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Human natural killer (NK) cell development is a step-by-step process characterized by phenotypically identified stages. CD161 is a marker informative of the NK cell lineage commitment, whereas CD56, CD117, and CD94/NKG2A contribute to define discrete differentiation stages. In cells undergoing in vitro differentiation from CD34+ umbilical cord blood (UCB) progenitors, LFA-1 expression allowed to discriminate between immature noncytolytic CD161+CD56+LFA-1− and more differentiated cytolytic CD161+CD56+LFA-1+ NK cells. CD161+CD56+LFA-1− NK cells produce large amounts of CXCL8 after phorbol myristate acetate (PMA) or cytokine treatment. Remarkably, CXCL8 mRNA expression was also detected in fresh stage III immature NK cells isolated from tonsils and these cells expressed CXCL8 protein on PMA stimulation. Within in vitro UCB-derived CD161+CD56+LFA-1− NK cells, CXCL8 release was also induced on antibody-mediated cross-linking of NKp44 and CD161. Such unexpected activating function of CD161 was confined to the CD161+CD56+LFA-1− subset, because it did not induce cytokine release or CD107a expression in CD161+CD56+LFA-1+ cells or in mature peripheral blood NK cells. Anti-CXCL8 neutralizing antibody induced a partial inhibition of NK cell differentiation, which suggests a regulatory role of CXCL8 during early NK cell differentiation. Altogether, these data provide novel information that may offer clues to optimize NK cell maturation in hematopoietic stem cell transplantation.

In multiple myeloma (MM), the addition of thalidomide or bortezomib to the standard oral melphalan/prednisone combination significantly increased response rate and event-free survival. In this multicenter phase 1/2 trial, dosing, safety, and efficacy of the 4-drug combination, bortezomib, melphalan, prednisone, and thalidomide (VMPT) was determined. Bortezomib was administered at 3 dose levels (1.0 mg/m2, 1.3 mg/m2, or 1.6 mg/m2) on days 1, 4, 15, and 22; melphalan was given at a dose of 6 mg/m2 on days 1 through 5 and prednisone at 60 mg/m2 on days 1 through 5. Thalidomide was delivered at 50 mg on days 1 through 35. Each course was repeated every 35 days. The maximum tolerated dose of bortezomib was 1.3 mg/m2. Thirty patients with relapsed or refractory MM were enrolled; 20 patients (67%) achieved a partial response (PR) including 13 patients (43%) who achieved at least a very good PR. Among 14 patients who received VMPT as second-line treatment, the PR rate was 79% and the immunofixation-negative complete response rate 36%. The 1-year progression-free survival was 61%, and the 1-year survival from study entry was 84%. Grade 3 nonhematologic adverse events included infections (5 patients), fatigue (1), vasculitis (1), and peripheral neuropathy (2); no grade 4 toxicities were recorded. Initial results showed that VMPT is an effective salvage therapy with a very high proportion of responses. The incidence of neurotoxicities was unexpectedly low.

The expression of a cellular receptor for the blood-clotting protease factor Xa, designated effector cell protease receptor-1 (EPR-1), was investigated in lymphoma. Immunohistochemical analysis demonstrated prominent reactivity of monoclonal antibodies to EPR-1 with Reed-Sternberg cells in 30 of 35 cases of nodular-sclerosis, lymphocyte-depletion, and mixed-cellularity Hodgkin's disease (HD). In contrast, several non-Hodgkin's lymphomas, or the nonneoplastic cellular components of HD, did not react with anti-EPR-1 monoclonal antibodies. A single molecular species of approximately 62 kD, consistent with the size and structural organization of EPR-1, was immunoblotted by an anti-EPR-1 monoclonal antibody from tissue samples of HD, but not from normal lymph nodes. Expression of EPR-1 transcripts in Reed-Sternberg cells was demonstrated by in situ hybridization with an antisense EPR-1 riboprobe, and by amplification of reverse-transcribed HD RNA with EPR-1-specific primers. These findings identify the factor Xa receptor, EPR-1, as a novel marker of Reed-Sternberg cells, and suggest its potential role in the histopathogenesis of HD.

