bluuebunny/crossref_metadata_2025 huggingface dataset:
Six Courses of Intensified CHOP Plus G-CSF Compared to Eight Courses of Standard CHOP in Patients with Intermediate-Risk Aggressive Non-Hodgkin Lymphoma Results of a Prospective Randomized HOVON Trial.
Authors: Verdonck Leo F. , van Imhoff Gustaaf W. , Raemakers John M.M. , Verhoef Gregor , Kramer Mark H.H. , Ossenkoppele Gert J. , Doorduijn Jeanette K. , Meulendijks Ine A.M.H. , van Wieringen Wessel N. , Sonneveld Pieter
Published: April 2020

Between November 1994 and February 2004 a total of 513 untreated patients with aggressive intermediate-risk NHL, aged 16–65 years, were recruited to the randomized HOVON-26 trial comparing intensified (I)CHOP plus G-CSF (given every 2 weeks for 6 courses) with standard CHOP (given every 3 weeks for 8 courses). I-CHOP included doxorubicin (D) at 70 mg/m2 and cyclophosphamide (C) at 1000 mg/m2 together with standard dose vincristine and prednisone. So, dose intensity (mg/m2/week) for both D and C was doubled in the I-CHOP arm compared to CHOP, whereas the cumulative dose of D and C was similar in both treatment arms. Intermediate-risk aggressive NHL was defined (HOVON criteria) as stage II disease with increased LDH and stage III/IV disease with normal LDH, corresponding to the low-intermediate (L-i) and high-intermediate (H-i) risk groups of the age-adjusted (aa) IPI score. The endpoints were CR rate, overall (OS), disease-free (DFS) and event-free survival (EFS).

Of 513 patients, 477 were eligible of whom 239 were randomized to CHOP and 238 to I-CHOP. Patients were well balanced for all risk factors. According to the aa-IPI score, in the L-i group 161 patients received CHOP and 137 I-CHOP, whereas in the H-i group 68 patients received CHOP and 80 patients I-CHOP.

As of July 1st 2005, median follow-up of all patients still alive is 50 months. Ninety-nine patients in the CHOP arm died and 85 patients in the I-CHOP arm died. NHL was the main cause of death in both treatment arms (76 and 61 patients, respectively). Treatment-related mortality (8%) was similar in both arms, however WHO grade 4 side-effects occurred twice as often (20 vs 10 events) in the I-CHOP arm. I-CHOP did result in better CR rate and 5 years estimates of OS, DFS, and EFS, but the differences were not significant (table 1). However, although not originally planned, subgroup analysis showed improved OS, DFS, and EFS in the low-intermediate group (table 2).

Although I-CHOP was not significantly better than CHOP in intermediate-risk aggressive NHL patients, subgroup analysis indicates a possible improvement in outcome for patients with low-intermediate risk aggressive NHL. Another advantage of I-CHOP might be that therapy duration is markedly reduced.

Table 1. CR rate and 5 yrs OS, DFS and EFS of intermediate-risk patients (HOVON criteria) CR rate OS DFS EFS CHOP 49% 57% 52% 25% I-CHOP 53% 62% 63% 32% p-value (log-rank) 0.41 (chi-square) 0.19 0.19 0.17 Table 2. 5 yrs OS, DFS and EFS in low- intermediate and high-intermediate risk groups (aa-IPI score) Low-intermediate aa IPI High-intermediate aa IPI OS DFS EFS OS DFS EFS CHOP 60% 48% 24% 52% 54% 25% I-CHOP 68% 66% 36% 52% 53% 28% p-value (log-rank) 0.06 0.03 0.03 0.78 0.85 0.84
Transplant Outcome in Acute Myeloid but Not Acute Lymphocytic Leukemia Following T Cell Depleted Allogeneic Stem Cell Transplantation May Be Favorably Influenced by Early Recovery of NK Cells.
Authors: Savani Bipin N. , Montero Aldemar , Mielke Stephan , Rezvani Katayoun , Shenoy Aarthi , Childs Richard , Barrett A. John
Published: April 2020

Between 05/1994 and 07/2005, 78 patients with acute leukemia (AML=49; ALL=29) received a T cell depleted stem cell transplant from an HLA-identical sibling, with 1–2 infusions of donor T cells on days 30–100. The conditioning regimen included 12–13.5 Gy total body irradiation (TBI) and cyclophosphamide (Cy) with or without fludarabine. Cyclosporine (CSA) was given as the sole GVHD prophylaxis after transplant. The stem cell source was peripheral blood in 75 and bone marrow in 3. The T cell dose ranged between 0.2 to 2 × 105CD3+ cells/kg. AML and ALL patients were distributed equally among transplant regimen and T cell dose variations. The median CD34 cell dose was 5.4 × 106 cells/kg, (5.4 for AML, and 5.5 for ALL; p=0.50). Actuarial disease free survival (DFS), transplant related mortality (TRM) and relapse rate was 42±8, 20±6 and 40±9 % for AML and 22±10; 35±19 and 64±11% for ALL patients. The median day 30 post-transplantation lymphocyte count (LC30) for AML patients was 418/μl (31–2105) and 778/μl (265–3295) for ALL patients; p=0.063). In multivariate analysis, LC30 emerged as an independent risk factor for transplant outcome in AML but not ALL patients (see figure). In AML patients LC30 above median was associated with superior DFS, lower TRM and lower relapse. CD34 dose and LC30 were highly correlated for AML and ALL analyzed together or separately. Lymphocyte subset analysis of 20 post-transplant samples on day 30 identified a large proportion of CD56+16+ natural killer (NK) cells (median 44%, range 15–86%). Since early post-transplant NK cells are immature and unregulated by KIR-expression they may exert anti-host reactivity. The more favorable outcome for AML but not ALL patients achieving higher lymphocyte counts early post-transplant recapitulates the myeloid leukemia-restricted benefit from superior NK cell recovery observed in AML but not ALL recipients of mismatched T cell depleted stem cell transplants reported by Ruggeri et al (Blood.1999;94:333–9).

