Search

Supplementary Figure from Circulating Tumor and Immune Cells for Minimally Invasive Risk Stratification of Smoldering Multiple Myeloma

Supplementary Data from Circulating Tumor and Immune Cells for Minimally Invasive Risk Stratification of Smoldering Multiple Myeloma

Supplementary Table from Circulating Tumor and Immune Cells for Minimally Invasive Risk Stratification of Smoldering Multiple Myeloma

Purpose:Early intervention in smoldering multiple myeloma (SMM) requires optimal risk stratification to avoid under- and overtreatment. We hypothesized that replacing bone marrow (BM) plasma cells (PC) for circulating tumor cells (CTC), and adding immune biomarkers in peripheral blood (PB) for the identification of patients at risk of progression due to lost immune surveillance, could improve the International Myeloma Working Group 20/2/20 model.Experimental Design:We report the outcomes of 150 patients with SMM enrolled in the iMMunocell study, in which serial assessment of tumor and immune cells in PB was performed every 6 months for a period of 3 years since enrollment.Results:Patients with >0.015% versus ≤0.015% CTCs at baseline had a median time-to-progression of 17 months versus not reached (HR, 4.9; P 20% BM PCs had no prognostic value in a multivariate analysis that included serum free light-chain ratio >20, >2 g/dL M-protein, and >0.015% CTCs. The 20/2/20 and 20/2/0.015 models yielded similar risk stratification (C-index of 0.76 and 0.78). The combination of the 20/2/0.015 model with an immune risk score based on the percentages of SLAN+ and SLAN− nonclassical monocytes, CD69+HLADR+ cytotoxic NK cells, and CD4+CXCR3+ stem central memory T cells, allowed patient’ stratification into low, intermediate-low, intermediate-high, and high-risk disease with 0%, 20%, 39%, and 73% rates of progression at 2 years.Conclusions:This study showed that CTCs outperform BM PCs for assessing tumor burden. Additional analysis in larger series are needed to define a consensus cutoff of CTCs for minimally invasive stratification of SMM.

Previous studies showed antitumor activity for plitidepsin plus dexamethasone (DXM) in relapsed/refractory multiple myeloma (r/r MM), and in vitro synergism with bortezomib (BTZ) or DXM against MM cells. This phase I trial evaluated plitidepsin (3-h intravenous infusion Day 1 and 15), BTZ (subcutaneous bolus Day 1, 4, 8, and 11), and DXM (orally Day 1, 8, 15, and 22), every 4 weeks in 36 r/r MM patients. Twenty-two patients were treated using a standard dose escalation design (10 at the recommended dose [RD] cohort), and 14 additional patients were treated to expand the RD cohort. No dose-limiting toxicities (DLTs) occurred during dose escalation. The highest dose level evaluated (plitidepsin 5.0 mg/m2 , BTZ 1.3 mg/m2 , DXM 40.0 mg) was the RD for phase II studies. Results shown herein are focused on this RD. Two patients had DLTs (grade 3 diarrhea, and grade 3 nausea/vomiting refractory to antiemetic therapy). Grade ? 3 hematological toxicity (thrombocytopenia 46%, anemia 33%, and neutropenia 17%) was manageable and did not result in treatment discontinuation. Transient and manageable grade 3 ALT increase (26%) was the most common biochemical abnormality. At the RD cohort, overall response rate was 22.2% (95%CI, 6.4%-47.6%), including one stringent complete response, one very good partial response, and two partial responses in r/r patients to BTZ and/or lenalidomide. The clinical benefit rate was 77.8% (95%CI, 52.4-93.6%). No major pharmacokinetic drug-drug interaction was found. In conclusion, the triple combination of plitidepsin, BTZ, and DXM showed an acceptable safety profile and had moderate activity in adult patients with r/r MM. FUNDING: The study was funded by Pharma Mar, S.A. ACKNOWLEDGMENTS: The authors thank the patients, their families, and investigators teams for their participation in this phase I clinical trial.

