bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effect of Bone Marrow Involvement on the Efficacy of Pegylated Liposomal Doxorubicin + Bortezomib Vs Bortezomib Alone in Patients with Relapsed/Refractory Multiple Myeloma
Authors: Shah Jatin , Blade Joan , Miguel Jesus San , Sonneveld Pieter , Luo Donghan , Lantz Keith C , Lowery Colin , Harousseau Jean-Luc , Orlowski Robert Z.
Published: April 2020

Background: Patients with relapsed/refractory multiple myeloma (RRMM) often have a large tumor burden with significant bone marrow involvement, conferring a poor prognostic feature. Those patients with significant bone marrow involvement may benefit from combination regimens with increased activity. Orlowski et al. reported a phase III randomized trial with pegylated liposomal doxorubicin (PLD) + bortezomib (B) vs. B alone in RRMM, which demonstrated improved time to progression (TTP) with the combination of PLD+B. (Orlowski, JCO 2007) We investigated the effect of bone marrow involvement on overall response rates and time to progression (TTP) of PLD+ B versus B alone in patients with RRMM.

Methods: Eligible patients were randomized to bolus IV B 1.3 mg/m2 on days 1, 4, 8, and 11 of each 3-week cycle (n=322) or to the same B regimen plus IV PLD 30 mg/m2 on day 4 (n=324) of each cycle. The subsets of patients with bone marrow involvement and those with >30% involvement were analyzed retrospectively.

Results: A total of 379 patients had bone marrow involvement and 349 patients had >30% bone marrow involvement compared with 259 with no bone marrow involvement. Time to disease progression (TTP) and complete response (CR) + partial response (PR) rates for PLD+B vs B alone in patients with or without any bone marrow involvement or >30% bone marrow involvement are listed in the table.

PLD+B B P valueb Hazard Ratioc a. Based on Kaplan-Meier product-limit estimates. b. Based on stratified Log-rank test. c. A hazard ratio >1 indicates an advantage for PLD+B. d. Cochran-Mantel-Haenszel test controlling for Beta2 microglobulin and response to initial treatment. No bone marrow involvement, N 132 127 TTP, Median days (95% CI)a 251 (212, 282) 198 (162, 227) 0.0134 1.69 (1.11, 2.57) Total CR+PR, n/evaluable (%) 49/118 (42) 43/124 (35) 0.2887d Any bone marrow involvement, N 188 191 TTP 311 (264, 354) 197 (169, 222) .0001 1.91 (1.37, 2.67) Total CR+PR 94/183 (51) 88/182 (48) .5187d >30% bone marrow involvement, N 172 177 TTP 276 (221, 415) 183 (158, 211) .0005 1.83 (1.30, 2.58) Total CR+PR 82/168 (49) 69/170 (41) .1446d

Regardless of bone marrow involvement or extent of bone marrow involvement, there was a statistically significant benefit in TTP for PLD+B vs. B alone. Safety profiles for the 2 regimens in the subsets of patients with or without bone marrow involvement were consistent with the known toxicities of the 2 agents.

Conclusions: These data suggest that the combination of PLD+B is an active salvage therapy and superior to B alone in patients with RRMM, regardless of the presence or extent of bone marrow involvement. Patients with significant bone marrow involvement and associated poor risk due to increased tumor burden may benefit from the addition of PLD.
Six Years of Experience and Outcome with Maintenance Therapy with Very Low Dose Thalidomide after Auto-SCT in Multiple Myeloma
Authors: Mettivier Vincenzo , Pezzullo Luca , Rocco Stefano , Finizio Olimpia , Musto Pellegrino , Nunziata Giuseppe , Bene Lucia , Mettivier Laura
Published: April 2020

New drugs and high dose therapy with auto-transplantation (auto-SCT) has improved prognosis of multiple myeloma (MM). New drugs are promising in upfront therapy while the role of maintenance is still debated. Thalidomide (thal) is an active drug in the treatment of myeloma, and is been investigated as first line therapy, the limit of this drug is the toxicity dependent dose and this determines a poor compliance. It could be useful in the control of minimal residual disease. We used low dose of thal as maintenance after autologous transplantation in patient with MM from January 2002 and here we bring our experience after six years of observation. From January 2002 to August 2008 17 patients (8 males and 9 females) with MM have been treated in our institution. Median age was 59,5 years (range 48–72). 10 were IgG, 3 IgA, 3 light chains and 1 plasma-cell leukaemia. Treatment was 4 cycles of VAD regimen followed by auto-SCT. 4/17 performed double auto-SCT. Three months after SCT these patients has begun the maintenance with thal 50 mg/die, to start thal maintenance 9 patients were in CR, 5 in PR and 3 in resistant disease and the median somministration of thal has been of 12 months (range 3–24 months). Median follow up from the beginning of maintenance therapy was 40 months (range 4–76) with 11/17 (64%) patients in CR or stable disease, with progression free survival (PFS) and overall survival (OS) projected at 75 months respectively of 53% and 51% from to start thal. In our experience we have observed a neurological toxicity (grade I–III) in the 65% of the patients but only 4 have had to suspend the treatment; a haematological toxicity of grade I in the 55% of the patients that have not behaved interruption of the treatment and finally in any case we have documented thrombotic episodes. Finally we have compared this group of patients with another group (18 patients) with the same clinical characteristics that we have observed in the same period but that have not effected maintenance with thal. In this last group 13/18 patients (72%) relapsed with median follow-up of 36 months (range 14–75) and median PFS and OS of 16 and 30 months respectively. The difference between the 2 groups is statistically significant for PFS (p: 0.003) and OS (p: 0.04). The median overall survival observed after progression, in the two groups, has been of 13 months in thal group and 17 months in the group of patients that have not effected the maintenance, this difference is not statistically different (p:0.06). In conclusion in 6 years of observation our experience has shown, even if the number of the patients is small, that maintenance with low doses of thal, after auto-transplantation, it not only has a good compliance but it improves the PFS and OS in this cohort and it doesn’t worsen the OS from the relapse.
Successful Salvage and Hematopoietic Reconstitution with Bortezomib and Melphalan in Relapsed and Refractory Multiple Myeloma
Authors: Mazumder Amitabha , Jagannath Sundar , Vesole David H.
Published: April 2020