Figure Figure
Definition of Cytochrome P450 1A2 as a Target Autoantigen in the Hepatitic Variant of Hepatic Chronic Graft-Versus-Host Disease.
Authors: Mullighan Charles G. , Bogdanos Dimitrios , Vergani Diego , Bardy Peter G.
Published: April 2020

Chronic graft-versus-host disease (cGVHD) of the liver mimicking autoimmune hepatitis is an increasingly well recognized complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). The target antigens and pathogenesis of this complication are poorly understood. We describe a case of hepatitic cGVHD in which cytochrome P450 1A2 (CYP1A2) was identified as a target antigen. A 47 year old male received an HLA matched sibling transplant for acute myeloid leukemia with normal cytogenetics following 3 cycles of idarubicin, cytarabine and etoposide remission induction therapy. Conditioning was with fludarabine 50mg/m2 and cyclophosphamide 120 mg/m2. Both donor and recipient were CMV IgG positive. Standard cyclosporin and short course methotrexate GVHD prophylaxis was used. Due to mixed lymphoid chimerism, cyclosporin was tapered from D94, and a donor leukocyte infusion (DLI) given at D124. One month following DLI, concomitant with attainment of 100% donor lymphoid chimerism, the patient developed florid oral changes of cGVHD and striking elevation of hepatic transaminases (ALT and AST peaked at 3604 (R 0–55 IU/L) amd 2241 (R 0–45 IU/L) respectively) with relatively minor elevation in hepatic ductal markers (GGT 327, R 0–60 IU/L). Liver biopsy was consistent with autoimmune hepatitis. Multiple recipient serum samples at this time demonstrated the presence of a high titre anti-liver microsomal (LM) antibody (titre 1:1280 by indirect immunofluorescence) reminiscent of hydralazine-induced hepatitis and immunofixed an antigen of approximately 51 kDa by immunoblotting using human liver homogenate. Neither pre-transplant recipient nor donor sera were reactive. Immunoblotting for a panel of 13 CYP450 antigens demonstrated that the antibody was specific for CYP1A2. This specificity was confirmed with blocking and inhibition experiments. Sequencing of the entire coding region of the CYP1A2 gene did not identify any coding variants present in the recipient but not the donor. Oral prednisolone resulted in brisk improvement in hepatitis and oral cGVHD, and was ceased at 12 months post transplant. The patient remains well with no further GVHD over 2 years post transplant. This is the first description of a target antigen in hepatitic cGVHD. The mechanism is unclear, but does not appear to be a CYP1A2 polymorphism acting as a minor histocompatibility antigen. Induction of hepatic autommune-like responses have been described in liver transplant recipients receiving calcineurin inhibitors, and it is possible that cyclosporin facilitated an autoimmune-type response in this case. Characterisation of autoantibody responses in additional cases of hepatitic cGVHD will be of great interest.

Figure Figure
Measuring Multiple Symptoms and Inflammatory Cytokines Related to Acute GVHD in AML/MDS Patients under Allogeneic BMT.
Authors: Giralt Sergio , Cleeland Charles S. , Reuben James , Lee Baing-Ning , Williams Lori , Mobley Gary , Wang Xin S.
Published: April 2020

The objective of this study was to prospectively assess the relationship between symptom severity and changes in inflammatory cytokines around the time of diagnosis of acute GVHD in AML/MDS patients during the first 100 days of allo-BMT. Methods: Weekly symptom assessment with the M. D. Anderson Symptom Inventory was done for 30 patients over time of BMT. A panel of inflammatory cytokines (IL-1ra, IL-6, IL-8, IL-10, IL-12, TNF-a) was assayed at multiple time points (pre BMT, conditioning, BMT day, Day 1, nadir, Day 7, 14, 21, 28, 60, 90–100, also including days 1, 3, 5, 10 and 15 after diagnosis of a GVHD for the 26 patients who developed it). Results: Of the 26 patients, 69% had grade 1 GVHD, 27% had grade 2, and 4% had grade 3. 42% were treated with 2mg/kg of steroids, 31% with topical steriods. Figure 1 displays Loess curves that show the multiple symptom pattern for the 26 GVHD patients. These nonspecific symptoms reached their highest levels before the day of diagnosis. Immediate symptom reduction after diagnosis was most likely caused by administration of steroids. The Lowess curves in Figure 2 show the changes in serum cytokines levels pre- and post-GVHD (log based 10). IL-8 was significantly correlated with some symptoms before and after GVHD (Table 1). Conclusion: During development of GVHD in the acute phase of allo-BMT, there is an observed relationship of certain symptom severity and increased or decreased inflammatory cytokine levels in this pilot study. Further study is warranted for a causal link between general sickness and specific cytokines during human GVHD development.

Table 1. Correlations between symptoms severity and cytokines Levels before and after the Development of GVHD in the First 100 Days after Allo-BMT IL-8 Pre-GVHD Shortness of breath r = −0.708, P<.01 Dry mouth r = 0.667, P<.05 Sadness r = 0.631, P<.05 Post-GVHD Sleep Disturbance r = 0.564, P<.05 Drowsiness r = −0.565, P<.05 Vomiting r = −0.558, P<.05 Figure Figure Figure Figure
Gene Signatures Predictive of Outcome in Higher Risk Childhood Acute Lymphoblastic Leukemia (ALL).
Authors: Moskowitz Naomi P. , Bhojwani Deepa , Kang Huining , Min Dong-Joon , Potter Jeffrey , Harvey Richard , Seibel Nita L. , Raetz Elizabeth , Sather Harland N. , Hunger Stephen P. , Willman Cheryl L. , Carroll William L.
Published: April 2020