Cancers of the same histotype may show profound differences in clinical behavior because they can arise from cells having acquired multiple mutations, which presumably occur over many years.[1][1] In contrast, it is well known that cancers with different morphologies and phenotypes, and those that

Background The 2/20/20 model to predict risk of transformation in SMM is established at diagnosis and not periodically re-evaluated, mainly because plasma cell (PC) quantification requires invasive bone marrow (BM) aspirates. It could be hypothesized that dynamic monitoring of tumor biomarkers could improve risk-stratification of SMM patients, but this requires minimally invasive methods to replace those performed in BM samples. Such methods should also monitor immune profiles to identify patients with stable tumor burden/genetics, but at risk of progression due to lost immune surveillance. Aim Compare the prognostic value of PC quantification in BM vs the evaluation of circulating tumor cells (CTCs) in peripheral blood (PB), and to define immune signatures predictive of time-to progression (TTP) in SMM. Methods 300 SMM patients are planned to be enrolled in the iMMunocell study that includes 26 sites across 8 European countries. PB samples are collected every 6 months during three years for next-generation flow (NGF) monitoring of CTCs and immune profiling of the T, NK, B and myeloid cell compartments. Cutoffs for risk stratification according to CTCs were calculated using maxstat. A total of 245 SMM patients were enrolled, and we report here a pre-planned interim analysis on the first 150. Results Thus far, 28/150 (19%) patients progressed to MM. Presence of ≤20% vs >20% PCs in BM by morphology failed to predict different TTP(19% vs 32% progressions at 2 years, respectively; p=0.14). Patients with ≤0.73 CTCs/µL had lower risk of transformation vs those with >0.73 CTCs/µL (15% vs 67% progressions at 2 years, respectively; HR: 5.1, p=4.9e-05). Patients with >0.73 CTCs/µL had median TTP of 17.5 months. Patients with low, intermediate and high-risk SMM according to the 2/20/20 model showed significantly different TTP (p=5.14e-06), but those with high-risk had median TTP not reached. Risk stratification using a new 0.7/2/20 (where 0.7 stands for >0.73 CTCs/µL) achieved identical statistical significance (p=5.14e-06), and patients with high-risk SMM had median TTP of 21 months. By applying the gradient boosting algorithm to the T cell dataset, we identified six subsets with an exhausted phenotype CD4+CD28negTIGIT+CD127lo, CD4+CD28+TIGIT+CD127+, CD4+CD28+TIGIT+CD127+CD25+, CD8+CD28negCD127+, CD8+CD28negTIGIT+ and CD8+CD28negTIGIT+PD1+ whose increased frequency was associated with inferior TTP. Patients showing an expansion of these T cell subsets had higher risk of transformation to active MM (HR: 7.33; p=0.002). Conclusions This is the first study performing CTC and immune monitoring every 6 months in PB samples from patients with SMM. Our results suggest that CTC numbers have greater prognostic value than BM PC counts, and that a new 0.7/2/20 model could be dynamically assessed to identify SMM patients at risk of developing active MM. Beyond CTC numbers,this study is also uncovering key immune cell types associated with disease progression.

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). Methods: An observational, retrospective and multicenter study has been performed including all patients who received at least one dose of BM under the expanded access program in the region of Madrid (Spain) from Nov 2019 to Jun 2021. Hematology centers provided data from the medical records and entered them in a case report form distributed to the sites. Primary endpoint was overall response rate (ORR). Secondary endpoints were progression free survival (PFS), overall survival (OS) and the incidence of treatment emergent adverse events (TEAEs), with a major focus on ocular and hematologic toxicity. Results: A total of 33 patients (pts), from 14 different centers, were included from February 2020 till May 2021. Median age was 70 (46-79) years. 55% of the pts were women. Median time from diagnosis was 71 (10-858) months. 30.3% were high-risk cytogenetic features. Median of prior therapy lines was 5 (3-8) and at least 88% of the pts were triple class refractory. The median number of BM doses per patient was 3 (1-16) and the median follow-up was 11 months (95%CI 6.34-15.66). ORR was 42.2%, and 18.2% achieved ≥VGPR. Median PFS was 3 months (95%CI 0.92-5.08). Median PFS for patients who achieved ≥PR was 11 months (HR 0,26; 95% CI 0,10-0,68). No significant differences were found in PFS according to age, cytogenetic risk and prior therapy lines. OS was 424 days (95% CI 107-740). The incidence of non-hematological TEAEs was 57.6% and the most common of which was ocular toxicity (45.5%). The incidence of ≥G3 non-hematological TEAEs was 30.3%. 51.5% of the pts were diagnosed of keratopathy and 21.2% was ≥G3. 30.3% of the pts showed a reduced visual acuity, but this event was resolved in 92.9% of the pts. The most common symptoms were blurry vision (30.3%, n=10) and dry eye (24.2%, n=8). The incidence of ≥G3 hematological TEAEs was 18.2% and thrombocytopenia was the most frequent (21.2%). Dose reductions of BM were required in 30.3% of the pts and delayed in 36.4% due to TEAEs. Main causes for treatment discontinuation (81%, n=27) were disease progression (54.5%, n=18), toxicity (15.2%, n=5), death (6.1%, n=3) and due to patient's decision (3%, n=1). Conclusion: Compassionate use of BM in heavily pretreated RR MM pts showed a relevant anti-myeloma activity with a manageable safety profile.These results are similar to those observed in the DREAMM-1 and DREAMM-2 clinical trials. Disclosures No relevant conflicts of interest to declare.

