bluuebunny/crossref_metadata_2025_split huggingface dataset:
Calr Is up-Regulated in Patients with Essential Thrombocythemia Independently from Calr and JAK2 Mutations
Authors: Brown Lilia , Graham Ciaren , Rinaldi Ciro 
Published: October 2019

Abstract
Somatic mutations in exon 9 of calreticulin (CALR) gene were recently discovered in patients with essential thrombocythemia (ET) and primary myelofibrosis (PMF) lacking JAK2 and MPL mutations, and absent in patient with polycythemia vera (PV). Among patients with ET or PMF with un-mutated JAK2 or MPL, CALR mutations were detected in 67% of those with ET and 88% of those with primary PMF. Several types of insertions or deletions were identified and all resulted in a frameshift in exon 9 generating a novel C-terminal peptide in the mutant CALR protein. Over expression of the most frequent CALR deletion caused cytokine-independent growth in vitro owing to the activation of signal transducer and activator of transcription 5 (STAT5) by means of an unknown mechanism. Patients with myeloproliferative neoplasms carrying CALR mutations present with higher platelet counts and lower haemoglobin levels than patients with mutated JAK2. Studies suggest these patients have a lower risk of thrombosis and longer overall survival than patients with mutated JAK2.
We analysed by Real Time PCR, CALR expression in peripheral blood (PB) of 38 patients affected by ET, 17 JAK2 mutated (45%), 4 CALR (10.5%) mutated, 1 MPL mutated (3%) and 14 with no apparent molecular abnormalities. These were compared with a cohort of healthy volunteers. We found a significant over expression of CALR (median 5.15; range 1.13-270.08) comparing with controls (median 0.38, range 0.18-1). CALR mRNA expression is independent from the CALR mutational status. No significant difference was found comparing CALR expression in CALR mutated (median 4.9, range 1.51-37.14) and CALR/JAK2 un-mutated patients (4.68, range 1.51-28.71). CALR up-regulation is not mutually exclusive with JAK2 mutations; no difference was seen in CALR mRNA between JAK2 mutated (median 5.09, range 1.13-270) and wild type ET patients (median 5.08, range 1.51-37). There was no significant difference when we correlated CALR expression with PLT counts, spleen size or type of cytoreductive therapy. A larger cohort of patients is required to confirm these preliminary findings.

Disclosures
No relevant conflicts of interest to declare.
Mutually Exclusive Recurrent Somatic Mutations in MAP2K1 and BRAF Support a Central Role for ERK Activation in LCH Pathogenesis
Authors: Chakraborty Rikhia 
Published: October 2019

Abstract
Langerhans Cell Histiocytosis (LCH) is a myeloproliferative disorder characterized by lesions composed of pathologic CD207+ dendritic cells (DCs) with an inflammatory infiltrate. BRAFV600E remains the only recurrent mutation reported in LCH. In order to evaluate the spectrum of somatic mutations in LCH, whole exome sequencing (WES) was performed on matched LCH and normal tissue samples obtained from 41 patients. Lesions from other histiocytic disorders, juvenile xanthogranuloma (JXG), Erdheim-Chester disease (ECD), and Rosai-Dorfman disease (RDD) were also evaluated. All of the lesions from histiocytic disorders were characterized by an extremely low overall rate of somatic mutations. Notably, 33% (7/21) of LCH cases with wild-type BRAF and none (0/20) with BRAFV600E harbored somatic mutations in MAP2K1 (six in-frame deletions and one missense mutation) that induced ERK phosphorylation in vitro. Single cases of somatic mutations of the MAPK pathway genes ARAF and ERBB3 were also detected. The ability of MAPK pathway inhibitors to suppress MEK and ERK phosphorylation in cell culture and primary tumor models was dependent on the specific LCH mutation. The findings of this study support a model in which ERK activation is a universal endpoint in LCH arising from pathologic activation of upstream signaling proteins.

Disclosures
No relevant conflicts of interest to declare.
Angiogenesis Associated with Alterations of the Bone Marrow Microenvironment Predicts Multiple Myeloma Progression to Symptomatic Disease in Mice and Humans
Authors: Calcinotto Arianna , Ponzoni Maurilio , Ria Roberto , Grioni Matteo , Cattaneo Elena , Villa Isabella , Bertilaccio Maria Teresa Sabrina , Chesi Marta , Rubinacci Alessandro , Tonon Giovanni , Bergsagel Peter Leif , Vacca Angelo , Bellone Matteo 
Published: October 2019