The use of risk adapted therapy has improved outcome for children with ALL and assessment of early response (ER) has been used to identify a subset of patients who benefit from augmented treatment. In spite of these advances, the majority of treatment failures occur in a “good risk” subset indicating the critical need for better predictors of outcome. We performed oligonucleotide microarrays on 99 B-precursor patients treated on CCG 1961 for high risk ALL to identify: 1) genetic pathways that play a role in early response to therapy (42 day 7 rapid (R)ER, 40 slow (S)ER) and 2) a signature that would predict event free survival (28 pts > 4 yr CCR, 31 relapses < 3 yrs). RNA was extracted and amplified before hybridization to Affymetrix U133 Plus 2.0 microarrays representing over 47,000 transcripts. Data were normalized, filtered and then analyzed by both unsupervised and supervised methods. We identified a robust signature that predicted SER at day 7 (79% of cases in a test set) but not RER thus limiting its utility in risk stratification. Furthermore, we also identified a highly significant set of genes that correlated with SER (M2/M3) at day 14. Importantly, expression of 47 probe sets were significantly different in patients who were in CCR for more than 4 years compared to cases that experienced a relapse within the first 3 years (FDR ≤0.05%). Logistic regression indicated that each of the 47 probe sets has significant prognostic value beyond that contained in the clinical covariates (including gender, age and WBC count). A major subset of these genes were involved in signal transduction (MAP3K3, YWHAZ, PCTK2, TGFBR1, PTPRE, IFNGR2, HSPA8). In a separate validation analysis, subsets of genes were selected using logistic regression with backward, forward and stepwise variable selection and the models proved to be significantly predictive of response in the independent expression data set of 220 patients generated previously using a smaller probe set with U95Av2 microarrays (Mosquera-Caro et al, Blood 2003, 102(11)). Interestingly, the genetic models superseded clinical co-variates such as age, WBC and gender. Finally, to assess further the predictive accuracy of these gene signatures, we randomly divided the data set into a training set (2/3 cases) that was used to develop a signature predictive of relapse that was tested for accuracy in the remaining cases (1/3 cases). This was repeated three times. On average, 77% of patients in CCR and 73% of patients who relapsed were accurately classified in the test set. With further validation these signatures of outcome (not early response) will allow for more precise identification of patients who are at very high risk for relapse and may provide a mechanistic understanding of why patients fail therapy.
VH Gene Usage, Frame Coding and Mutational Status Analysis in Adults with Acute Lymphoblastic Leukaemia (Adult ALL).
Authors: Rai Lena , Casanova Anouska , Hoffbrand A.Victor , Foroni Letizia
Published: April 2020

Analysis of immunoglobulin gene rearrangement for minimal residual disease as a prognostic parameter is a well established test in a variety of leukaemias such as ALL and CLL. The aim of this study was: 1) to analyse VH gene usage, occurrence of in-frame (IF) and out-of-frame (OF) sequences and rate of mutation; 2) to correlate these parameters with overall clinical outcome in adult ALL patients, as this may identify a novel prognostic marker in ALL, especially in the absence of other prognostic indicators.

Patients and methods: DNA from 82 de novo adult ALL patients (31F/51M) was obtained from PB or BM diagnostic samples. Using primers for the FR1 and JH segments, 102 clonal rearrangements were identified. Median WBC was 13.5 × 109/I (range 1.1–550 × 109/I). Sixty-three patients are, to date, in complete clinical remission (median follow up: 7 months; range: 3 weeks–82.5 months), 15 patients have relapsed (median relapse 5.7 mo; range: 2.1–22 mo) and 2 patients had resistant disease. Patients carrying the t(9;22) translocation and patients who received an allogeneic stem cell transplantation were excluded when calculating relapse free survival to avoid bias towards bad and good performance, respectively.

Results: A heterogeneous representation of VH gene usage was demonstrated. Recurrent rearrangements showed preferential use of VH3-66 (4 cases), VH1-2 (7 cases) and VH6 (8 cases; predominant in MLL-AF4 cases), when compared to the non-leukaemic controls (Mortuza et al, 2002; JCO, 20:1094). JH4 and JH6 were found in approximately equal proportions both in control and leukaemic groups, while JH5 (32 cases) was favoured in the leukaemic group. A similar trend was observed with D2-2 (23 cases) when compared to the other D segments in the latter.

There was a predominance of OF sequences (66 alleles; 64%; from 53 patients), although no statistical difference in relapse free survival between IF and OF was observed.

Mutations >1% were observed in 18/82 (22%) patients (18/102 alleles; 18%). These were twice as common in the IF sequences (11/18 patients; 61%) compared to OF sequences (in 6/17 pts, 35.3%). However mutations had no impact on overall survival (p=NS) and neither did immunophenotype (pre B Vs common ALL) (p=NS).

Conclusion: Unique to ALL patients, a considerable proportion of cases carry out-of-frame immunoglobulin rearrangements while a smaller proportion are in-frame, without any apparent clinical impact. In contrast to observations made in other leukemias, such as CLL, occurrence of mutation does not appear to affect clinical outcome in our data set, although a longer follow-up period may be required for this prognostic factor to be fully evaluated.
Clinical Implication of Bone Marrow Angiogenesis in Childhood Acute Lymphocytic Leukemia.
Authors: Won Sung Chul , Rha Sun Young , Oh Seung Hwan , Kim Hwang Min , Lyu Chuhl Joo
Published: April 2020

Purpose: Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are associated with increased angiogenesis, growth, and metastasis in solid tumors. But, until today, the importance of theses factors on leukemia, especially childhood acute lymphocytic leukemia (ALL) has received limited attention. Therefore, this study examined the bone marrow plasma VEGF and bFGF levels in ALL patients and normal controls.