The management of relapsed and refractory multiple myeloma (RRMM) patients who have failed older conventional and more recent novel agents remains challenging. Patients typically have short response duration, develop prolonged cytopenias and are usually not eligible for clinical trials due to inadequate bone marrow reserve. Furthermore, these patients often have inadequate numbers of stem cells (<3 × 10(6) CD34/kg) remaining for a myeloablative transplant. In patients who have remaining stem cells in storage, we have developed an algorithm to enable the reconstitution of hematopopiesis to allow enrollment in clinical trials. In addition, our algorithm tests the synergy of bortezomib and non-myeloablative doses of melphalan with stem cell boost support. We have treated 6 patients with RRMM who have failed novel agents (including bortezomib) and chemotherapy (dexamethasone, cytoxan, etoposide, cisplatin, and doxil) and who have had limited duration of response previously with high dose melphalan. All patients were cytopenic (ANC < 1000 and/or platelets < 50,000/mcl) and thus not eligible for existing clinical trials. The melphalan was administered intravenously at 30–50 mg/m2 on days 1 and 4, dependent on the patient age and performance status. Bortezomib was administered 1.3 mg/m2 by intravenous push prior to the melphalan on the same schedule. The patients then received 1.5 to 2.2 × 10(6) CD34+ cells/kg. (depending upon the amount of remaining stem cells). All patients were treated in the outpatient clinic. 3 patients required hospitalization for neutropenic fever; the median hospitalization duration was 3 days (range 0 to 6). The overall response (MR, PR, VGPR) was 100% including: 5 PR and 1 VGPR.. More importantly, all patients achieved improved hematopoiesis with WBC > 2000 and platelets > 50,000/mcl. The median duration of response exceeded 2 months. All patients were able to proceed on to new clinical trials. We conclude that the combination of bortezomib and melphalan produces an effective salvage strategy, with hematopoietic reconstitution in RRMM. This non-myeloablative approach serves as a bridge for disease control and hematopoietic reconstitution to allow for subsequent eligibility on clinical trials. We recommend that during stem cell collection that sufficient hematopoietic stem cells be stored to support future myeloablative and similar non-myeloablative chemotherapy regimens.
Clinical Experience and Prognostic Factors with Bortezomib-Combined Regimen in 40 Patients with Multiple Myeloma
Authors: Li Juan , Zhao Ying
Published: April 2020

To explore the medical effects, prognostic factors relating to the effects and side effects of bortezomib-combined regimen in treatment of multiple myeloma (MM), and evaluate the safety of this regimen in patients with special conditions.

Methods: Forty newly diagnosed and relapsed or refractory MM treated with the regimen of combination of bortezomib and dexamethasone in a cycle of 3 weeks. All of the patients received a median of 3 (1–9) cycles of the treatment. Response to bortezomib was evaluated according to the criteria of the European Group for Blood and Marrow Transplantation (EBMT) and adverse events were graded according to the National Cancer Institute Common Toxicity Criteria.

Results: Total response rate was 75.0% (30/40), and the rate of CR+nCR was 42.5% (17/40). Eighty percent patients (24/30) showed effects after the first cycle, although the median time of best effect was 2 cycles. Compare to the other types, light-chain type had higher total response rate (100.0% vs. 65.5%, P=0.038)and CR rate (36.4% vs. 6.9%, P=0.039), and also responded earlier with 81.8% (9/11) patients reached the best effect after the first cycle. The factors which showed no relationship to the effects were age (<65 years vs. >65 years, P=0.081), sex (P=0.696), DS staging (P=1.0), ISS staging(P=0.969), newly or retreated (P=0.731), using thalidomide or not(P=0.338), renal function damage or not(P=0.401), PLT level(P=1.0), hypercalcemia or not(P=0.306), Hb level(P=0.70)and the ratio of tumor cells in bone marrow(P=0.693). Grade III~IV adverse effects in this VD regimen was low, including leucocytopenia, thrombocytopenia, diarrh ea and debility, could be relieved by symptomatic treatment or delay the chemotherapy. The rate of infection was high and which was one of the important cause of death. Renal function improvement could be showed in all of the 10 patients who had renal inadequacy, and compared with normal patients, the incidences of side and adverse effects had no statistical differences.

Conclusion: The Bortezomib combination regimen has significant effect in MM treatment. It shows more significant in light-chain type patients, and show earlier effect onset. This regimen can be tolerant in most patients, and is also safe in patients with renal inadequacy.
Clinical Efficacy of Pegylated Liposomal Doxorubicin, Vincristine and Dexamethasone in Patients with Newly Diagnosed Multiple Myeloma
Authors: Wu Wen , Gao Xiaodong , Xu Lan , Yan Hua , Shen Zhixiang , Hu Jiong
Published: April 2020

Object: To investigate the short-term and long-term efficacy and toxicity of pegylated liposomal doxorubicin, vincristine and dexamethasone (DVD) in patients with newly diagnosed multiple myeloma (MM).

Methods: Twenty-five patients (13 males, 12 females, median age 55 years) with newly diagnosed multiple myeloma were treated with pegylated liposomal doxorubicin 40 mg/m2 and vincristine 2 mg intravenously on day 1 plus dexamethasone 40 mg intravenously or orally on days 1–4 (DVD) for median 4.5 (2–8) cycles. Treatment was repeated every 4 weeks. Response was evaluated according to the International Uniform Response Criteria for Multiple Myeloma (2006) before initiation of each course. Adverse events were graded according to the National Cancer Institute Common Toxicity Criteria, version 3.0.