Abstract
While multiple myeloma (MM) is almost invariably preceded by asymptomatic monoclonal gammopathy of undetermined significance (MGUS) and/or smoldering MM (SMM), the alterations of the bone marrow (BM) microenvironment that establish progression to symptomatic disease are circumstantial. Our aim was to identify changes in the BM microenvironment eliciting angiogenesis in MM.
We have found that in Vk*MYC mice harboring oncogene-driven plasma cell proliferative disorder, the progression from asymptomatic to symptomatic MM was characterized by substantial modifications of the BM microenvironment, including a switch from a T helper type 1 (Th1) to a Th2 response, and accumulation of CD206+Tie2+ macrophages, which eventually favored angiogenesis and increased in microvessel density (MVD). Notably, MVD was also increased at diagnosis in the BM of MGUS and SMM patients that subsequently progressed to MM when compared with MGUS and SMM that remained quiescent.
These findings suggest a multistep pathogenic process in MM, in which the immune system may contribute to angiogenesis and disease progression. They also suggest investigating MVD in asymptomatic patients as prognostic factor for the progression and outcome of this disease.

Disclosures
No relevant conflicts of interest to declare.
Synergistic in Vitro Activity of MOR202, a Human CD38 Antibody, in Combination with Pomalidomide
Authors: Endell Jan , Boxhammer Rainer , Steidl Stefan 
Published: October 2019

Abstract
Background: MOR202 is a fully human anti-CD38 antibody currently being tested in a Phase I/IIa clinical trial in multiple myeloma (MM). It mediates antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP) in MM cells with high potency (EC50 ~200 pM) representing a possible promising new therapy for MM patients. In this in vitro study, we evaluated the synergistic potential of MOR202 and pomalidomide (POM), a newly approved IMiD® immunomodulatory agent in MM therapy.
Methods: Using flow cytometry analysis, POM was evaluated in relation to its effects on several parameters anticipated to be relevant for anti-tumor activity when combined with MOR202. This included the induction of direct cytotoxicity and CD38 upregulation in several MM cell lines, as well as the activation of human immune effector cells derived from peripheral blood mononuclear cells of healthy donors. On a functional level the interaction of MOR202 and POM was assessed using FACS-based ADCC assays. Different incubation schemes prior to the ADCC assays were evaluated in order to distinguish the influence of POM on ADCC activity when pre-incubated for 72 hours either on target or effector cells or on both in parallel. The observed combination effects were analyzed for synergistic potential. Experiments were carried out in triplicate and mean values (±SEM) were calculated.
Results: POM as a single agent showed cytotoxic effects on MM cell lines with high potency (EC50 ~150 nM) and additionally induced an up to 2.7-fold upregulation of CD38 (EC50 ~20 nM) on CD38-expressing MM cell lines. Both effects were maximal at the last tested time point of 72 hours and strongest on cell lines with comparably lower CD38 expression levels. In combination with the observed activation of effector cells these POM-mediated mechanisms lead to a synergistically enhanced cytotoxic activity of MOR202. This synergistic benefit ranged between 1.2-fold and 3.1-fold above theoretical additivity depending on the cell line used and was most pronounced in the case of strong CD38 upregulation.
Figure 1 Exemplary ADCC dose-response curves for the comparably lower CD38 expressing cell line AMO-1 after POM pre-treatment of effector cells, target cells or both. Figure 1. Exemplary ADCC dose-response curves for the comparably lower CD38 expressing cell line AMO-1 after POM pre-treatment of effector cells, target cells or both.
Conclusions: The cytotoxic activity of MOR202 on MM cells was enhanced and synergized when combined with the immunomodulator agent POM via multiple mechanisms, namely direct cytotoxicity, CD38 upregulation and activation of effector cells. These results provide a mechanistic rationale for combining MOR202 and POM and warrant further evaluation in the clinical setting.

Disclosures
Endell: MorphoSys AG: Employment, Patents & Royalties. Boxhammer:Morphosys AG: Employment, Patents & Royalties. Steidl:MorphoSys AG: Employment, Patents & Royalties.
Evaluation of the Bacterial Etiologies of Infections in Patients Undergoing Hematopoietic Stem Cell Transplantation
Authors: Hizmali Lokman , Tapan Umit , Eraksoy Haluk 
Published: October 2019

