bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Research: Treatment with Warfarin Followed By Hydroxyurea for Thrombosis Caused By JAK2V617F-Positive Essential Thrombocythemia Associated with Mutation in the Thrombomodulin Gene and Deficiency of Protein S
Authors: Sun Xingfu , Fu Jianxin , Li Jianyong , Lu Hua 
Published: November 2020

JAK2V617F-positive essential thrombocythemia is rarely associated with mutation in the thrombomodulin gene and deficiency of protein S without being reported in the literature. A case of a previously healthy 50-year-old Chinese female presenting with superior mesenteric vein thrombosis and portal thrombosis showed by enhanced computed tomography(CT) scan is reported. Blood platelet count showed 644×109/L. Molecular testing for genetic mutation found a JAK2V617F mutation by 11089 copies per 100ng DNA revealed by real-time polymerase chain reaction (RT-PCR), with the mutation rate being over ten percent. Level of free protein S decreased to 30.4%(reference:70%-140%). Heterozygous missense mutation(c.404C&gt;G, i.e.nucleotide 404 of encoding region was mutated from cytosine to guanine) in exon 1 in the thrombomodulin gene was detected in peripheral blood, which caused p.Pro135Arg(i.e.amino acid No.135 was changed from proline to arginine). The chromosomal position of thrombomodulin gene was at chr20:23029738. Family investigation showed that a similar mutation in the thrombomodulin gene was detected in peripheral blood in her daughter. Low molecular weight heparin was given at the beginning of treatment, and bleeding was monitored. In the meantime, warfarin was administered orally and the dosage of which was adjusted to maintain the international normalized ratio of prothrombin time at the range of 2 to 3. Adjust the dosage of hydroxyurea according to blood cell count. A month later, no new thrombosis was found in enhanced CT scan. It was an amazing feat that the patient was in a state of remission without new thrombosis being found for a year. JAK2V617F-positive essential thrombocythemia with mutation in the thrombomodulin gene and deficiency of protein S tends to be incurable. Warfarin alone with hydroxyurea may serve as an effective option to those cases.

Disclosures
No relevant conflicts of interest to declare.
Late Splenic Sequestration: A Consequence of Early Hydroxyurea Use?
Authors: Raghunandan Sharmila , Goubeaux Derrick L , Hollenkamp Christine R , Blaylark Rae M , Nelson Stephen C 
Published: November 2020

Background: Acute splenic sequestration is a potentially life-threatening complication in patients with sickle cell disease. It is the second leading cause of death in young children with hemoglobin SS/Sβ0 thalassemia (SCD). The incidence approaches 35% during the first two years of life in SCD. Although splenic sequestration can occur in patients with any sickle phenotype, patients with SCD tend to be significantly younger when compared to patients with hemoglobin SC disease. Splenic function may be preserved in children treated with hydroxyurea. We recommend hydroxyurea for all SCD patients diagnosed via newborn screening, with a goal of starting therapy before 12 months of age. With this aggressive approach and potential for maintaining some splenic function, this study evaluates acute splenic sequestration in this young cohort and compares this to institutional and historical controls
Methods: We performed a retrospective search of electronic medical records at Children's Minnesota from 1/1/1999 through 12/31/2019 to identify patients with SCD and splenic sequestration. We used a broad array of ICD-9 an ICD-10 codes in order to capture all appropriate patients. Following this initial search, in-depth chart review was performed to identify patients with SCD and confirmed splenic sequestration. Data collected includes age, sex, hydroxyurea use, splenectomy, and hematologic results including hemoglobin F percentage, if available. Student t-tests for independent samples were performed to compare cohorts. This study was granted exemption by our Institutional Review Board.
Results: Over this 21 year period, the initial search revealed 151 patients. Of those, 74 had SCD. In-depth chart review found 38 (24M, 14F) of these children were confirmed to have splenic sequestration. Twelve patients started hydroxyurea before 2 years of age at a mean age of 12.6 months, median 11 months (cohort A). Of these, six started hydroxyurea during infancy at a mean and median age of 6.5 months (cohort B). The remaining 26 patients were not on hydroxyurea at the time of splenic sequestration (cohort C). Children in cohorts A and B were significantly older at the time of splenic sequestration compared to patients in cohort C (mean 58 and 45.8 months vs. 14.9 months, p&lt; 0.0001).
Conclusions: Children with SCD who start hydroxyurea at a young age may develop splenic sequestration much later in life than previously expected. More study is needed to confirm this finding. It will be important to educate families and healthcare providers about this potential unintended consequence of early hydroxyurea use.