Results: After 4.5 (2–8) courses of the median cycles, clinical response was observed in 20 patients (80%), including complete response in 4 (16%), very good partial response in 3 (12%), partial response in 10 (40%), minimal response in 3 (12%) and stable disease in 2 (8%). The median time to initial response was 1.2 months and the median time to best response was 4 months. After 25 (2–50) months of median follow-up, the median progression-free survival was 20 months, while the median overall survival has not yet been reached. The overall survival rate was 72% (18/25). The most common adverse events were gastrointestinal symptoms (nausea and vomiting in 10, constipation in 9 patients), neutropenia (7 patients), anemia (6 patients) and thrombocytopenia (4 patients). DVD was associated with more hand-foot syndrome (4 patients) and mucitis (2 patients).

Conclusions: DVD scheme is an effective therapy with a high response rate and manageable toxicities for patients with newly diagnosed multiple myeloma.
Modulation of Soluble Interleukin 2 Receptor (sIL-2R) Levels in Patients with HIVWaldenstrom Macroglobulinemia (HIV-WM) Treated with Biaxin-Pentoxifylline-Dexamethasone Followed by Prednisolone (BPD-Pred)
Authors: McPherson Eugene , Tassy P.
Published: April 2020

Serum levels of sIL-2R represents the total amount of activated T-lymphocytes in tumor infiltrating lymphocytes of cancer tissues and metastatic organ sites. Most malignant diseases overexpress sIL-2R in comparison to non-malignant controls. Immunomodulatory effect of IL-2 therapy on interferon (IFN), tumor necrosis factor alpha (TNF-a) production, nuclear factor kappa–b (NFkb) in vivo and in vitro and on the expression of sIL-2R in Human Immunodeficiency Virus – Waldenstrom Macroglobulinemia (HIV-WM) patients with IgM is not well defined. We present a 42 year old female with HIV, CD4-343/cu mm, viral load (VL) of >100K (persistently elevated VL), hypercoagulable state, serum viscosity >3 centpoises, IgM level of >1 gram with serum free light chain kappa/lambda ratio elevated as well as IgG and IgA on immunofixation studies. She was treated initially with BPD (Biaxin, Pentoxifylline, Dexamethasone) with partial response and with poor paraprotein parameter response. Her HAART regimen consisted of ritonavir base therapy. When dexamethasone was weaned off and prednisolone therapy started, greater than 50% reduction of paraprotein (IgM) occurred over 4–6 weeks. We measued serum sIL-2R, C-reactive protein (CRP), throughout treatment over 12 weeks. The serum sIL-2R level significantly decreased from 21,513.274 pg/ml to 5,424.779 pg/ml (normal 3,592.9 – 9,734.5 pg/ml). The C-reactive protein normalized and IgM was now 0.211 gram and within normal limits as well as serum viscosity.

CONCLUSIONS: The suppression and modulation therapy of CRP gene transcription with manipulation of NFkb/IKKb with ritonavir based HAART and BPD-Pred with normalization of sIL-2R, CRP, and IgM paraprotein in HIV-WM patients may have an independent effect on prognosis and maintenance therapy. Serum sIL-2R levels may serve as a useful marker in evaluating HIV-WM disease, stage for stage and monitoring disease progression.
BAALC and WT1 Transcript Amount Discriminates Secondary or Reactive Eosinophilia from Idiopathic Hypereosinophilic Syndrome or Chronic Eosinophilic Leukemia
Authors: Cen Jiannong , Chen Zixing , Qi Xiaofei , Yao Li , He Jun , Qiu Qiaocheng , Qiu Huiyin
Published: April 2020

Idiopathic hypereosinophilic syndromes (HES) or chronic eosinophilic leukemia (CEL) comprise a spectrum of indolent to aggressive diseases characterized by persistent hypereosinophilia. Hypereosinophilia can result from the presence of a defect in the hematopoietic stem cell giving rise to eosinophilia, it can present in many myeloproliferative disorders or alternatively it may be a reactive form, secondary to many clinical conditions. The fusion gene FIP1L1-PDGFR alpha was identified in a subset of patients presenting with HES/CEL. In spite of this, the majority of HES/CEL patients do not present detectable molecular lesions and for many of them the diagnosis is based on exclusion criteria and sometimes it remains doubt. CD34-positive progenitor cells from bone marrow (BM) express BAALC and WT1. Overexpression of BAALC and WT1 were seen in patients with AML and ALL. In a subset of AML it marked poor prognosis, suggesting a role for BAALC or WT1 overexpression in acute leukemia. To explored the possibility to distinguish between HES/CEL and reactive hypereosinophilia based on the measurement of BAALC and WT1 transcript amount. Twenty-two patients with hypereosinophilia were characterized at the molecular level and analyzed for BAALC and WT1 expression. The transcription of FIP1L1-PDGFRalpha fusion gene was detected by nested RT-PCR. The relative transcript amount of BAALC and WT1 were determined by real time PCR analyses. The FIP1L1-PDGFRalpha fusion gene expressed has been identified in bone marrow mononuclear cells of 4 cases. The relative expression level of BAALC and WT1 in these 4 cases with positive FIP1L1-PDGFRalpha fusion gene expression were 2.27(0.27–6.8) and 0.39(0.002–0.90), respectively. Whereas the relative amount of transcripts of BAALC and WT1 in 18 patients with negative FIP1L1-PDGFRalpha fusion gene were 0.069(0.015–0.11) and 0.054(0–0.34) respectively. The relative amount of transcripts of BAALC and WT1 in patients with HES/CEL were 32 times and 7 times than that in those with negative FIP1L1-PDGFRalpha fusion gene, respectively. These results clearly demonstrates that BAALC and WT1 quantitative assessment allows to discriminate between HES/CEL and reactive eosinophilia and represents a useful tool for disease monitoring especially in the patients lacking a marker of clonality.
Frequent Reduction or Absence of Detection of the JAK2-Mutated Clone in JAK2V617F-Positive Patients within the First Years of Hydroxyurea Therapy
Authors: Girodon François , Schaeffer C.éline , Cleyrat C.édric , Mounier Morgane , Lafon Ingrid , Santos FréD.éric Dos , Duval Aurélie , Maynadie Marc , Hermouet Sylvie
Published: April 2020

Objective: To analyse the effect of hydroxyurea (HU) on the JAK2-V617F allelic ratio (%JAK2-V617F) of patients with Polycythaemia Vera (PV) and Essential Thrombocythaemia (ET).