Abstract 3879 Purpose. The use of early (interim) positron emission tomography (PET) restaging during front-line therapy in Hodgkin's lymphoma (HL) has considerably increased in clinical practice as an early recognition of treatment failure allows patients to be addressed to more intensive treatment regimens. Patients and Methods. Between June 1997 and June 2009, 304 newly-diagnosed Hodgkin's lymphoma patients (147 early-stage and 157 advanced-stage) were treated with the ABVD regimen at two Italian institutions. Patients underwent to a PET staging and restaging at baseline, after 2 cycles of therapy and at the end of the treatment. Results. 53 patients showed a positive interim PET and only 13/53 (24.5%) achieved a complete response (CR), whereas 251 patients showed a negative PET and 231/251 (92%) remained in CR. Comparison between interim PET-positive and interim PET-negative patients indicated a significant association between PET findings and 9-year progression-free survival (p=0.0000) and 9-year overall survival (p=0.0000), with a median follow-up of 31 months. Among the early-stage patients, 19 had a positive interim PET and only 4 (21%) achieved a CR; among the 128 negative interim PET patients, 122 (97.6%) obtained a CR. In the advanced-stage subset, 34 patients showed a persistently positive PET (with only 9/34, 26.4% in CR), whereas 123 showed a negative interim PET, with 109 (88.6%) remaining in CR. Conclusions. Our results confirm the role of early PET as a significant step forward for the management of both early and advanced-stage HL patients, offering the potential for an immediate switch to high-dose treatments, if required. Disclosures: No relevant conflicts of interest to declare.

Abstract 1659 Poster Board I-685 Background Hodgkin lymphoma (HL) is a curable malignancy with a long-term survival of around 80%. FDG-PET is a noninvasive imaging modality widely used in lymphoma patients. Early PET assessment of response to therapy is a routine part of management in HL patients, and an independent, strong predictor of progression-free survival. Patients and Methods 178 patients, with a diagnosis of HL, underwent to an early PET evaluation during their course of chemotherapy and were considered eligible for the study. 85 patients (48%) were male and 93 (52%) female; the median age at diagnosis was 33 (13-78) years. 6 patients (3%) had stage I disease; 106 patients (60%) stage II; 34 (19%) stage III and 32 (18%) stage IV (bone marrow involvement in 5 cases). B-symptoms were detected in 81 patients (46%). A mediastinal bulk was detected in 54 cases (30%). The majority of patients (173, 97%) underwent to ABVD as first line therapy; 5 received BEACOPP chemotherapy (3%). Early PET evaluation was performed after the second course of therapy. Results were classified into complete response (CR), partial response (PR), stable disease (SD) or progressive disease (PD) according to International Workshop standardized response criteria. PET scan was performed again at the end of the first-line treatment. 44 patients have been addressed to a second-line therapy, in presence of PR, PD or relapsing disease; in particular, 39 patients received an autologous stem-cells transplantation (ASCT), and 3 an allogeneic bone marrow transplantation (ABMT). Results At a median follow up of 41,85 (5,23-141,77) months, 152 patients are alive and in CR; 7 in PR; 3 alive with SD and 7 present a PD. 9 patients have died. 150 patients presented with a negative PET after 2 cycles, and 28 with a positive one (26 in PR, 1 with SD and 1 with PD). More specifically, of the 178 initial patients, 150 (84%) had a negative early PET and 28 (16%) a positive early PET. Of those with a negative PET, 135 (90%) experienced a continuous CR, while among those with a positive early PET, none obtained at least a stable CR. Of this unfavourable group of patients, 9 (32%) reached, and still maintain, a CR after ASCT. Conclusions Our experience indeed confirms the highly predictive value of a negative early PET during the therapy for HL. Moreover we may suggest the potential role of ASCT in inducing a CR in around one-third of those unfavorable patients with a positive early interim PET. Disclosures No relevant conflicts of interest to declare.