Disclosures
No relevant conflicts of interest to declare.

OffLabel Disclosure:
Hydroxyurea use in children with sickle cell disease &lt; 2 years of age
Developing a Simple Algorithm Based on Multiparameter Flow Cytometry for Fast Screening of Acute Promyelocytic Leukemia
Authors: Ceni Vitória Ceni , Pagnano Katia B , Duarte Gislaine B O , Pellegrini Marina DB , Duarte Bruno Kosa , Metze Konradin , Lorand-Metze Irene 
Published: November 2020

Introduction: Acute promyelocytic leukemia (APL) is a genetically and molecularly well-defined type of acute leukemia that is curable but has a frequent early mortality due to bleeding. So, there is a need for a fast diagnostic screening in order to start appropriate therapy. Multiparameter flow cytometry (MFC) is usually performed in all types of acute myeloid leukemias (AMLs) but only few features have been described as characteristic of APL.
Aim: to develop a diagnostic algorithm based on the intensity of expression of several antigens examined by MFC in AML that could reliably discriminate between APL and the other types of AML.
Material and Methods: Consecutive newly diagnosed AMLs treated in our Institution during the last 2 years entered the study. Immunophenotyping was included in the diagnostic workup. An 8-color platform based on the Euroflow recommendations was used. The mean fluorescence intensity (MFI) of each antigen tested was assessed and those best discriminating between APL and all other types of AML were obtained by a discriminant analysis. Phenotypic characteristics of normal myeloblasts taken from examinations of bone marrow (BM) MFC performed for the diagnosis of cytopenias were used as controls.
Results: 24 cases of APL and 56 cases of other primary AML entered the study. Median age: 39 (23-56) and 62(26-81) years respectively. Concerning ELN risk groups of non-APL cases, 13 were favorable risk, 26 were intermediate and 09 were adverse risk. In 8 cases risk assessment was not possible due to the absence of cytogenetics. Moreover, among APL patients, 7 cases had a FLT3-ITD mutation. Among non-APL AMLs, 4 had FLT3-ITD mutation, 4 had NPM1 and 10 had FLT3-ITD and NPM1mutation.
Concerning antigen expression, CD34 was expressed in only 1/24 APL samples, and in 18/56 samples from non-APL AMLs. The following flow features were differentially expressed in both groups: SSC (p &lt;0.0001), CD45 (p=0.02), CD13 (p=0.001), CD64 (p=0.004), HLA-DR (p&lt;0.0001) and CD33 (p&lt;0.0001) (Table 1). In the discriminant analysis, MFI CD34 and MFI HLA-DR were able to accurately classify APL and non-APL AML in only 62.5%. However, after the addition of the ratio of SSC between blasts and lymphocytes, these 3 parameters were able to differentiate APL from non-APL AML in 91.2% of the cases.
Conclusion: MFC was adequate for a fast screening of APL in most cases. Expression of CD34 was not very useful, as many AMLs do not express this antigen, similar to APL, but SSC, together with HLA-DR could discriminate both types of leukemia in most cases.

