-anemia-ABI group. A multivariate Cox regression analysis showed that anemia was an independent prognostic factor of PFS in the upfront ABI group (hazard ratio, 4.66; P = 0.014). Patients in the non-anemia-ABI group were determined to have a significantly longer PFS than those in the non-anemia-ADT/CAB group (n = 68) (P < 0.001). However, no significant difference was observed in the PFS between patients in the anemia-ABI and the anemia-ADT/CAB groups (n = 63). Multivariate analyses showed that upfront ABI could significantly prolong the PFS of patients without anemia (hazard ratio, 0.17; P < 0.001), whereas ABI did not prolong the PFS of patients with anemia. Conclusion Pretreatment anemia was a prognostic factor among mHSPC patients who received upfront ABI. Although the upfront ABI significantly improved the PFS of mHSPC patients without anemia, its efficacy in patients with anemia might be limited.

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PET Evaluation Before Autografting Has a Strong Prognostic Impact in High-Risk, Relapsed Follicular Lymphoma Patients

Blood ◽

10.1182/blood.v118.21.1612.1612 ◽

2011 ◽

Vol 118 (21) ◽

pp. 1612-1612

Author(s):

Loic Ysebaert ◽

Jehan Dupuis ◽

Michel Meignan ◽

Anne Julian ◽

Christian Recher ◽

...

Keyword(s):

Stem Cell ◽

Follicular Lymphoma ◽

Conditioning Regimen ◽

Tumor Burden ◽

Prognostic Impact ◽

High Tumor Burden ◽

Richter Syndrome ◽

The Impact ◽

Cell Harvest

Abstract Abstract 1612 INTRODUCTION: in follicular lymphoma (FL) patients with high tumor burden (as defined by GELF criteria), R-CHOP is the standard upfront immunochemotherapy. Results from the PRIMA study suggest that a positive PET after induction therapy predicts for earlier relapses, even despite maintenance with rituximab (RTX) for two years [Trotman J, 2011]. For patients under 65y, who relapse after R-CHOP+/−RTX maintenance with high tumor burden, R-chemo+autografting is a recommended option. In a retrospective cohort of 43 relapsed FL pts in two French University Hospitals, we explored the role of such a strategy on outcome - in the era of new RTX based modalities - and also evaluated the impact of PET results before autografting. PATIENTS AND METHODS: Patients with relapsed FL after R-CHOP, with at least 1 GELF criteria at relapse (high tumor burden), and who received R-chemo before autografting were eligible. IWC+PETresponse criteria (Cheson 2007) were used, after R-CHOP frontline, and after salvage (before autografting). Patients with Richter transformation at relapse were excluded from this study. OS was calculated from date of salvage to date of death or last follow-up; progression-free survival (PFS) and time to next treatment (TTNT) were calculated from completion of salvage to date of FL relapse or next chemotherapy, respectively. RESULTS: 43 pts (60% males) younger than 65y were identified: they received either FCR-based (2 cycles of FCR, 1 cycle of R-DHAP then stem cell harvest, 2 last cycles of FCR, n=25) or R-DHAP-based (4 cycles of R-DHAP or DHAOx (oxaliplatin replacing cisplatin), and stem cell harvest, n=18). Characteristics at salvage: median age was 54 (range 28–62), with median GELF score of 1 (1–4). Thirty % had progression within 6 mo of R-CHOP (refractory);median PFS was 12mo (range 1–40mo) andmedian TTNT was 15mo. RTX maintenance in 12/43 pts did not significantly increased PFS (15 vs 9 mo, p=0.1). FLIPI1 was low in 39%, Int 36%, high 25% of pts, and FLIPI2 was low in 22%, Int 62.5%, high 15.5% of pts. 1/43 pt had CD20- relapse (who received RTX maintenance), and received FC without RTX. Results: response to FCR/R-DHAP included CR+CRu 68/72%, PR 24/28% respectively, PET evaluation was found negative in 23.5/36% respectively (p=ns). Median stem cell harvest (median 2 leukaphereses) was 4.37 vs 7.8.106 CD34+/kg in FCR vs R-DHAP pts (Mann-Whitney p=0.09), without failure (only one patient required plerixafor). Four patients did not receive the planned autologous transplant (2 failures after FCR (including one Richter transformation), 1 hepatitis before conditioning regimen, 1 withdrawal of consent). Conditioning regimen for the remaining 39 pts was BEAM in 16 (4 FCR+12 R-DHAP), Zevalin-BEAM in 23 pts (9 FCR+14 R-DHAP), including 9 pts who further received RTX maintenance post-autografting. At a median follow up of living pts of 18mo, 9 pts had relapsed, of which 8 needed additional therapy. At time of analysis (June 2011), 37 pts were alive. Causes for death included 1 pancytopenia and infection, 1 Richter syndrome, 1second cancer, 2 complications of allografting (received later on). Late complications were common: pancytopenia or prolonged grade III-IV neutropenia (>3mo) after procedure occurred in 5/43 pts (3 FCR, 2 R-DHAP, without evidence of MDS/AML in these cases), Richter syndrome occurred in 2 patients (1 death), second cancer in 3 pts (1 Hodgkin, 1 womb sarcoma, 1 lung cancer). On an intend-to-treat basis, 41 pts are evaluable. For FCR/R-DHAP pts, 2y PFS was 68/68%, 2y TTNT 80/70%, and 2y OS 73/100%, respectively (logrank p=ns). Two years TTNT tended to be higher in pts receiving Z-BEAM (55 vs 83%), or with PET negativity before autografting (70 vs 85%), both without reaching significance (logrank p=0.1). PET negativity before autografting was the only variable statistically associated with superior OS in our series (logrank p=0.0003). CONCLUSIONS: Our data suggest that PET negativity before consolidative autografting is an achievable and desirable goal in FL salvaged after R-CHOP. Disclosures: Ysebaert: Roche: Membership on an entity's Board of Directors or advisory committees, Research Funding.

