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Endothelial cells (ECs) lining arteries and veins have distinct molecular/functional signatures. The underlying regulatory mechanisms are incompletely understood. Here, we established a specific fingerprint of freshly isolated arterial and venous ECs from human umbilical cord comprising 64 arterial and 12 venous genes, representing distinct functions/pathways. Among the arterial genes were 8 transcription factors (TFs), including Notch target HEY2, the current “gold standard” determinant for arterial EC (aEC) specification. Culture abrogated differential gene expression in part due to gradual loss of canonical Notch activity and HEY2 expression. Notably, restoring HEY2 expression or Delta-like4–induced Notch signaling in cultured ECs only partially reinstated the aEC gene signature, whereas combined overexpression of the 8 TFs restored this fingerprint more robustly. Whereas some TFs stimulated few genes, others boosted a large proportion of arterial genes. Although there was some overlap and crossregulation, the TFs largely complemented each other in regulating the aEC gene profile. Finally, overexpression of the 8 TFs in human umbilical vein ECs conveyed an arterial-like behavior upon their implantation in a Matrigel plug in vivo. Thus, our study shows that Notch signaling determines only part of the aEC signature and identifies additional novel and complementary transcriptional players in the complex regulation of human arteriovenous EC identity.

Endothelial cells (ECs) lining arteries and veins have distinct molecular/functional signatures. The underlying regulatory mechanisms are incompletely understood. Here, we established a specific fingerprint of freshly isolated arterial and venous ECs from human umbilical cord comprising 64 arterial and 12 venous genes, representing distinct functions/pathways. Among the arterial genes were 8 transcription factors (TFs), including Notch target HEY2, the current “gold standard” determinant for arterial EC (aEC) specification. Culture abrogated differential gene expression in part due to gradual loss of canonical Notch activity and HEY2expression. Notably, restoring HEY2expression or Delta-like4–induced Notch signaling in cultured ECs only partially reinstated the aEC gene signature, whereas combined overexpression of the 8 TFs restored this fingerprint more robustly. Whereas some TFs stimulated few genes, others boosted a large proportion of arterial genes. Although there was some overlap and crossregulation, the TFs largely complemented each other in regulating the aEC gene profile. Finally, overexpression of the 8 TFs in human umbilical vein ECs conveyed an arterial-like behavior upon their implantation in a Matrigel plug in vivo. Thus, our study shows that Notch signaling determines only part of the aEC signature and identifies additional novel and complementary transcriptional players in the complex regulation of human arteriovenous EC identity.

Kwon: Gilead: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria; Jazz: Consultancy, Honoraria. Duarte:Incyte Corporation: Other: Has received speaker and advisor fees. Hernandez-Rivas:Rovi: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees; Abbvie: Membership on an entity's Board of Directors or advisory committees; Roche: Membership on an entity's Board of Directors or advisory committees; AstraZeneca: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees; Celgene/BMS: Membership on an entity's Board of Directors or advisory committees. Jimenez Yuste:NovoNordisk: Consultancy, Honoraria, Research Funding; Roche: Consultancy, Honoraria; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Pfizer: Consultancy, Honoraria, Research Funding; Grifols: Honoraria, Research Funding; Bayer: Honoraria; Sobi: Consultancy, Honoraria, Research Funding; CSL: Honoraria; Octapharma: Honoraria.

Peter Paschka and Hartmut Döhner contributed equally.

COVID-19 is a severe infectious complication in patients with underlying medical conditions such as having undergone hematopoietic stem cell transplantation (HCT). This prospective survey reports outcome on 272 COVID-19 patients from 19 countries having undergone allogeneic (n = 175) or autologous (n = 97) HCT reported to the EBMT registry or to the GETH. All patients had the diagnosis of SARS-CoV-2 documented by PCR. Patients were included in this analysis if COVID-19 diagnosis was before April 10, 2020. The overall survival was estimate by using the Kaplan Meier methods, considering the death due to any cause as an event and the time from COVID-19 infection to the latest follow-up as survival time; difference between groups were tested by the log-rank test. Univariate and multivariate risk factor analysis for overall survival were performed with the Cox regression model.

Duarte: Incyte Corporation: Other: Has received speaker and advisor fees. Kwon:Jazz: Consultancy, Honoraria; Gilead: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria. Mielke:Novartis: Honoraria, Other: received via my institution, Speakers Bureau; Celgene/BMS: Honoraria, Other: received via my institution , Speakers Bureau; Bellicum: Honoraria, Other: received via my institution, Speakers Bureau; Kite/Gilead: Honoraria, Other: received via my institution , Speakers Bureau; Miltenyi: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: received via my institution , Speakers Bureau; KIADIS Pharma: Honoraria, Other: received via my institution , Speakers Bureau; DNA Prime: Honoraria, Other: received via my institution , Speakers Bureau. López Jiménez:MSD: Speakers Bureau; Roche: Research Funding, Speakers Bureau; Takeda: Speakers Bureau; Janssen: Research Funding, Speakers Bureau; Abbvie: Research Funding, Speakers Bureau; Gilead: Research Funding, Speakers Bureau.

