Laura Percy

APRIL promotes cell cycle progression in primary multiple myeloma cells: influence of D-type cyclin group and translocation status

A proliferation-inducing ligand (APRIL) promotes survival and drug resistance in multiple myeloma (MM) cell lines. We studied the effect of APRIL on cell-cycle behavior in primary MM cells and correlated our findings with D-type cyclin expression by immunohistochemistry and/or Western blotting. In MM cases, expressing cyclin D2 APRIL significantly increased the percentage of CD138(+) cells in S + G(2)/M phase (from 8.4% ± 1.9% to 14.3% ± 2.6%, n = 15, P < .01), whereas a lesser effect was seen in cases expressing cyclin D1 (n = 18). Cell-cycle response to APRIL was most marked for cyclin D2-expressing cases with IgH translocations (P < .01) and was accompanied by increased expression of cyclin D2, CDK4, CDK6, and phospho-retinoblastoma protein. Cell-cycle proteins in cyclin D1(+) cells were not modulated by APRIL. Surface expression of B-cell maturation antigen and transmembrane activator and calcium-modulating cyclophilin ligand interactor was not significantly different between cyclin D1(+) and D2(+) MM cells. We observed activation of nuclear factor-κB and PI3-kinase pathways in response to APRIL in both cyclin D1(+) and D2(+) MM cells. In conclusion, APRIL stimulates G(1)/S progression in cyclin D2(+) MM cells bearing IgH translocations but has minimal effect on cyclin D1(+) cells, suggesting MM cells from different cyclin D/translocation classes rely on different mechanisms for cell-cycle re-entry.

CD20-positive multiple myeloma - differential expression of cyclins D1 and D2 suggests a heterogeneous disease

CD20 expression is found in approximately 15–20% of multiple myeloma (MM) patients, although the significance of such expression remains unclear (Kapoor et al, 2008). Overexpression of one of cyclins D1, D2 or D3 is ubiquitous in MM and several studies have shown an association between CD20 expression, cyclin D1 expression, and the t(11;14) translocation in particular. In one study, based on flowcytometry of bone marrow aspirates, the t(11;14) translocation was found in 83% of CD20+ cases (10/12), a finding confirmed in a larger study in 2005 (Robillard et al, 2003; Mateo et al, 2005). Furthermore, immunohistochemical studies of MM bone marrow biopsies (Ely et al, 2005) and gene expression profiling experiments (Zhan et al, 2006) have also found an association between cyclin D1, t(11;14) and CD20 expression. Although an early study noted a poorer outcome for CD20+ MM cases (San Miguel et al, 1991), the t(11;14) translocation has subsequently been shown to be associated with a better prognosis (Moreau et al, 2002), thus the prognostic significance of CD20 expression in MM is unclear. In contrast to cyclin D1 expressing CD20+ MM cases, little is known about the biological or clinical behaviour of CD20+ MM cases expressing cyclin D2. Therefore, we aimed to characterise CD20+ MM cases associated with cyclin D2 expression by immunohistochemistry. MM trephine biopsies where CD20 immunohistochemistry was performed were retrieved from departmental records and immunohistochemical findings were correlated with clinical data. CD20, cyclin D1 and cyclin D2 expression were determined by immunohistochemical staining of MM bone marrow biopsies. Samples were deemed to be positive for CD20 if >20% CD138+ cells in a biopsy had detectable expression of CD20. Intensity of CD20 staining and spatial

Weekly intravenous bortezomib is effective and well tolerated in relapsed/refractory myeloma.

Bortezomib is an effective antimyeloma therapy, but clinical benefits can be limited by neurotoxicity. In newly diagnosed, older patients, modification of the biweekly dosing schedule to weekly regimens improves tolerability whilst maintaining efficacy. There is less information on the efficacy and tolerability of weekly bortezomib regimens in the relapsed/refractory setting. Here, we report our experience of weekly intravenous bortezomib in clinical practice in relapsed/refractory patients.

RAS mutation status and bortezomib therapy for relapsed multiple myeloma.

A.M. Schut participated in the design of the present study, helped to coordinate the study, performed experiments, analysed and interpreted data and wrote the manuscript. A. Venemans-Jellema performed data analysis, interpreted data and revised the manuscript. J. C. M. Meijers supervised some of the experiments, interpreted data and revised the manuscript. S. Middeldorp and F.R. Rosendaal participated in the study design, interpreted data and revised the manuscript. P. G. de Groot and M. Roest supervised some of the experiments, interpreted data and revised the manuscript. T. Lisman participated in the design and supervision of the present study, interpreted data and wrote the report. S.C. Cannegieter participated in study design, supervised data analysis, interpreted data and revised the manuscript.

The bone marrow stromal compartment in multiple myeloma patients retains capability for osteogenic differentiation in vitro: defining the stromal defect in myeloma

Defects in bone repair contribute to multiple myeloma (MM) bone disease. It is unknown whether this reflects failure of osteogenic differentiation from mesenchymal stromal cells (MSC), inherent stromal defects or mature cell dysfunction. We quantified the number of fibroblast colony‐forming units (CFU‐f) and osteoblast colony‐forming units (CFU‐ob) in freshly isolated bone marrow (BM) from healthy individuals (N = 10) and MM patients (N = 54). CFU‐f and CFU‐ob were present in MM BM, at comparable frequency to normal subjects, irrespective of disease stage, and the presence of bone disease. Adherent cultures from MM BM are able to differentiate into osteoblasts, as indicated by the early upregulation of RUNX2, SP7, AXIN2 and DLX5, and the production of alkaline phosphatase and calcium. Coculture with MM cells failed to prevent osteogenic differentiation of adult human MSC. On the other hand, MM cells induced cell cycle progression in resting MSC in a cell contact dependent manner. This effect was confirmed using both primary CD138+ cells and MM cell lines, and was not seen with B or T cell lines. Our data confirm the presence of osteoblast progenitors and the preservation of osteogenic function in MM, however dysregulation of cell cycle control may contribute to the loss of normal bone homeostasis that ultimately results in osteolytic bone loss.

Cohort analysis of FISH testing of CD138(+) cells in relapsed multiple myeloma: implications for prognosis and choice of therapy.

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Improved response with post-ASCT consolidation by low dose thalidomide, cyclophosphamide and dexamethasone as first line treatment for multiple myeloma.

The use of consolidation or maintenance to improve disease response, and hence clinical outcome, following autologous stem cell transplantation (ASCT) remains the subject of intense clinical research. We carried out a single‐arm study to assess the toxicity and efficacy of a short block of consolidation therapy with cyclophosphamide, low dose thalidomide and dexamethasone (CTD) in patients within 6 months following ASCT, as part of frontline therapy for symptomatic multiple myeloma. Forty‐five patients who had not progressed were enrolled on the study, and 43 completed treatment on protocol. This regimen was well tolerated soon after ASCT, with only grade 1/2 toxicity apart from neutropenia, and no long‐term sequelae. Importantly, CTD consolidation improved the depth of response in treated patients, increasing the complete/very good partial response rate from 44% at 3 months, to 72% at 12 months, which was significantly higher compared with a historical group of control patients (P = 0·002). There was a trend to longer progression‐free survival that favoured the study group. Consolidation therapy did not adversely affect subsequent disease response to salvage therapies at relapse. We conclude that CTD consolidation may be a useful, non‐toxic and cost‐effective strategy to deepen disease response following ASCT, and deserves further study in a randomized trial.

A new prognostic model for myeloma patients relapsing from upfront autologous transplantation based on ISS and PFS1

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Re-infused lymphocyte dose does not influence disease control following upfront autologous stem cell transplantation for multiple myeloma.

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