Methods: Bone marrow plasmas at diagnosis from 33 ALL patients (median age 5.97 years; range 1.8–13.9 years) were used for analysis. All ALL patients are divided into 3 groups; one group is standard risk ALL, other group is high risk ALL who did not experience relapse, and the other group is high risk ALL who have experienced relapse during treatment. The bone marrow levels of bFGF and VEGF were determined by enzyme-linked immunosorbent assay (R & D Systems) and compared with the bone marrow levels of 7 healthy control subjects (median age 11.98 years; 6 months–13.6 years).

Results: The VEGF levels were higher in high risk ALL with relapse (N=7, 171.95 ± 202.54 pg/mL) compared with standard risk (N=9, 9.11 ± 21.47 pg/mL), high risk ALL without relapse (N=17, 37.09 ± 38.77 pg/mL) or normal control (N=7, 72.9 ± 68.22 pg/mL)(P=0.006).

Figure Figure

The bFGF levels were also significantly higher in high risk ALL patients who experienced relapse (high risk with relapse; 37.26 ± 35.73 pg/mL, standard risk ALL; 7.90 ± 12.99 pg/mL, high risk ALL without relapse; 7.27 ± 10.93 pg/mL, normal control; 6.39 ± 6.13 pg/mL) (P=0.003).

Figure Figure

The Pearson correlation coefficient between VEGF and bFGF was 0.775 (P < 0.01). Three patients who showed high levels of both angiogenic markers (VEGF; 366.80 ± 141.29 pg/mL, bFGF; 70.27 ± 5.57 pg/mL) are relapsed and died due to disease progression. The other relapsed patients except these 3 patients are alive without disease after salvage chemotherapy.

Conclusion: Our data suggest that the increased levels of VEGF and bFGF in bone marrow may play an important role in estimating prognosis of childhood ALL.
The Repertoire of Gene Rearrangements in Acute Lymphoblastic Leukemia.
Authors: Morley Alexander A. , Brisco Michael J. , Sykes Pamela J. , Latham Sue , Hughes Elizabeth , Wilczek Vicki , Kuss Bryone J. , Waters Keith
Published: April 2020

Rearrangements of the immunoglobulin and T-cell receptor genes provide molecular markers for clones in acute lymphoblastic leukemia (ALL). Determination of the repertoire of gene rearrangements in ALL aids in understanding the clonal biology of the disease and provides molecular markers which can be used to quantify minimal residual disease (MRD).

We have developed a sensitive PCR-based method for analysing the repertoire of immunoglobulin heavy chain (IgH) rearrangements in ALL. Multiple parallel quantitative PCR’s are performed in microplates using different segment-specific primers in different wells in order to determine the individual V (D) and J segments utilised by each rearrangement. The number of rearrangements detected in 18 children and 10 adults with ALL is shown in the table:

VDJ rearrangements DJ rearrangements No. of rearrangements 1 2 3 4 1 2 Childhood ALL 0 11 3 2 1 1 Adult ALL 7 2 0 1

Since each PCR well contained only 2 ng of DNA, more sensitive repertoire analysis was also performed in samples from 10 of the children and 4 of the adults by using 100 ng of DNA in an initial preamplification, which involved a multiplexed PCR containing primers for all leader and J sequences of the IgH gene and which thus amplified all immunoglobulin sequences. The IgH repertoire of the amplified material was then analysed. This two-step approach should theoretically enable detection of clones which comprise down to approximately 10−4 the leukemic population. It detected all rearrangements previously detected by one-step repertoire analysis and, in addition, it detected 0–5 (mean 1.2) rearrangements marking small clones in childhood ALL and 0–3 (mean 1.0) rearrangements marking small clones in adult ALL. Sequencing showed that most, but not all, small clones had a lineage relationship to the dominant clone present in the leukemic population.

Repertoire analysis of IgH rearrangements is a promising technique for identifying molecular markers for measurement of MRD in B-ALL, particularly childhood ALL, since:

–it is conceptually simple and relatively quick –it detects IgH rearrangements with high efficiency, probably higher than that of current techniques. –IgH rearrangements are the best markers to use for measurement of MRD owing to the specificity and sensitivity that they provide –an enhanced ability to identify markers for both large and small leukemic clones may improve the identification of patients prone to relapse.
Value of Molecular Monitoring for Minimal Residual Disease Preceding Autologous SCT Following Diagnosis of B Cell Acute Lymphoblastic Leukemia in Patients Treated with the MRC UKALL12 Protocol.
Authors: Foroni Letizia , Mitchell Wayne , Rai Lena J. , Casanova Anouska , Patel Bella , Richards Sue , Fielding Adele , Goldstone Anthony H.
Published: April 2020

Introduction and aims. Molecular monitoring of minimal residual disease has provided an independent and prognostically significant parameter in evaluating outcome in adult and childhood acute lymphoblastic leukaemia (ALL). The aim of our study was to assess the impact of MRD measured in bone marrow samples collected at time of harvest or prior to harvest in adult B cell receiving autologous stem cell transplant and correlate MRD with overall clinical outcome.

Patients and Methodology. Patients were selected as de novo adult ALL of B cells. All patients were negative for the t(9;22) or (4;11) and had received chemotherapy followed by an autologous SCT (A-SCT). All patients had at least one molecular marker which was tested by quantitative or semi-quantitative PCR with sensitivity ≥1E4. End points were either clinical relapse or CCR with follow up ≥12 months except for two patients who died in CR following transplant, due to infections. Time point for molecular evaluation was a test preceding A-SCT or harvested BM. Nineteen patients were evaluable prior to/or at time of harvest. Nine were females and nine were males. Median age was 25.2 yrs (range: 15.2–52 yrs), total WCC was 7.1 (range: 2.3–68.9×109/l) at time of diagnosis with common ALL (14 pts) as the predominant phenotype. Patients received an A-SCT at a median period of 6 months from diagnosis (range: 5–18 months). Eight patients relapsed (median period to relapse: 22 mo; range: 8–35 mo) and 8 were in continuous clinical remission (CCR) (median follow up 33 mo; range: 8–139 mo). Median interval between Auto-SCT and relapse was 13.7 mo (range: 3–29 mo).