Methods: Thirty-six patients were examined sequentially prior to and after on-set of (HU) therapy (8 PV, 17 ET), or while remaining untreated (2 PV, 9 ET). For all patients, the %JAK2-V617F was determined in purified blood granulocytes using sensitive allele-specific, quantitative PCRs. In a second study, two distinct groups of patients were examined at a single time point at the time of diagnosis (99 PV, 178 ET) or while receiving HU (36 PV, 98 ET).

Results: HU therapy (median duration: 15 months) reduced the %JAK2V617F by >30% in 13/25 patients (4 PV, 9 ET). For 3 patients, JAK2V617F remained undetectable for 3–27 months. There was no significant change in %JAK2-V617F for the 11 patients who received no treatment. In the single time point study of two large cohorts, the mean %JAK2-V617F was lower for patients with HU therapy (median duration: 32 months) than for patients at diagnosis (24% vs. 33%, p<0.01). However, when analysed by gender, the decrease in %JAK2-V617F was significant only in female patients; this was true for both PV (39% vs. 52%, p=0.0249) and ET (12% vs. 17%, p=0.0364).

Conclusion: Reduction and even the disappearance of the JAK2-V617F-mutated clone can be obtained with HU therapy in PV and ET patients, most likely within the first two years of treatment.
A Retrospective Literature Review of Erythromelalgia and Myelorpoliferative Disorders
Authors: Milani Cannon , Pascual Sheila , Colvin Gerald A.
Published: April 2020

Cutaneous manifestations of myeloproliferative disorders (MPD) herald an uncommon diagnostic clue preceding a diagnosis of Essential Thrombocytosis (ET) and Polycythemia Vera (PV). If left untreated, they may progress towards acrocyanosis and even peripheral gangrene. This clincal entity can present months to years prior to an official diagnosis of a MPD. These atypical skin finding in ET and PV are referred to as Erythromelalgia, characterized by red, congested distal extremities secondary to microvascular complications. The pathophysiology of this skin abnormality involves spontaneous activation and aggregation of hypersensitive thrombocythaemic platelets in the end-arterial microvasculature, involving the peripheral, cerebral, and coronary circulation of thrombocythemia patients. The microvascular disturbances are frequently preceded or followed by atypical transient neurologic, ocular or coronary ischemic events. Thus these atypical skin findings can signal potential limb ischemia and necrosis.

We performed an extensive retrospective case review of the entire Engish language medical literature. The first reported case of Erythromelalgia was in 1872 by Weir-Mitchell. By 1938, Smith and Allen advocated the use of acteylsalicyclic acid as they reported pronounced pain relief from the use of 0.65 grams, up to five times daily. In our search, the number of cases reporting signs and symptoms of Erythromelalgia specifically preceding ET or PV, number less than twenty. We wanted to determine from review of the literatue if treatment with aspirin at the time of diagnosis of Erythomelalgia led to improved outcome.

The overriding conclusion of our review was that aspirin induced prompt relief of painful symptoms and seemed to prevent thrombotic sequelae in this patient cohort. Clinicians need to consider the underlying presence of a myeloproliferative disorder in patients manifesting vasculopathy associated skin findings, as it may be the harbinger of potentially fatal ischemic events. Earlier diagnosis and treatment may lead to improved outcome.
ALK Activates a Non-Canonical Beta-catenin Pathway in Anaplastic Large Cell Lymphoma
Authors: Ysebaert Loic , Lamant Laurence , Espinos Estelle , Giuriato Sylvie , Delsol Georges , Laurent Guy
Published: April 2020

Molecular profiling of Anaplastic Lymphoma Kinase (ALK)-positive versus negative cell lines and patients’ samples has unravelled critical roles for transcriptional factors (TF) (C/EBPbeta, Bcl-6) in Anaplastic Large Cell Lymphomas (ALCL), suggesting that targeting these TF with ALK tyrosine kinase inhibitors should yield better results (Lamant L et al., Blood 2008). In this latter work of our group, beta-catenin and its transcriptional partner T Cell Factor 4 (TCF4) were also found upregulated at a transcriptional level in ALK+ vs ALK− patients’ samples and cell lines. We sought to determine the relevance of such a finding. At the protein level, ALK+ cell lines (SU-DHL-1, Karpas299 and COST) and ALK− cell line (FEPD) express detectable beta-catenin. TCF-4 is also expressed at similar levels among cell lines, whatever ALK status. Since beta-catenin is mainly regulated through post-transcriptional mechanism, we assessed its phosphorylation status. ALK+ (SU-DHL-1 and Karpas299), but not ALK− cell lines displayed increased tyrosine phosphorylation at both Tyr142 and Tyr654 residues, as well as association between beta-catenin and ALK (in immuno-precipitation assays). Moreover, both total beta-catenin and phosphoTyr-beta-catenin were found associated with TCF4, and therefore transcriptionally active. Using the tetracycline system to allow conditional expression of NPM-ALK in MEF cell line (murine embryonic fibroblasts), we found that beta-catenin phosphorylation became barely detectable upon loss of NPM-ALK expression. Based on these findings, we investigated the functional consequences of the disruption of beta-catenin/TCF4 complexes using small molecules such as PKF115–584 and CGP049090 (a generous gift from Novartis). Interestingly, both compounds induced dissociation of beta-catenin/TCF4 complexes at 0.5μM for 24h, and also induced apoptosis in SU-DHL-1 cells. But, since compounds could not dissociate beta-catenin/ALK complexes, they should be combined to ALK inhibitors to fully exert their anti-lymphoma effects. Moreover, the pool of beta-catenin linked to Glycogene Synthase Kinase 3beta (regulated by external Wnt-dependant signals) is neither affected by small compounds, indicating a specific beta-catenin/TCF4 disruption with these drugs. To conclude, this study shows that, in ALCL, ALK activates a Wnt-independent pathway, which appears to be critical for cell survival. This study offers a rationale for investigating the potential of molecules designed to interfere with beta-catenin/TCF/LEF proteins and currently evaluated in colon carcinomas, alone or in combination with ALK tyrosine kinase inhibitors.
Clinical Implications of Immunophenotyping in Staging Diffuse Large B-Cell Lymphoma
Authors: Talaulikar Dipti , Dahlstrom Jane E , Shadbolt Bruce , McDonald Anne
Published: April 2020