© 2018 The Authors., [Background]: The present prospective, multinational, noninterventional study aimed to document and describe real-world treatment regimens and disease progression in multiple myeloma (MM) patients. [Patients and Methods]: Adult patients initiating any new MM therapy from October 2010 to October 2012 were eligible. A multistage patient/site recruitment model was applied to minimize the selection bias; enrollment was stratified by country, region, and practice type. The patient medical and disease features, treatment history, and remission status were recorded at baseline, and prospective data on treatment, efficacy, and safety were collected electronically every 3 months., [Results]: A total of 2358 patients were enrolled. Of these patients, 775 and 1583 did and did not undergo stem cell transplantation (SCT) at any time during treatment, respectively. Of the patients in the SCT and non-SCT groups, 49%, 21%, 14%, and 15% and 57%, 20%, 12% and 10% were enrolled at treatment line 1, 2, 3, and ≥ 4, respectively. In the SCT and non-SCT groups, 45% and 54% of the patients had received bortezomib-based therapy without thalidomide/lenalidomide, 12% and 18% had received thalidomide/lenalidomide-based therapy without bortezomib, and 30% and 4% had received bortezomib plus thalidomide/lenalidomide-based therapy as frontline treatment, respectively. The corresponding proportions of SCT and non-SCT patients in lines 2, 3, and ≥ 4 were 45% and 37%, 30% and 37%, and 12% and 3%, 33% and 27%, 35% and 32%, and 8% and 2%, and 27% and 27%, 27% and 23%, and 6% and 4%, respectively. In the SCT and non-SCT patients, the overall response rate was 86% to 97% and 64% to 85% in line 1, 74% to 78% and 59% to 68% in line 2, 55% to 83% and 48% to 60% in line 3, and 49% to 65% and 36% and 45% in line 4, respectively, for regimens that included bortezomib and/or thalidomide/lenalidomide., [Conclusion]: The results of our prospective study have revealed great diversity in the treatment regimens used to manage MM in real-life practice. This diversity was linked to factors such as novel agent accessibility and evolving treatment recommendations. Our results provide insight into associated clinical benefits., Writing support during the development of our report was provided by Laura Mulcahy and Catherine Crookes of FireKite, an Ashfield company, a part of UDG Healthcare plc, which was funded by Millennium Pharmaceuticals, Inc, and Janssen Global Services, LLC. The EMMOS study was supported by research funding from Janssen Pharmaceutical NV and Millennium Pharmaceuticals, Inc.

Mediterranean environments are of most interest to study pea adaptability to terminal drought conditions especially in the current context of global climate change. In our work we have tested nine pea cultivars in five South European and North African locations, characterised by different agro climatic conditions within the Mediterranean climate. Data were processed through the additive main effects and multiplicative interaction analysis. Grain yield, aboveground biomass and flowering date were assessed within each mega-environment with parametric and non-parametric methods, establishing ranks for the genotypes within each condition attending to their stability parameters and mean values. The field analysis revealed HR1 as a wide-adapted genotype, whereas others such as Desso showed the best adaptation in South Mediterranean areas. Our results also highlighted the potential interest of these genotypes and others (i.e. Messire and ZP108) in breeding programs and further studies on drought tolerance., Financial support by Spanish project AGL2014-52871, Andalusian project AGR-2883, Serbian project TR-31024 and the FP7 ERANet-ARIMNET-Medileg is acknowledged.

Powdery mildew caused by Erysiphe pisi DC is an important pea disease which causes high yield and quality losses. The most efficient way to control this disease is the use of resistant cultivars. Three genes of resistance are known so far in pea, namely er1, er2 and Er3. The most widely deployed gene in pea cultivars worldwide is er1. Resistance conferred by er1 is complete and considered to be durable, being recently demonstrated that er1 is in fact a member of the mlo gene family. Resistance is caused by colony abortion albeit the responsible cellular mechanisms causing this colony abortion are not known. In this study, the presence of different mechanisms related with colony abortion, such as callose apposition, protein cross-linking and hypersensitive response was quantified in a resistant pea er1 line compared to a susceptible check. Our histological studies showed that protein cross-linking, plays a determinant role in the penetration resistance conferred by er1, whereas callose apposition or hypersensitive response played little role., Financial support by project AGL2011-22524 co-financed by FEDER is acknowledged.