Abstract

Objectives: Bacterial infections increase morbidity and mortality in patients undergoing hematopoietic stem cell transplantation (HSCT). The characteristics of the bacteria causing infections in these patients undergo dynamic changes both globally and locally during the transplantation process. The etiology of bacterial isolates and analysis of changes in the antibiotic sensitivities are crucial for the selection of appropriate prophylactic and empiric therapies. In this study we aimed to evaluate the first episode of bacterial infection in patients undergoing allogeneic or autologous hematopoietic stem cell transplantation and to identify bacterial isolates and the resistance profile of causative bacteria.
Materials and Methods: In this retrospective study, the charts of 195 patients who underwent allogeneic or autologous hematopoietic stem cell transplantation between January 2010 and December 2013 were reviewed. Ninety-six patients who had microbiologic evidence of bacterial infection were included in the study. The culture results from the first infectious episode during the transplantation process were evaluated. Patients were grouped according to the type of transplantation, and categorized according to the transplantation process, presence of neutropenia and infection status. Microbiological examination of samples obtained for culture and sensitivity analysis were performed according to standards of the National Committee for Clinical Laboratory Standards Institute (CLSI).
Results: Gram-negative and Gram-positive bacteria were identified in 54.2% and 44.8% of the infectious episodes, respectively. Poly-microbial etiology was identified in only 1.0% of these episodes. E. coli and coagulase-negative staphylococci (CoNS) were the most frequently isolated pathogens. In the early stage infections, Gram-positive organisms were more frequently isolated (72.1%) (p=0.042) and a significantly larger number of patients had undergone autologous stem cell transplantation (p<0.001). This situation was explained by the absence of antimicrobial prophylaxis in the autologous group, intensive induction chemotherapy, presence of catheter and mucosal damage. The frequency of extended-spectrum β-lactamase (ESBL) was found to be very high in the allogeneic group compared to the autologous group (36.4% vs. 13.3%). All of the staphylococcal isolates in the allogeneic group were noted to be methicillin resistant.
Conclusions: Treatment and prophylaxis have become increasingly more challenging with the infections caused by resistant pathogens. It is imperative to carefully monitor causative agents of bacterial infections and to plan prophylactic and empiric treatments according to characteristics of the individual patients.

Disclosures
No relevant conflicts of interest to declare.
Defibrotide for Prophylaxis of Hepatic Veno-Occlusive Disease in Autologous, Non-Purged Peripheral Stem Cell Transplantation for High Risk Neuroblastoma Patients
Authors: Unal Emel , Tacyildiz Nurdan , Yavuz Gulsan , Dincaslan Handan , Tanyildiz Gulsah , Ayyildiz Erol , Ozcan Muhit 
Published: October 2019

Abstract
High-dose chemotherapy (HDC) represents the standard of treatment for high-risk neuroblastoma(NBL), hepatic veno-occlusive disease (VOD) is a common, 10-50% and serious complication of haematological stem cell transplantation (HSCT), with up to 90% mortality rates. We planned study to assess whether the use of prophylactic defibrotide in paediatric patients who were heavily treated with chemo-radiotherapy before transplant, and then underwent autologous HSCT.
Seventeen patients who underwent autologous, unpurged peripheral stem cell transplantation PBSC with a high risk of developing VOD, between January 2003-July 2014, were given Defibrotide prophylaxis 25mg/kg/day for 30 days, commencing on -1 of conditioning regimen. All patients were stratified by INSS stage, age, N-MYC status. All were treated with six cycles of induction chemotherapy, myeloablative intensification, surgery for primary site, radiation therapy to the primary tumour site plus metastatic sites i.e bone metastases including skull. Meta-iodobenzylguanidin treatment as targeted radiotherapy was given on -21 prior to myeloablative chemotherapy. Oral 13-cis retinoic acid was employed on day +90 post-transplant. CD34+ cell mobilization and PBSC collection was carreid out after two-four cycles of induction cycles. Conditioning regimen and stem cell infusion was done following four-six weeks of last chemotherapy cycle, in order to reduce the toxicitiy. Conditioning regimen CEM consisted Carboplatin, Etoposide, Melfalan. There were no toxic deaths. All of the patients receieved antimicrobial prophylaxis and total parenteral nutrition support when was needed. Myeloid engraftment on day+13, erythroid engraftment on day+18 and thrombocyte engraftment was achieved on day +23.
Defibrotide prophylaxis seems to reduce incidence of VOD and is well tolerated. VOD incidence and severity was reduced in the defibrotide group which suggests that defibrotide might be effective in preventing and treating VOD. Sufficiently powered randomised trials are now required to definitively test the role of defibrotide in this setting.