Diffuse Large B-cell Lymphoma (DLBCL) is the commonest subtype of Non-Hodgkin Lymphoma (NHL). Staging bone marrow (BM) biopsies in NHL are conventionally examined using histology alone, although the use of immunophenotyping to aid diagnosis is increasing. This study addresses the clinical impact of routine use of flow cytometry (FL) and immunohistochemistry (IHC) in 156 histologically proven DLBCL cases at initial diagnosis. The median age of the patient cohort was 62 years (range 20–86 years) and the gender ratio was 1.53 in favor of males. Nine patients were treated with palliative intent and were excluded from all survival analyses. The rest were treated with anthracycline based chemotherapy regimens; 36 patients received concomitant Rituximab. Median overall survival (OS) was 6.1 years (95% CI: 3.8, 8.4). Histological involvement was noted in 30 cases (19.2 %) using standardized Cheson criteria [Cheson BD, Horning SJ, Coiffier B et al. J Clin Oncol1999; 17:1244]. FL and IHC (using T-cell, B-cell, and light chain markers) detected an additional 17 cases (10.9%) each with 4 cases detected by both methods. Thirty (19.2 %) patients were upstaged to stage IV with the use of these investigations (stage I: 6, stage II: 12, stage III: 12). Kaplan Meier curves demonstrated that positivity on FL, IHC and both (FL + IHC) resulted in a significantly worse OS and progression free survival (PFS) compared to negative cases. Cox proportional hazards models were used to determine the additional benefit of FL and IHC in predicting OS over histological involvement, after adjusting for age, performance status and use of Rituximab. The first analysis compared FL with histology and showed that FL added significant independent prognostic value [Histology: HR = 2.1, 95% CI 1.0, 4.3, p=0.05; FL: HR = 2.0, 95% CI 1.0, 3.8, p=0.04]. The second analysis compared histology with IHC and showed that the two had co linearity in their prediction of survival with IHC adding significantly more than histology alone [Histology: HR = 1.3, 95% CI 0.5, 3.0, p=0.6; IHC: HR = 2.3, 95% CI 1.1, 5.0, p = 0.03]. In the final model, we examined the interactive effect of FL and IHC over and above histological diagnosis and found the interaction to be the stronger predictor of OS [Histology: HR = 1.8, 95% CI 0.8, 3.7, p =0.1; FL/IHC: HR = 2.7, 95% CI 1.2, 6.2, p=0.02]. In conclusion, our results show that FL and IHC performed routinely can improve detection of BM involvement in DLBCL and upstage ~20 % patients to stage IV disease. We found that FL added significant independent prognostic value while IHC showed greater co linearity with routine histological diagnosis. The interactive effect of FL/IHC had additional benefit over routine histology in predicting survival. These results suggest that staging BM in DLBCL should have FL and IHC performed routinely at initial diagnosis.
Non-Hodgkin Lymphoma and Smoking: A Meta-Analysis of Case-Control Studies
Authors: Milani Cannon , Pascual Sheila , Mitri Joanna , Castillo Jorge
Published: April 2020

Background: The incidence of non-Hodgkins lymphoma (NHL) has followed a sustained increasing pattern for the last few decades. Potential factors implicated in developing an increased risk for NHL include immunosuppression, bacterial, and viral infections. The pathological heterogeneity of NHL suggests multifactorial etiologies. Several reports around the world have linked lifestyle habits to the development of NHL, but thus far, the relationship between smoking and NHL has been largely inconclusive.

Objective: To perform a meta-analysis of published case-control studies to clarify the potential relationship between smoking and the development of NHL.

Methods: An English literature search was conducted using Pubmed of case-control studies investigating the relationship between smoking and development of NHL. Prospective studies, case reports, editorials and letters were not included. Data were gathered including the following variables: country of origin, source of cases and controls, number of female/male cases and controls, adjusted odds ratio (OR) with 95% confidence intervals (CI) for former and current smokers, divided by male/female when available, and adjustments. Effect measures were obtained trough fixed and random-effects models to assess for heterogeneity.

Results: Eighteen case-control studies were included in the meta-analysis. A total of 22,226 cases and 30,792 controls were finally analyzed. Based on our meta-analysis, former smokers had an OR of 1.02 (95% CI, 0.92 – 1.12) when compared to never smokers; heterogeneity between studies was evident (p < 0.001; I2 = 75.3%). Current smokers, on the other hand, had an OR of 1.06 (95% CI, 1.00 – 1.14); some degree of heterogeneity was observed (p = 0.04; I2 = 41.1%).

