bluuebunny/crossref_metadata_2025_split huggingface dataset:
Germline JAK2 Mutations In The Kinase Domain Are Responsible For Hereditary Thrombocytosis and Are Resistant To JAK2 and HSP90 Inhibitors
Authors: Marty Caroline , Saint-martin Cecile , Pecquet Christian , Grosjean Sarah , Saliba Joseph , Mouton Celine , Emilie Leroy , Harutyunyan Ashot S. , Abgrall Jean-François , Favier Remy , Solary Eric , Kralovics Robert , Constantinescu Stefan N. , Najman Albert , Vainchenker William , Plo Isabelle , Bellanné-Chantelot Christine 
Published: October 2019

Abstract
The main molecular basis of essential thrombocytemia and hereditary thrombocytosis are acquired and germline activating mutations affecting the thrombopoietin signaling axis, respectively. We have identified two families with hereditary thrombocytosis presenting novel heterozygous germline mutations of JAK2. One family carries the JAK2 R867Q mutation located in the kinase domain, while the other presents two JAK2 mutations, S755R/R938Q, located in cis in both the pseudokinase and kinase domains. Expression of JAK2 R867Q and S755R/R938Q induced spontaneous growth of Ba/F3-MPL but not of Ba/F3-EPOR cells. Interestingly, both Ba/F3-MPL cells expressing the mutants and platelets from patients displayed a TPO-independent phosphorylation of STAT1. JAK2 R867Q and S755R/R938Q, compared to JAK2 V617F, had significantly longer half-lives correlating with an increased MPL cell surface expression. Moreover, these mutations conferred a resistance to JAK2 and HSP90 inhibitors compared to JAK2 V617F. Our results suggest that the mutations in the kinase domain of JAK2 may confer a weak activation of signaling specifically dependent on MPL, while inducing resistance to clinically available JAK2 inhibitors.

Disclosures:
No relevant conflicts of interest to declare.
Stereotyped Subset #4 In Chronic Lymphocytic Leukemia Is Associated With Distinct Gene and Microrna Transcriptional Profile
Authors: Maura Francesco , Mosca Laura , Cutrona Giovanna , Matis Serena , Lionetti Marta , Fabris Sonia , Carlotta Massucco , Reverberi Daniele , Gentile Massimo , Recchia Anna Grazia , Monica Colombo , Bossio Sabrina , Negrini Massimo , Tassone Pierfrancesco , Rossi Davide , Gaidano Gianluca , Di Raimondo Francesco , Morabito Fortunato , Ferrarini Manlio , Neri Antonino 
Published: October 2019

Abstract
Chronic lymphocytic leukemia (CLL) is a lymphoproliferative disorder characterized by the monoclonal accumulation of B lymphocytes and a variable clinical course. Specific B-Cell Receptor (BCR) utilized by leukemic cells may influence disease progression and outcome. Highly homologous BCR, “stereotyped BCR”, are expressed in a recurrent fraction of patients with CLL and in some cases they were associated with distinct biological and clinical features. Stereotyped subset #4 have been reported to exhibit a favorable clinical course and to be the most frequent stereotyped BCR among the IGHV mutated (IGHV-M) cases. In this study we performed a comprehensive clinical, biological and molecular characterization of leukemic cells from 16 patients utilizing stereotyped subset #4 BCR (IGHV4-34) among a representative prospective cohort of 462 Binet stage A CLL patients enrolled in O-CLL1 protocol (clinicaltrial.gov identifier NCT00917540). In all cases, biological and molecular analyses were performed in peripheral CD19+ B-cells. All subset #4 patients were characterized by lower CD38 expression, unique IGHV-M configuration and absence of NOTCH1 and SF3B1 mutations. None subset #4 patients showed unfavorable cytogenetic deletions (i.e del11q23 and del17p13). Gene expression profiling (GEP) analysis was performed on 217 patients, including 9 subset #4 cases for whom RNA material was available. Supervised analysis comparing subset #4 vs all other patients (208) revealed 14 differentially expressed genes. Furthermore subset #4 patients were characterized by a significant downregulation of WDFY4, MEF2A and upregulation of PDGFA, FGFR1 and TFEC genes when compared with the remaining IGHV-M patients. miRNA profiles were analyzed in 229 patients including 10 subset #4 patients for whom RNA material was available. A specific miRNA expression pattern involving the upregulation of miR-497 and miR-29c was found in subset# 4 cases. Furthermore, we demonstrated that transfection of the miR-497 mimic in primary leukemic CLL cells induces, after 48 and/or 72 h, a downregulation of BCL2, known to be a validated target in different solid cancers. Our data provide a contribution to the biological definition of CLL patients with specific stereotyped IGHV4-34 BCR and identify for the first time distinct gene and miRNA expression profiles associated with this subset, providing further evidence of the putative leading role of HCDR3 conformation in CLL.