Disclosures
Pagnano: Astellas: Other: Advisory Board and lecture; Novartis: Other: Advisory Board; Pintpharma: Other: Lecture; EMS: Other: Lecture. Duarte:Janssen: Other: Lecture; Astellas: Other: Lecture.
Use of Ppmx-T003 As a Potent Inhibitor of Erythrocytosis in Polycythemia Vera
Authors: Masubuchi Nami , Araki Marito , Shioiri Kaho , Kitazawa Sota , Kihara Yoshihiko , Ohira Yuta , Nomura Fumiko , Matsuura Tadashi , Komatsu Norio 
Published: November 2020

Iron incorporation through transferrin (Tf) and its receptor, TfR1, plays a critical role in erythropoiesis. We hypothesized that blockade of TfR1 may be a promising strategy to suppress erythrocytosis observed in polycythemia vera (PV). To examine this possibility, we developed PPMX-T003, a monoclonal antibody against human TfR1, through phage display screening. PPMX-T003 was observed to bind to TfR1 with a high affinity(Kd= 3.56×10-10)and presented strong ability to suppress the growth of PV model cell lines such as HEL (EC50= 0.07 nM). PPMX-T003 induced TfR1 internalization but blocked the recycling of the receptor presumably because it was strongly bound, even in the endosome, which switched the fate of TfR1 from recycling to degradation. Consequently, the PPMX-T003 treatment greatly reduced the intracellular iron concentration, along withcell cycle arrest in the G2/M phase in HEL cells. In contrast, PPMX-T003 failed to block cell proliferation in human umbilical vein endothelial cells that expressed a subtle level of TfR1. These findings suggest that PPMX-T003 is a potent inhibitor for cells whose growth is dependent on iron incorporation by TfR1. This led us to examine whether PPMX-T003 blocked the endogenous erythroid colony (EEC) formation in erythroid progenitor cells in PV. We observed that PPMX-T003, at a concentration of 40 ng/mL, exhibited complete blockade of EEC formation inJAK2 V617F-positive PV patients' cells. Although hydroxycarbamide (HU) and phlebotomy are well-established treatment strategies for the management of hematocrit in PV patients, there is a risk of secondary leukemia associated with HU treatment and side effects such as fatigue and restless leg syndrome due to phlebotomy-induced iron deficiency. Therefore, we suggest PPMX-T003, which may preferentially suppress erythrocytosis without changing the iron level in peripheral blood, as an alternative treatment for PV patients.

Disclosures
Ohira: PPMX: Current Employment, Current equity holder in private company. Nomura:PPMX: Current Employment. Matsuura:PPMX: Current Employment. Komatsu:Takeda Pharmaceutical Co., Ltd, Novartis Pharma KK, Shire Japan KK: Speakers Bureau; AbbVie: Other: member of safety assessment committee in M13-834 clinical trial.; PPMX: Consultancy, Research Funding; Meiji Seika Pharma Co., Ltd.: Patents & Royalties: PCT/JP2020/008434, Research Funding; Otsuka Pharmaceutical Co., Ltd., PharmaEssentia Japan KK, AbbVie GK, Celgene KK, Novartis Pharma KK, Shire Japan KK, Japan Tobacco Inc: Consultancy; Otsuka Pharmaceutical Co., Ltd., Shire Japan KK, Novartis Pharma KK, PharmaEssentia Japan KK, Fuso Pharmaceutical Industries, Ltd., Fujifilm Wako Pure Chemical Corporation, Chugai Pharmaceutical Co., Ltd., Kyowa Hakko Kirin Co., Ltd., Takeda Pharmaceutica: Research Funding.
A Novel t(9;22;11) Translocation Variant Involving 11q23 in a Patient with Chronic Myeloid Leukemia
Authors: Yuehui Lin , Zheng Qinglong , Wu Tong , LIU DAN 
Published: November 2020