Conclusions: Despite the limitations of this meta-analysis, current smokers seem to have small but statistically significant increase in the risk of developing NHL. Clinically speaking, smoking may be a non-negligible risk factor for developing NHL. Although a causal relationship will be hard to prove, further research is needed to clarify the potential role of smoking in lymphomagenesis.
Clinical Outcome of 119 Children with Non-Hodgkin Lymphoma Treated in a Single Center in China
Authors: Zhang Yonghong , Jin Ling , Yang Jing , Duan Yanlong , Zhou Chunjv , Gao Zifen
Published: April 2020

One hundred and nineteen children with non-Hodgkin lymphoma were treated between February 2003 and December 2006 in Beijing Children’s Hospital on BCH-2003-NHL protocol. The diagnosis was made by histopathology of the biopsied tissue and/or bone marrow, and disease was classified according to WHO-2001 pathologic classification. We applied modified LMB89 protocol to cases with B-cell lymphoma; modified BFM90-ALL protocol for lymphoblastic lymphoma and cutaneous T-cell/NK cell lymphoma; and modified BFM90-ALCL protocol for anaplastic large-cell lymphoma (ALCL). There were 50 cases (42%) of B cell lymphoma including 32 cases of Burkitt¡’s lymphoma, 10 cases of Burkitt-like lymphoma and 8 cases of diffuse large B cell lymphoma; 44 cases (37%) of lymphoblastic lymphoma; 19 cases (16%) of ALCL; and 6 cases (5%) of cutaneous T-cell/NK cell lymphoma. The 85 boys and 34 girls (ratio, 2.5:1) ranged in age from 2 to 15 years (median, 7.8 years) at diagnosis. B cell lymphoma typically presented as abdomen mass and acute abdomen; nasopharynx and tonsil were also common sites of involvement. Lymphoblastic lymphoma generally presented with mediastinal mass and bone marrow involvement. There was no typical presentation for ALCL. According to the St. Jude staging system, 19 cases had stage I–II, and 94 cases stage III–VI diseases (exclude 6 cases of cutaneous T-cell/NK cell lymphoma). Seven cases had CNS involvement and 25 cases involved bone marrow. The treatment duration was 2 to 8 months for B-cell lymphoma, 2.5 to 3 years for lymphoblastic lymphoma and 1 to 1.5 years for ALCL. The follow-up rate was 100% and median observation period was 23 months. The overall survival (OS) at 3 years was 90.7% and the 3-year event-free survival (EFS) estimate was 82.3%. For B-cell lymphoma, 3-year OS was 88.68% and 3-year EFS was 81.8%. For lymphoblastoma lymphoma, the rates were 89.3% and 69.4%, respectively. All cases of ALCL are alive with on undergoing treatment for relapse. Patients with ALCL achieved the best 3-year OS (100%) and had 3-year EFS of 94.2%. Grade 3 or 4 bone marrow suppression occurred in 97.5% of patients with B-cell lymphoma, 100% of those with lymphoblastic lymphoma and 89.5% of cases with ALCL. As of to date, 11 patients have died, the causes of death include infection (n=4), abandonment of therapy (n=6) and relapse (n=1). Univarate analysis showed that stage IV disease, failure to achieve complete remission after 3 months of treatment, and bulky mass are were associated with poor prognosis £all P values <0.05£©. In summary, we have achieved excellent treatment results using modified international protocols. Infection and financial problem remained the main reasons of treatment failure.
Abcb10 Physically Interacts with Mitoferrin1 to Enhance Its Stability for Heme Synthesis in the Erythroid Mitochondria
Authors: Chen Wen , Li Liangtao , Langer Nathaniel B. , Paradkar Prasad N. , Schultz Iman J. , Hyde Brigham B. , Shirihai Orian S. , Kaplan Jerry , Paw Barry H.
Published: April 2020

We previously described a zebrafish mutant, frascati (frs), with hypochromic anemia and erythroid maturation arrest. Using positional cloning, we identified mitoferrin1 (mfrn1, slc25a37) as the gene disrupted in frs mutants (Shaw GC, et al. 2006 Nature 440:96–100). Mfrn1, the principle iron-importer in the mitochondria, is essential for heme and iron-sulphur (Fe/S) cluster syntheses in erythroblasts, and is required for primitive and definitive erythropoiesis. The biochemistry for Mfrn1-mediated iron acquisition into the mitochondria of developing erythroblasts, however, is poorly understood. In order to identify Mfrn1-associated proteins involved in mitochondrial iron homeostasis, we employed the strategy of in vivo epitope-tagged affinity purification and mass spectrometry (MS). A “bait protein,” Biotag-mouse Mfrn1 (BT-Mfrn1), was engineered to affinity-purify the associated proteins. A series of control experiments were first conducted to demonstrate that the BT-Mfrn1 protein properly targeted to the mitochondria and functionally complemented the anemia in frs embryos. A two-dimensional Blue Native gel followed by SDS-PAGE and western blot showed Mfrn1 forms higher-order protein complexes with interacting proteins in the mitochondrial fraction. A stable mouse erythroleukemia (MEL) clone expressing BT-Mfrn1 protein was derived as a source for protein purification. Affinity-purified BT-Mfrn1 protein complexes were analyzed by MS in triplicate runs in comparison to control MEL cells. Abcb10, a GATA-1 induced ATP-binding cassette transporter highly expressed in hematopoietic tissues and involved in heme biosynthesis (Shirihai OS, et al. 2000EMBO J.19:2492–2502), was found as one of the proteins which physically associated with Mfrn1 during MEL differentiation. The Abcb10:Mfrn1 interaction was confirmed by immunoprecipitation (IP) and western analysis with endogenously expressed proteins in MEL cells and proteins expressed by transient transfection in heterologous cells. Mfrn1 was previously shown to have longer protein half-life in differentiated MEL cells compared to undifferentiated cells (Paradkar PN, et al. submitted). To test our hypothesis that the Abcb10:Mfrn1 interaction may enhance Mfrn1 protein stability, a pulse-chase assay using 35S-methionine labeled cells was performed, followed by IP. Abcb10 was found to stabilize Mfrn1 protein in COS7 cells co-transfected with Abcb10 and Mfrn1 compared to control cells transfected with Mfrn1 alone. Similar results were obtained when total steady-state Mfrn1 protein was evaluated by western analysis comparing lysates from Abcb10:Mfrn1 cotransfected and Mfrn1 transfected cells. Our data suggest that Abcb10 physically interacts with Mfrn1 to enhance its stability and promote Mfrn1-dependent mitochondrial heme biogenesis.
Meningeal Dissemination (MD) in Primary CNS Lymphoma (PCNSL): Multimodal Diagnosis, Presentation and Outcome in a Large Monocenter Patient Cohort
Authors: Kiewe Philipp , Korfel Agnieszka , Fischer Lars , Martus Peter , Thiel Eckhard
Published: April 2020