Results. In seven patients residual disease was demonstrated in the BM prior to A-SCT and 6 of them relapsed. In 12 patients no residual disease was detected at time of harvest or prior to transplant and 10 are at present in CCR. The association between MRD positivity and relapse and MRD negativity and CCR was statistically significant in this cohort of patients (p=0.002)(Figure 1).

Conclusions. Molecular monitoring of MRD can provide a useful tool for the monitoring of residual disease in BM harvest prior to SCT and correlates with outcome. It is therefore important that all BM harvests are tested for residual disease in future clinical trial that may use MRD for patients’ stratification.

MRD tests prior to Auto-SCT and relapse FS MRD tests prior to Auto-SCT and relapse FS
Inhibition of the B Cell Associated Tyrosine Kinase SYK as a Potential Therapeutic Target in Aggressive Lymphomas.
Authors: Bertoni Francesco , Rinaldi Andrea , Sasso Anna , Uccella Silvia , Martin Vittoria , Napoli Sara , Pruneri Giancarlo , Zucca Emanuele , Capella Carlo , Catapano Carlo V. , Tibiletti Maria Grazia , Cavalli Franco
Published: April 2020

By using a combination of genomic and expression profiling (Affymetrix GeneChip Mapping 10k Xba131 and U133 set), we had previously analysed a series of 26 mantle cell lymphoma (MCL) samples to identify regions containing genes that might be relevant to MCL pathogenesis and that could represent new therapeutic targets (Rinaldi et al, Ann Oncol2005; vol. 16 suppl. 5, abstract 447). Of interest, one MCL cell line showed a genomic amplification of 9q22 with concomitant over-expression of SYK, mapped within the region. SYK is a tyrosine kinase involved in B cell receptor signaling, a pathway that is believed to play a major role in B cell lymphoma growth and survival. Western blotting experiments and FISH analysis confirmed the over-expression of SYK and the presence of extra-copies of SYK, respectively. Immunohistochemistry on twelve aggressive lymphomas [MCL and diffuse large B cell lymphomas (DLBCL)] confirmed that SYK is over-expressed in a subset of lymphomas. Based upon all these data, we treated seven established human lymphoma cell lines (four MCL, three DLBCL) with increasing doses of piceatannol (Sigma), a SYK inhibitor. All, but one, cell lines showed arrest of cell proliferation. The JeKo-1 and SU-DHL-6 (MCL and DLBCL) cell lines showed an IC50 of less than 10 μM after 72 hr of drug exposure; four cell lines had an IC50 between 30–50 μM. The two cell lines with a high-sensitivity to SYK inhibition had a constitutively high expression of SYK due to DNA gain, as shown by Western blotting and FISH. A high percentage of necrotic and apoptotic cells (39%) has been shown in JeKo-1 by Annexin V staining. Further, in vitro analyses and more extensive immunohistochemistry and FISH on MCL and DLBCL clinical samples (using tissue microarrays) are under way to elucidate the mechanism of action of cytotoxicity and the clinical relevance of SYK DNA amplification. In conclusion, our in vitro data indicate SYK inhibition as a new therapeutic target for the treatment of a subset of aggressive lymphomas, over-expressing SYK. SYK inhibitors are already in clinical development for treatment of asthma. Work partially supported by the Krebsforschung Schweiz and the Swiss Group for Clinical Research (SAKK).
The Humanized Anti-CD40 Monoclonal Antibody SGN-40 Targets Hodgkin’s Disease Cells through Multiple Mechanisms.
Authors: Law Che-Leung , McEarchern Julie A. , Cerveny Charles G. , Smith Leia M. , Nesterova Albina , Gordon Kristine A. , Grewal Iqbal S. , Wahl Alan F.
Published: April 2020

CD40 is a member of the TNF receptor superfamily expressed by multiple lineages within the hematopoietic system that carries out essential functions in the maturation of antigen-presenting cells, cross-priming of cytotoxic T cells, initiation of T cell-dependent antibody responses, and Ig class-switching. CD40 is widely expressed by lymphoid malignancies including B-cell precursor acute lymphoblastic leukemia (B-ALL), non-Hodgkin’s lymphoma (NHL), Hodgkin’s disease (HD), and multiple myeloma (MM). SGN-40, a humanized version of the murine anti-CD40 monoclonal antibody S2C6, mediates antibody-dependent cellular cytotoxicity (ADCC) against both NHL and MM cells, directly induces apoptosis in NHL lines, and inhibits the proliferative response of MM cells toward autocrine IL-6 through down-modulation of the IL-6 receptor. Based on these activities, SGN-40 is currently in phase I clinical trials for relapsed and/or refractory MM and NHL. In this study, we examined the expression of CD40 on primary tumors isolated from 93 independent patients with HD. Seventy nine of these 93 cases (85%) demonstrated expression of CD40 by both the Reed-Sternberg cells as well as the infiltrating mononuclear cells. In a 1+ to 3+ scoring system, 35% of cases showed intense 3+ staining, 24% were positive (2+), and 26% were weakly positive (1+). Flow cytometry revealed that 8 of 9 HD cell lines expressed surface CD40, with receptor numbers ranging from 2,000 to > 370,000. SGN-40 induced CD40-positive HD cell lysis through ADCC in a dose-dependent manner, with IC50 values < 10 ng/mL. Fc-dependent effector function of SGN-40 was further exemplified by its ability to enhance macrophage mediated antibody-dependent cellular phagocytosis (ADCP). Specific ADCP against CD40-positive target cells was detected when SGN-40 was present at concentrations > 10 ng/mL. Furthermore, direct anti-proliferation signaling by SGN-40 in HD cells was also observed. In the L-1236 cell line, SGN-40 at doses ≥ 100 ng/ml reduced DNA synthesis to 25% of the untreated cells as measured by 3H-thymidine incorporation assays. Taken together, these data suggest that HD may be a suitable therapeutic target for SGN-40.
Cucurbitacins, a Family of Natural Compounds That Effectively Disrupt Stat3 Signaling in Antigen-Presenting Cells (APCs) Are Promising Agents To Overcome Tumor-Induced CD4+ T-Cell Tolerance.
Authors: Cheng Fengdong , Wang Hongwei , Suarez Alfonso , Horna Pedro , Sebti Said , Sotomayor Eduardo M.
Published: April 2020