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Circulating Mir-153 Participates in Occurrence of Acute Graft-Versus-Host Disease By Down-Regulating Indoleamine-2,3-Dioxygenase

Blood ◽

10.1182/blood.v124.21.2428.2428 ◽

2014 ◽

Vol 124 (21) ◽

pp. 2428-2428

Author(s):

Xiaosu Zhao ◽

Yinuo Wang ◽

Jun Wan ◽

Meng Lv ◽

Lanping Xu ◽

...

Keyword(s):

Mirna Expression ◽

Roc Analysis ◽

Bioinformatics Analysis ◽

Conflicts Of Interest ◽

Luciferase Assay ◽

Regulatory Function ◽

Indoleamine 2,3 Dioxygenase ◽

Potential Biomarker ◽

Significant Difference ◽

Peking University

Abstract Background Indoleamine-2,3-dioxygenase (IDO) is able to catalyze the first and rate-limiting step in the catabolism of tryptophan, which is the essential amino acid for T cell proliferation. Many previous studies have proved that IDO is able to play important roles in immunosuppression-associated with tumor immunity, autoimmunity and chronic infection. Since donor T cells will be activated, proliferate, differentiate and finally lead to tissue injury, growing research paid more attention to the immunoregulatory role of IDO in the process of aGVHD. Our previous work also demonstrated that plasma IDO level was correlated with the severity of aGVHD. Increased IDO expression would serve as a protective molecular during aGVHD. However, the exact regulatory mechanism of IDO is still unclear. MicroRNAs (miRNAs) are a class of small non-coding RNA that negatively regulate gene expression by translational repression or induction of mRNA degradation. Recently, circulating miRNAs have been reported to be the promising biomarkers for various kinds of diseases. We speculated that miRNA that might regulate those GVHD-related molecules/cytokines expression including IDO, and the circulating miRNA that directed at IDO would serve as new potential biomarker for aGVHD. This study is to find the potential regulatory miRNA of IDO and investigate the correlation between plasma miRNA expression and aGVHD. Methods The potential regulatory miRNA for IDO was identified through bioinformatics analysis using TargetScan and Miranda. The regulatory function of target miRNA on IDO protein and gene was confirmed by Western Blot analysis and luciferase assay. Human plasma samples from 70 patients who underwent allo-HSCT from Sep 2012 to Jun 2013 were prospectively collected at +7 day(d), +14d, +21d, +30d, +45d, +60d, +90d and the occurrence of aGVHD from Peking University Institute of Hematology. All patients provided informed consent. This study was approved by the institutional review board at the Peking University Institute of Hematology (Protocol number 2013-42). Real-time quantitative PCR (RQ-PCR) analysis was performed to examine the miRNA expression of plasma at different time points posttransplant. Receiver operating characteristic (ROC) analysis was employed to determine the optimal cut-off value for predict the occurrence of aGVHD. Results Through bioinformatics analysis, we found the predicted target sites in its 3’UTR binding miRNA-153. The intracellular IDO protein would be reduced when miRNA-153 was transfected into interferon-gamma-treated Hela cells. Surprisingly, the mRNA expression level of IDO was also affected in miRNA-153 transfected cells obviously (P<0.001). In addition, luciferase assay in 293T cells showed that miRNA-153 was able to bind directly to the promoter region of IDO gene and affect its transcription. Luciferase activity dropped around 44% when wild-type construct was co-transfected into 293T cells with miRNA-153 (P=0.009). However, when the potential binding sites were mutated, this phenomenon could not be observed. This indicates that the IDO 3’UTR is normally targeted by endogenous miR-153. Among the 70 patients who underwent allo-HSCT, 35 patients developed into aGVHD and 35 patients did not. A significant difference in miRNA-153 expression levels between the two groups was confirmed at +7d (P<0.001). Furthermore, we used ROC analysis to evaluate the diagnostic accuracy. Plasma levels of miRNA-153 at day 7 could predict an impending aGVHD with a sensitivity of 38.71% and a specificity of 100%, and the area under the curve (AUC) was 0.898 (95%CI: 0.821-0.974, P<0.0001). Conclusions Our study suggested that miRNA-153 might participate in occurrence of aGVHD by down regulating IDO and might be a putative target for novel aGVHD therapy. The plasma level of miRNA-153 at +7d after allo-HSCT would be a good biomarker for predicting the occurrence of aGVHD. Disclosures No relevant conflicts of interest to declare.