Disclosures:
No relevant conflicts of interest to declare.
YM155, a Survivin Suppressant, Induces Cell Death Via Suppression Of c-Myc Expression In Multiple Myeloma Cells
Authors: Ito Shigeki , Asahi Maki , Sasaki Ryousei , Oyake Tatsuo , Tamura Hideto , Ishida Yoji 
Published: October 2019

Abstract
Survivin is a member of the inhibitor of apoptosis protein (IAP) family with its dual roles in mitosis and apoptosis, and emerges as an attractive target for cancer therapy. Recent reports have demonstrated that survivin overexpression is associated with drug resistance and poor outcome in hematological malignancy including multiple myeloma (MM). YM155, a novel molecular targeted agent, suppresses survivin through the inhibition of transcription. However, the effect of this agent on MM cells has not been elucidated. In this study, we investigated the effect of YM155 on proliferation and survival of five human MM cell lines. YM155 inhibited the proliferation of these cells in a dose- and time-dependent manner (IC50 = 10 nM in 3 and 100 nM in 2 cell lines, respectively). Annexin V assay showed that YM155 induced apoptosis in these cells. To better understand these effects of YM155 on MM cells, we evaluated the intracellular signaling and apoptosis-associated protein status. Immunoblot analyses showed that YM155 reduced not only survivin but also myeloid cell leukemia sequence 1 (Mcl-1) and X-linked inhibitor of apoptosis protein (XIAP) expression. We also observed the activation of caspase-3 and poly(ADP-ribose) polymerase in YM155-treated cells, indicating that YM155 induces caspase-dependent apoptosis. YM155 did not affect phosphorylation status of Erk1/2 and STAT3. Interestingly, we found that YM155 suppressed c-Myc and interferon regulatory factor 4 (IRF4) expression, both of which are recognized as an important oncogene in the pathogenesis of MM. In addition, c-Myc and IRF4 protein levels were reduced at 6 and 12 hours after treatment with YM155, respectively. As IRF4 and c-Myc form a positive feedback loop in myeloma cells, this observation indicates that c-Myc inhibition by YM155 treatment might lead to subsequent inhibition of IRF4 expression, and thus raises the possibility of YM155 target for c-Myc rather than IRF4. We next examined the mechanism of downregulation of c-Myc in RPMI8226 cells. Real-time quantitative RT-PCR assay showed that YM155 treatment reduced c-Myc mRNA level. On the other hand, proteasome inhibitor did not prevent the suppression of c-Myc expression by YM155 treatment. These Results suggest that YM155 transcriptionally at least in part represses c-Myc in RPMI8226 cells. In conclusion, YM155 suppresses cell proliferation and survival in MM cells in part via not only inhibiting anti-apoptotic proteins such as survivin, Mcl-1 and XIAP but also repressing c-Myc oncogene. Further study is needed to clarify the molecular mechanism of downregulation of c-Myc induced by YM155. Our Results may provide a platform for clinical trials of YM155 in MM.

Disclosures:
No relevant conflicts of interest to declare.
Survival In Adult Acute Lymphoblastic Leukemia Survival By Age, Gender and Ethnicity
Authors: Ghimire Krishna B , Shah Binay K. 
Published: October 2019

Abstract

Background
Studies have shown improvement in survival in adult acute lymphoblastic leukemia (ALL) with the use of risk-directed therapy pediatric-inspired regimens. We analyzed US Surveillance, Epidemiology and End Results (SEER) cancer registry database to evaluate whether survival of adult ALL patients has improved in general population.