Signal transducer and activator of transcription 3 (Stat3) is a key mediator of several cytokines and growth factors signaling pathways. On myeloid cells, activation of Stat3 to its phosphorylated form (pStat3) has been shown to negatively regulate inflammatory responses. Recently, we have unambiguously demonstrated that Stat3 signaling in APCs also play a central role in the decision leading to immune activation versus immune tolerance of antigen-specific T-cells1. In spite of these advances, there is however a paucity of therapeutic strategies targeting this signaling pathway in immune cells.

Using a high throughput cytoblot screening for phospho-Stat3 inhibition, we have recently identified a family of natural compounds known as Cucurbitacins that effectively disrupt Stat3 signaling at different levels2. Three compounds have been identified, Cucurbitacin A (CuA) that inhibits phospho-JAK-2, Cucurbitacin I (CuI) a dual inhibitor of p-JAK2 and p-Stat3 and Cucurbitacin Q (CuQ) a selective inhibitor of p-Stat3. In vitro treatment of peritoneal elicited macrophages (PEM) and bone marrow-derived dendritic cells (DCs) with increasing concentrations of CuA or CuI resulted in inhibition of p-Stat3 and enhanced antigen presentation to naive CD4+ T cells specific for a MHC class II restricted epitope of influenza hemagglutinin (HA). Indeed, these clonotypic T cells displayed increased antigen-specific proliferation and IL-2 production as compared to clonotypic T cells encountering cognate antigen on untreated APCs. Furthermore, unlike untreated PEM or DCs, which are unable to trigger IFN-gamma production by CD4+ T-cells, Cucurbitacin-treated APCs efficiently trigger the production of this cytokine by naïve CD4+ T-cells in response to cognate antigen. Given the above results, we explored next whether inhibition of Stat3 signaling in B-cell lymphomas by Cucurbitacins might increase the intrinsic antigen-presenting capabilities of these malignant B-cells. Reminiscent of our findings with bone marrow derived APCs, Cucurbitacin-treated A20 lymphoma cells also display enhanced antigen-presenting cell function leading to increased proliferation, IL-2 and IFN-gamma by naive antigen-specific CD4+ T-cells. More importantly, tolerant CD4+ T-cells (isolated from lymphoma bearing mice) exposed to Cucurbitacin-treated A20 B-cells regained their ability to proliferate and produce significant amounts of IL-2 and IFN-gamma in response to cognate antigen stimulation.

Taken together, the ability of Cucurbitacins to inhibit p-Stat3 in normal APCs as well as in malignant B-cells make these natural compounds a promising agents to overcome the remarkable barrier that tolerance to tumor antigens has imposed to cancer immunotherapeutic strategies.
Nongastric Marginal Zone B-Cell Lymphoma: Analysis of 247 Cases - Clinical Presentation and Treatment Outcomes of Nongastric Marginal Zone B-Cell Lymphoma.
Authors: Oh Sung Yong , Ryoo Baek-Yeol , Kim Won Seog , Park Yeon Hee , Kim Kihyun , Kim Hyun Jung , Kwon Jung Mi , Lee Jeeyun , Ko Yong H. , Ahn Yong Chan , Oh Suk Jung , Lee Soon , Kim Hyo Jin , Kwon Hyuk Chan , Bang Soo-Mee , Kim Jung Han , Park Jinny , Lee Seung-Sook , Kim Ho Yong , Park Keunchil
Published: April 2020

Purpose: Nongastric marginal zone B-cell lymphoma (NG-MZL) is a relatively uncommon indolent lymphoma. We report a retrospective analysis of 247 patients with .NG-MZL, presenting their clinical features and therapeutic outcomes.

Methods: From 1990 to 2005, a total of 247 patients with histologically confirmed NG-MZL were analyzed.

Results: The median age was 49 years (range, 13–89 years). The study involved 129 males (52.2%) and 118 females (47.8%) The most common involving site was orbit and ocular adnexa (48.6%) followed by lymph node and lymphatic organs (17.8%), bowel (9.3%), lung (6.1%), thyroid gland (4.9%), salivary gland (4.5%) in the decreasing order of frequency. Ann Abor stage I/II disease was present in 78%(167 out of 215). BM involvement was less than 10%(19 out of 211). B symptom was observed in only 2%. One and eighty-six patients out of 208 were in low or low-intermediate risk group (89%) according to international prognostic index(IPI). Eighty percents (172/215) were in low risk group in follicular lymphoma international prognostic index (FLIPI). Patients were treated with a variety of therapeutic strategies. Complete and partial remissions were achieved in 139(92%) and 8(5.3%) of the 151 stage I/II patients, respectively, with an overall response rate of 97.3%. Especially, radiation containing treatment achieved 96% CR rate (108 out of 113). In 38 patients with stage III/IV, CR and PR were achieved in 17(44.7%) and 11(28.9%). The estimated 5-year overall survival (OS) and progression free survival (PFS) were 93.8% and 70.1%, respectively. Although anthracyclin contaning regimen could achieve higher CR rate, it did not improve PFS. Stage III/IV, low hemoglobin, high/high-intermediate IPI, poor risk FLIPI and nodal MZL were poor prognostic factors for PFS in multivariate analysis.