Disclosures
No relevant conflicts of interest to declare.
HLA and Kir Mismatch Transplant Affect Cytopenia Beyond Day 28 after Thalassemia Transplant: A Primary Manifestation of Chronic Graft Versus Host Disease
Authors: Zhou Xiaohui , Li Chunfu , Liao Jianyun , Liu Xiangjun 
Published: October 2019

Abstract
Background Cytopenia beyond day 28 post-transplant (CB-28PT) following hematopoietic stem cell transplantation (HSCT) with β-thalassemia major (TM) rarely was reported. The exact mechanism for the development of CB-28PT is not well known.
Aim To find out causes of CB-28PT cytopenia.
Method We retrospectively analyzed data (HLA mismatch status, HLA antibody status of patients, KIR gene mismatch status, KIR-ligand matching status, donor/patient CMV status, donor/patient age and sex) of 93 TM patients underwent HLA 8/8 fully matched or 7/8 matched unrelated donor HSCT. All the patients used sole NF-08-TM protocol with median follow-up time of 19 (r: 2-44) months.
Results Results show a significant association between DRB1 mismatch and CB-28PT (P = 0.012). In addition, presence of Class I HLA antibody in the patient’s sera seems increase the chance of CB-28PT. Finally, the matching between inhibitory KIR2DL1 and their corresponding ligand HLA-C2 has a protective effect for CB-28PT.
Conclusion We propose that CB-28PT may be a primary manifestation of cGVHD in pediatric TM patients undergoing HSCT positive influenced by HLA DRB1 mismatch, HLA class I antibody and negatively affected by KIR-ligand match.

Disclosures
No relevant conflicts of interest to declare.
Generating Chimeric Antigen Receptors Utilizing Novel Anti-CD3 Nanobeads
Authors: Schultz Liora M. , Czerwinski Debra K , Fu Aihua , Levy Shoshana , Levy Ronald 
Published: October 2019

Abstract
The processes of ex vivo transduction of T cells to express chimeric antigen receptors (CARs) and of CAR+ T cell expansion influence the phenotype, function and ultimate fate of the final CAR+T cell product infused into patients. CAR constructs, despite expression of endogenous activation signals, require exogenous T cell activation during CAR transduction to allow optimal lenti-viral or retroviral-mediated integration of the CAR gene of interest into T cells. Clinical CAR therapy trials utilize anti-CD3 antibody-mediated activation or combined CD3 and CD28 stimulation using CD3, CD28 specific magnetic beads. We introduce novel magnetic nanoparticle beads generated from iron oxide nanoparticles conjugated to streptavidin and bound to biotinylated T cell activating antibodies for the purpose of CAR transduction. The small size of these nanobeads confers the advantage of decreased steric hindrance and enhanced capability of bead surface antibodies to access T cell surface antigen for binding and stimulation. We achieve efficient CAR transduction using anti-CD3 nanobead-mediated T cell stimulation and demonstrate CD19 specific CAR-mediated cytotoxicity of CD19+ tumor using an annexin V and 7AAD cytotoxicity assay. Evaluation of T cell phenotype following anti-CD3 nanobead-mediated T cell activation demonstrates preferential activation of naïve T cells as compared to central and effector memory cells. Addition of anti-CD28 costimulation is not necessary to achieving or inhibiting this preferential naïve T cell activation. Naïve T cells exhibit greater replicative capacity and anti-tumor function as compared to both effector and central memory T cells for adoptive transfer. We anticipate that preferential generation of naïve T cell derived CAR+ T cells achieved by introducing anti-CD3 nanobead stimulation can further improve the outcomes of clinical trials using CAR therapy.

Disclosures
Fu: NVIGEN Inc.: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties.
Retrospective Observational Study on Multiple Myeloma Cases Admitted to Kfsh – Dammam Between 1st of June 2006 till the End of December 2013
Authors: Abduljalil Omar Zeid , Mohiuddin Abid , Al Hashmi Hani Hassan 
Published: October 2019

Abstract
Introduction: Multiple myeloma (MM) is characterized by the neoplastic proliferation of a single clone of plasma cells producing a monoclonal immunoglobulin. It accounts for approximately 1 percent of all cancers and slightly more than 10 percent of hematologic malignancies in the United States. There is a scarcity on the information of myeloma in the kingdom of Saudi Arabia.Hence was this observational study from a single center in Dammam, Saudi Arabia. . Method:
Retrospective chart review of consecutive patients Diagnosed with Multiple myeloma in the period between 1st of May 2006 until end of December 2013 at King Fahd Specialist Hospital in Dammam (KFSHD). Purpose:
to describe the presenting characteristics of MM patients in our region, treatment patterns prescribed with the outcome of therapy that include progression free survival and overall survival. Result:
63 patients with multiple myeloma were diagnosed and treated during this period. The percentage of patients with ECOG (≥2) is 52.3 % . 60 % of the patients had stage III international staging system (ISS). Only 17.4% received Triple agents as primary therapy as most of our patients received duplet therapy. 46% received High Dose therapy and Autologous stem cell transplantation. the PFS was 41.1 % and the OS of 69.1% at 95 months. Conclusion:
Our results showed that we had rather younger patient population with a median age of 60 compared to the average western myeloma patient population. Despite the relatively young age those patients unfortunately had relatively poor performance status and high ISS stage which correlate with worse clinical outcome. However, the response to therapy and the outcome of our patients were superior to the outcome of any cohort of patients with similar disease characteristic. This observation worth further studying which could be due to an unknown disease or treatment related factors that might affect positively the patient’s outcome.