Stem cells must proliferate and differentiate to generate the lineages that shape mature organs; understanding these 2 processes and their interaction is one of the central themes in current biomedicine. An intriguing aspect is asymmetric division, by which 2 daughter cells with different fates are generated. Several cell fate determinants participate in asymmetric division, with the endocytic adaptor Numb as the best-known example. Here, we have explored the role of asymmetric division in thymocyte development, visualizing the differential segregation of Numb and pre-TCR in thymic precursors. Analysis of mice where Numb had been inhibited by expressing a dominant negative revealed enhanced pre–T-cell receptor (TCR) signaling and a smaller thymus. Conversely, Numb overexpression resulted in loss of asymmetric division and a larger thymus. The conclusion is that Numb determines the levels of pre-TCR signaling in dividing thymocytes and, ultimately, the size of the pool from which mature T lymphocytes are selected.

Stem cells must proliferate and differentiate to generate the lineages that shape mature organs; understanding these 2 processes and their interaction is one of the central themes in current biomedicine. An intriguing aspect is asymmetric division, by which 2 daughter cells with different fates are generated. Several cell fate determinants participate in asymmetric division, with the endocytic adaptor Numb as the best-known example. Here, we have explored the role of asymmetric division in thymocyte development, visualizing the differential segregation of Numb and pre-TCR in thymic precursors. Analysis of mice where Numb had been inhibited by expressing a dominant negative revealed enhanced pre–T-cell receptor (TCR) signaling and a smaller thymus. Conversely, Numb overexpression resulted in loss of asymmetric division and a larger thymus. The conclusion is that Numb determines the levels of pre-TCR signaling in dividing thymocytes and, ultimately, the size of the pool from which mature T lymphocytes are selected.

Background: Heavily pretreated relapsed and refractory multiple myeloma (RR MM) constitutes a specific and unmet medical need. Median survival ranges from as little as 6 to 9 months, and responses to treatment are characteristically short (Richardson et al. 2007). Belantamab Mafodotin (BM), a novel anti-BCMA antibody conjugated to microtubule-disrupting agent monomethyl auristatin F, showed single-agent activity in the phase 1 DREAMM-1 and phase 2 DREAMM-2 studies in heavily pre-treated patients with RRMM (Lonial et al, 2019 & 2021). We aim to assess efficacy and safety of BM treatment administered via the expanded access compassionate care program for triple class MMRR patients in the region of Madrid (Spain).

No relevant conflicts of interest to declare.

Linker for activation of T cells (LAT) is an adaptor protein required for organization of the signaling machinery downstream of the platelet collagen receptor, the glycoprotein VI (GPVI). Here, we investigated the effect of LAT mutations on specific signaling pathways and on platelet functions in response to GPVI triggering by convulxin (Cvx). Using mice containing tyrosine to phenylalanine mutations of the adaptor, we show the crucial role played by the tyrosine residues at positions 175, 195, and 235 in the phosphorylation of LAT and in the whole pattern of protein tyrosine phosphorylation in response to Cvx. These 3 C-terminal tyrosine residues are important to recruit the tyrosine kinase Fyn, which may be involved in LAT phosphorylation. Efficient phosphoinositide 3-kinase (PI3K) activation requires the 3 C-terminal tyrosine residues of LAT but not its tyrosine 136. Interestingly, single mutation of the tyrosine 136 results in the loss of phospholipase C γ2 (PLCγ2) activation without affecting its PI3K-dependent membrane association, and is sufficient to impair platelet responses to Cvx. Thus, activation of PLCγ2 via GPVI is dependent on 2 complementary events: its interaction with the tyrosine 136 of LAT and its membrane location, which itself requires events mediated by the 3 C-terminal tyrosines of LAT.

Linker for activation of T cells (LAT) is an adaptor protein required for organization of the signaling machinery downstream of the platelet collagen receptor, the glycoprotein VI (GPVI). Here, we investigated the effect of LAT mutations on specific signaling pathways and on platelet functions in response to GPVI triggering by convulxin (Cvx). Using mice containing tyrosine to phenylalanine mutations of the adaptor, we show the crucial role played by the tyrosine residues at positions 175, 195, and 235 in the phosphorylation of LAT and in the whole pattern of protein tyrosine phosphorylation in response to Cvx. These 3 C-terminal tyrosine residues are important to recruit the tyrosine kinase Fyn, which may be involved in LAT phosphorylation. Efficient phosphoinositide 3-kinase (PI3K) activation requires the 3 C-terminal tyrosine residues of LAT but not its tyrosine 136. Interestingly, single mutation of the tyrosine 136 results in the loss of phospholipase C γ2 (PLCγ2) activation without affecting its PI3K-dependent membrane association, and is sufficient to impair platelet responses to Cvx. Thus, activation of PLCγ2 via GPVI is dependent on 2 complementary events: its interaction with the tyrosine 136 of LAT and its membrane location, which itself requires events mediated by the 3 C-terminal tyrosines of LAT.

No relevant conflicts of interest to declare.