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Prognostic Implications of Oligoclonal Bands in Patients with Multiple Myeloma:Its Development per Se did Not Confer the Prologation of Survival

Blood ◽

10.1182/blood.v124.21.4721.4721 ◽

2014 ◽

Vol 124 (21) ◽

pp. 4721-4721 ◽

Cited By ~ 1

Author(s):

Manabu Fujisawa ◽

Yasuhito Suehara ◽

Keisuke Seike ◽

Kota Fukumoto ◽

Masafumi Fukaya ◽

...

Keyword(s):

Light Chain ◽

Conflicts Of Interest ◽

Free Light Chain ◽

Oligoclonal Bands ◽

Prognostic Impact ◽

Myeloma Protein ◽

Myeloma Cells ◽

Response To Chemotherapy ◽

Significant Difference ◽

Clinical Records

Abstract Background: Multiple myeloma (MM) is characterised by the production of monoclonal immunoglobulin (Ig) and clonal proliferation of neoplastic plasma cells in bone marrow. The emergence of oligoclonal bands (OB) in serum and/or urine immunofixation electrophoresis (IFE) that is different to that observed at diagnosis has been reported with varying frequency in patients with stem cell transplantation (SCT) or favourable response to chemotherapy, although its prognostic relevance remains unclear. Here, we retrospectively analysed clinical records and results serial serum and/or urine IFE to determine the frequency, clinical characteristics and prognostic impact of OB that developed after treatment. We also analysed the effects of OB on the results of free light chain (FLC) assay that may affect the stringent CR (sCR) criteria proposed by the IMWG. Patients: We retrospectively reviewed 180 patients with MM admitted to the Department of Hematology/Oncology at Kameda Medical Center, Kamogawa, Japan, from January 2006 to May 2014.Seventeen patients were excluded from the analysis due to lack of appropriate follow-up data. Method: An OB was defined as the presence of a serum and/or urine IFE monoclonal spike that was different from the original myeloma protein in heavy and/or light chains, as well as a different IFE migration pattern. IFE was performed at least every three months after obtaining very good partial response (VGPR) and then until the disease progression or relapse was confirmed. Overall survival (OS) and progression free survival (PFS) were analyzed in 173 MM patients by the Kaplan–Meier method, and differences between the curves were calculated by two-sided log-rank test. Free light chain (FLC) and minimal residual disease measurement by multicolour flowcytometory (MFC) were performed to evaluate the response to treatment. Results: Myeloma response more than VGPR and CR were achieved in 87 (53.3%) and 54 (33.1%) patients, respectively. None of the patients with less than PR developed OB, and OB developed in 36.3% (12/33) and 51.9% (28/54) of the patients with VGPR and CR, respectively. Among the 30 patients who received SCT, 18 patients (60.0%) developed OB, whereas only 12 of 133 patients who did not receive SCT developed OB. The emergence of OB was significantly higher in patients receiving SCT (P < 0.001). The median PFS of the patients with and without OB were 79.4 months and 19.2 months (P = 0.028), respectively, and those of OS did not reached and 41.1 months (P < 0.001), respectively (Figure 1). However, if patient response was limited to CR, median PFS and OS of patients with and without OB were not significantly different between the two groups (79.4 months vs. 80.1months, respectively, P = 0.646; and not reached and not reached, respectively, P = 0.776; Figure 2). OB was observed in 60% of patients after SCT, and in 36.6% of patients more than VGPR without SCT (P<0.001). In the patients who received SCT, the presence of OB was not associated with either PFS or OS (20.0 months vs 44.7 months, P=0.543 and not reached vs not reached, P= 0.739, respectively; Figure3). We also analyzed the association between residual myeloma cells assessed by the MFC in the patients who achieved CR according to the development of OB. Although the patients with OB showed slightly lower levels of residual myeloma cells than those without OB (2.49 x10-4vs 4.15 x10-4), there was no significant difference between the two groups (P= 0.054). As the presence of OB may result in an abnormal serum FLC kappa/lambda ratio, we examined the association of FLC kappa/lambda ratio and development of OB in 28 patients who achieved CR. Abnormal FLC kappa/lambda ratio was observed in 10 (35.7%) and 4 (15.3%) patients with and without OB, respectively, among those with CR (P = 0.118). Conversely, OB was detected in 10 of the 14 patients with an abnormal FLC ratio (71.4%) versus 18 of 40 (45.0%) with a normal serum FLC ratio (P = 0.164) among those with CR. Conclusions: In conclusion, we showed that the emergence of OB occurred exclusively in patients with favourable myeloma responses (more than VGPR) and was associated with prolonged survival. Patients with OB appeared to have fewer residual myeloma cells compared to those without OB. However, its development did not confer an additional survival benefit among the patients who achieved CR. In addition, it is possible that the emergence of OB affects the sCR response criteria proposed by the IMWG. Figure 1 Figure 1. Figure 2 Figure 2. Figure 3 Figure 3. Disclosures No relevant conflicts of interest to declare.