Methods
We analyzed US Surveillance, Epidemiology and End Results (SEER) 18 registries using survival session to calculate relative survival rates (RS) during 1973-1989, 1990-1999 and 2000-2009 among adult (≥20 years) ALL patients. We used Z-test in the SEER*Stat software to compare the relative survival rates in several cohorts categorized by race, gender, and age groups (20-39, ≥ 40 years).

Results
There were 7,435 adult ALL patients reported in SEER 18 registries during 1973-2009. The majority of patients were Caucasian (85%) and Male (57%).
The relative survival rates for adult ALL patients between 1973-2009 were 55.8±0.6% and 25.0±0.5% at 1 year and 5 years respectively. The relative survival rates improved significantly for each successive time periods, with an improvement from 45.4%, to 51.6%, to 60.6% at 1 year and, from 15.3%, to 23.7%, to 29.0% at 5 years for time periods 1973-1989, 1990-1999 and 2000-2009 respectively. For younger patients (age 20-39 years), 1- and 5-year RS rates improved from 1973-1989 to 1990-1999 but not from 1990-1999 to 2000-2009. For older patients (age ≥40 years) RS improved for each successive time periods.
The 1- and 5- year relative survival rates improved significantly for both male and female patients during successive time periods. Interestingly, although there was significant improvement in survival rates at 1- and 5- years for Caucasians, improvement was not seen for AA adult ALL patients during subsequent time periods examined.

Conclusions
Although adult ALL survival rates have improved for most cohorts examined, the survival of African American patients has not improved. Similarly, survival rates of younger patients (20-39 years) has not improved from 1990-1999 to 2000-2009.

Disclosures:
No relevant conflicts of interest to declare.
Access To Therapeutic Surgical Procedures For Pathologic Fractures In Multiple Myeloma Patients: Analysis of Nationwide Inpatient Sample (NIS)
Authors: Shah Nishi N , Nooka Ajay K. , Lonial Sagar 
Published: October 2019

Abstract

Introduction
Myeloma patients that experience a pathological fracture have 44% increased risk of mortality compared to their counterparts without pathological fractures. Given the negative impact of pathological fractures on survival, we have explored if access to obtain a therapeutic procedure influences the survival. We analyzed the nationwide inpatient cohort to evaluate for access to procedures and other hospitalization variables for myeloma patients admitted to the hospital for the diagnosis of pathologic fractures.

Methodology
Myeloma patients admitted in the hospital for the primary diagnosis of pathologic fractures from 2001-2010 were analyzed from the Nationwide Inpatient Sample (NIS). Procedures and diagnoses were identified using ICD-9-CM and NIS CCS codes. Variables of therapeutic procedures, length of stay (LOS), in-hospital mortality (IHM) and hospital charges were explored using multivariate logistic regression. Costs are derived from total hospital charges using cost-to-charge ratios based on hospital accounting reports from the Centers for Medicare and Medicaid Services and reflect the actual costs to produce hospital services.

Results
Of the 5154 myeloma patient admitted for pathological fractures, 2422 patients (47%) underwent a therapeutic procedure. The mean LOS for the primary diagnosis was 7.23 days (SE 0.14); cost for the hospitalization was $19344 and the IHM is 2.32%. The probability of getting a therapeutic surgical procedure and the cost of hospitalization is lower in patients >65, but had similar LOS and IHM compared to patients <65. Similar probability of not getting a surgical procedure was seen in medicare patients; but longer LOS may have contributed to the similar cost. However, in the medicaid patients, the odds of getting a surgical procedure was 66% low; the LOS and the cost are significantly high compared to the private insurance. IHM did not differ significantly across all groups.

Conclusions
This analysis addresses the question that older myeloma patients (age >65) and those that have the payer status of medicare and medicaid are significantly associated with the probability of not getting a therapeutic surgical procedure when they are admitted with pathological fracture. However, this did not result in increased in-hospital mortality suggesting that probably pathological fractures may reflect the biology of the underlying disease with negative impact on survival or the morbidity associated with a pathological fracture may impede the long term survival in myeloma patients.