Background:Defining the extent of disease is essential for determining the prognosis and management of patients with Hodgkin (HL) and non-Hodgkin lymphoma (NHL) as the presence of advanced disease, including involvement of an extralymphatic site, is associated with inferior outcomes (Cheson et al,2014). 18F-fluorodeoxyglucose positron emission tomography (FDG PET-CT) has an excellent diagnostic performance for evaluation of bone marrow infiltration (BMI) in patients with lymphoma but it is expensive and not always available at low and middle-income health care facilities. Unilateral bone marrow biopsy (BMB) is a more accessible test and has been the standard to identify lymphomatous BMI. Unfortunately, it is an unpleasant procedure for the patient and can be limited by technical constraints. The aim of this single-center retrospective cohort study was to evaluate the diagnostic performance of FDG PET-CT for BMI in lymphoma patients, comparing it with BMB as the gold standard. Also, we evaluated for discordant results between both studies along with its effect on patient treatment.

Loscertales: Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Gilead Sciences: Honoraria, Membership on an entity's Board of Directors or advisory committees; Roche: Honoraria, Speakers Bureau. Alegre:Janssen: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees.

Ibrutinib is a first-in-class Bruton kinase inhibitor (BTK) registered for the treatment of Chronic Lymphocytic Leukemia (CLL), mantle cell linphoma (MCL) and Waldenström Macroglobulinemia (WM).

Alegre: Celgene Corporation: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees; Amgen: Membership on an entity's Board of Directors or advisory committees.

Introduction:We have recently reported that the expression of Krüppel-like factor 4 (KLF4) is upregulated in pediatric Burkitt lymphoma and it was suggested to be a prognostic biomarker for survival (Valencia-Hipolito et al.,2014)*. In addition, preliminary findings have demonstrated that KLF4 is also a resistant factor for drug-induced apoptosis.

Introduction

No relevant conflicts of interest to declare.

Alegre: Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Jansen: Membership on an entity's Board of Directors or advisory committees, Research Funding.

Alegre: Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Jansen: Membership on an entity's Board of Directors or advisory committees, Research Funding.

BACKGROUND:Peripheral T/NK–cell lymphomas (PTCL) are a heterogeneous group of neoplasms with a median survival of 2-3 years. Currently, there is not standard treatment although consolidation of response with autologous stem cell transplantation (ASCT) is recommended as front line therapy, except in anaplastic ALK+ and T/NK nasal localized lymphomas.

Introduction:Conventional radiography remains the “gold standard” technique for bone involvement assessment in patients with multiple myeloma (MM). Newer imaging modalities such as whole-body Magnetic Resonance (MR) and 18FDG-PET/CT have emerged as more sensitive techniques than routine skeletal survey in the detection of bone involvement in the diagnostic and follow up of patients with MM. The advantages and disadvantages of MR and 18FDG-PET/CT are discussed.

Bendamustine is a dual alkylating agent with demonstrated efficacy in de novo multiple myeloma (MM) and in relapsed/refractory (R/R MM). We present the preliminary results from the experience of Spanish compassionate use registry of this agent in different treatment schedules of R/R MM, promoted by the GEM/PETHEMA Group (Benda-MMRR-11). This study has been approved by local ethical committee, CEIC, HUP, Madrid, Spain.72 patients, 42 males and 30 females, with advanced R/R MM after several lines of previous treatment received a schedule containing Bendamustine (Levact®, Mundipharma, UK). Median age was 66 (35–86), with a median of 4 previous salvage lines (1–11). Bendamustine dose used ranged between 60 and 120 mg/m2 iv on days 1 and 2 for each 28 days cycle. The median of cycles was 2 (1–9). The combinations used were: Bendamustine + Prednisone in 27 patients, Bendamustine + Thalidomide + Prednisone in 11 cases, Bendamustine + Dexamethasone in 14 patients, Bendamustine + Bortezomib + Dexamethasone in 7 patients, Bendamustine + Bortezomib + Prednisone in 3 patients, Bendamustine + Thalidomide + Dexamethasone in 2 cases, Bendamustine monotherapy in 5 patients, Bendamustine + Thalidomide in 1 case, Bendamustine with Caelyx in 1 case and Bendamustine with Rituximab in 1 case.In the 69 evaluable patients, the response was: overall response rate (ORR) 30.24%%, with complete response (CR) 11.5%, partial response (PR) 13.04%. Minimal response (MR) was 5.7%, and stable disease (SD) was 17.39%. Progression was documented in 52.1% of patients. In general, treatment was well tolerated; the most common adverse effect was hematological toxicity with grade 3–4 neutropenia in 31.8%, grade 3–4 thrombocytopenia in 30.4% and grade 3–4 anemia in 17.3%. Other toxicities included grade 3–4 infections in 27% of patients and grade 3–4 asthenia in 11.5%. Clinical results will be updated with more details regarding TTP, OS and efficacy and toxicity with each schedule.Bendamustine is an effective salvage therapy in patients with advanced R/R MM. Our results are consistent with previous published data in larger series of patients. More experience is needed with this agent to define the best combination and to assess its grade of efficacy that will be probably superior in earlier stages of myeloma evolution.Off Label Use: compassionate use.