Abiotic stresses caused by adverse environmental conditions are responsible for heavy economic losses on pea crop, being drought one of the most important abiotic constraints. Development of pea cultivars well adapted to dry conditions has been one of the major tasks in breeding programs. The increasing food requirements drive the necessity to broaden the molecular basis of tolerance to drought to develop pea cultivars well adapted to dry conditions. We have used a shotgun proteomic approach (nLC-MSMS) to study the tolerance to drought in three pea genotypes that were selected based on differences in the level of water deficit tolerance. Multivariate statistical analysis of data unraveled 367 significant differences of 700 identified when genotypes and/or treatment were compared. More than half of the significantly changed proteins belong to primary metabolism and protein regulation categories. We propose different mechanisms to cope drought in the genotypes studied. Maintenance of the primary metabolism and protein protection seems a strategy for drought tolerance. On the other hand susceptibility might be related to maintenance of the homeostatic equilibrium, a very energy consuming process. Data are available via ProteomeXchange with identifier PXD004587., María-Ángeles Castillejo was funded by a postdoctoral fellowship from the Spanish Ministry of Education, through the Mobility Program R-D + I 2008–2011, Code 2010-0236. Rebeca Iglesias-García was funded by the excellence Project 07-AGR-34730 (Andalusian Regional Government).

Bendamustine is a bifunctional alkylating agent with proven activity in myeloma. In this study 60 newly diagnosed myeloma patients were given bendamustine plus bortezomib and prednisone in a regimen consisting of one cycle of bortezomib twice weekly for 6 weeks (1.3 mg/m2 on days 1, 4, 8, 11, 22, 25, 29, and 32), plus bendamustine (90 mg/m2 on days 1 and 4) and prednisone. The following cycles included bortezomib once weekly. Patients who were transplant candidates proceeded to stem cell collection after four cycles and the transplant was performed after six cycles. Patients who were not candidates for transplantation received up to nine cycles. Forty-two patients were transplant candidates and after six cycles, 50% achieved at least a very good partial response, with 24% having complete responses; 35 proceeded to a transplant, and the complete response rate was 54%. Seventeen patients continued up to nine cycles, and 57% achieved at least a very good partial response, including 26% with complete responses. The 2-year progression-free survival and overall survival rates were 62% and 86%, respectively. The safety profile was manageable, but stem cell mobilization was compromised in 35% of patients. In summary, this combination is effective in untreated patients, with an acceptable toxicity profile, but given the introduction of second-generation novel agents and monoclonal antibodies, the combination will probably be better reserved for relapsing patients, in whom stem cell collection is not needed, while cost-effective combinations with non-cross-resistant drugs continue to represent a medical need. This trial was registered with ClinicalTrials.gov, number NCT01376401.

Pea (Pisum sativum L.) is an important grain legume whose yield stability and production is constrained by drought stress in most environments. However, little is known on availability of drought adaptation sources and its genetic control in pea. The aim of this work was to study the genetic of drought adaptation in pea and identify the genomic regions controlling the trait. Towards this objective, in this work, we assessed drought symptoms and relative water content in soil (RWCS) and leaves (RWCL) along a time course of water stress on a pea Recombinant Inbreed Lines (RILs) population from two parents known to segregate for drought adaptation. Drought adaptation in this population was a quantitative trait. QTL analysis using composite interval mapping (CIM) and multiple interval mapping (MIM) allowed us to identify ten quantitative trait loci (QTLs) associated with the traits explaining individually from 9 to 33 % of the phenotypic variation depending on the variable assessed and altogether from 20 to 57 %. A set of reproducible markers linked to these QTLs (A6, AA175, AC74, AD57, AB141, AB64, Psblox2, PsAAP2_SNP4, and DipeptIV_SNP1) were identified. These markers can be used to select the individuals harbouring the desired QTLs in pea breeding programs for drought adaptation., This work was supported by EU project FP7-KBBE2013.1.2-02-613551 “LEGATO”.

Purple urine bag syndrome (PUBS) is an uncommon but particularly striking phenomenon characterised by a chemical reaction involving the urine, plastic and certain enzymes from some sulphatase- and phosphatase-producing bacteria, including Proteus mirabilis, Escherichia coli and Morganella morganii, amongst others. Following this reaction, the catheter and the bag may be stained red, blue or purple. This phenomenon tends to occur in patients with multiple pathology and with urinary catheters, as part of a urinary tract infection. We describe two clinical cases of PUBS in institutionalised patients with permanent urinary catheters.