Background:The progression of Philadelphia chromosome positive chronic myeloid leukemia (CML) is frequently accompanied by cytogenetic evolution, commonly unbalanced chromosomal changes. but balanced chromosomal translocations are very rare in CML, especially translocations involving the 11q23. The few reported cases with blast phase (BP) of CML carrying a 11q23 rearrangement results in insufficient responses to tyrosine kinase inhibitors (TKIs) and possess a poor prognosis.
Methods:Cytogenetic analysis , fluorescence in situ hybridization (FISH), RNA sequencing (RNA-seq), targeted genomic sequencing and multi-parametric flow cytometry analysis were performed to identify the chromosome translocations and pathogenic gene alterations in a 36-year-old female with myeloid BP of CML.
Results: In BP, the bone marrow (BM) aspiration showed 61% myeloid blasts; Multi-parametric flow cytometry analysis revealed the abnormal myeloid blasts expression of the following antigens: CD117, CD13, CD33, CD38, partially expressed CD15, CD64. Chromosome analysis revealed a t(11;22)(q23;q11) translocation in addition to the t(9;22)(q34;q11). Fluorescence in situ hybridization (FISH) test confirmed that the t(11;22)(q23;q11) involved the mixed lineage leukemia(MLL) gene on 11q23 and RNA sequence revealed MLL-SEPT5 and BCR/ABL1(p210) fusion transcripts positive. Mutations on 339 commonly mutated genes in hematologic malignancies were analyzed by targeted next-generation sequencing showed ASXL1 p.G949Vfs*2 mutation.
The patient failed to respond to both imatinib and dasatinib despite the absence of resistance-associated mutations in the BCR/ABL1 gene and she had a myeloid blast crisis at 19 months after initiation of first- and second-generation TKI treatment. After BP, she received ponatinb, a third-generation TKI with chemotherapy. Regretly she didn't achieve a complete remission(CR) and was in the process of salvage transplantation at present.
Conclusions:The presence of 11q23 rearrangements in BC of CML is rare and most likely accounts for the adverse clinical outcome. We first report a patient who diagnosed CML with t(11;22)(q23;q11) and MLL-SEPT5 fusion gene positive in BP of CML. The clinical course was aggressive, and therapy was poorly tolerated.

Disclosures
No relevant conflicts of interest to declare.
Activating NOTCH1 Mutations Alter T-ALL Chemosensitivity By PREX2-AKT Pathway
Authors: Yu Hui , DU Yuxin , Xu Ji , Sun Zhenchang , Fu Xiaorui , Young Ken H. , Zhang Mingzhi 
Published: November 2020

Mutations on NOTCH1 gene are the most commonly found mutations in T-cell acute lymphoblastic leukemia (T-ALL) and they are reported to be favorable indicators for T-ALL patients' prognosis. However, the effects of activating NOTCH1 mutations on T-ALL's chemosensitivity have not been studied. We reported that NOTCH1 inhibition by γ-secretase inhibitors or shRNA knockdown in MOLT-3 cells could reduce the T-ALL cells' sensitivity to chemotherapeutic drugs. However, this effect was absent in Jurkat and CUTLL cells. We further demonstrated that NOTCH1 inhibition could activate the PI3K-AKT pathway in a cell specific pattern similar as their effects on chemosensitivity. RNA-seq revealed that Phosphatidylinositol-3,4,5-Trisphosphate Dependent Rac Exchange Factor 2 (PREX2) is a target gene of NOTCH1 and may mediate the effects of activating NOTCH1 mutations on chemosensitivity. Consistently, we proved that overexpression of PREX2 could mimic the effects of NOTCH1 inhibition on chemosensitivity. Our study has highlighted the effects of activating NOTCH1 mutations on T-ALL's chemosensitivity by altering PREX2-AKT pathway, which may explain the favorable effects of NOTCH1 mutations on T-ALL patients' prognosis, as well as provided potential targets to alter T-ALL cells' chemosensitivity.