Abstract 4035

Treatment of frail or elderly patients with relapsing symptomatic/active Multiple Myeloma (MM) is very difficult due to concomitant diseases, impaired bone marrow reserve, systemic toxicity, relatively decreased renal function and general problems of old age. Dexamethasone and new agents (thalidomide, lenalidomide, bortezomib and bendamustine) have been used in this setting, in most cases with doses adapted to the clinical situation.To retrospectively analyze the management of frail and/or very elderly MM patients with relapsed and active disease treated with reduced doses of the aforementioned agents in five hospitals in Madrid, Spain.The files of this group of MM patients were studied. The most common treatment has been the combination of low doses of lenalidomide (len) and of dexamethasone (dex), whereas treatment with reduced doses of other agents has been anecdotal; therefore we analyzed the results of len/dex combinations. Len and dex have been used in lower than standard doses, adapted to the individual initial situation of the patients and tailored according to effect and toxicity throughout treatment. There was no specific protocol and the management of the patients has depended exclusively on the practice and criteria of the treating physicians. Patient risk was stratified following the Salmon and Durie (S&D) score and the International Staging System (ISS). Response was assessed with the IMWG criteria. The study has been approved by the Ethics Committee of Hospital Ramon y Cajal, as coordinating center.38 patients were included in the study. Mean age was 79 years (range 68–90). 30 pts (79%) were older than 75 years and 10 pts had over 85 years. More than half of the patients (21) had two or more comorbidities. Patients had previously received 1 to 5 (m=1. 8) different treatment modalities, including steroids, melphalan (25), bortezomib (20), thalidomide (6) (or their combinations), and others or even APBSCT (3). 23 pts (60%) had IgG (m=4087 mg/dl, range 868–13000); 13 (34%) IgA (m=2115, range 355–4930) and 2 (5%) only light chains. 22 had κ and 15 λ light chains. 19 (50%) had BJ proteinuria. Mean Hemoglobin level was 10. 7 gr/dl (7. 5–14. 1) and mean creatinine level 1. 3 mg/dl (0. 4–12. 9); 28 (74%) had bone disease. 3 pts had S&D stage I, 22 stage II, and another 13 stage III. 13 pts had ISS stage I, 17 had stage II and 7 stage III. Patients received between 1 and 30 cycles of len/dex (m= 8). Median initial Len dose was 10 mg, the majority between 5 and 15mg, although 4 received 25 mg that were rapidly reduced. Mean initial dex dose was 20mg/day for 4 days. 4 pts (10. 5%) achieved Complete Remission (CR) (3 with negative IF), 27 (71%) Partial Remission (PR) (5 with VGPR) and 2 (5%) a significant, but lesser than 50%, reduction of the M-component (Stable Disease, Std). Altogether, overall response (CR+PR+Std) occurred in 33 pts (86%). The best response occurred after 2 to 9 cycles (m=4) of len/dex. Treatment was stopped in 15 patients due to neurological (4) or hematological (1) toxicity, pulmonary embolism (1), unrelated causes (4) and after achieving a plateau response (5). Time to next treatment was 1–30 months, (m=8 mo). 7 pts relapsed after 3–21 months (m=7). 10 patients died, 5 of related (disease progression, cardiac amyloidosis, renal progression to ESRF) and 5 of unrelated (cancer, sepsis, myocardial infarction, congestive heart failure) causes. Grade III-IV bone marrow toxicity occurred in 9 pts and neurological toxicity (PNP) in 5 (all of them had previously been treated with bortezomib or thalidomide).Personalized low doses of len/dex have been the most common treatment for frail/very elderly patients with relapsed MM in our centers and it is an active and tolerable option in this setting. The haematological toxicity was expectable and manageable, but prior treatments with bortezomib or thalidomide were associated with limiting neurotoxicity.No relevant conflicts of interest to declare.

Abstract 5049

Stem cells (SC) show a potential of cellular therapy in diffuse coronary disease with controverted results. We present here an analysis of a group of patients treated with TMLR plus ABMC. (Caiber Program Advanced Therapy Proyect FIS 2009).19 patients with DCD were included. Age 66 (range: 52–78). ABMC were obtained by aspiration and processed using HARVEST-TM system -TALEX (Harvest Technologies Inc. GMBH, Munich) to obtain 20 ml. Surgery consisted of anterior thoracotomy incision at the fourth and fifth intercostal space. Between 15–30 transmyocardial channels were realized with TLMR PHOENIX-TM (Cardiogenesis, Irvine, CA, USA) and CrystalFlex 1 mm optical fibre. 0.5–2 cc of MO were administered in each channel. Total number CMN were determined by automatic cell counter and flow cytometry and were typified specific cell populations such as CD34+ and CD133+.The duration of stem cell collection was less than 30 minutes. All the procedure was performed on surgery in 1 step. Any complications were observed with this technique. Correlation between the cellular count and demographic data was not observed. After a follow up of 22 months (5-36) only one patient has died (20m) of DCD. Initial clinical results are satisfactory with a significant reduction in the functional grade of angina.Our data show that TMLR plus ABMC is a safe method with clinical efficacy in patients with DCD that are not candidate to other therapy options. The described method offers efficiency and simplicity to obtain cells. More cases and follow-up are required, as well as later functional evaluations.Alegre: Celgene: Honoraria. Off Label Use: Lenalidomide is not approved for the treatment of smoldering multiple myeloma.