Disclosures
No relevant conflicts of interest to declare.
Interventions to combat Vitamin D Deficiency in patients undergoing Hematopoietic Stem Cell Transplantation
Authors: Pereira Andrea Z , Piovacari Silvia MF , Lucio Fabiana , Tanaka Marcia , Barrere Ana Paula N , Silva Juliana Bernardo , Ribeiro Andreza Alice Feitosa , Hamerschlak Nelson 
Published: October 2019

Abstract
Introduction: Patients undergoing Hematopoietic Stem Cell Transplantation(HSCT) may have low vitamin D (VD) level because of decreased exposure to sunlight, the major cause of VD Deficiency (VDD), from prolonged hospital stays, limited outdoor activity, and sunscreen use, and decreased oral intake caused by gastrointestinal treatment toxicity. Besides that gastrointestinal graft-versus-host disease (GVHD) limit absorption of VD. Some medications received during the HSCT can increased the VD catabolism, and alterate renal and kidney function.
Objectives: To evaluate the reduction of number of patients with VD Deficiency in patients undergoing HSCT after educational classes for the multi-professional team (physicians, dietitians and nurses).
Methods: We analyzed 72 patients undergoing HSCT May 2012 to January 2014 in the Hematology-Oncology and Bone Marrow Transplantation Center at Albert Einstein Hospital in São Paulo, Brazil. The serum levels of vitamin were measured in the first day of hospitalization of the patients adults (>= 18 years) who would be undergoing HSCT. All types of HSCT patients were included.We used in our study the VDD was defined and recommended by the Institute of Medicine as a 25(OH)D <=20 ng/ml, VD insufficiency of 21-29 ng/ml, and VD normal >=30 ng/ml.In the 2012 the multi-professional HSCT Team had 3 classes about VD and, everyone were informes about the VD research protocol.
Results: 72 adult patients were observed in this study, aged between 18 and 74 years, with the majority (77.8%) with less than 65 years. Of the total, 59.7% were men and 41.7% had normal body mass index. 100% of the lymphoma patients had VDD. In 2012(n:33), 60% patients had VDD and in 2013 (n:39), 40% (p<0,05).
Conclusions: When all of members of HSCT team were informed about the benefits of high VD levels in patients undergoing HSCT by classes and research protocol, we can reduce VDD.

Disclosures
No relevant conflicts of interest to declare.
Concurrent Tumors in Patients with Hematologic Malignancies
Authors: Sumoza Luis David , Messinger Marina , Sumoza Patricia , Aguirre Roxana , Harsha Poola , Avila Dayra 
Published: October 2019

Abstract
Background: Secondary tumors has been described in a variety of patients with either hematologic malignancies or solid neoplasm, and most of the time is related to previous chemotherapy and radiation therapy exposure, but little is found in the literature about synchronous or metachronous neoplasm that can be found in patient with a newly diagnosis of hematologic malignancy; we report 45 cases, presented in a single institution from 2007 to present.
Methods: A retrospective review of records using our tumor registry data, from patient with hematologic malignancies at John H. Stroger Hospital of Cook County, was performed and 45 patients with either synchronous or metachronous neoplasm were identified.
Results: Lung cancer was the most common malignancy representing 22.2%, Colorectal cancer 20%, Prostate cancer 17.7%, Breast cancer 11.1%, Urothelial cancer (Kidney and Bladder) 8.8%, Myelodisplastic syndrome, Pancreatic cancer, Thyroid cancer, Non-Hodgking lymphoma, Acute Myeloide Leukemia, Vulvar cancer, Testicular cancer and Skin Cancers 2.2% each one respectively.
Results: We found that in our heterogeneous population of patient with hematologic malignancies, the incidence of synchronous or metachronous neoplasm practically follows a very similar pattern of the general population, despite that many of this patients have been exposed to chemotherapy and radiation therapy.

Disclosures
No relevant conflicts of interest to declare.