Disclosures:
Lonial: Millennium: Consultancy; Celgene: Consultancy; Novartis: Consultancy; BMS: Consultancy; Sanofi: Consultancy; Onyx: Consultancy.
Long-Term Sequelae and Late Effects In Childhood Acute Lymphoblastic Leukemia (ALL) Survivors: A Single AIEOP Center Experience
Authors: Parasole Rosanna , Petruzziello Fara , De Matteo Antonia , Mangione Argia , Sepe Gaia , Iuliano Annunziata , Pinto Annamaria , Ferrante Maria Rosa , Vitale Virginia , Poggi Vincenzo 
Published: October 2019

Abstract
The advent of more intensive chemotherapy and the improvement of supportive cares have dramatically changed the natural history of childhood acute lymphoblastic leukemia (ALL), with current estimated 5-year overall survival of about 80%. The increased survival rate and the establishment of follow-up survey for long term survivors (LTS) have allowed the identification of late chemo-radiotherapy adverse effects on psychological and general health. We retrospectively evaluate the incidence and type of sequelae and / or late effects in a cohort of 301 childhood ALL LTS, followed in a single pediatric AIEOP center.
From June 1986 to June 2013, 301 LTS (154 male and 147 female), aged <18 years at ALL diagnosis, were followed-up by a multidisciplinary team. The surveillance protocol is summarized in Figure 1. The timing of follow-up (FU) was modified, case by case, in relation to the appearance of adverse events or organ diseases. Survivors' results were compared with chronic medical and psychological conditions of siblings (n=89).
Mean FU time (time from the stop therapy to the last control) was 6 years (range 1.8-26.8 years). The majority of LTS were teenager or young adults : 35% ranged between 15 and 20 years; 19.7% was more than 21 years old and the 45.3% was less than 14 years old. During FU, 16 late recurrences (5.3%) were identified and 3 secondary malignancies (0.99%) such as one mesenteric paraganglioma and two Acute Myeloid Leukemia in second complete remission ALL LTS. 40 patients (13.3%) received cranial radiotherapy during treatment. 39 LTS (13%) reported at least one sequelae. The most frequent sequelae were neurological and orthopedic (6% and 3% respectively) as summarized in Figure 2 151 LTS (50.17%) presented at least one late effect as showed in Figure 3.
In our experience at least one late effect occurred in 50.17% of LTS; these late complications affect negatively the quality of life of survivors. Endocrine-metabolic events are the most frequent late effect (34,5%). 13% of LTS have at least one sequelae mainly neurological and orthopedic. Prevention and/or early identification of complications during follow-up survey of LTS are crucial in order to decrease the long-term health risks associated with curative treatment for childhood ALL.

Disclosures:
No relevant conflicts of interest to declare.
Radiation Therapy In Peripheral T-Cell Lymphoma Of The Head and Neck
Authors: Fernandes Annemarie T. , Svoboda Jakub , Nasta Sunita D. , Wilson Lynn D. , Plastaras John P. 
Published: October 2019

Abstract

Introduction
The role of radiation therapy in peripheral T-cell lymphoma (PTCL) is not well established. While the NCCN guidelines recommend combined modality treatment with consolidative radiation therapy after chemotherapy for localized (stage I and II) disease, the data supporting this recommendation is lacking and is extrapolated from B-cell lymphoma.

Methods
This is a retrospective analysis of outcome in patients with non-cutaneous peripheral T-cell lymphoma of the head and neck from the Surveillance, Epidemiology and End Results (SEER) database diagnosed between 1981-2010. Survival was estimated by Kaplan-Meier estimates using the log-rank test. Univariate and multivariable analyses were performed using Cox regression analysis.