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Phase II Study of Interferon-Alpha and DA-EPOCH+/-R in Lymphomatoid Granulomatosis

Blood ◽

10.1182/blood-2018-99-116707 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 785-785 ◽

Cited By ~ 1

Author(s):

Christopher Melani ◽

Mark Roschewski ◽

Stefania Pittaluga ◽

Jeffrey Cohen ◽

Andrea Nicole Lucas ◽

...

Keyword(s):

Conflicts Of Interest ◽

Histologic Grade ◽

Lymphomatoid Granulomatosis ◽

Common Disease ◽

Low Grade ◽

Primary Therapy ◽

High Grade ◽

Cns Involvement ◽

Cell Counts ◽

Significant Difference

Abstract Background: Lymphomatoid granulomatosis (LYG) is a rare EBV-driven B-cell lymphoproliferative disorder characterized by a reactive T-cell infiltrate that is angioinvasive and angiodestructive. Dysregulated immune surveillance of EBV is thought to contribute to the pathogenesis of LYG and grading of disease is based on the density and number of EBV+ large atypical B-cells. Grade I-II (low-grade) disease is typically polyclonal/oligoclonal and immune-dependent whereas grade III (high-grade) disease is typically monoclonal and immune-independent. Herein, we report additional and extended results from an on-going prospective study at the National Cancer Institute on the treatment of patients with low and high-grade LYG using Interferon-α (INF-α) and/or DA-EPOCH+/-R, respectively. Methods: Pts with histologically confirmed LYG by the Laboratory of Pathology, NCI, were eligible. LYG of any grade or stage was included as were pts with untreated or previously treated disease. Baseline tests included laboratory studies, body CT and FDG-PET, CT or MRI brain, echocardiogram, LP with flow cytometry/cytopathology, and bone marrow aspiration/biopsy. Grade 1-2 LYG pts received primary therapy with INF-α initially at 7.5 MIU TIW which was dose-escalated every 1-2 wks. as tolerated until best response and then continued for 1 yr. Pts with grade 3 LYG received combination chemotherapy with DA-EPOCH+/-R for up to 6 cycles. Pts with progression after primary INF-α therapy and those who progressed after or failed to achieve CR after DA-EPOCH+/-R could cross over to the other treatment. Restaging CT was performed every 4 wks. until stable dose of INF-α and then every 3 mos. and after cycles 4 and 6 of DA-EPOCH+/-R. Surveillance CT was performed every 3, 4, 6 and 12 mos. for post-treatment yrs. 1, 2, 3, and 4-5, respectively, and then as clinically indicated. Results: 70 LYG pts were enrolled between Jan. 1991 and Feb. 2018. Characteristics included; male sex in 45 (64%) pts, median (range) age of 46.2 yrs. (14.9-67) and histologic grade I in 19 (27%), II in 24 (34%), and III in 27 (39%) pts. Twenty-three (33%) pts were untreated, and 32 (46%) and 21 (30%), respectively, had received steroids and/or chemotherapy+/-R. The most common disease sites included lung (100%), CNS (37%), skin (33%), kidney (17%), and liver (17%). Peripheral blood at baseline showed median (range) CD4, CD8 and NK-cell counts of 510 (80-1864; Normal: 359-1565), 138 (8-1245; Normal: 178-853), and 96 (25-1567; Normal: 126-729) cells/uL, respectively, and a median (range) EBV viral load of 100 (1-13,950) copies/mL and 3.48 (0-4.48) Log 10 IU/mL. With a median potential follow-up of 12.9 yrs., 5-yr. PFS and OS for all 67 treated pts was 41% and 70.8%, respectively (Fig.1). There was no significant difference in PFS or OS associated with baseline histologic grade (p=0.47 and p=0.23, respectively) or CNS involvement (p=0.67 and p=0.98, respectively). Of 49 INF-α treated pts, 47 were evaluable for response with an ORR of 60% (CR 55%) and a median (range) TTR of 6.7 mo. (1.4-40) and 5-yr. DOR of 66.6% (95% CI: 45.4-81.1%). ORR was similar [60% (CR 55%)] in the 20 INF-α treated pts with baseline CNS involvement. Overall, in 49 INF-α treated pts, 5-yr. PFS and OS were 46.2% and 72.5%, respectively (Fig.1). Median (range) dose and duration of INF-α was 20 MIU (7.5-40) and 7.9 mo. (0.5-48.6), respectively. Nineteen pts received secondary therapy with DA-EPOCH+/-R for progression during/after INF-α with an ORR of 72% (CR/CRu 56%) in 18 evaluable pts. In 13 evaluable pts refractory to INF-α, ORR was 92% (CR/CRu 69%) with DA-EPOCH+/-R. Of 18 DA-EPOCH+/-R treated pts, 17 were evaluable for response with an ORR of 76% (CR/CRu 41%) and a median (range) TTR of 4.1 mo. (2.6-7.4) and median DOR of 1.5 yrs. (95% CI: 0.3-9.2). Overall, in 18 DA-EPOCH+/-R treated pts, 5-yr. PFS and OS were 27.8% and 66.2%, respectively (Fig.1). Eight pts received secondary INF-α therapy for progression during/after or failure to achieve CR after DA-EPOCH+/-R with an ORR of 75% (CR/CRu 63%). Conclusion: INF-α and DA-EPOCH+/-R result in prolonged remissions in a significant subset of pts with low and high-grade LYG, respectively. Progression to high-grade disease and relapse with low-grade disease following INF-α and DA-EPOCH+/-R treatment, respectively, occurs frequently due to the continued immune dysregulation of EBV and can be successfully treated through cross over to the alternative treatment modality. Disclosures No relevant conflicts of interest to declare.