8006Background: Despite recent progress, MM remains incurable, hence the need for agents with novel mechanisms of action. Marine-derived plitidepsin targets eEF1A2, a protein overexpressed in MM. P...

Purple urine bag syndrome (PUBS) is an uncommon but particularly striking phenomenon characterised by a chemical reaction involving the urine, plastic and certain enzymes from some sulphatase- and phosphatase-producing bacteria, including Proteus mirabilis, Escherichia coli and Morganella morganii, amongst others. Following this reaction, the catheter and the bag may be stained red, blue or purple. This phenomenon tends to occur in patients with multiple pathology and with urinary catheters, as part of a urinary tract infection. We describe two clinical cases of PUBS in institutionalised patients with permanent urinary catheters.

Objective TO report a paradoxical reaction of Raynaud phenomenon following the repeated administration of iloprost in a patient with diffuse cutaneous systemic sclerosis with vascular involvement. Case Summary In January 2006, a 40-year-old male was diagnosed with diffuse cutaneous systemic sclerosis with pulmonary, esophageal, cutaneous, and vascular involvement (Raynaud phenomenon, with digital ulcers on his hands). In December 2008, treatment with iloprost was started due to worsening disease. Nine cycles of iloprost were administered at a rate of 0.5–1 ng/kg/min (6 hours per day, for 5 days every 6–8 weeks); the patient tolerated this treatment well. However, on the fourth day of cycles 10 and 11, the patient developed paradoxical Raynaud phenomenon in the hand with perfusion when the infusion was increased to 1 ng/kg/min, requiring treatment to be stopped. Treatment was continued during cycles 12 and 13 at 0.5 ng/kg/min; the patient tolerated the treatment well, although paradoxical Raynaud phenomenon occurred when the rate of infusion was increased. Discussion Raynaud phenomenon is extremely common in patients with scleroderma, and often is severe. Iloprost has vasodilating, antiplatelet, cytoprotective, and immunomodulating properties, and has been found to be an efficacious alternative to nifedipine for the treatment of Raynaud phenomenon in patients with scleroderma. The Naranjo probability scale indicated that iloprost was the probable cause of the paradoxical Raynaud phenomenon in this patient. Conclusions This case demonstrates a probable relationship between the rate of infusion of iloprost and the paradoxical reaction of Raynaud phenomenon.

Prostatic lymphoma is an exceptional pathology, that usually is diagnosed because its prostatic symthomatology or as consequence of its invasion by an extraprostatic lymphoma. We present a case of a patient affected by a prostatic lymphoma and we perform a review of the literature in order to establish the diagnostic and therapeutic steps.

Resumen El linfoma prostatico es una patologia excepcional que suele diagnosticarse como consecuencia de su sintomatomatologia prostatica o como consecuencia de su invasion por un linfoma extraprostatico. Presentamos un caso de un paciente afecto por un linfoma prostatico y realizamos una revision de la literatura, para establecer las pautas diagnosticas y terapeuticas.