Abstract 1181

Aim: To evaluate the efficacy of Lenalidomide (Len) as compassionate use in relapsed or progressive Multiple Myeloma (MM) up to its approval in Spain. Patients and Methods: GEM-PETHEMA designed a transverse and retrospective, multicenter analysis of MM cases which Len compassionate use was requested until December, 2007. The decision to treat these patients was previous and independent from the decision to conduct the present analysis and depended only on the clinical criteria of the responsible doctors. At least, one response assessment was a must for efficacy analysis. Results: 111 MM patients have been included. Eligible patients for efficacy were 103. Mean age was 65.7 (38–85); 53 m, 50 f. Previous median lines of therapy were 3 (1–8): 92 (89.3%) have received bortezomib; 37 (35.9%) autologous PBSCT and 26 (25.2%) thalidomide. Extramedullary plasmocitomas were present in 25 (24.3%). Mean Len dose was 22.8 mg (± 4.9): 82 (79.6%) received the standard dose and schedule (25 mg d for 3 w every 4 w) and 21 (20.4%) received less dose and/or different schedule. 92 (89.3%) received Dexametasone (mean dose 58.33 mg/w) [± 35.8]). Response: 4 (3.9%) sCR, 11 CR (10.7%), 12 VGPR (11.7%), 41 PR (39.8%), 20 SD (19.4%), 13 PD (12.6%), 2 NE (1.9). Among groups, response equal or superior to PR was observed in all settings: 64.1% in prior exposed to bortezomib, 46.1% in prior exposed to thalidomide and 40.0% in patients with extramedullary plasmocitomas. Previous transplant, the number of previous lines received, renal failure, age, or cytogenetic did not affect significantly the overall response rate. Median duration of treatment was 7.7 m (1–21); median TTP was 8.3 m and median global survival since starting Len therapy was 11 m (6–22). Median survival since diagnostic was 49 m (40–60). At the time of analysis, 46 (45.5%) patients were still on Len therapy, and 72 (79.6%) were alive. Toxicity: ≥ grade II: neutropenia, 51 (46.4%); thrombocytopenia, 39 (35.5%); DVT, 5 (4.5%); rash, 3 (2.7%); neutropenic fever, 8 (7.3%); others, 13 (11.8%). DVT/PE prophylaxis was used in 89 (86.4%) patients: LWMH in 43.8 % and low dose aspirin in 48.3 %. No PE was reported. Conclusions: Lenalidomide is effective in this heavily pre-treated MM population-progressive or refractory to standard therapy-even in different clinical settings. The most frequent association to Len was intermediate dose of Dex. Although response rate was superior in patients exposed previously to bortezomib, no differences on duration and survival were observed. Patients with extramedullary plasmocitomas showed also response. Toxicity, mainly myelosupression was predictable and manageable with dose adjustments and cytokine support.

Introduction: Stem cell (SC) based therapy in ischemic heart disease is actually in development with the aim of improving cardiac regeneration. The way of administration, source of SC and the mechanisms of its clinical effects are yet not well defined. We present preliminary results of a group of patients treated with laser revascularization and autologous bone marrow (BM) cells administration, evaluating cells immunophenotype and clinical results. Patients and methods: Fourteen patients (57–78 y) with diffuse coronary disease and medically refractory class III/IV angina were prospectively evaluated for cell-based therapy combined with laser transmyocardial revascularization (TMR) PHOENIX-TM (Cardiogenesis, Irvine, CA, USA). BM mononuclear cells (MC), were obtained by BM aspiration (120 mL) and processed for concentration using HARVEST-TM system (Harvest Technologies Inc. GMBH, Munich). Immunophenotypic characterization of the final infused product was performed by standard four-color flow cytometry with monoclonal antibodies directed against the human surface antigens CD29, CD49a, CD105, CD106, CD117, CD34, CD133 and CD45, to evaluate different cell populations. Results and conclusions: Patients received a median of 56.8×106 (43.7–79) BMMC/ mL 0.5×106 (0.05–1) CD34+ cells/mL and 0.23×106 (0.001–0.7) CD133+ cells/mL. All the patients are alive after this treatment: median of 9 months, (1–15), with improvement of their cardiological clinical status and QoL. As observed (Table 1) the BMMC used in the therapy contained a significant proportion of CD133+ and CD34+ hematopoietic SC. In addition, the samples contained cells with CD105+ CD45− or CD29+ CD45− CD34− phenotypes which have been described as mesenchymal stem cells (MSC). Bone marrow CD106+ positive cells represent a multipotent subpopulation of MSC including endotelial. Correlation between the relative quantity of the different cell subsets and clinical results of the patients will be presented. TABLE 1 CD34 CD133 CD117 CD29 CD49a CD105 CD106 Mean 3.53 2.42 7.37 2.23 0.75 6.75 0.33 Std. Deviation 1.72 1.68 2.60 0.97 0.75 3.34 0.23 25% Percentile 1.95 0.85 4.95 1.25 0.15 3.10 0.2 Median 3.20 2.15 6.70 2.30 0.40 7.05 0.25 75% Percentile 5.45 4.25 10.45 3.15 1.70 10.10 0.55

Aim: To evaluate the efficacy of Lenalidomide (Len) as compassionate use in relapsed or progressive Multiple Myeloma (MM) up to its approval in Spain.

