bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti-CD 19 and Anti- CD 20 Chimeric Antigen Receptor-Modified T Cells for B-Cell Malignancies: A Systematic Review and Meta-Analysis
Authors: Riaz Irbaz Bin , Husnain Muhammad , Kamal Muhammad Umar , McBride Ali , Hua Anh , Geoge Laeth , Hamadani Auon Abbas , Rahman Bushra , Ali Zeeshan , Sipra Qurat-ul-Ain Riaz , Zahid Umar , Raina Ammad , Saleh Ahlam , Anwer Faiz
Published: October 2019

Background:

Chimeric antigen receptor (CAR) modified T cells targeting CD19 and CD20 have shown activity in Phase I, II trials of patients with hematological malignancies. CD19 and CD20 targeted CAR constructs from several different institutions have demonstrated consistently high anti-tumor efficacy in children and adults with relapsed B-cell acute lymphoblastic leukemia (B-ALL), chronic lymphocytic leukemia (CLL) and B-cell non-Hodgkin lymphoma (B-NHL). We conducted a systematic review and meta-analysis of all published clinical trials studying the role of efficacy as well as safety of CD-19 and CD-20 CAR-T therapy for B-cell hematologic malignancies.

Methods:

Literature search was performed using MEDLINE (Ovid SP and PubMed), EMBASE, The Cochrane Library, Scopus and Web of Science. We conducted meta-analysis using random effects model using Comprehensive Metaanalysis 3.0. Heterogeneity was assessed using Q-statistic and it was quantified using I2statistic.

Results:

After a comprehensive literature search 4476 studies were identified and finally 15 eligible studies involving 121 patients were included in the final systematic review. The infused CAR T-cell dose was in the range of 0.76 x 106 to 3 x 107 cells/kg in seven trials and in the range of 0.8 x 107 cells to 3.3 x 109 cells/m2in 6 trials. Among 106 patients, the overall response to treatment after CAR T-cell infusion were 33% complete remission (CR), 30 % partial remission (PR), 21% stable disease (SD), and 8% progressive disease (PD). Three studies described response to immunotherapy as minimal residual disease negative (MRD-) and as no evidence of disease (NED) and after including it, the overall CR was 38%. Major adverse events included fever (38%), hypotension (23%), chills (29%), rigor (28%), fatigue 17% and dyspnea 10%. One patient died from cytokine release syndrome, which is potentially serious complication. Neurological symptoms require prompt recognition and management.

Conclusion:

CAR T cell therapy is a promising treatment for refractory and relapsed hematological malignancies. The durable responses of CAR T cell therapy help integrate it in standard treatment protocols. Patients who are refractory to standard salvage chemotherapy or relapse after allo - HSCT have overall poor prognosis and can potentially achieve remission again with CAR T cell therapy.

Disclosures

McBride: Sanofi: Research Funding. Anwer:Seattle Genetics: Other: Advisory Board Participant; Incyte: Speakers Bureau.
TIM3 Dynamically Expressed on Peripheral Blood NK Cells in Acute Myeloid Leukemia Patients: The Expression Decreased When Untreated, and Restored While Disease Improved
Authors: Li Caixia , Chen Xiaochen , Gu Caihong , Wang Tong , Zou Qiu , Yang Dan , Wu Depei
Published: October 2019

Objective: Preliminary study the expression and clinical significance of TIM3 on peripheral blood NK cells in patients with acute myeloid leukemia (AML). Methods: We investigated the expression characteristics of TIM3 on the peripheral blood NK cells of newly diagnosed AML patients and its clinical significance. Peripheral blood was obtained from 50 patients with newly diagnosed AML before intervention, with peripheral blood from 30 cases of healthy volunteers collected as normal control. Expression levels of TIM3 on the peripheral blood NK cells and subsets were assayed with flow cytometry. Then the expression of gamma interferon in Tim3+CD56+NK cells and Tim3-CD56+NK cells were analyzed by intracellular immunofluorescence labeling and flow cytometry. Results: Compared with the normal control, in the newly diagnosed AML patients, the NK cells percentage and the proportion of CD56dimNK/CD56+NK reduced, while the proportion of CD56brightNK/CD56+NK increased significantly, and the proportions of Tim3+CD56+NK/CD56+NK and Tim3+CD56dimNK/CD56dimNK decreased significantly. However, in AML patients with complete remission after chemotherapy, the proportion of peripheral blood NK cells and cell subsets recovered. Furthermore, compared with Tim3-CD56+NK cells, IFN-γsecretion level of Tim3+CD56+NK cells in the newly diagnosed AML patients is increased significantly. Conclusion: The dynamic expression of TIM3 on NK cell and its cell subsets before and after chemotherapy in AML patients suggest that Tim3 and NK cells may play an important role in the occurrence and development of AML.