Background. Recently, gene expression profiling (GEP) indicated histone-deacetylases (HDAC) as potential therapeutic targets in peripheral T-cell lymphomas (PTCL) not otherwise specified (NOS), the commonest PTCL type. Consistently, phase II trials demonstrated the efficacy of some HDAC inhibitors (HDACi), including SAHA, which was approved for cutaneous T-cell lymphomas (CTCL) treatment. Aims and methods. We investigated the anti-tumour effects of ITF2357 (Italfarmaco, Italy), a novel hydroxamic acid HDACi, on PTCL primarily-cultured cells and cell lines (HH and FEDP), and in a xenografted mouse-model of CTCL. Cultured cells were incubated with different dosages of ITF2357 and SAHA (ranging from 0.5 to 2.5 mM). Cell viability, assessed by trypan-blue exclusion assay, cell-cycle progression, assessed by bromodeossiuridine assay, and apoptotic rate, determined by flow-cytometry analysis of annexin-V binding populations were determined at 48, 72 and 120 hours. Nude mice, injected with HH cells, received ITF2357 (10–20mg/Kg, per os) for 14 days. Micro-PET scan was adopted for disease measurement and treatment response evaluation. Finally, GEP of cell lines exposed to ITF2357 and SAHA were generated to elucidate their mechanisms of action. Results. Cell viability of HH cells treated with ITF2357 ranged from 50% (0.5 mM, at 48 h), to Conclusion. Taken together, these data demonstrate that ITF2357 is effective against PTCLs ex vivo and in vivo, by nominating it for clinical evaluation in this setting.

FDG-PET role in the assessment of lymphoma patients is well established but only few papers evaluated the usefulness of FDG-PET during follow up. Aim: to prospectively investigate the value of serial FDG-PET scans in the follow up of lymphoma patients in complete remission. All lymphoma patients who achieved a complete remission were prospectively enrolled in the study and scheduled for serial FDG-PET scans at 6, 12, 18 and 24 months; further scans were then carried out on annual basis (overall 421 pts, 160 pts with Hodgkin’s Disease (HD) and 261 pts with non-Hodgkin Lymphoma (NHL) were studied). All patients had a final assessment using other imaging procedures and/or biopsy and/or clinical evolution. FDG-PET findings were reported as positive, indeterminate or negative for relapse; after comparison with all available data, PET results were categorized as true positive (TP), true negative (TN), false positive (FP), indeterminate turned out to be relapse (I+) and indeterminate turned out to be complete remission (I-). Results: PET documented relapse in 42 cases at 6 mo (14 HD (8.8%) and 28 NHL (10.7%); in 31 cases at 12 mo (14 HD (9.5%) and 17 NHL (7.3%); in 27 cases at 18 mo (6 HD (4.5%) and 21 NHL (3.2%); in 9 cases at 24 mo (3 HD (2.4%) and 6 NHL (3.2%); and in 5 cases at > 36 mo (2 HD (2.8%) and 3 NHL (6.5%). Out of 125 scans reported as positive for relapse, 109 turned out to be TP (PPV of 87%); no false negative scan was recorded, and in the great majority of cases PET detected the presence of relapse before clinical evidence. Our results confirm that FDG-PET is a valid tool for lymphoma patients follow-up. The higher incidence of relapse occurred in both HD and NHL quite early after complete remission (at 6 and 12 months for HD and at 6, 12 and 18 months for NHD), thus confirming the usefulness of performing FDG-PET scans at these times in order to identify recurrence. The role of serial PET at later times (after 18 months for HD and 24 months for NHL) was found less relevant.