Background and Objectives: Recently, knowledge of the specific genetic markers responsible for cancer malignancies and their associated signaling pathways have generated many new targets that promise to increase drug efficacy while reducing side effects such as hematotoxicity. Although hematotoxicity is widely assumed to be the result of depleting hematopoietic progenitors, in particular myeloid progenitors, there are no assays to test new compounds in bone marrow samples to investigate this effect. Because these effects are patient-specific, an assay that identifies those patients risk of severe hematotoxicity by certain treatments could help personalizing patient treatment. Here, we show the ability of our flow cytometry-based automated Exvitech© platform to measure depletion analysis of different subsets of CD34+ progenitors and other cell subsets to potentially establish a new assay to screen drug candidates and combinations for hematotoxicity, as well as personalizing therapy to the individual sensitive patient at risk. So that in multiple myeloma (MM) we can identify at the same time CD34+ cells and pathological plasma cells; we could actually measure depletion of both malignant cells and progenitor cells on the same patient sample. Patients and methods: 16 normal bone marrow (NBM) and 4 MM samples were studied. For a proof of concept to test the hypothesis, we have selected two known cytotoxic drugs (cytarabine and clofarabine: N=10NBM) and two novel drugs with low expected cytotoxicity (ruxolotinib and volasertib: N=6NBM). The whole sample was plated into 96-well assay plates containing 8 concentrations of each drug and incubated for 48-hours for NBM and 12h with Bortezomib for MM samples. A multiple staining (CD45v450/Anexin-FITC/CD117-PE/CD34PerCP/CD38APC/CD19APCya7) was used to distinguish between both populations. Drug response was evaluated as a depletion survival index of each cell population relative to the average of 6 control wells in each plate. Results: As expected, nucleoside induces hematotoxicity in most of the studied NBM samples, but not all. Results reflect that cytarabine has similar activity than clofarabine in terms of efficacy (Ymax: 30% vs 23%) but with less potency (EC50: 6µM vs 0.2µM) in the immature population (N=10; Figure 1). This reflects a lower hematological toxicity which is consistent with clinical practice. Interestingly, for both drugs there is a large range of inter-patient variability inside this population in terms of efficacy (cytarabine, range Ymax: 4%-75% and clofarabine, range Ymax: 10%-37%) and potency (cytarabine, range EC50: 1µM-14µM and clofarabine, range EC50: 0.01µM-2µM) suggesting that in a subsets of vulnerable patients, drug doses could be tailored. By contrast, ruxolitinib and volasertib had little effect (Ymax: 80% vs 73%) in the immature population with minimal interpatient variability confirming the low toxicity expected for these novel drugs even at very high concentrations never achieve in vivo (Figure 1). Figure 2 shows bortezomib activity in MM bone marrow samples measuring both the dose response on malignant and myeloid progenitor cells. For Patient 1 the drug depletes myeloid precursor at lower doses than malignant cells, suggestive of severe hematotoxicity before therapeutic benefit can be achieved. Patient 2 shows the opposite case, where bortezomib depletes malignant cells completely without depleting myeloid precursors, suggestive of a good therapeutic index for this individual patient. Conclusions: These preliminary results show that Vivia Exvitech© platform is able to measure hematopoietic progenitors depletion in addition to other cell populations for novel drugs or before patientxs treatment that could contribute to a more selective drug development or a better clinical management of patients. This approach enables screening for hematotoxicty potential new discovery compounds, new drug candidates, and their synergistic combinations, thus supporting drug discovery and development. This assay could be helpful to both hematological and solid tumor drugs. The platform can measure both malignant and progenitor cells in bone marrow samples of MM and Non Hodgkin's Lymphoma. This simultaneous analysis shown for bortezomib could help guiding the clinical response and possible hematological toxicities associated to drug treatments. Figure 1: Figure 1:. Figure 2: Figure 2:. Disclosures Primo: Vivia Biotech: Employment. Ballesteros:Vivia Biotech: Employment. Robles:Vivia Biotech: Employment. Gorrochategui:Vivia Biotech: Employment. Garcia:Vivia Biotech: Employment. Hernandez:Vivia Biotech: Employment. Martinez:Vivia Biotech: Membership on an entity's Board of Directors or advisory committees.

Patients with severe and persistent bleeding have high mortality rates despite standard therapy, including blood transfusion support. The aim of the study was to evaluate the role of rFVIIa in the management of severe bleeding, refractory to other treatments.All cases with severe bleeding and failure of previous treatments who received rFVIIa (n = 21) at one single center were retrospectively included in the study.A response after the administration of rFVIIa was reported in 16 of 21 patients (76.2%). Hemorrhage was completely halted in 14 cases, and 12 patients (57.1%) were alive 30 days after treatment. The use of rFVIIa was associated with a normalization of coagulation tests, especially the prothrombin time (p = 0.001). A marked reduction in blood requirements, red cell units (p = 0.003), fresh frozen plasma (p = 0.009) and platelets (p = 0.017) was also observed.rFVIIa may have an important role in the achievement of an adequate hemostasis and reduces blood requirements in patients with severe bleeding, emerging as an alternative to blood transfusion.

Fundamento y objetivo Los pacientes con hemorragias graves y persistentes presentan una elevada mortalidad pese a los tratamientos convencionales, entre ellos el soporte transfusional. El objetivo del estudio ha sido evaluar el papel del factor VII activo recombinante (rFVIIa) en el tratamiento de hemorragias graves, refractarias a otros tratamientos. Pacientes y metodos En el estudio se incluyeron retrospectivamente todos los casos (n = 21) tratados en un unico centro con rFVIIa debido a hemorragias graves, en las que previamente habian fracasado otras medidas terapeuticas. Resultados La tasa de respuesta al tratamiento fue del 76,2%, cediendo completamente la hemorragia en 14 pacientes. La supervivencia a los 30 dias tras la administracion del farmaco fue del 57,1% (12 de 21 pacientes). La utilizacion de rFVIIa se asocio con una mejoria de los parametros de la coagulacion, especialmente del tiempo de protrombina (p = 0,001), asi como con una marcada reduccion de los requerimientos transfusionales, concentrados de hematies (p = 0,003), plasma fresco (p = 0,009) y plaquetas (p = 0,017). Conclusiones El rFVIIa consigue una adecuada hemostasia y disminucion de requerimientos de hemoderivados en hemorragias graves, por lo que representa una alternativa a la transfusion en estos pacientes.