Disclosures

No relevant conflicts of interest to declare.
Prognostic Impact of FLT3-ITD and DNMT3A R882 Double Mutations in Patients with Acute Myeloid Leukemia
Authors: Tang Shanhao , Tang Xiaowen , Shen Hongjie , Xue Shengli , Wang Tingjing , Lu Jing , Yin Jia , Wu Depei
Published: October 2019

Background: FLT3-ITD and DNMT3A R882 mutations are both independent factors for poor prognosis of acute myeloid leukemia (AML). Allogeneic hematopoietic stem cell transplantation (allo-HSCT) has been confirmed to improve the outcome of AML with either of gene mutations. However, the clinical characteristics of AML with both of FLT3-ITD and DNMT3A R882 mutations and their outcome post allo-HSCT had been rarely evaluated.

Objective: To investigate the clinical characteristics and transplantation outcomes of AML patients with the double mutations.

Methods: A total of 241 patients were included in this retrospective analysis. Molecular genetic mutation sets in bone marrow (including FLT3-ITD, DNMT3A, C-kit, CEBPA, FLT3-TKD, and NMP1) of 241 AML patients were detected at diagnosis using direct sequencing method. And the clinical data of these patients were analyzed retrospectively.

Results: (1)According to the genetic mutation types, the 241 AML patients were divided into four groups: FLT3-ITD+DNMT3A R882+, FLT3-ITD+DNMT3A R882-, FLT3-ITD-DNMT3A R882+ and FLT3-ITD-DNMT3A R882- groups, which consisted of 19, 38, 21 and 163 patients, respectively. (2) Patients with FLT3-ITD+DNMT3A R882+ have high white blood cell count and a low complete remission rate after first induction chemotherapy (52.6%), which are common in M5、M2、M4 with normal chromosome karyotype, higher early cumulative incidence of relapse(6m-CIR 36.8±1.3%、12m-CIR 49.7±1.6%) and cumulative mortality rate (6m-CMR 26.3±10.1%、12m-CMR 48.5±11.8%) . (3) 2-year CIR of the FLT3-ITD+DNMT3A R882+ was (72.2±2.7%), which was significantly higher than that of the FLT3-ITD+DNMT3A R882- group (38.6±0.7%) and FLT3-ITD-DNMT3A R882+ group (36.8±1.6%). While the 2-year overall survival (OS) rate and 2-year leukemia-free survival (LFS) rate of double mutation group were (30.9±13.3% ) and (11.3±10.2%), respectively, which were considerably lower than those of the FLT3-ITD+DNMT3A R882- and FLT3-ITD-DNMT3A R882+ groups (2y-OS: 67.5±7.8%, 61.4±12.4%: 2y-LFS: 47.9±8.4%, 56.8±12.5%). Meanwhile, the 2-year CIR rates of the FLT3-ITD+DNMT3A R882- and FLT3-ITD-DNMT3A R882+ groups were significantly higher than that of the FLT3-ITD-DNMT3A R882- group (16.3±0.1%), while their 2-year OS rate and 2-year LFS rate were significantly lower than those of the FLT3-ITD-DNMT3A R882- group (2y-OS: 82.5±3.1%、2y-LFS: 80.9±3.2%). (3) Univariate and multivariate analyses revealed that disease status prior to transplantation, grade III-IV aGVHD, FLT3-ITD, DNMT3A R882 and FLT3-ITD+DNMT3A R882+ were independent factors for poor prognosis post transplantation.

Conclusion: AML patients with FLT3-ITD and DNMT3A R882 double mutations still present a very poor prognosis after transplantation with low OS and LFSas well as a high CIR rate. Therefore, it is necessary to explore prevention strategy for relapse post transplantation.