Osteonecrosis of the jaw (ONJ) is a frequent complication in bisphosphonate-treated multiple myeloma (MM) patients. The pathogenesis is unclear, and major risk factors are duration of bisphosphonate treatment and dental procedures. The histology of osteonecrotic bone shows osteomyelitis and inflammatory infiltrates, and, in most cases, presence of Actynomycetes. Since dental procedures are a major risk factor for ONJ development and oral microflora can be involved in the pathogenesis of the disease, we conducted a retrospective observational trial comparing ONJ occurrence and related risk factors in two groups of MM patients, who received zoledronic acid treatment at two Italian hematological centres. In one centre all patients systematically received as antibiotic prophylaxis amoxicillin-clavulanate 1 gm bid or levofloxacin 500 mg once a day starting from one day before to 3 days after any dental procedure (group A, 52 patients), while in the other centre patients did not receive any prophylaxis (group B, 61 patients). Dental procedures were categorized according to their invasivity and their supposed probability to cause ONJ. Extractions, implants, and professional cleanings were considered at high risk, while fillings were considered low risk procedures. Thirty-three group A patients (63%) and 32 group B patients (52%) received high risk procedures; 4 group A patients (8%) and 5 group B patients (8%) received low risk procedures, while 15 (29%) and 24 (39%) patients, respectively, had a denture. The duration of zoledronic acid exposure differed significantly between the two groups, with a median of 26 months for A patients and 12 months for B patients (p

Defibrotide (DF) is a novel orally bioavailable polydisperse oligonucleotide with anti-thrombotic and anti-adhesive effects, which has been shown to be active in various micrangiopathies, including the treatment and prophylaxis of veno-occlusive disease. While DF has minimal inhibitory effect on multiple myeloma (MM) in cell isolates, it showed single agent activity on human MM xenografts in SCID/NOD mice and markedly increased responsiveness of MM to cytotoxic agents, including melphalan, cyclophosphamide and dexamethasone in the same models. DF might thus enhance the response rate of Melphalan, Prednisone and Thalidomide, while protecting against the prothrombotic state seen with this combination in the treatment of MM. In this multicenter, open label, non-randomised phase I/II trial, dosing safety and efficacy of melphalan, prednisone, thalidomide, and DF (MPTD) were determined in pts with relapsed/refractory MM. Between March 2006 and July 2006 we enrolled 14 MM pts in first or second relapse. Primary refractory or pts receiving therapeutic anticoagulation were excluded. Oral melphalan was administered at 0.25 mg/Kg on D1–4, oral prednisone at 1.5 mg/kg on D 1–4, thalidomide was delivered at 50 mg on D1–35 on cycle 1 and at 100 mg from cycle 2 to cycle 6. DF dose levels were: Level + 1 DF = 17 mg/Kg i.v. or 2.4 g p.o. D1–4, followed by 1.6 g p.o. through D 35 Level + 2 DF = 34 mg/Kg i.v. or 4.8 g p.o. D 1–4, followed by 3.2 g p.o. through D 35 Level + 3 DF = 51mg/Kg i.v. or 7.2 g p.o. on D 1–4, followed by 4.8 g p.o. through D 35. Each course was repeated every 35d for a total of 6 courses and no DVT prophylaxis was used. Eleven pts received at least one MPTD-course and were evaluated. 9 pts were in 1st relapse (5 after autologous transplant, 4 conventional chemotherapy), 2 pts in 2nd relapse (autologous transplant, conventional chemotherapy, thalidomide based regimen). In the first 8 pts who received DF at Level +1 one DLT was observed: G3 ileus, which resolved. G3 or 4 hematological toxicities were reported in 7 pts (63%), which were manageable. Two pts (18%) experienced G 3 non-hematologic toxicity: 1 pt ataxia and motor neuropathy and 1 pt with G 3 ileus described above. G 1–2 non-hematological toxicities included fatigue (5 pts), neuropathy (4), infections (3), constipation (2), urinary frequency (1), hyperglycemia (1), rash (2), CVS (hypotension and bradycardia) (2) and lower limb edema (2). There were no DVTs reported, only 1 episode of G 1 bleeding and no G4 non-hematological toxicity. M-protein reduction was evaluated according to the EBMT/IBMTR criteria, after 1 and 3 cycles of MPTD see table. After 3 cycles the overall response rate was 80%. Initial results suggest that MPTD is an effective oral salvage therapy with manageable toxicity and no DVT seen to date in pts with relapsed, refractory MM. Accrual continues to define maximal tolerated dose. An update of these results will be presented. M-protein reduction % 1 cycle (n=11) 3 cycle (n=5) 75–99 9.5% 60% 50–74 27% 20% 25–50 27% Stable disease 27% Progression disease 9.5% 20%