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Impact of Missing KIR Ligands in HLA-Matched Unrelated Donor HSCT.

Blood ◽

10.1182/blood.v114.22.1185.1185 ◽

2009 ◽

Vol 114 (22) ◽

pp. 1185-1185

Author(s):

Aining Sun ◽

Weiyang Li ◽

Wu Depei ◽

Jun He ◽

Xiaojing Bao ◽

...

Keyword(s):

Conflicts Of Interest ◽

Chronic Gvhd ◽

Hla Class I ◽

Unrelated Donor ◽

Prognostic Impact ◽

Matched Unrelated Donor ◽

Hematopoietic Stem ◽

Immunoglobulin Receptor ◽

Significant Difference ◽

Hla Genotype

Abstract Abstract 1185 Poster Board I-207 Objective: To analyze the prognostic impact of missing ligands for inhibitory killer-immunoglobulin receptor(KIR) in HLA-matched hematopoietic stem cell transplantation(HSCT) using unrelated donor. Methods: HLA genotype of 51 patients (ALL 22 cases, AML 13 cases, CML 14 cases, MDS 1 cases and HAL 1 cases) and their matched unrelated donors was determined by polymerase chain reaction sequence oligonucleotide probes(PCR-SSOP) and sequence specific primers (PCR-SSP). The KIR genotype was determined by PCR-SSP. Results: Patients were divided into those with(n=37) and those without(n=14) missing 1 or more HLA class I ligands for donor inhibitory KIR. The period of platelet reconstruction was shorter in patients [ 13 d(10d∼27d)] with missing KIR ligands than those[14d(12d∼21d)] without missing KIR ligands(P=0.046). There was no significant difference in neutrophil recovery, ≥II° acute GVHD(13.5% vs 35.7%, P>0.05) and extensive chronic GVHD (16.2% vs 28.6%, P>0.05) between the two groups. The 3-year continuous complete remission(CCR) rate for patients with and without missing KIR ligands was 73.6% and 37.4%, respectively(P=0.183). The 3-year overall survival(OS) rate for the two groups was 77.5% and 52.4%, respectively(P=0.533). Conclusions: In HLA-matched unrelated donor HSCT, missing KIR ligands may be associated with enhanced engraftment, decreased severe GVHD, improved CCR and OS. Disclosures: No relevant conflicts of interest to declare.

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Preliminary Results of a Study of the Platelet Dense Granule and Serotonin Storage Pool in Autistic Children and Their Families.