Disclosures

No relevant conflicts of interest to declare.
CD30 Expression and Its Correlation with Clinicopathologic Features in De Novo Diffuse Large B-Cell Lymphoma
Authors: Qian Shenxian , Zhao YiFei
Published: October 2019

Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease disease with 40% of patients presenting as refractory/relapsing disease following R-CHOP treatment. Recently, CD30 was reported as a useful predictor with favorable clinical outcome. However, CD30 expression patterns and the clinicopathologic features of CD30 positive DLBCL are not well described thus far, Here, we studied149 patients with de novo DLBCL to investigate clinicopathologic features of CD30-positive DLBCL by immunohistochemistry. useing a >0% cut-off,CD30 was expressed in approximately 13% (19)patients with DLBCL, showing a lower frequency of MYC/BCL2 co-expression. The clinic features were very similar between the CD30+ and CD30- groups,such as age, sex, B symptoms, staging, ECOG PS, LDH level, extranodal site involvement, IPI, GCB or non GCB type, bone marrow involvement. By univariate analysis Patients with CD30-positive DLBCLs showed better progression-free survival and overall survival compared with patients with CD30-negative DLBCLs, although the superior outcome of CD30 positivity had minimal effects on DLBCL with MYC/BCL2 co-expression. Epstein-Barr virus (EBV) was identified in 8(5.3%) cases, all of which were almost exclusively positive for CD30 expression (8/8) (P < 0·001). We conclude CD30 is expressed in a substantial proportion of DLBCL and CD30 immunohistochemistry may be a useful prognostic marker in R-CHOP treated GCB-DLBCL. The significant association of CD30 with EBV-positive DLBCL suggests a distinct pathobiology for these cases. We concluded that CD30 may be useful as a prognostic marker in rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) treated DLBCLs, indicating favorable outcomes

Disclosures

No relevant conflicts of interest to declare.
Co-Expression of MYC, BCL2 and PD-L1 in Primary Central Nervous System Diffuse Large B Cell Lymphomas ( PCNSL)
Authors: Saad Nibal , Anand Ankit , Naous Rana , Neerja Vajpayee , Zhang Shengle , Gentile Teresa
Published: October 2019

Background

Increased protein expression and gene translocations of MYC and BCL2/BCL6promote increased cell growth and inhibit apoptosis. They are associated with inferior prognosis in systemic diffuse large B cell lymphomas (DLBCLs). Their role in primary PCNSLs however is less clear. The immune checkpoint molecules, programmed death 1 (PD1) and its ligand PD-L1 are involved in inhibitory pathways to maintain immune tolerance and it is now clear that tumor cells interfere with these pathways to promote escape from immune surveillance. Antibody blockade of these immune checkpoints has been shown to enhance antitumor immunity and is emerging as an important target for cancer immunotherapy. We examined the co expression of these proteins on tumor samples from patients with PCNSL .

Methods

Immunohistochemistry (IHC) for MYC, BCL2 and PDL-1 and florescence in situ hybridization ( FISH) for MYC (8q24), BCL2 ( 18q21) and BCL6 (3q27) were performed on tumor samples from 14 patients with PCNSL.

Results

Eleven patients were positive for PDL-1 (79%) by IHC. Of these eleven patients, six (55%) were also positive for both MYC and BCL2 by IHC and negative by FISH. Eight patients (57%) were positive for MYC and BCL2 by IHC but negative for PD-L1. One patient was positive by FISH for MYC, BCL2, and BCL6 but negative for PDL-1.

Conclusions

Our results, although in a small number of patients, demonstrate that a large percentage of patients with PCNSL who express MYC and BCL2 also express PD-L1.It has been suggested that the frequent over expression of MYC, BCL2, and BCL6 noted in PCNSL may underlie the inferior prognosis as compared to systemic DLBCL. We suggest that the co-expression of MYC/ BCL2 with PD-L1 may contribute to this poor prognosis by engaging the complimentary mechanisms of increasing cell growth, inhibiting apoptosis and evading immune surveillance. The use of PD-L1 inhibitors in hematologic malignancies is limited but has demonstrated clinical responses in relapsed/refractory disease following multiple lines of therapy and warrants further investigation in PCNSLs.

Table Table.