Background. Lenalidomide (Revlimid® ) is a novel, orally active immunomodulatory drug, effective in multiple myeloma (MM) patients. In newly diagnosed patients the addition of Thalidomide to the standard oral melphalan and prednisone (MP) significantly increase response rate and event free-survival compared with MP. No data are available on the clinical use of Revlimid® in combination with MP. In this multicenter phase I/II trial, we evaluate the dosing, safety and efficacy of the combination Revlimid® , melphalan and prednisone (R-MP). Methods. Patients (pts) with newly diagnosed symptomatic MM older than 65 years were treated with 9 courses of Revlimid® (5–10 mg/day for 21days every 4–6 weeks) plus MP (melphalan 0.18–0.25 mg/kg and prednisone 2 mg/kg for 4 days every 4–6 weeks) followed by maintenance therapy with Revlimid® alone (10 mg/day for 21days every 4–6 weeks). Four different dose-levels were tested: 1.melphalan 0.18 mg/kg + Revlimid® 5 mg/day; 2.melphalan 0.25 mg/kg + Revlimid® 5 mg/day; 3.melphalan 0.18 mg/kg + Revlimid® 10 mg/day; 4.melphalan 0.25 mg/kg + Revlimid® 10 mg/day. Each cohort included 6 pts, with additional 15 pts enrolled at dose level 3 and 4. All pts received ciprofloxacin and aspirin (100 mg/day) as prophylaxis. Results. Between January and October 2005, 54 pts (median age 71) were enrolled in the study. No DLTs were observed in the first 2 dose-levels. In dose-level 3, one pt experienced DLT (grade 4 neutropenia lasting>7 days). In dose-level 4, three pts showed DLTs (neutropenic fever, grade 3 cutaneous toxicity, pulmonary embolism, delay in the start of cycle 2) during the first cycle. The MTD was defined at dose-level 3 (melphalan 0.18 mg/kg+Revlimid® 10 mg/day). In the dose-levels 3 and 4, after a median of 7 cycles, all patients showed at least a minimal response and 85.4% of patients showed at least a partial response (PR); 41.5% of patients achieved at least a very good partial response (VGPR) and 17.1% of patients reached immunofixation negative complete remission (CR). In the dose-level 3, defined as MTD, 85.6% of patients showed at least a PR, including 52.3% of patients who achieved at least a VGPR and 23.8% who showed immunofixation negative CR. After a median follow up of 9.6 months, the progression free survival (PFS) was 87% at 16 months. FISH informations on chromosome 13q deletion were available in 42 patients (79%): no difference in response rate and PFS was observed between patients with or without 13q deletion. Toxicity was manageable, and occurred more frequently during early cycles. Major grade 3–4 adverse events consisted of hematological toxicities (neutropenia 66%, thrombocytopenia 34% and anemia 17%); major grade 3–4 non-hematological toxicities were cutaneous eruption (10%) and febrile neutropenia (8%). Three cases of tromboembolic events occurred: two of them after aspirin discontinuation, at cycle 7 and during maintenance. Conclusions. R-MP induced a high proportion of responses and appeared to overcome the poor prognosis of patients with chromosome 13q deletion. It was well tolerated, toxicities were predictable and manageable. An update of these data will be presented.

BACKGROUND: In newly diagnosed patients, the addition of thalidomide or bortezomib (Velcade™) to the standard oral melphalan/prednisone combination significantly increased response rate and event-free survival. In this multicenter, open-label, non randomized, phase I/II trial, the safety/efficacy profile of the 4 drug combination, bortezomib (Velcade™), melphalan, prednisone, and thalidomide (VMPT) was evaluated in patients with relapsed/refractory myeloma. METHODS: Bortezomib was administered by IV bolus on days 1, 4, 15, 22 at three dose levels: in the first cohort (10 patients) at 1.0 mg/m2; in the second cohort (10 patients) at 1.3 mg/m2and in the third cohort (10 patients) at 1.6 mg/m2. Oral melphalan was administered at 6 mg/m2 on days 1–5, oral prednisone at 60 mg/m2 on days 1–5. Thalidomide was delivered at 50 mg on days 1–35. Each course was repeated every 35 days for a total of 6 courses. RESULTS: Thirty patients, median age 66 years (range 38–79), with relapsed or refractory myeloma were enrolled. Fourteen patients received VMPT as second line of therapy, 16 as third line. Twenty patients received prior autologous transplant, 10 conventional chemotherapy and 9 thalidomide-based regimens. After a median of 6 courses, 20 patients (67%) achieved a partial response (PR) including 13 patients (43%) who achieved at least a very good partial response (VGPR). Among patients who received VMPT as second line treatment, the PR rate was 79%, and the immunofixation negative complete response rate 36%. The 1-year progression-free survival was 61%, and the 1-year survival from study entry was 84%. Grade 3 non-hematological adverse events included: infections (5 patients), fatigue (1), vasculitis (1) and peripheral neuropathy (2); no grade 4 toxicities were recorded. CONCLUSIONS: Initial results showed that VMPT is an effective salvage therapy with a high proportion of responses. Toxicities were manageable. The incidence of neurotoxicities was unexpectedly low.