Introduction: The frequency of meningeal dissemination (MD) in primary central nervous system lymphoma (PCNSL) reported in the literature differs. The optimal management and the prognostic impact of MD have not been defined.

Methods: Data on MD from all 92 immunocompetent patients primarily diagnosed with PCNSL at our institution between January 1994 and February 2007 were retrospectively analysed for MD evaluation. All patients received cranial MRI or CT, and in 71 patients CSF was obtained by lumbar puncture. High-dose methotrexate was the intended treatment in all patients.

Results: Cerebrospinal fluid (CSF) was obtained by lumbar puncture in 71 patients (median age 62 years) of a total of 92 patients with PCNSL. Cytomorphologic CSF examination was performed in 63, immunophenotyping in 32 and PCR of the CDR3 region in 37 patients. Eight of 63 patients (13%) had positive cytomorphology, one of 32 (3%) positive immunocytology and 6 of 37 (16%) had monoclonal B-cells on PCR analysis. Evidence of MD on MRI or CT was found in three of the 71 patients (4%). Altogether, 11 (16%) patients had evidence of MD diagnosed by either of the methods used. Median CSF cell count was 5/μl (range, 0–237; n=66), and median protein concentration 76 mg/dl (range, 93–4117; n=55). An elevated cell count (>5/μl) was found in 27 (41%) patients, an elevated protein (>45 mg/dl) in 44 (80%). No significant correlation was found between proof of MD and CSF pleocytosis or elevated protein. Patients with MD did not differ from other patients with respect to age, gender, clinical presentation, performance status, CSF cell count and protein concentration, number of intracerebral lesions, lesion location or histology. After a median follow-up of 56 months, median EFS was 26 (95% CI, 4.3 to 47.7) months for MD patients and 34.1 (95% CI, 26.2 to 42) months for those without MD, p=0.26 (log-rank). Of 21 patients with relapse after initial treatment response, 16 had isolated cerebral relapse, one isolated meningeal relapse, two systemic relapse (one with cutaneous, and one with testicular manifestation), and one concomitant systemic (lung/thoracic wall) and cerebral relapse. Of the four MD patients with relapse after initial response, three had cerebral relapse, and one had systemic (cutaneous) relapse. The median overall survival (OAS) of patients with MD was 45.5 months (95% CI, 16.6 to 74.4), of those without MD 42.5 months (95% CI, 35.8 to 49.2) (p=0.27).

Conclusions: The frequency of MD detected in our cohort was low despite the use of highly sensitive diagnostic methods. No impact on either EFS or OAS was seen for MD.
CD3-Negative CD4+ T-Cells: A Useful Diagnostic Tool with High Specificity in Angioimmunoblastic Lymphadenopathy (AILD)-Type T-Cell Lymphoma
Authors: Singh Anju , Flörcken Anne , van Lessen Antje , Dörken Bernd , Pezzutto Antonio , Westermann Jörg
Published: April 2020

Angioimmunoblastic lymphadenopathy (AILD)-type T-cell lymphoma is one of the common T cell lymphomas in Western countries. Many patients present with symptoms of a systemic disease and diagnosis can often be challenging. Particularly in cases in which histological confirmation cannot be easily achieved flowcytometry of peripheral blood can give important clues for the differential diagnosis of AILD. We have previously reported that CD4-negative CD3+ T-cells in peripheral blood are a characteristic immunophenotypic finding in AILD patients. Gene scan analysis for the TCR gamma chain and intracellular CD3 expression clearly showed that these CD3-negative CD4+ cells are of T cell origin. In this study we further evaluated the phenotype of the CD4− CD3+ T cells and we compared their frequency in AILD patients with the frequency in patients with other histologically confirmed leukemic T cell lymphomas. 14 patients with AILD (clinical stage III/IV) were compared with 26 patients with other subtypes of leukemic T cell lymphomas (6 T-large granular lymphocyte leukemias (T-LGL), 4 T-chronic lymphocytic leukemias/prolymphocytic leukemias (T-CLL/PLL), 5 anaplastic large cell lymphomas (ALCL), 1 adult T-cell leukemia/lymphoma (ATLL), 1 mycosis fungoides and 9 peripheral T-cell lymphomas. Flowcytometric analysis of peripheral blood was performed in a lymphocyte gate using fluorochrome-labeled antibodies against CD3, CD2, CD4, CD5, CD7, CD8, CD16, CD56, CD57 and TCR. In 14/14 AILD patients a small but distinct population of CD3-negative CD4+ T cells was found (mean percentage of CD3-negative CD4+ T cells in the gate: 12,0 ± 17,6 %, range 0,05 – 51,8 %). In contrast, CD3-negative CD4+ T cells could be detected in only 1/26 patients with other leukemic T cell lymphomas. This patient had been diagnosed with mycosis fungoides. Further immunophenotypic analysis showed that the aberrant T cells in AILD also express pan T cell markers, such as CD2, CD5 and partially CD7, surface TCR expression could not be detected. In conclusion our comparative study shows that flowcytometric detection of CD3-negative CD4+ T cells in peripheral blood is a characteristic feature of AILD with a high specificity. Therefore, flowcytometry is particulary useful in the differential diagnosis of AILD, even if the aberrant T cell population has a very low frequency. Although AILD is mostly CD3+ on immunohistochemistry, the presence of low numbers of CD3-negative CD4+ T cells in peripheral blood appears to be a typical feature of AILD. Cytoplasmic CD3-expression is one possible explanation for this discrepancy. Since complete or partial loss of pan T cell antigens is a characteristic feature of many T cell lymphomas, the aberrant CD3-negative CD4+ T cells might also represent just a subfraction of lymphoma cells in AILD. Further biological characterization of this subpopulation should be performed.
Cholesterol Rich-Domains Regulate VEGFR-1 (FLT-1) Expression and Signaling in Acute Leukemia Cells
Authors: Casalou Cristina , Gomes Ana , Carvalho Tania , Dias Sergio
Published: April 2020