Disclosures

No relevant conflicts of interest to declare.
Molecular Predictors of Treatment Failure in ALK-Negative Anaplastic Large T-Cell Lymphoma
Authors: Gorenkova Liliya , Kravchenko Sergey Kirillovich , Mkrtchyan Asmik , Musyirin Vsevolod , Misyurina Anna Evgenevna , Savchenko Valeriy G.
Published: October 2019

ALK-negative anaplastic large T-cell lymphoma (ALK-negative ALTCL) is a rare aggressive disease, constituting 12% of all T-cell lymphomas of adults. When using CHOP/CHOEP chemotherapy overall and event-free survival is less than 50%. Factors of poor prognosis include IPI (3-5), CNS and skin involvement as well as molecular predictors.

The aim of current study was to detect rearrangements (R) of DUSP22 gene, t(6;7)(p25.3;q32.3), inv(3)(q26q28), involving TP63 gene using fluorescence in situ hybridization and ТР63 mutation by Sanger sequencing.

23 patients with ALK-negative ALTCL were involved in current study: 14 males, 9 females. Advanced stage of disease was diagnosed in 70 % of patients. All patients were treated with high-dose chemotherapy.

In 20% of patients were revealed DUSP22-R, 13% - TP63 abnormalities, 67% of patients were triple-negative (without ALK-R, DUSP22-R, TP63-R).

There weren't detected any mutations in DNA-binding domain of TP63 gene.

Comparing data of molecular studies and clinical data it was revealed that presence or absence of above-mentioned gene rearrangements did not affect disease stage: III stage was diagnosed in 30% of cases with DUSP22-R and in 33% of triple-negative cases, IV stage in 100% cases with TP63-R, in 67% with DUSP22-R, in 40% with triple-negative variant. Majority of patients had IPI (3-5) in all three cytogenetic groups. In 100% of cases with DUSP22-Rwere revealed more than 2 extranodal site of involvement.

Overall (OS) and progression-free survival (PFS) depending on molecular profile were 100% and 100%, respectively, in case with DUSP22-R. In case of TP63-R OS and PFS were 8% and 10%, respectively, in triple-negative cases - 60% and 39%, respectively.

Thus, detection of DUSP22, TP63 gene rearrangements has great practical value in ALK-negative ALTCL. Statistical significance should be confirmed by further study.

Disclosures

No relevant conflicts of interest to declare.
Nivolumab Induces Impressive Complete Responses in Heavily Pretreated Classical Hodgkin Lymphoma (cHL): Immune System Reloaded in Case Series
Authors: Dada Reyad , Malas Al-Beirouti Bassim T.
Published: October 2019

Background: cHL is a histopathologically distinguish cancer type with an interesting microenvironment. The number of malignant cells is small in relation to the dominant T lymphocytes that have lost their antitumor activity through different mechanisms. Therefore, reactivation of host antitumoral immune response is promising approach to convert cHL in amenable disease for the immune system. Nivolumab is a novel antibody that binds to program death receptor-1 (PD-1) and prevents immune tolerance. Recently, several published clinical trials confirmed the clinical efficacy of single agent nivolumab in patients with few cancer types. Publications on nivolumab in cHL are very scarce. The available literature is limited to only one phase 1 (Ansell et al., N Engl J Med 2015;372:311-9) and another non-randomized phase 2 (Younes et al., Clin Oncol 34, 2016 (suppl; abstr 7535) clinical study that included 23 and 80 patients with relapsed/refractory cHL respectively.

Case series: We report on three patients with heavily pretreatedcHL who failed several lines (mean 6.3, range: 3-9) of chemotherapy. Table 1 illustrates the patients' characteristics. All patients had stage IV disease at initiation of nivolumab. Two patients had very poor performance status (ECOG 3 and 4) attributed to progressivecHL and were oxygen dependent. One patient was wheelchair bound and the other was bedridden.

The patients were started on single agent nivolumab 3 mg/kg every two weeks. Impressive clinical responses were observed in all patients after just one dose of nivolumab. In particular, significant rapid clinical improvement was obtained in both patients with the worst performance status. Both patients became mobile and self-dependent within 2-4 days after the first dose. This pattern of clinical response resembles that seen on the 1stday of immune response mediated recovery from viral respiratory infection.

All patients reached complete metabolic response after 4 doses with continuous improvement of clinical condition. Both oxygen dependent patients became oxygen independent. No treatment related significant side effects were observed.

Conclusion: Pre-treatedcHL is amenable to novel immunotherapy. Nivolumab induces impressive clinical and radiological responses with excellent tolerance. The drug enriches our treatments options by reloading the immune system response against cancer. Further clinical studies are needed to determine the effectiveness on large patients' cohort.