Conclusion: NG-MZL is an indolent disease. For localized disease radiation achieved excellent local control. Anthracyclin containing chemotherapy cannot improve outcome. Both IPI and FLIPI can be applied to predict the prognosis.
Adenovirus-Interferon-g (TG1042) Demonstates Clinical Activity in Relapsing Primary Cutaneous Lymphoma (PCL): A Phase I/II Multicentric Trial.
Authors: Urosevic Mirjana , Luger Thomas E. , Sterry Wolfram , Dreno Brigitte , Dereure Olivier , Baudard Marion , Bagot Martine , Bleuzen Pascal , Rossi Jean-Francois , Dummer Rheinard
Published: April 2020

PCL has been successfully treated with interferons (IFNs), which can offset the Th2 dominance associated with PCL. Intratumoral (IT) injection of TG1042 (a non-replicating recombinant adenovirus with a human IFN-γ cDNA insert) allows high local levels of IFN-γ without severe toxicity induced by systemic delivery. We undertook a multicentric, phase I/II, open-label, trial of repeated, IT injection of TG1042 in patients with advanced primary T cell (CTCL) and multilesional B cell (CBCL) cutaneous lymphoma. The designated lesions were injected on day 1, 8 and 15 with no injection in the fourth week (1 cycle) and thereafter up to 4 cycles. Immunohistochemistry and qPCR were performed on injected lesions biopsied at baseline and after each cycle. In the phase I step, undertaken on a monocentric basis, 9 patients were enrolled in 3 successive cohorts at the doses of 3×109 viral particles (vp), 3×1010 and 3×1011 vp injected in only one lesion and 9 additonal patients were treated at the doses of 3×1011 vp divided in up to 3 lesions. In the phase II step, 18 additional patients have been planned to be treated on a multicentric basis at 3×1011 vp injected in up to 3 lesions. To date, the 36 planned patients (30 CTCL and 6 CBCL) have been enrolled and received a total of 280 injections, 26 patients have completed the treatment. Treatment is well tolerated with only 3 grade III toxicity. Injection site reaction and flue like syndrom are the most commonly observed adverse events. Local clinical response has been observed in 13 (7 complete responses) out of 27 evaluable patients. Disappearance of non-injected lesions was also observed and led to 12 overall responses (6 complete responses) out of 26 evaluable patients. Histology demonstrates pronounced differences in infiltrate patterns following treatment, with signs of vasculitis and increased numbers of eosinophils, neutrophils, CD8 and TIA-1+ve cells. CD4/CD8 ratio decreased in most tumors. Transgene-IFN-γ mRNA was detectable in injected lesions. Gene expression analysis of biopsies and PBMC shows up-regulation of IFN-γ and various immune response-associated genes. Final clinical and immunological data will be presented. In conclusion, these data show that the administration of TG1042 is well tolerated and results in clinical responses associated with in situ immunological changes. These encouraging results demonstrate a potential significant benefit of TG1042 for the treatment of primary cutaneous lymphoma.
No Role for Chemoradiotherapy When Compared with Chemotherapy Alone in Elderly Patients with Localized Low Risk Aggressive Lymphoma: Final Results of the LNH93-4 GELA Study.
Authors: Fillet Georges , Bonnet C. , Mounier N. , Ganem G. , Molina T. , Thieblemont C. , Fermé C. , Quesnel B. , Martin C. , Gisselbrecht C. , Tilly H. , Reyes F.
Published: April 2020

Chemoradiotherapy is standard treatment for localized aggressive lymphoma (Miller et al., NEJM, 1998). Because previously published series were heterogeneous with regard to prognostic factors such as age, we aimed to determine the optimal therapy for elderly patients with low risk localized lymphoma. From March 1993 to June 2002, 576 patients (pts) over 60y of age with aggressive lymphoma (WF histology: F, G, H or Kiel anaplastic) and without any adverse factor of the age-adjusted International Pronostic Index were randomly assigned to a chemoradiotherapy arm (299 pts) consisting of 4 cycles of CHOP (doxorubicin 50 mg/m2 d1, cyclophosphamide 750 mg/m2 d1, vincristine 1.4 mg/m2 [up to 2 mg] d1, prednisone 60 mg/m2 d1-5) given every 3 weeks followed by 40 Gy involved-field radiotherapy or to a chemotherapy-alone arm (277 pts) consisting of 4 cycles of CHOP given every 3 weeks. Randomization was stratified according to center and the presence of bulky disease (≥ 10 cm). Principal characteristics were: median age, 68y; male gender, 52%; stage I, 63%; bulky disease, 8%; extranodal involvement, 56%; diffuse large B-cell histology, 80%. Complete response at the end of treatment was similar in both groups (89% and 90 % respectively). Treatment-related death occurred in 1% of pts in each group. On an intent-to-treat basis and with a median follow-up of 6.8y, event-free survival (EFS) and overall survival (OS) did not differ significantly between the two treatment groups (p= 0.7 and p = 0.6, respectively). 5y-EFS rates were 68% for pts treated with chemotherapy alone as compared to 66% for those receiving chemoradiotherapy; 5y-OS rates were 72% and 68% respectively. 66%, of deaths resulted from lymphoma progression. In the subgroup of 247 pts over 70y of age, the 5y-OS was higher in those treated by chemotherapy alone (70% as compared to 58%) but this trend did not reach significance (p=0.1). In a multivariate analysis of the 576 pts, EFS and OS were affected by stage II (p<0.0001), male gender (p=0.02), not by tumor bulk.