Vascular endothelial growth factor receptors -1 (FLT-1) and -2 (KDR) are expressed by subsets of acute and chronic leukemias, where they signal in paracrine and/or autocrine manner to induce cell survival, proliferation and migration. We have previously shown that acute lymphocytic leukemia migration in response to VEGF via FLT-1 modulates the onset of extramedullary disease, and thus has clinical predictive value (Fragoso et al, Blood 2006). Acute leukemia cell (AML) migration, induced by PlGF/VEGF activation of FLT-1 results in the formation of actin membrane protrusions with concomitant increased ERK1/2 and P38 phosphorylation and activation of Rho-GTPases (Casalou et al, 2007). Since we have found an in vitro association of FLT-1 with caveolin-1, actin and HSp90, we hypothesised that cholesterol-rich domains might regulate FLT-1 mediated survival, proliferation or migration of acute myeloid (AML) and lymphoid (ALL) leukemias. First we found by FACS and RQ-PCR that FLT-1 expression is up-regulated by increased cholesterol/HDL levels in vitro. As shown by sucrose gradient fractionation and western blotting, PlGF/VEGF stimulation of AML cells results in re-localization of FLT-1 to cholesterol-rich domains. Accordingly, FLT-1 localization within cholesterol-rich domains is abrogated by exposing leukemia cells to b-methyl-cyclodextrin (MbCD) which removes intracellular cholesterol. Additionally, FLT-1 phosphorylation is abolished by treatment of AML cells with MbCD or Nystatin, an inhibitor of lipid raft endocytosis. Functionally, AML cells exposure to high levels of total cholesterol/HDL for 24 hours exerted a protective effect from actinomycin D-induced apoptosis and promoted PlGF/VEGF-induced AML migration in transwell migration assays. Together, these results show that on subsets of acute leukemias cholesterol/HDL cellular-content regulates FLT-1 expression and signalling, resulting in decreased apoptosis and induction of cell migration. In vivo, we show that cholesterol-rich diet significantly increases bone marrow VEGF levels in mice; inoculation of FLT-1 expressing acute leukemias into mice fed with cholesterolrich diet significantly accelerated disease progression and worsened disease outcome. Taken together, our data show the molecular basis by which cellular and systemic cholesterol regulates VEGF and VEGFR-1 signalling on subsets of acute leukemias, modulating cell migration and survival and thereby regulating disease progression.
The Transmembrane Domain of PDGFR-β Plays An Important Role in ETV6-PDGFR-β Activation
Authors: Toffalini Federica , Demoulin Jean-Baptiste
Published: April 2020

The ETV6-PDGFRβ hybrid protein (EPβ) is found in a subset of patients with chronic myelomonocytic leukemia (CMML) associated with eosinophilia. EPβ is the archetype of a larger group of hybrid receptors that are produced by chromosomal translocations of PDGFR genes and cause atypical myeloproliferative neoplasms. In EPβ, the N-terminal portion of the ETV6 transcription factor replaces the PDGFRβ ligand-binding domain and induces the activation of the chimera through its pointed domain (PNT domain, also called helix-loop-helix or SAM domain). This domain mediates ligand-independent EPβ oligomerization, resulting in constitutive tyrosine kinase activity. In addition, we showed that EPβ and other hybrid receptor tyrosine kinases are much more stable than wild-type receptors and that the deletion of the PNT domain induced EPβ protein degradation, suggesting a link between the clustering and the stabilization of the EPβ protein. The PDGFRβ transmembrane domain (TM domain) is retained in EPβ and in most PDGFRβ hybrid proteins that have been described. We observed that the deletion of the TM domain (EPβ-ΔTM mutant) strongly impaired the ability of EPβ to sustain growth factor-independent proliferation of Ba/F3 and 32D cells. The phosphorylation of the mutant protein was also markedly reduced. We confirmed that EPβ is not inserted in membranes but resides in the cytosol, indicating that the PDGFRβ TM domain does not act as a transmembrane domain in EPβ but has a completely different function. The EPβ-ΔTM mutant retained the ability to self-associate in co-immunoprecipitation experiments, but showed a decreased level of polymerization when using cross-linking agents, suggesting that this domain is required for optimal clustering of EPβ. In line with our findings on the PNT domain, the EPβ-ΔTM protein was less stable and, as a result, was expressed at a lower level. In conclusion, we demonstrate that the TM domain plays a role in EPβ activation by promoting the clustering of the protein and by preventing its degradation in cooperation with the PNT domain.