Disclosures

No relevant conflicts of interest to declare.
Synergistic Lethality with Gemcitabine and Arsenic Trioxide Against Human T Cell Lymphoma Jurkat Cells
Authors: Zhang Mingwan , Xu Xiaojun , Ye Yongbin , Liu Qifa
Published: October 2019

Abstract There is still lack suitable agent that significantly changes the poor prognoses of T-cell non-Hodgkin's lymphoma (NHL), so that part of the patients who are far beyond the standard of first-line could not received effective treatment, therefore, it is urgent that seeking for a salvage treatment for patients with refractory malignant lymphoma, our destination is combination strategy aimed to higher anti-tumor activity with lower toxicity.

GEM is a deoxycytidine analog with anti-metabolite activity in tumors through destroying the replication of cells and inhibiting ribonucleotide reductase as a prodrug. Recently, many clinical trials show that gemcitabine alone or in combination with other chemotherapy drugs can improve overall response rate and survival outcomes in malignant lymphoma patients compared with other chemotherapy regimens, such as vinorelbine and liposomal doxorubicin, busulfan and melphalan, which are active agents in all types of malignant lymphoma.

ATO as a traditional medicine has been evaluated its good efficacy in a variety of hematologic malignancies, including myeloproliferative diseases, multiple myeloma, lymphoma. Currently, many clinical trials represented that the addition of ATO to standard treatment regimens can improve survival outcomes and may allow a reduction in cytotoxic chemotherapy exposure in other malignancies. The basis of related research found that ATO play killing effect on the cancer cells through multiple apoptotic pathways and many mechanisms. ATO was shown to bear relevant cytotoxic activity on lymphoma cell lines. The safety profile of ATO for the effective treatment and its toxicity is easily under control, which make it a preferred solution for combined chemotherapy scheme.

Thus, in this study we found that the combination of ATO and GEM can lead to synergistic the cytotoxic effect, exert an enhanced effect on inhibiting Akt signaling and Bcl-2, up-regulating cleaved caspase-3, and inhibit the cell proliferation and self-renewal ability, and induce G0/G1 cell cycle arrest and apotosis in Jurkat cells, which provide preclinical evidence and a theoretical basis for combination therapy with ATO and GEM as a salvage therapy in treating relapsed or refractory aggressive non-Hodgkin's lymphoma.

Disclosures

No relevant conflicts of interest to declare.
Prx II(Peroxiredoxin II) Activity Has Important Roles As a New Mechanism to Control ABL Tyrosine Kinase Activity in STIs(Signal Transduction Inhibitor) Treated CML Patients
Authors: Yoo Eun-Sun , Mun Yeung-Chul , Ahn Jee-Young , Huh Jung-Won , Seong Chu-Myong
Published: October 2019

Purpose: In this study, we investigated the roles of PRX II, one of 3 critical peroxidases besides catalase and GPXs, in CML primary cells at diagnosis and remission while patients were treated with STI(Signal Transduction Inhibitor) and tested the same roles in Imatinib(IM) sensitive Ph+ cell lines and resistant cell lines as well.

Methods: Newly diagnosed CML cells, IM resistant K562 cells and parental K562 cells were treated STIs and analyzed western blot assay to detect BCR-ABL, phosphorylated BCR-ABL and PRX2 protein expression level. We added N-Acetylcysteine (0-5mM, 6hr) to K562 cells to show antioxidant effect of imatinib and analyzed DCF-DA detection for intracellular ROS level and western blot for PRX2 protein level. MTT assay was performed to detect cell death by NAC time-dependent treatment of 5mM NAC(0, 24, 40, 48hr). Imatinib resistant K562 cells were established by treatment of gradual increment of imatinib. We also repeatedly investigated the effects of IM therapy using PRXII overexpressed K562 cells by transfection.

Results: At diagnosis of CML, ROS level was elevated and PRX II was either absent or significantly suppressed. As Ph chromosomes were decreased with STIs, suppressed or absent PRXs levels were restored to the level of normal individuals. These findings were also inversely correlated with the level of Ph chromosomes in the cases of disease progression and re-remission with further treatment. When STI were treated in Ph positive cell line, we found deceased cell survival and ROS level by MTT assay and DCF-DA methods respectively, but elevated PRX II by western blot. By the treatment of NAC into Ph+ cell lines, the level of DCF-DA was decreased and MTT level was down, but PRX II level was elevated. Interestingly, the level of BCR-ABL oncogene were decreased in PRX II tranfected cells. Meanwhile, we observed that PRX II restoration was mild or weak in Imatinib resistant K-562, which we established in our lab.