Background Bortezomib-based combinations, including alkylating agents (VMP or CyBorD) or immunomodulatory drugs (VTD or RVD) have been established as regimens widely used in newly diagnosed MM patients. Bendamustine is a bifunctional alkylating agent effective in relapsed and/or refractory MM patients, and approved in Europe in combination with prednisone for elderly newly diagnosed MM. Since bendamustine may be more efficient than other alkylators, an attractive possibility would be to explore it in combination with bortezomib and prednisone (BVP) in newly diagnosed MM patients both transplant and non transplant candidates. Patients and Methods 60 newly diagnosed MM patients were included in the trial. The first cycle consisted on bendamustine at 90 mg/m2 given IV on days 1 and 4, in combination with bortezomib at 1,3 mg/m2 given IV on days 1, 4, 8, 11, 22, 25, 29 and 32 and prednisone at 60 mg/m2 given PO on days 1 to 4. In the following cycles, bendamustine was given on days 1 and 8, and bortezomib on days 1, 8, 15 and 22 (weekly schedule), and prednisone as it was previously described. Patients younger than 65 years proceeded to peripheral blood stem cell collection (PBSC) using growth factors alone after 4 cycles; HDT-ASCT was performed after 6 cycles. Patients older than 65 years received up to nine 28-day cycles. Results Between May 2011 and July 2012 enrollment was completed (60 pts). Median age was 61 years (range 38-82; 18 pts ≥65), 67% had ISS stage II/III, and 67% had unfavorable cytogenetics: t(4;14), t(14;16), del 17p or 1q gains by FISH. After a median of 6 cycles (2-9), 75% of patients achieved at least PR, including 16% of sCR, 9% CR and 28% of VGPR. Although the differences were not statistically significant, there was a trend to higher CR rate in the group of patients Forty patients proceeded to stem cell collection after a median of 4 cycles of BVP. Upon using G-CSF alone, 14 pts (35%) failed to collect a minimun of 2 x 106 CD34+ cells/Kg. An ammendment was done and plerixafor was recomended for poor mobilizers (peripheral CD34 cell count inferior to 10/μL on day 4); all patients but 2 achieved, with G-CSF plus plerixafor, the minimum of CD34+ cells required to proceed to ASCT. These 2 patients successfully collected CD34+ cells using chemotherapy plus G-CSF and plerixafor. Of the 31 patients who received HDT-ASCT, sCR and CR rate before transplant was 18% and 13%, respectively, upgrading up to 39% of sCR and 13% CR after transplant. 7 pts (22%) achieved immunophenotypic CR. After a median f/u of 12 months (5-25), 8 pts have progressed, resulting in a 15-m TTP of 85%. Concerning OS, 89% of patients remained alive at 15 months. None of patients achieving sCR and CR have progressed and all of them are alive at 15 months. Regarding cytogenetic abnormalities, although there were not significant differences, one patient progressed in standard risk group and five in the high risk subgroup resulting in a 15 m-TTP of 93% vs 85%. No significant differences have been observed in terms of 15 m-OS between standard and high risk cytogenetic subgroup (100% vs 92%, respectively). As far as toxicity is concerned, hematologic toxicities included: G3/4 anemia (11%), neutropenia (23%), and thrombocytopenia (14%). The most common G3/4 non-hematologic toxicities were: asthenia (10%), infections (9%), and peripheral neuropathy (4%). Conclusions In patients candidates to HDT-ASCT, response rates obtained before and after transplant are comparable to other three drug bortezomib-based combinations, such as VTD or CyBorD. However, growth factors alone for stem cell collection after four BVP cycles as induction resulted in a 35% of poor mobilizers who were rescued with plerixafor. In the elderly population, although the number of patients included was small, BVP seems not superior to VMP in response rates. Disclosures: Mateos: Janssen, Mundipharma: Honoraria. Off Label Use: bendamustine plus bortezomib and prednisone is not an approved combination for first line of therapy. Oriol:Celgene: Consultancy. Ocio:Onyx: Consultancy, Research Funding; Novartis: Consultancy; Array Biopharma: Consultancy, Research Funding; Bristol Myers Squibb: Consultancy; Celgene: Consultancy, Research Funding. Alegre:Celgene: Membership on an entity’s Board of Directors or advisory committees, Research Funding; Janssen: Membership on an entity’s Board of Directors or advisory committees, Research Funding. Bladé:Janssen, Mundipharma: Honoraria. San Miguel:Janssen, Mundipharma: Membership on an entity’s Board of Directors or advisory committees.