Conclusion: mature data of the LNH93-4 trial indicate that CHOP plus radiotherapy does not provide any advantage over CHOP alone for the treatment of low risk localized aggressive lymphoma in elderly pts. To note, although the majority of patients had stage I disease, local control by irradiation did not decrease the overall relapse rate. A similar conclusion was recently reached in young adult pts treated by chemotherapy alone (Reyes et al, NEMJ 2005). Taking these results together, we consider that local irradiation following chemotherapy has no role in the treatment of pts with low risk localized aggressive lymphoma. On the basis of its previous published results (Coiffier et al, NEJM, 2002), the GELA recommends to treat such elderly pts with rituximab plus CHOP.
Lyposomal Doxorubicin (Caelyx), Cyclophosphamide - Rituximab (DC-R) Plus GM-CSF as a Salvage Therapy in B-Diffuse Large Cell Lymphoma (B-DLCL) after Autologous Stem Cell Transplantation (ASCT) Failure.
Authors: Olivieri Attilio , Montanari Mauro , Capelli Debora , Scortechini Ilaria , Gini Guido , Trappolini Silvia , Leoni Pietro
Published: April 2020

B-DLCL patients after ASCT failure have a very poor prognosis, salvage chemotherapy or a second transplantation cannot substantially modify the very poor outcome which is characterized by a median overall survival (OS) of 3 months. We recently demonstrated the feasibility of chemoimmunotherapy after ASCT by associating a CHOP-modified schedule+Rituximab and GM-CSF, but often the antracycline retreatment in these patients can be hampered by the risk of severe cardiac damage or by a reduced organ reserve. For this reason we adopted a modified version of this protocol for patients receiving a previous Doxorubicin cumulative dose>300 mg/sm, with the substitution of conventional antracycline with a Lyposomal Pegylated Doxorubicin (DC-R schema); the other inclusion criteria were: diagnosis of B-DLCL CD20+, P.S (WHO)= 0–2, age < 75 years, relapse or progressive disease (PD) after ASCT, measurable disease, absence of severe organ dysfunction and CNS involvement. Seventeen patients were eligible for this salvage protocol; the median age was 47 (28–73) years; the P.S. (WHO) was 0–1 in 11 patients and 2 in six; the disease status after ASCT was: untested relapse in 14 cases and PD in 3 cases. The DC-R + GM-CSF schema (every three weeks) consisting in: R 375 mg/sm day 1 and 15, Caelyx 30 mg/sm and cyclophosphamide 750 mg/sm day 1, GM-CSF 150 mg/day from day 5 until neutrophils recovery. Patients showing disease progression after two courses were excluded while the responders received two more coursesof DC-R+GM-CSF; patients achieving complete remission (CR) after 4 courses did not receive any further treatment. All the17 patients received the planned treatment and were evaluable for response: the overall response rate (ORR) was 58.8% with 10 CR (58.8%), 7 patients showed a PD after 1–2 courses. The toxicity (WHO) consisted in: grade III–IV neutropenia in 11 patients (64.7%) and thrombocytopenia in 2 patients (23.5%), grade I–II infections in 2 patients and grade IV (pneumonia) in one patient. With a median follow up of 15 (1–74) months, 8 out of 10 responders patients are alive and in CR with a median disease-free survival of 24.5 (8–70) months, one patient is alive and in relapse at + 23 months and 1 patients died for infection months after while in CR. Our experience shows that DC-R + GM-CSF is an effective salvage treatment for B-DLCL after ASCT failure; indeed the follow up > 12 months in 5/17 (29.4%) patients in CR suggests a chance of cure.
The Role of Antiviral Treatment in Hepatitis C Virus (HCV)-Related Non-Hodgkin Lymphoma: An Update of a Multicenter Study.
Authors: Vallisa Daniele , Bernuzzi Patrizia , Arcaini Luca , Lazzaro Antonio , Trabacchi Elena , Anselmi Elisa , Arcari Anna Lisa , Moroni Carlo , Berte Raffaella , Malingher Alessandro , Lazzarino Mario , Cavanna Luigi
Published: April 2020

HCV is largely, although not homogenously, diffuse in North-western Europe and U.S.A. It has been shown to play a role both in hepatocellular carcinoma and in B-cell non-Hodgkin lymphoma (B-NHL). Up to now the exact biological mechanisms that could explain the lymphomagenic role of the virus are unknown. We have previously published a series of 13 patients, affected by low grade B-cell NHL and characterized by an indolent course (i.e. doubling time less than 1 year, no bulky disease), who underwent antiviral treatment only with peghilated interferon and ribavirin (peghilated interferon 50–70 microgram weekly, ribavirin 1000–1200 mg daily). Now we report an update of this study. Up to now we were able to evaluate 16 patients with a mean follow up of 15,1± 7,6 months (range 2–28 months). Eight patients experienced complete or good partial haematological response that has lasted up to now with a mean follow up of 16,5 months, among them 3 splenic marginal lymphomas, 2 nodal marginal, 1 follicular lymphoma, 1 plasmocytoid and 1 marginal extranodal lymphoma. Three other patients achieved a long lasting partial response.

The only one relapse (marginal nodal lymphoma) occurred about one year after the end of treatment, hematological relapse happened together with viral relapse, the lymphoma reappeared as highly chemo resistant high grade lymphoma, and two months later the patient died. Interestingly complete and good partial responses were more likely to be seen in viral genotype 2 (p=0.035) and were strictly related to the decrease of viral load under treatment (p=<0.001). Toxicity causes the stop of the treatment in 3 patients; however one of them was able to achieve complete hematological response. Time to achieve hematological response was quite long (mean 8± 4,5 months).

This kind of experience strongly provides a role for antiviral treatment in patients affected by HCV related low grade B-cell NHL. Especially viral genotype 2 infection may be considered a good prognostic marker for hematological response as well as decrease of viral load under treatment. Toxicity in our hands was however significant and further experiences are warranted in order to better modulate antiviral therapy doses.