Disclosures
No relevant conflicts of interest to declare.
Anti CD19/22 Cocktail CAR T-Cell Therapy Can Improve the Outcomes of Patients with TP53-Mutated Relapsed/Refractory B-Cell Lymphoma
Authors: Wei JIA , Xiao Min , Mao Zekai , Wang Na , Li Chunrui , Cao Yang , Sun Weimin , Xiao Yi , Gu Chaojiang , Zhang Shangkun , Zhang Yicheng , Zhang Tongcun , Zhou Jianfeng , Huang Liang 
Published: November 2020

Mutations of the Tp53 gene occur in a subset of patients with refractory/relapse B-cell lymphoma and confer an exceedingly adverse prognosis. Chimeric antigen receptor-modified (CAR) T-cell immunotherapy represents a novel promising treatment and has achieved impressive anti-tumor responses in patients with refractory or relapsed B cell malignancies. Whether CAR T cell therapy can improve the outcomes of refractory/relapse B-cell lymphoma with Tp53 mutations or high-risk functional classification of Tp53 mutations has not been fully investigated yet.
In phase 2 trial, we evaluated the anti-CAR 19/22 cocktail therapy in patients with relapsed or refractory B-cell lymphoma with Tp53 mutations. The primary endpoint was the percentage of patients with an objective response (complete and partial response). The high-risk Tp53 mutations were defined as missense mutations, 'disruptive' mutations and the 'high-risk group of evolutionary action (EAp53) score. The outcome impact of CAR T cell therapy on high-risk Tp53 mutations were also assessed.
Between September 2016 to January 2020, a total of 30 patients [median age, 44.0 years (range: 28.0 to 61.0)] were enrolled. The primary efficacy analysis showed that 76.7 % (95%CI: 61% to 93%) patients have best objective response(Figure 1A), while 60.0 % (95%CI: 41% to 79%) patients at month three maintained the objective response(Figure 1B). At a median follow-up of 7.95 months (range: 1.13 to 42.76), the 12-months progression-free survival and overall survival rate were 43.33% and 45.33% respectively(Figure 1C-D). The therapy led to similar serious and life-threatening toxic effects with those reported with other CAR T-cell therapies. For all 30 enrolled patients, total 32 Tp53 Mutations were detected in this cohort including 23 missense, 4 splice site mutations, 4 insertions, 2 deletions and 1 nonsense mutation. Two Tp53 mutations were found in 2 patients. 21 patients were assessed with Tp53 FISH and 14 patients (66.66%) had 17p-. There is no difference on ORR,OS and PFS in patients based on three different Tp53-specific scoring systems. The ORR at three months predicts the long-term survival of this group of patients (Figure 1E-F).
CAR19/22 T-cell cocktail therapy can improve the outcome of r/r B-cell lymphoma with high-risk of functional classification of Tp53 mutation. The patients who achieved overall response at month three can greatly benefit from CAR19/22 T-cell cocktail therapy with long-term favorable outcome.
Trial registration: ChiCTR-OPN-16008526.
Key words: B cell lymphoma; CAR T-cell therapy; Tp53 mutation; outcome
Figure 1

Disclosures
No relevant conflicts of interest to declare.
The Cytogenetic Aberrations of Lymphoplasmacytic Lymphoma/ Waldenström's Macroglobulinemia in China
Authors: Wenjie Xiong , Yi Shuhua , Liu Huimin , Wang Tingyu , Liu Wei , Zou Dehui , Qiu Lugui 
Published: November 2020