Invasive fungal infections (IFI) remain as an important cause of morbidity and mortality in adult patients with acute leukemia (AL). The incidence of proven or probable mold and yeast infections ranges from 5–24% among this selected high-risk patients. Prophylaxis is a commonly used treatment strategy, because the diagnosis of IFI is difficult and often delayed. A meta-analysis showed that itraconazole was more effective than fluconazole as prophylaxis of IFI, and a recent randomized trial demonstrated that in patients undergoing chemotehrapy for acute myelogenous leukemia (AML) posaconazole prevented IFI more effectively than did either fluconazole or itraconazole. Voriconazole (Vor) is a second-generation triazole with an extended spectrum of activity and is the drug of choice for invasive aspergillosis, the main mould that causes IFI among patients with acute leukemia. The drug is available for intravenous or oral administration and is well tolerated with transient visual disturbances, liver enzyme abnormalities and skin rashes being the most common adverse events reported, that rarely lead to treatment discontinuation. In this pilot study we evaluated Vor as prohylaxis treatment for IFI in 70 consecutive cases that underwent induction chemotherapy treatment for acute leukemia in two Clinical Units of Hematology in Spain. Methods:Oral prophylaxis with VOR at a fixed dose of 200mg/12h after two loading doses of 400mg was started on the first day of chemotherapy and continued until recovery from neutropenia or until ocurrence of an IFI. Patients who were unable to tolerate orally could receive VOR intravenously. Galactomanan was measured twice a week. HR-CT were performed in selected cases. The primary efficacy end point was the incidence of proven/probable IFI. Secondary end points included treatment success (vs failure) and the ocurrence of emergent mould infections. Treatment failure was defined as the ocurrence of proven/probable IFI; receipt of any other systemic antifungal agent for suspected IFI; discontinuation of the study drug for an adverse event; or withdrawal from the study with no additional follow-up. Results:From Jan 06 to Feb 07, 70 consecutive cases of AL that received induction chemotherapy for AML(58) or ALL(12) were included in the study. Median age was 57 (20–75). Median days with 0.1x109/L and 0.5x109/L were 16(10–35) and 19(12–38) respectively. Neutropenic fever ocurred in 62 cases (88,5%) and in 53% of them (33/62) a documented bacteria was isolated from the blood cultures. Galactomanan was possitive in only two cases (2,8%). HR-CT findings consisting with IFI were observed in 3 cases. Proven or probable IFI ocurred in 3 cases (A, flavus, A. fumigatus and Fusarium) (4,2%). Treatment failure as defined ocurred in 10 cases (16%). Conclusions: Prophylaxis with voriconazole was an effective and well tolerated regimen for IFI prevention in adult patients with acute leukemia.

Background Many proteins with tandem repeats in their sequence have been described and classified according to the length of the repeats: I) Repeats of short oligopeptides (from 2 to 20 amino acids), including structural cell wall proteins and arabinogalactan proteins. II) Repeats that range in length from 20 to 40 residues, including proteins with a well-established three-dimensional structure often involved in mediating protein-protein interactions. (III) Longer repeats in the order of 100 amino acids that constitute structurally and functionally independent units. Here we analyse ShooT specific (ST) proteins, a family of proteins with tandem repeats of unknown function that were first found in Leguminosae, and their possible similarities to other proteins with tandem repeats. Results ST protein sequences were only found in dicotyledonous plants, limited to several plant families, mainly the Fabaceae and the Asteraceae. ST mRNAs accumulate mainly in the roots and under biotic interactions. Most ST proteins have one or several Domain(s) of Unknown Function 2775 (DUF2775). All deduced ST proteins have a signal peptide, indicating that these proteins enter the secretory pathway, and the mature proteins have tandem repeat oligopeptides that share a hexapeptide (E/D)FEPRP followed by 4 partially conserved amino acids, which could determine a putative N-glycosylation signal, and a fully conserved tyrosine. In a phylogenetic tree, the sequences clade according to taxonomic group. A possible involvement in symbiosis and abiotic stress as well as in plant cell elongation is suggested, although different STs could play different roles in plant development. Conclusions We describe a new family of proteins called ST whose presence is limited to the plant kingdom, specifically to a few families of dicotyledonous plants. They present 20 to 40 amino acid tandem repeat sequences with different characteristics (signal peptide, DUF2775 domain, conservative repeat regions) from the described group of 20 to 40 amino acid tandem repeat proteins and also from known cell wall proteins with repeat sequences. Several putative roles in plant physiology can be inferred from the characteristics found.