Results
Of the 307 patients analyzed, 130 (43%) underwent radiation therapy. The median age was 59 years old, 60.3% were male, 73.9% were Caucasian and the median year of diagnosis was 2004. The majority of patients had PTCL, Not Otherwise Specified (74.3%), followed by anaplastic large cell histology (22.2%). Patients were grouped as stage I (47.2%), stage II (29.9%) and stage III/IV (22.8%). Head and neck sites included the nasal cavity (28.3%), oropharynx (25.7%), oral cavity (21.2%), salivary gland (11.7%), nasopharynx (7.8%) and larynx/hypopharynx (5.2%).
The median follow-up was 54 months for survivors. Radiation therapy was associated with a higher 5-year overall survival in all patients (56.7% vs. 38.4%, p=0.001), patients with stage I disease (63.4% vs. 53.4%, p=0.036) and patients with stage II disease (60.8% vs. 36.3%, p=0.034), see Figure 1. Radiation therapy was not associated with a difference in overall survival in patients with stage III/IV disease (p=0.91).
Univariate analysis demonstrated that age, stage, radiation therapy and marital status were predictive of overall survival, but sex, and race were not. The year (p=0.93) or decade of treatment (p=0.67) did not impact overall survival. On multivariable analysis, increasing age (HR: 1.02, p<0.001), increasing stage (HR: 1.42, p=0.004), radiation therapy (HR: 0.60, p=0.011) and marital status of not married (HR: 1.49, p=0.035) remained statistically significant predictors of overall survival.
An additional analysis was performed excluding patients who survived less than 3 months to account for mortality related to disease progression or chemotherapy toxicity. After excluding these 45 patients, radiation therapy was still associated with increased overall survival in all patients (p<0.001) and in patient with limited stage disease (p=0.021).

Conclusions
The integration of local radiation therapy to the treatment of PTCL of the head and neck may improve overall survival in patients with limited stage disease.

Disclosures:
No relevant conflicts of interest to declare.
C5a Complement Depletion Suppresses In Vivo B Lymphoma Progression and Enhances Development Of Cellular Anti-Tumor Immune Response
Authors: Veeramani Suresh , Weiner George J. 
Published: October 2019

Abstract

Background
Proteins within the complement system have complex effects on cellular immune responses. In previous studies, we found that active complement components, especially C5a, can dampen the development of antigen-specific immune responses following vaccination with a model antigen, in part by promoting generation of APC-induced T regulatory (Treg) cells. These studies also demonstrated that B lymphoma cell lines exposed to complement can induce Treg generation in vitro. The current study was designed to address whether depletion of C5a could enhance development of a cellular anti-lymphoma immune response in vivo.

Methods
Immunocompetent Balb/C mice were inoculated subcutaneously with syngeneic A20 B lymphoma cells mixed with either 10 μg of rat anti-mouse C5a monoclonal antibody (mAb) or 10 μg of isotype-matched Rat IgG2a control mAb. Tumor growth was followed. In select experiments, mice were sacrificed and analyzed for the percentage and activity of tumor-infiltrating T cells and A20-specific splenic T cell responses.

Results
1. Tumor progression. Lymphoma grew more slowly in mice treated with anti-C5a mAb compared to mice treated with control mAb (p<0.05) {Fig. 1).
2. Intratumoral T cells. Tumors from mice treated with anti-C5a mAb had higher CD8+ T cell infiltration compared to mice treated with control mAb (p=0.002) (Fig. 2). Tumor-infiltrating CD8+ T cells showed a trend towards higher intracellular IFNg production in mice treated with anti-C5a mAb compared to control mAb (p=0.051).
3. Splenic T cells. Splenic T cells from mice treated with anti-C5a mAb produced IFNg to a greater degree than did splenic T cells from control mice when splenocytes were cultured with irradiated A20 cells in vitro (p=0.041) (Fig. 3). There was a trend towards decreased numbers of splenic CD4+CD25highFoxp3+ Tregs in C5a-depleted mice compared to control mice.

Conclusions
Depletion of C5a at the site of tumor inoculation slows tumor growth and increases the number of tumor infiltrating CD8 T cells in a syngenic immunocompetent model of lymphoma. A trend towards enhanced production of IFNg in the tumor infiltrating T cells, increased numbers of tumor-specific splenic T cells, and reduced numbers of splenic Tregs, suggests intratumoral C5a depletion can enhance tumor-specific immune responses both within the tumor and systemically. Ongoing studies are exploring the molecular mechanisms involved in C5a-promoted tumor progression and the use of C5a depletion as a novel strategy to improve anti-tumor immunity.

Disclosures:
No relevant conflicts of interest to declare.