Blood ◽

10.1182/blood.v114.22.4456.4456 ◽

2009 ◽

Vol 114 (22) ◽

pp. 4456-4456

Author(s):

Nicole Zadzilka ◽

Lisa Rinker ◽

Victor S. Flauta ◽

William T. Gunning

Keyword(s):

Migraine Headache ◽

Selective Serotonin Reuptake Inhibitors ◽

Conflicts Of Interest ◽

Autism Spectrum ◽

Dense Granule ◽

Autistic Children ◽

Storage Pool ◽

Potential Biomarker ◽

Significant Difference ◽

Abstract Submission

Abstract Abstract 4456 The etiology of autism is thought to be neurobiological, but the mechanisms involved are not known. One possible clue into this disorder can be found in multiple clinical studies which have established that at least 30-40% of individuals diagnosed with autism have significantly elevated levels of serotonin (5HT) in their blood. Serotonin transport by platelets and 5HT plasma levels have been found to be altered in patients with multiple neurobiological disorders, including bipolar disorder, schizophrenia, depression, aggression, autism, and migraine headache. Serotonin has numerous functions, including regulation of vasoconstriction, vasodilatation, and coagulation. A review of the literature reveals few studies of coagulopathy related to autism other than attempts to associate vaccination or infections with the etiology of autism. One recent study, of a limited number of individuals with autism spectrum disorders (ASD) and their families, reported a significant incidence of thrombophilia. All children evaluated (10/10) and 15 of 16 family members in the study were found to be at risk for thrombophilia disorders. We have been evaluating patients for PL dysfunction for many years and recently, have included ELISA tests to determine the 5HT concentration in PLs of patients suffering from migraine headache or syncope. All of our investigations to date have evaluated patients for a deficiency of the PL storage pool (DG SPD) including a decrease of PL DGs and their stored biochemical constituents; we have never conducted a study related to potentially increased numbers of DGs or the concentration of biochemicals contained in these granules. As at least one report exists in the literature suggesting the potential for thrombophilia in patients diagnosed with an autism spectrum disorder and their families, we decided to assess the platelet storage pool in autistic patients and their families as a potential model of platelet dysfunction due to elevated DGs and/or 5HT levels and/or as a potential biomarker for autism. As the major storage pool of 5HT in the circulation is known to exist in platelet PL DGs, it is possible that a PL DG storage pool could be an important mediator of ASD. Our hypothesis is that autistic children may have elevated numbers of DGs and increased levels of PL 5HT. Peripheral blood samples were obtained from children diagnosed with autism as well as their parents and siblings. Platelets were obtained via centrifugation and DG content determined by electron microscopy and an ELISA assay was utilized to determine PL 5HT concentration for all subjects. A questionnaire was also used to evaluate family bleeding history and drug therapy for the autistic children. At present, 4 families have been recruited for the study. Autistic children (n = 5) have an average of 5.42 ± 0.69 DG/PL and a mean 5HT concentration of 447.6.4 ± 85.6 ng/109 PL (PL 5HT normal range = 215-850 ng/109 PL). Their parents have an average of 5.00 ± 0.54 DG/PL and a mean 5HT concentration of 427.6.4 ± 82.66 ng/109 PL and one five year old sibling was found to have 8.17 DG/PL and 2233 ng/109 PL. Control subjects (n=21) have an average of 4.16 ± 0.09 DG/PL and a 5HT concentration of 589.6 ± 56.25 ng/109. There is no statistically significant difference between controls, parents or autistic subjects by ANOVA. Our preliminary data does not support our hypothesis. However, with serotonin being the immediate precursor of melatonin, we do not discount the fact that all of our subjects were receiving drug therapies including melatonin supplementation (n=2) or selective serotonin reuptake inhibitors (n=3) which diminish the PL 5HT concentration. Thus, the therapeutic regimens for these subjects could account for their normal 5HT levels. This project had been initiated just prior to the time of abstract submission; we are currently recruiting study participants and will present additional data at the meeting. Disclosures: No relevant conflicts of interest to declare.

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Influence of the fusion gene ETV6-RUNX1 in the prognosis of the Pediatric B- Precursor Acute Lymphoblastic Leukemia

Blood ◽

10.1182/blood.v118.21.2515.2515 ◽

2011 ◽

Vol 118 (21) ◽

pp. 2515-2515

Author(s):

Alejandro Contento-Gonzalo ◽

Antonio Jimenez-Velazco ◽

Alcala-Peña Magdalena ◽

Manuel Barrios ◽

Katie Hurst ◽

...

Keyword(s):