Introduction:Stem cell (SC) based therapy in ischemic heart disease is actually in development with the aim of improving cardiac regeneration. The way of administration, source of SC and the mechanisms of its clinical effects are yet not well defined. We present preliminary results of a group of patients treated with laser revascularization and autologous bone marrow (BM) cells administration, evaluating cells immunophenotype and clinical results.

Introduction Intensification therapy in MM with autologous PBSCT is widely indicated for younger patients with favourable results regarding response, OS and PFS. However, there is no clear plateau in the survival curve. To improve these results and to prolong remission duration, various maintenance treatments have been proposed. Interpheron alpha2b s.c, at low dose, steroids on alternate days and most recently, low dose oral thalidomide, have demonstrated improvements in EFS and OS times but these results needs confirmation. Recently a new formulation of interpheron alpha2b (Pegintron®) is available conjugated with polietilenglicol with the advantage of being administered on a unique weekly dose. This new formulation of Interpheron alpha has been tested scarcely in MM. We present the preliminary results of a spanish, phase II, non comparative, multicentric study (PI-MM-01) with Pegintron® (Schering-Plough) as maintenance treatment after autologous PBSCT. Patients y Methods From May 2003 to March 2007, 30 patients were included in this study, 64% female and 36% male, with a median 56 years, received Pegintron® once a week subcutaneously as maintenance treatment after favourable response post-HDT with autologous PBSCT when the engraftement was stable and complete. The initial dose was 15 mg/week × 2 week and this dose was escalated to 25 mg/week and then 35 mg/week. The final dose was adopted according clinical and hematological tolerance. The maintenance treatment was continued at least 5 years postransplant or until toxicity, relapse or progressión. Results 30 patients were evaluable for this preliminary analysis. The median time from transplant to IFN treatment was 3.8m. The median dose of Pegintron was 15 mg/week. 9 patients have suspended the treatment (30%). 5 cases (16%) due to progression, 2 (4%) for toxicity and 2(4%) for other reason. The remainder 21 patients (76%) continue the treatment with clinical response, with median duration of 16 months (2–42). At present all patients are alive. Astenia, “pseudo-flu” symptoms and thrombopenia and neutropenia were the most common adverse effects observed, that limited the dose escalation, being 15 mg/week the best tolerated and mos frequent dose (70%). One patient suspended the treatment due to dermatological reaction with pruritus. Conclusions and Comments Although these results need completion and further analysis, maintenance treatment with a weekly dose of Interpheron-a2b conjugated with Polietilenglicol (Pegintron®) is well tolerated after autologous PBSCT in MM. No major adverse effects were observed and no relevant negative impact was observed on the autologous graft. Pegintron® sc could be an alternative to standard interpheron sc with the main advantage of therapy simplification with only one dose weekly. More experiences and longer follow up are needed to evaluate the role of this strategy in the global treatment of MM and new approaches for maintenance have to be investigated, including association and comparison with steroids or new drugs as thalidomide, bortezomib or lenalidomide.

Introduction and Objectives. Lenalidomide is a new inmunomodulatory drug which has shown effectiveness, in combination with dexamethasone, in relapsed or progressive Multiple Myeloma (MM). In our country, little experience outside clinical trials exists since lenalidomide is only currently available in the compassionate use program, for patients with advanced MM. With the purpose of sharing the clinical practical experience with this drug, we considered it interesting to gather the outstanding data on the use of lenalidomide in a multicentric registry, throughout the Spanish scope. We present the preliminary results of this registry. Patients and methods. We designed a registry, canalized through GEM-PETHEMA, in which main clinical data of MM patients relapsed or progressive treated with lenalidomide within the compassionate use program, were collected. The decision to treat these patients was prior to and independent of the decision to make this registry and was only based on clinical criteria. The selected patients had at least one response assessment. Results. 19 valid questionnaires have been received, 14 patients (pts.) being assessable for response. The treatment was started between 03/06 and 04/07. Average age: 63 years (40–77); 9 men and 5 women. Type of MM: IgG 9 pts, IgA 3 pts, 1 non-secretory, 1 Bence Jones K. Average M-protein 3.25 g/dL(SD: 3,32); average creatinine 1,05 (SD: 0.36); average bm plasma cells infiltration: 36%. Median lines of previous treatments: 4 (2–8). 13 pts had received bortezomib, 8 pts had received thalidomide, and 7 had undergone autologous PBSCT. 7 pts showed extramedullary plasmocytomas.2 patients received the standard dose (25 mg daily, P.O. 21 days every 4 weeks). 2 patients received lower doses. All the patients, except one, received Dexamethasone simultaneously (average 56 mg per week). Prophylaxis for DVT: 7 pts with LWMH and 3 pts with salicylates. 4 patients did not receive prophylaxis. At the time of data collection 10 patients where still on the treatment. Lenalidomide was discontinued in 2 patients due to progression (median time to progression 6 months (1–8); 1 pts underwent PBSCT after reaching PR and discontinued the treatment due to stable disease after 6 months. Response: 2 CR (14%), 1 VGPR (7%), 6 PR (43%), 3 SD (21%). Among the 9 pts with a favorable response (64%): 8 had previously received bortezomib, 4 had received thalidomide, and 4 had undergone PBSCT - Toxicity: Neutropenia G3: 5 pts/G4: 2 pts; 5 patients needed G-CSF. Thrombocytopenia G3: 3 pts/G4: 3 pts. Skin rash in 1 pt. Neither TVP/TEP nor neutropenic fever were observed. Conclusions. Although a more detailed analysis and a longer follow-up is needed the results of our registry are consistent with other published clinical trials: lenalidomide associated with dexamethasone induces durable objective responses in a high percentage of patients with relapsed or progressive MM, after several treatment lines. The tolerance of this drug is acceptable, being the mielosuppression the most relevant toxicity. The role of this drug at low dose for long term maintenance have to be investigated.