Conclusions: The importance of the roles of ROS and its PRX II, antioxydant enzymes in CML is further extablished by our work. Our finding may contribute to find a new pathway on which TKIs are working besides the mechanisms of ATP binding competitively, blocking the binding of ABL-BCR kinase and substract resulting apoptosis of Ph+ cells. Furthermore, our finding may be useful to overcome the STIs resistant CML in the clinics.

Disclosures

No relevant conflicts of interest to declare.
Factors Related to the Development of Acquired Von Willebrand Syndrome in Patients with Essential Thrombocythemia and Polycythemia Vera
Authors: Rottenstreich Amihai , Kleinstern Geffen , Krichevsky Svetlana , Varon David , Lavie David , Kalish Yosef
Published: October 2019

Objective: We characterized acquired von Willebrand syndrome (AVWS) among essential thrombocythemia (ET) and polycythemia vera (PV) patients.

Methods: A review of patients with ET or PV evaluated for AVWS.

Results: Of 116 patients with ET, 64 (55%) developed AVWS; of 57 with PV, 28 (49%) developed AVWS. Median platelet counts of ET and PV patients who developed AVWS were 920 X 109/L and 679 X 109/L, respectively (P=0.01). Of patients who developed AVWS, 69.5% had platelet counts below 1000 X 109/L. Bleeding was more common in patients with AVWS, among both ET and PV patients (P<0.001). VWF:RCo levels and VWF:RCo/VWF:Ag ratio were lower among JAK2 V617F positive- vs. JAK2 V617F negative- ET patients (P=0.02 and P=0.002, respectively); whereas VWF:Ag levels were comparable (P=0.96). ET patients harboring the JAK2 V617F mutation were more likely to develop AVWS than were calreticulin-positive patients (70.3% vs. 45.7%, P=0.02), despite lower platelet counts (median 773 vs. 920 X 109/l, P=0.05). In multivariable analysis, age (β=0.26, P=0.002), platelet count (β=-0.38, P<0.001), hemoglobin level (β=-0.22, P=0.01) and JAK2 V617F mutation (β=-0.23, P=0.01) independently predicted VWF:RCo, among ET patients; whereas only platelet count predicted VWF:RCo among PV patients (β=-0.49, P<0.001).

Conclusion: Among ET and PV patients, AVWS was common and associated with higher bleeding rates and higher platelet count; nonetheless, most AVWS patients had platelet counts under 1000 X 109/L. Thus, AVWS screening should be included in routine assessment of ET and PV patients. Among ET patients, JAK2 V617F was a main driver for the development of AVWS.

Disclosures

No relevant conflicts of interest to declare.
Influence of JAK2V617F Allele Burden on Hematological Response in the Treatment of Polycythemia Vera
Authors: Shikhbabaeva Dzhariyat , Shuvaev Vasily , Golovchenko Regina , Udaleva Vera , Fominykh Mikhail , Zotova Irina I , Polushkina Lyubov , Martynkevich Irina , Abdulkadyrov Kudrat
Published: October 2019

Background. The presence of the JAK2V617F mutation is a major criterion in diagnostic of Polycythemia Vera (PV). The role of JAK2V617F allele burden in the progression of PV is not completely understood. The significance of this variable for the progression of the disease, complications development and response to therapy are still a big question mark.

Aim. To estimate the significance of the level of JAK2V617F allele burden in the treatment of PV.

Methods. Seventy-nine patients (pts), 48 females and 31 males, were included in the study. PV was diagnosed from 1980 to 2016. Median age was 61 years (range, 28 - 85), median of the observation period was 4.9 year (0,2 - 35 years). JAK2V617F allele burden was studied in all pts. Patients received therapy with phlebotomy (erythrocytapheresis), hydroxyurea, interferon-alpha or combined therapy. Hematological response to the therapy was evaluated according to ELN criteria[1]. Differences between the groups were assessed with the Mann-Whitney U test.