Allogeneic approaches employing low dose TBI nonmyeloablative conditionings reported a dramatic reduction of transplant-related mortality (TRM) compared to conventional high dose regimens in newly diagnosed multiple myeloma (MM) (Maloney et al, Blood, 2003). The role of allografting, however, compared to autologous HCT remains to be determined. From September 1999 to July 2004, 241 consecutive MM patients, up to the age of 65, were diagnosed at five academic Italian Institutions. Overall, 194/241 had natural siblings (Table 1): 158/194 (81%) were HLA typed, while 36/194 (19%) were not typed for the following reasons: patients not eligible for high dose chemotherapy (n. 14); siblings not eligible for peripheral hematopoietic cell (PHC) donation (n.11); patient refusal to high dose chemotherapy (n. 9), unknown (n.2). Seventy-six/158 (48%) with a matched donor were offered a tandem autologous- nonmyeloablative allogeneic HCT approach. Eventually, 56/76 (73%), the “auto-allo group”, were enrolled while 20 did not enter the tandem program as 5 siblings (5/76, 7%) were not eligible for PHC donation, 5 patients refused an allogeneic HCT, and 10 patients preferred allografting as a possible salvage treatment. Of 102 patients without matched donors or after refusal to allografting, 73, “double-auto group”, underwent a standard double autologous transplant while 29 received less intense treatments because of clinical conditions or patient preference. Table 1 Newly diagnosed pts 241 With sibs/without sibs 194 /47 (total 241) HLA typed /not HLA typed 158 /36 (total 194) Matched sibs /No matched sibs 76 /82 (total 158) Auto-Allo”/“Double Auto”/Other”“ 56 /73 /29 (total 158) After induction chemotherapy, patients of both groups underwent G-CSF mobilised autografting with high dose melphalan (200 mg/m2). In the “auto-allo” group, the autologous HCT was followed, 2-4 months later, by low dose (2.0 Gy) TBI, allogeneic PHC infusion, and post transplant mycophenolate mofetil and cyclosporin. In the “double-auto group”, patients received a second autologous HCT. Patients characteristics were as follows: age: 54 (range 34–65) vs 53 (range 33–64) (p=ns); stage III myeloma: 77% vs 64% (p=0.03); beta 2 microglobulin > 2,5 mg/dl: 75% vs 59% (p=0.005), for the “auto-allo group” and for “the double-auto group”, respectively. At the time of this analysis, 56/56 of the “auto-allo group” and 55/73 of “the double-auto group” had completed the transplant programs. After median follow up of 3 years (range 11–80 months), TRM was 11% vs 4% (p=0.09); complete remission rates, defined as the disappearance of the monoclonal paraprotein by immunofixation, were 46% vs 16% (p=0.0001); overall survivals were 84% versus 62% (p=0.003); progression free survivals were 75% vs 41% (p=0.00008); event free survivals were 61% (34/56)% vs 38% (30/73) (p=0.006) in the “auto-allo group” and in the “double-auto” group, respectively. Longer follow up is needed, however data suggest that the “auto- non myeloablative allo” approach is not inferior to “double autologous” HCT in newly diagnosed MM.