Object:Lymphoplasmacytic lymphoma/ Waldenström's macroglobulinemia (LPL/WM) is a rare B-cell proliferative malignancy characterized by immunoglobulin M monoclonal gammopathy and /or bone marrow infiltration by lymphoplasmacytic cells. Recently, there are no large-scale studies to detect the clinical features, genetic background in patients with LPL/WM in China even Asia. The aim of this study is to explore the clinical and genetic differences, observe the impact of cytogenetic abnormalities on survival and prognosis in Chinese patients.
Methods:305 patients with LPL/WM admitted in our hospital from June 1998 to December 2018 were retrospectively analyzed. Baseline characteristics, laboratory detection and therapy were compared using Chi-square test and Kaplan-Meier methodology was undertaken for survival analysis.
Results:The median age of 305 patients in the study was 62 years (range 32-87) and male to female ratio was 2.45:1(216/89). The most common symptoms at diagnosis were fatigue (64.9%) and bleeding (14.1%).The patients in low, intermediate and high risk group of the international prognostic scoring system for WM (ISSWM) was 17.6%, 36.5% and 45.9%,respectively.The median hemoglobin was 8.5g/dl(range 2.4-18.7). About 41.3% patients had IgM higher than 4000mg/dl. The cytogenetic abnormalities were uncommon in LPL/WM patients. The chromosomal karyotypes were successful in 194 cases, of which 20 patients showed clonal abnormalities. 12 patients had two or more chromosome abnormalities. The most common chromosome abnormalities were deletion 6/6q(25%) and Y chromosome loss (20%). The median progression-free survival (PFS) and overall survival (OS) in patients with normal and abnormal chromosome karyotypes were similar. 194 patients had FISH detection, of which 33 patients (17%) had FISH abnormalities, 9 (4.6%) patients had two kind of abnormalities. The patients with more than one cytogenetic abnormality had lower survival than just had one abnormality or normal patients. The most common aberrations were 6q deletion and trisomy 12. 17p deletion was detected in 11 patients (11/184,6.0%) and it was a poor prognostic marker of PFS and OS. Single 13q14 deletion had not impact on PFS or OS, but when companied with other cytogenetic aberration, it could worsen the PFS and OS . Besides related to survival, cytogenetic abnormal was also related to the clinical features.
Conclusions:Despite the low incidence and abnormal cytogenetic detection rate of LPL/WM, the clinical and genetic features are unique in Chinese patients. The cytogenetic aberrations play an important role in predicting prognosis.

Disclosures
No relevant conflicts of interest to declare.
Consecutive Hypoalbuminemia Predicts Inferior Outcome in Patients with Diffuse Large B-Cell Lymphoma
Authors: Wei Xiaolei , Zheng Jingxia , Zhang Zewen , Liu Qiongzhi , Zhan Minglang , Huang Weimin , Liu Jianbo , Wei Qi , Wei Yongqiang , Feng Ru 
Published: November 2020

Albumin at diagnosis can be used to predict survival in diffuse large B-cell lymphoma (DLBCL). But whether consecutive albuminemia, which seen in DLBCL patients from time to time, could be more accuracy to predict outcome remains unknown.
We retrospectively analyzed 574 de novo DLBCL patients treated with R-CHOP from our and other 2 centers. All the patients were divided into a training cohort (n=278) and a validation cohort (n=296) depending on the source of the patients. Overall survival (OS) and progression-free survival (PFS) were analyzed by the method of Kaplan-Meier and the differences between groups were compared by the log-rank test. Multivariable analysis was conducted by Cox proportional hazard regression model.
The cutoff value of albumin for survival analysis was 39.2g/L with an area under the curve value of 0.589±0.034 (p=0.009) by receiver operating characteristic curve. In the training cohort, 163(58.6%) patients had low serum albumin at diagnosis and 80 of them were present with consecutive hypoalbuminemia. Patients with consecutive hypoalbuminemia showed inferior OS and PFS (p=0.010 andp=0.079, respectively). Similar survival differences were also observed in the independent validation cohort (p=0.006 andp=0.030, respectively). Multivariate analysis revealed that consecutive hypoalbuminemia was an independent prognostic factor OS (relative ratio [RR], 0.499; 95% confidence interval [CI], 0.317-0.786,p=0.003) and PFS (RR, 0.576; 95% CI, 0.413-0.804,p=0.001) in all entire DLBCL patients independent of IPI.
In conclusion, our study suggests that consecutive hypoalbuminemia is a simple and effective prognostic factor in DLBCL patients, allowing the identification of an inferior outcome subgroup in DLBCL patients, which may need to closely follow up.

Disclosures
No relevant conflicts of interest to declare.