Introduction Plasma Cell Leukemia (PCL) is characterized by a high number of plasma cells in peripheral blood (>2 × 109/L) and more than 20% of circulating plasmocytes on differential count. Primary PCL or secondary form, after previous multiple myeloma (MM), represent the most aggressive forms of clonal gammapathy with very poor outcome from both conventional chemotherapy and autologous or allogeneic transplant. For this reasons new effective treatment approaches are required. The proteasome inhibitor Bortezomib has been shown to be effective in advanced MM with a rapid response but few reports have been published regarding its potential role on PCL. We report here the clinical results of a serie of PCL patients that received proteasome inhibitor, Bortezomib (Velcade®), as induction therapy. Patients and Clinical Results We describe 10 patients with PCL treated with Bortezomib. Median age was 58 y (30–70). 4 cases were primary, de novoPCL and 6 cases were secondary forms of relapsed or progressive MM. Patients with secondary PCL had received a median of 3 previous lines of therapy including autologous transplantation (5 cases). Bortezomib at standard dose iv: 1.3 mg/m2 days 1,4, 8,11 every 21 days were used alone in 2 patients and in combination with high dose of Dexamethasone +/− Adryamicin in the rest. 3 cases recived the BDD scheme (Bortezomib iv, Pegylated Doxorrubicin iv and oral Dexamethasone). 9 patients were evaluable for drug efficacy with 7 cases showing some grade of favourable response with the first cycle, including 3 cases of CR. 4 patients have died: 1 during first weeks and 3 for progression, 2–3 months after initial response. 3 cases received allogeneic transplant with related donor and reduced conditioning and are alive 12, 14 and 30 months postransplant. The median follow up since PCL diagnosis of 6 alive patients is 14 months (3–39 months). 3 patients received maintenance therapy that included low dose thalidomide. Grade 2 thrombopenia, neutropenia and gastrointestinal symptoms were the main secondary toxicity. No complicated tumoral lysis syndrome were observed. Conclusions and Comments Bortezomib has been shown to be effective by several mechanisms in patients with MM resistant to multiple previous treatments and as induction therapy alone or in combination with other drugs. The favourable responses observed in these cases of PCL; including de novo patients, and the biological ex vivo results in PCL cell lines, suggest the potential efficacy of this agent combined with dexamethasone and doxorubicin. Although more experience is necessary these data support future trials with Bortezomib combined with other drugs for example BDD scheme in the induction treatment of PCL. The role of allogeneic intensification therapy has also to be investigated in young patients after control of PCL with this approach. For maintenance of response, the role of this drug or other new drugs as thalidomide or lenalidomide has to be investigated.

IntroductionIntensification therapy in MM with autologous PBSCT is widely indicated for younger patients with favourable results regarding response, OS and PFS. However, there is no clear plateau in the survival curve. To improve these results and to prolong remission duration, various maintenance treatments have been proposed. Interpheron alpha2b s.c, at low dose, steroids on alternate days and most recently, low dose oral thalidomide, have demonstrated improvements in EFS and OS times but these results needs confirmation. Recently a new formulation of interpheron alpha2b (Pegintron®) is available conjugated with polietilenglicol with the advantage of being administered on a unique weekly dose. This new formulation of Interpheron alpha has been tested scarcely in MM. We present the preliminary results of a spanish, phase II, non comparative, multicentric study (PI-MM-01) with Pegintron® (Schering-Plough) as maintenance treatment after autologous PBSCT.

Introduction and Objectives. Lenalidomide is a new inmunomodulatory drug which has shown effectiveness, in combination with dexamethasone, in relapsed or progressive Multiple Myeloma (MM). In our country, little experience outside clinical trials exists since lenalidomide is only currently available in the compassionate use program, for patients with advanced MM. With the purpose of sharing the clinical practical experience with this drug, we considered it interesting to gather the outstanding data on the use of lenalidomide in a multicentric registry, throughout the Spanish scope. We present the preliminary results of this registry.

IntroductionPlasma Cell Leukemia (PCL) is characterized by a high number of plasma cells in peripheral blood (>2 × 109/L) and more than 20% of circulating plasmocytes on differential count. Primary PCL or secondary form, after previous multiple myeloma (MM), represent the most aggressive forms of clonal gammapathy with very poor outcome from both conventional chemotherapy and autologous or allogeneic transplant. For this reasons new effective treatment approaches are required. The proteasome inhibitor Bortezomib has been shown to be effective in advanced MM with a rapid response but few reports have been published regarding its potential role on PCL. We report here the clinical results of a serie of PCL patients that received proteasome inhibitor, Bortezomib (Velcade®), as induction therapy.