Bortezomib (Velcade™) is an effective treatment for advanced multiple myeloma (MM). The association of Velcade™ to standard MP significantly increased response rate.When combined with Thalidomide, Velcade™ showed additive activity. Based on these observations, a phase II multicenter trial of Velcade™ plus Melphalan, Prednisone and Thalidomide (V-MPT) as salvage treatment was conducted. This is a dose escalation study with three sequential dose levels of Velcade™. The V-MPT regimen included 6 five-week courses of oral Melphalan (6 mg/m2 on days 1–5), Prednisone (60 mg/m2 on days 1–5), Thalidomide (100 mg continuously). In the first cohort Velcade™ at 1 mg/m2 was added on days 1, 4, 15, 22 of each course followed by a 13-day rest period. In the second and in the third cohort, Velcade™ was administered at 1.3 mg/m2 and 1.6 mg/m2, respectively. DLT was defined as the occurrence in the first 2 courses of any grade 3–4 non hematological toxicities, a grade 4 neutropenia ≥ a week, or any grade 4 hematological toxicity except neutropenia. Velcade™ dose increment was implemented whenever at least 3 patients (pts) completed 2 courses without any DLT. As of July 2005, 20 pts have been enrolled in this study, median age 65 years (range 38–73), 65% IgG, 20% IgA, 15% Bence Jones. The median b2 microglobulin was 3.83 mg/L (range 0.5–13.8). Nine pts received V-MPT as second line therapy, 11 as third line. Fourteen patients received prior autologous transplant, 5 conventional chemotherapy and 4 thalidomide-based regimens. In the first 10 pts cohort, 3 DLT were observed in the first 2 courses (grade 3 pneumonia at cycle 1, grade 3 febrile neutropenia at cycle 1 and grade 3 vasculitis at cycle 2). Treatment was discontinued in pts who experienced pneumonia and vasculitis. Velcade™ was reduced to 0.7 mg/m2 in patient who experienced febrile neutropenia. Grade 3 hematological toxicities included thrombocytopenia (3 pts) and neutropenia (5 pts). The most common grade 1–2 toxicities were: constipation, infections, rash. In the second cohort, five of the 10 pts completed at least 2 cycles. Two DLT in the first 2 courses were observed (grade 3 Herpes Zoster infections). Grade 3 hematological toxicities included thrombocytopenia (1 pt) and neutropenia (1 pt). The most common grade 1–2 toxicities were: constipation, infections, fatigue. After introduction of prophylaxis with acyclovir, no new HZV reactivation was observed. Among the 6 pts with baseline peripheral neuropathy before V-MPT treatment, 4 pts remained stable, and 2 worsened (grade 2). Treatment-related (grade 1) neuropathy developed de novo in 2 pts. After a median of 3 courses (range 2–6) and a median follow-up of 5 months, 10 pts (67%) had a response ≥ 50%: 2 complete responses (CR), 1 near CR, 7 partial responses. Two pts (13%) achieved minor response (MR) and one stable disease. One pt experienced progressive disease after a MR and one was refractory to treatment. Initial results showed that V-MPT is a promising regimen for advanced myeloma. Further investigation is ongoing to determine the maximum tolerated dose of this combination. Updated results will be presented at the meeting.