Acute Lymphoblastic Leukemia ◽

Conflicts Of Interest ◽

Lymphoblastic Leukemia ◽

Disease Free Survival ◽

Rank Test ◽

Prognostic Impact ◽

Excellent Outcome ◽

Log Rank Test ◽

Significant Difference

Abstract Abstract 2515 INTRODUCTION AND OBJECTIVES: The ETV6-RUNX1 (TEL-AML1) rearrangement comes from the translocation of two chromosomes t(12;21)(p12;q22), and represents one of the most frequently detected anomalies (15–30%) in the B-precursor Acute Lymphoblastic Leukemia (B-ALL). It must be identified by polymerase chain reaction (PCR) or fluorescent in situ hybridization (FISH) methods, since this translocation is not detected by conventional cytogenetic techniques. The prognostic value of ETV6-RUNX1 is still a matter of controversy. Recently, the group of the St Jude Children's Hospital reported an excellent outcome in patients carrying the translocation, whereas the BMF group did not find any significant difference in the survival of ETV6-RUNX1 positive patients when compared to ETV6-RUNX1 negative. The aim of our study has been to determine the prognostic impact of the ETV6-RUNX1 rearrangement in patients diagnosed of B-ALL in our Hospital, after a long period of follow-up and with the same Spanish treatment protocols (from PETHEMA and SHOP groups). PATIENTS AND METHODS: All patients with B-ALL diagnosis from January 1997 to May 2011 were included in the study: in total, 114 patients with a mean of age of 6 ys (0.3–14). The type of leukemia was ALL common (83 patients), pro-B (17 patients), pre-B (13 patients) and mature (1 patient). All the children over 1 yr received treatment according to PETHEMA group protocols, adjusted to the risk. Children under 1 yr were treated following SHOP group protocols. Seventeen patients received an allogeneic transplantation. The main clinical features of the positive and negative patients for ETV6-RUNX1 are detailed in table 1. ETV6-RUNX1 assay was performed in our laboratory by RT-PCR, according to the European BIOMED project methodology. RESULTS: ETV6-RUNX1 was found in 31 of the 114 patients (27.2%). These patients showed a significantly higher frequency of myeloid antigens (p<0.001), and were always positive for CD10 (p=0.006). All cases of positive ETV6-RUNX1 were over 2 years old. No significant differences between positive and negative ETV6-RUNX1 were obtained when complete remissions (100 vs 80%), relapse (16 vs 20%) or deaths (10 vs 13%) were analyzed. Furthermore, estimation of disease free survival (DSF) at 14 ys for both groups were similar: 80 ± 8% for positive vs 66 ± 7% for negative (p=0.21, log-rank test). And the same happened for overall survival (OS): 87 ± 7% for positive vs 83 ± 5% for negative (p=0.4, log-rank test). DISCUSSION: In our series, including patients with B-ALL treated with similar protocols with long periods of follow-up, we could not find differences between positive and negative ETV6-RUNX1 patients. It is well known that the intensity of the chemotherapy regimen and the age of inclusion in different protocols may influence the prognosis. Therefore, at present, it is still a matter of discussion if previous reported differences in the B-ALL ETV6-RUNX1 positive group could be explained by a different stratification in risk groups or by different chemotherapy regimens. This work has been funded by a grant from AECC, Carmen Lavigne Prize 2010 Disclosures: No relevant conflicts of interest to declare.

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Prognostic Interaction Between ASXL1 and TET2 Mutations in Chronic Myelomonocytic Leukemia

Blood ◽

10.1182/blood.v126.23.2864.2864 ◽

2015 ◽

Vol 126 (23) ◽

pp. 2864-2864

Author(s):

Mrinal M Patnaik ◽

Terra L. Lasho ◽

Pooja Vijayvargiya ◽

Christy Finke ◽

Curtis A. Hanson ◽

...

Keyword(s):