Introduction In spite of the favourable results observed after autologous PBSCT in MM there is no clear plateau in the survival curve and eventually most patients relapse with a median EFS between 40–50 months postransplant. To improve these results and sustain remission, various maintenance treatments have been proposed with minimal and discrepant benefits. Interpheron alpha2b s.c. at low dose and steroids on alternate days, have demonstrated modest improvements in EFS and OS times (5–12 months) after standad-dose therapya but its role in the setting of high dose therapy (HDT) needs confirmation. Recently a new formulation of interpheron alpha2b is available conjugated with polietilenglicol (Pegintron®), that need only one dose weekly and has not been evaluated in MM. We present the preliminary and interim results of a spanish, open study, no comparative, multicentric with Pegintron® (Schering-Plough) as maintenance treatment after autologous PBSCT. Patients y Methods 17 patients with MM received Pegintron® once wekly subcutaneously as maintenance treatment after favourable response post-HDT with autologous PBSCT when the engraftement was completed (7 CR and 10 PR). The initial dose was 15 mg/week x 2 week and this dose was escalated to 25 mg/week and then 35 mg/week. The final dose was adopted according clinical and hematological tolerance. Intermitent oral steroids were allowed. The maintenance treatment was continued until toxicity, relapse or progressión. Results Of the 17 patients included so far in the study, 2 patients suspended the treatment. One case by personal decision an the other for progression, 14 months after transplant. The remainder 15 patients follow the treatment with a median of 10.7± 5 months (1.5–18.6). Most patients received also zolendronate iv/months and oral low dose dexamethasone (20 mg x 4 x months) associated to Pegintron. Mild bone pain, “flu-lyke syndrome” and grade I thrombopenia and neutropenia, were the most common adverse effect.. The best tolerated dose was 15 mg/week. With the exception of one case, all patients maintain the response achieved with HDT with a median of 15 months (3–22) after transplant. Comments and Conclusions These prelimiar results show that maintenance treatment with a weekly dose of Interpheron-a2b conjugated with Polietilenglicol (Pegintron®) is effective and well tolerated after autologous PBSCT in MM. No major adverse effects were observed and no relevant negative effecst was presented on the autologous graft. Pegintron® could be an alternative to sc standar interpheron with the main advantage of therapy simplification (one dose weekly). More experience and longer follow up are needed to evaluate the role of this strategy in the global treatment of MM and new approaches for maintenance have to be investigated, including new drugs as bortezomib, thalidomide an lenalidomide, associated or not with bisphosphonates, Pegintron and steroids.

Introduction: Justification and Objectives There are little epidemiologic data about Myeloma Multiple in Spain. The heterogeneity and complexity of this pathology, and the several sociodemographic factors, justify the interest of this type of studies. On the other hand, this disease needs a high assignement of welfare and therapeutic resources, and besides new strategies have arisen for its treatment. The Spanish Leukaemia and Lymphoma Foundation (FLL)has analyzed the actual situation of Myeloma Multiple in Spain, compiling several epidemiologic and welfare parameters about the disease in a Multiple Myeloma “White Book” for Spain. Patients and Methods The period of analysis was 10 years, from 1.991 to 2.001. The information was compiled from a Hospital Based Survey about MM and the following official sources: International Agency for Research on Cancer (IARC): “Cancer incidence in Five continents” and “Incidence and Mortaliity for Cancer In Spain, Patterns and Tendences” Data Base of EUCAN and GLOBOCAN (Cancer Incidence, Mortality and Prevalence Worldwide) EUROCARE III study, (IARC Cancer Base Number 5, Lyon, IARCPress). “Natural Changes of Population and Demography. Spanish National Statistic Institute (INE) and National Spanish Center for Epidemiology (CNE.) Official Records for Mortality Rates by MM in Spain in Spanish Regions (CCAA) (death records Demograhy Records of INE). INE Hospital Case Rate Inquest. CMBD Data Base of Department of Health (Inmunoproliferative Neoplasm and Multiple Myeloma, CIE-9 Diagnosis Code: 203.0)The indicators used were: Deaths number, median of age, proportional mortality, mortality rates, mortality rates adjusted by age, gender and potential years of life lost due to multiple myeloma, (item 203 of ICD-10, OMS) during the last ten years in Spain. Results The incidence rates of MM, adjusted to the European population were: 3,54 cases by 100.000/year for men and 2,54 cases by 100.000/year for women. The global incidence rate were 4,44 cases by 100.000/year for men and 4,22 cases by 100.000/year for women. These rates were similar in all geographic regions. Performed predictions show a prevalence increase during the following five years, which means more than 2.400 cases in men and more than 2.100 in women MM cases per year. Regarding mortality, rates, myeloma is a very slightly frequent cause of death: 3.23 cases by 100.000/years of men and 2.3 cases of women. A whole an increase of mortality of 45.2 per cent was observed for the period of time between 1992 and 2001. Comments and Conclusions The MM incidence and mortality rates in Spain for this period were lower than expected in comparison with other European epidemiology studies. Nevertheless we observed that the MM mortality and prevalence rates present a continuous and uniform increment in the last years. Part of these increases can be due to the incorporation of new technologies, more sensitive for diagnosis, and to the increase of aging of the population. Furthermore certain occupational and chemical exposures and other environmental changes could explain these trends.

Introduction

Introduction: Justification and Objectives