Introduction: array-CGH is significantly impacting on cancer cytogenetic. We used this technique to perform pan-genomic screening in 15 patients with MM and 4 with de novo PCL. Patients and methods. Bone marrow samples were employed. If tumor contamination was below 20% plasma cells were purified with Myltenyi columns. Array-CGH was performed as follows: genomic DNAs, from both the tumor and normal reference cells, labeled with different fluorescent dyes were cohybridized to 1 Mb resolution arrays containing 2600 Bacteria Artificial Chromosome (BAC) clones (Spectral Genomics Inc, Houston, TX, USA) according to manufacturer’s recommendations. Variations in DNA sequence copy number for each BAC clone was assessed by relative fluorescence signal intensities, in a single hybridization, providing a locus-by-locus measure of DNA copy-number changes. Results: The assay was validated as follows: three normal DNAs were tested revealing no genetic imbalances. One normal male was tested against one normal female and sex chromosome (ch) imbalances were effectively detected. Array-CGH results for ch 13q14.3 deletions were matched with FISH results and concordance was seen in 87% of cases. The median number of lesions/patient observed in our panel was 17 (4–135) (fig 1a). Also the amount of the total genome affected by chromosomal imbalances was highly variable (median 3.9% range: 0.14%–27%) (fig 1a). The amount of involved genome did not correlate with the actual number of lesions (fig 1a). A good correlation was noticed between the amount of losses and gains in each patient (fig1b). Notably PCL do not have a more disrupted genome compared to MM patients as one might expects based on the highly malignant behavior (fig 1a). Interestingly two patients with a prolonged clinical history of MGUS prior to MM diagnosis had massive presence of losses ad gains. Of 2600 BACs 934 were never affected, 864 were targeted only in one patient (pt), 401 in two pts, 296 in 3–5 pts and only 105 were targeted in six pts or more (fig 1c). These 105 recurrent imbalances could be attributed to 9 different abnormalities. Among these we have identified five recurrent lesions (occurring in at least six patients) that have not been previously described. These are 19p13.2 (gain 9 pts, loss 1 pt), 14q12 (loss 3 pts, gain 4 pts), 16q12.1 (loss 6 pts) 11q24 (gain 6 pts), 9q23 (gain 6 pts). Conclusions: array-CGH allow effective pan-genomic screening of MM patients; the pattern of genetic disruption is highly heterogeneous with a majority of non-recurrent or uncommonly recurrent lesions; a number of highly recurrent lesions have been identified that will require assesment for prognostic impact; the overall amount of perturbed genome does not seem to correlate with more aggressive disease, and might be the reflection of alternative biologic features (f.e. a prolonged history of clonal disease). Figure Figure

PURPOSE: High-dose therapy followed by autologous stem cell transplantation (AT) is the standard treatment for newly diagnosed multiple myeloma patients. Which is the best treatment option for patients relapsing after AT has not been defined. To address this issues the efficacy of Thalidomide and Dexamethasone (TD), AT and Conventional Chemotherapy (CC) was evaluated. MATERIALS AND METHODS: We retrospective analysed the outcome of 90 multiple myeloma patients, median age 61, range 34–77, who received a first salvage treatment between January 1999 and September 2003. All patients received AT at diagnosis. Relapse was defined as introduction of therapeutic modality that was different from maintenance treatment. After a median time from diagnosis of 32 months, 43 patients were treated with TD (Thalidomide 100 mg/day associated with DEX 40 mg on days 1-4 each month), after a median time from diagnosis of 29 months, 28 patients were treated at first relapse with AT (86% single MEL100, 11% double MEL100, 4% single MEL200), and after a median time from diagnosis of 32 months, 19 patients were treated with CC (32% Doxorubicin, Cyclophosphamide, Etoposide, Cisplatinum combination chemotherapy, 26% Doxorubicin containing regimens, 26% Cyclophosphamide containing regimens, 11% other therapies). Data were analysed when the median follow-up from the start of salvage TD was 30 months (range 4.5–45), from the start of salvage AT was 18 months (range 3.5–24) and from the start of salvage CC was 21 months (range 2–19.5). End points of the study were response, progression free survival (PFS) from first relapse and overall survival (OS) from first relapse and from diagnosis. RESULTS: Patients characteristics were similar among different groups. At relapse the response rate after TD was: 19% near complete remission (nCR) (absence of M-Protein detected by electrophoresis), 28% partial response (PR) (M-Protein reduction 50–99%), 35% stable disease (SD) (M-Protein reduction 0–49%) and 19% progressive disease (PD). After AT was: 11% nCR, 71% PR, 11% SD, 7% PD; and after CC was: 16% PR, 32% SD and 53% PD. Response rate was significantly lower for patients receiving salvage CC in comparison with AT and TD (p The median PFS from relapse was 20.3 months for TD, 9 months for AT and 4.5 months for CC (p The multivariate analysis indicates that TD, β2 microglobulin and age were the only independent risk factors associated with improved outcome. CONCLUSION: In conclusion, TD improved PFS and OS in myeloma patients relapsing after autologous transplant.