Survival Data ◽

Prognostic Model ◽

Conflicts Of Interest ◽

Univariate Analysis ◽

Multivariable Analysis ◽

Chronic Myelomonocytic Leukemia ◽

Signal Pathways ◽

Prognostic Impact ◽

Significant Difference ◽

Myelomonocytic Leukemia

Abstract Background : Gene mutations are common (~90%) in patients with chronic myelomonocytic leukemia (CMML) and involve epigenetic regulators (TET2 ~ 60%, ASXL1 ~40%), spliceosome components (SRSF2 ~40%) and signal pathways (RAS ~30%). Of these, thus far, only ASXL1 mutations have been shown to adversely impact overall survival (OS). In the current study, we used a 27-gene panel assay to identify additional prognostically-relevant mutations in CMML and to also determine if number of mutations carries prognostic relevance. Methods : 175 patients with WHO-defined CMML were included in the study. All patients had bone marrow (BM) biopsies and cytogenetics performed at diagnosis. Targeted capture assays were carried out on BM DNA specimens obtained at diagnosis for the following genes; TET2, DNMT3A, IDH1, IDH2, ASXL1, EZH2, SUZ12, SRSF2, SF3B1, ZRSR2, U2AF1, PTPN11, Tp53, SH2B3, RUNX1, CBL, NRAS, JAK2, CSF3R, FLT3, KIT, CALR, MPL, NPM1, CEBPA, IKZF, and SETBP1. Paired-end indexed libraries were prepared from individual patient DNA using the NEBNext Ultra Library prep protocol on the Agilent Bravo liquid handler. Capture libraries were assembled according to Nimblegen standard library protocol. Base-calling was performed using Illumina's RTA version 1.17.21.3. Genesifter software was utilized to analyze targeted sequence data. Specific variants were deemed as mutations if they were associated with a hematological malignancy (as identified by COSMIC database), or if they were not associated with a dbSNP Results: Among the 175 study patients, 66% were males and median age was 70 years. 146 (83%) patients were subclassified as CMML-1. At a median follow-up of 23 months, 146 (83%) deaths and 25 (14%) leukemic transformations were documented. Median survivals were 24 months for CMML-1 and 16 months for CMML-2 (p=0.38). Mutational frequencies were; TET2 46%, ASXL1 45%, SRSF2 45%, SETBP1 19%, CBL 14%, RUNX1 14%, NRAS 12%, U2AF1 8%, SF3B1 6%, ZRSR2 6%, Tp53 5%, DNMT3A 5%, IDH2 5%, PTPN11 5%, SH2B3 5%, JAK2 4%, NPM1 3%, CSF3R 2%, IDH1 2%, EZH2 1%, SUZ12 1%, KIT 1%, FLT3 1%, CALR 1%. 172 patients (98%) had at least one mutation, 21 (12%) had 2, 24 (14%) had 3, 20 (11%) had 4, 9 (5%) had 5, while one (1%) patient had 6 concurrent mutations. Risk stratification was based on Mayo prognostic model: 25% high, 32% intermediate and 43 % low risk. In univariate analysis, presence of ASXL1 mutations (p=0.01), absence of TET2 mutations (p=0.005) and presence of DNMT3A mutations (p=0.02) were associated with inferior survival; in multivariable analysis, ASXL1 (p=0.01) and TET2 (p=0.03) mutations remained significant. In order to determine prognostic interaction between these two mutations, patients were stratified into four mutational categories: ASXL1wt/TET2wt (n =56), ASXL1mut/TET2wt (n =31), ASXL1mut/TET2mut (n =50) and ASXL1wt/TET2mut (n =38). Survival data in these four groups showed significant difference in favor of ASXL1wt/TET2mut (median survival 38 months; p=0.016), compared to those with ASXL1wt/TET2wt (19 months), ASXL1mut/TET2wt (31 months)and ASXL1mut/TET2mut (16 months); there was no significant difference in survival among the latter three groups (p=0.3) (Figure). The number of mutations per patient did not affect outcome (p=0.3). In multivariable analysis, presence of ASXL1 mutations (P=0.01) and absence of TET2 mutations (p=0.003) remained significant when risk factors used in the Mayo prognostic model (HB <10 gm/dl, AMC >10 x 10(9)/L, platelets <100 x 10(9)/L, circulating IMC) were added to the model; the same was true for ASXL1wt/TET2mut (p=0.036). In a separate multivariable analysis that included the Mayo prognostic model as a single variable along with presence of ASXL1 and absence of TET2 mutations; or absence of ASXL1wt/TET2mut mutational status, the respective hazard ratios were 1.4 (95% CI 1.07-2.1; p=0.012), 1.5 (95% CI 1.07-2.1; p=0.03) and 1.8 (95% CI 1.2-2.7; p=0.001). Leukemia-free survival was worse in ZRSR2 -mutated cases (p=0.03). Conclusions: Almost 100% of patients with CMML express one or more myeloid neoplasm-relevant mutations. The current study suggests a favorable prognostic impact from TET2 mutations, unless accompanied by ASXL1 mutations. Disclosures No relevant conflicts of interest to declare.

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Correlation between Lipid-Associated Sialic Acid and Tumor Burden in Melanoma

The International Journal of Biological Markers ◽

10.1177/172460089400900408 ◽

1994 ◽

Vol 9 (4) ◽

pp. 247-250 ◽

Cited By ~ 9

Author(s):

A.C. Buzaid ◽

A.B. Sandler ◽

C.L. Hayden ◽

J. Scinto ◽

W.-J. Poo ◽

...

Keyword(s):

Sialic Acid ◽

Positive Predictive Value ◽

Tumor Marker ◽

Metastatic Disease ◽

Predictive Value ◽

Clinical Evidence ◽

Tumor Burden ◽

High Tumor Burden ◽

Significant Difference ◽

The Relationship

Recent studies have suggested that lipid-associated sialic acid (LSA) may be a useful tumor marker for monitoring patients with melanoma, but the relationship between LSA and tumor burden has not been previously studied. We therefore examined LSA levels in 240 patients of whom 169 had no clinical evidence of disease (NED) and 71 had metastatic disease. There was a statistically significant difference in LSA levels in patients with NED compared with metastatic disease as well as those with high tumor burden compared with low or intermediate tumor burden. There was no difference between the groups with low and intermediate tumor burden. An LSA level of 25 mg/dl provided a positive predictive value of 70% and a negative value of approximately 80%. Our data show that LSA levels correlate with tumor burden in patients with melanoma.

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