Results. Out of the 79 patients, complete hematological response (CHR) was achieved in 8 (10%) pts, partial response (PHR) in 48 (60.8%) pts, and no response was achieved (no response - NR) in 23 (29,2%) pts. The mean level of allele burden in the group of patients with PHR was 68%(CI 57%-79%), in that without response - 65%(CI 43%-86%), and in the group with CHR was 35% (CI 19%-50%). The JAK2V617F allele burden was significantly different between the groups with CHR and PHR (p<0,03). We also observed statistical trends (p<0,09) regarding differences between the groups with CHR and NR. The JAK2V617F allele burden by groups is shown in Figure 1. Consequently, patients who achieved CHR on treatment, had lower JAK2V617F allele burden compared to patients with PHR or NR to the therapy.

Conclusion. The level of JAK2V617F allele burden can be a predictive marker of a response to treatment of PV. An individual approach to each patient, including a detailed study of molecular factors, is the key to successful prevention of complications and ensuring positive outcomes.

[1] Barosi G, Birgegard G, Finazzi G, et al. Response Criteria for Essential

Thrombocythemia and Polycythemia Vera: Result of a European LeukemiaNet

(ELN) Consensus Conference. Blood. 2009 Mar 10

Figure The JAK2V617F allele burden levels in patients groups by treatment response. Figure. The JAK2V617F allele burden levels in patients groups by treatment response.

Disclosures

Shuvaev: Novartis pharma: Honoraria; Pfizer: Honoraria; BMS: Honoraria. Fominykh:Novartis Pharma: Honoraria; BMS: Honoraria.
AMPK Expressed By Multiple Myeloma Cells Inhibited Metformin-Induced Myeloma Cells Proliferation
Authors: Zi Fuming , He Jingsong , Zhang Enfan , Yang Yang , Huang He , Yi Qing , Cai Zhen
Published: October 2019

Background

Multiple myeloma(MM) is a malignant plasma cells proliferative disease which is characterized by increased blood calcium level, renal insufficiency, anemia, and bone lesions(CRAB). MM is still uncured disease. To further explore the mechanisms of MM was an urgent problem needs to be solved. AMPK is an "energy sensor". Studies had demonstrated that metformin could activate AMPK and exert anti-tumor effect in many tumors. However, a paradoxical phenomenon was found that knockdown AMPK could enhance the anti-tumor effect of metformin. Recent studies had revealed that AMPK maybe a double-edged sword in tumors. Our previous study had discovered that metformin exert anti-myeloma effect without activating AMPK. In this study, we mainly focus on the expression of AMPK in MM cells and to explore whether the expression of AMPK in MM cells could block the anti-myeloma effect of metformin.

Methods

The expression of AMPK in MM cells was detected by Western blotting. Small interfering RNA(siRNA) was used to knockdown the expression of AMPK. Apoptotic cells of MM cells were detected by Annexin V-FITC/PI using flow cytometry analysis. Western blotting was used to elucidate the apoptotic protein of MM cells induced by metformin with or without knockdown AMPK.

Results

Our study revealed that AMPK was obviously expressed by MM cells(Fig.1). Metformin could not activate AMPK signaling pathway. Knockdown AMPK with siRNA could enhance the anti-myeloma effect of metformin(Fig.2 and Fig.3).

Conclusions

Taken together, our data indicated that AMPK was expressed by MM cells. Inhibit AMPK could enhance the anti-myeloma effect of metformin. AMPK maybe a new therapy target.

Disclosures

No relevant conflicts of interest to declare.
SLC30A10 Deficiency: A Novel Cause of Polycythemia and Hepcidin Deficiency
Authors: Bartnikas Thomas Benedict
Published: October 2019

Like iron, manganese is an essential yet potentially toxic metal. While inherited forms of iron excess were discovered long ago, the first disease of inherited manganese excess was only recently reported in 2012. Mutations in a metal transport protein, SLC30A10, were described in patients with manganese excess, polycythemia, liver failure and Parkinsonism. Manganese excess in these patients is attributed to an essential role for SLC30A10 in hepatobiliary manganese excretion. Polycythemia, liver failure and Parkinsonism are attributed to manganese excess although the underlying mechanisms have yet to be established. To explore the role of SLC30A10 in metal biology and disease, we have generated Slc30a10-deficient mice. Characterization of these mice indicates that murine Slc30a10 deficiency recapitulates the human disease. Given that manganese and iron share some transport pathways, we interrogated iron homeostasis in these mutant mice. Intriguingly, mice display hepcidin deficiency and tissue iron excess. While hepcidin deficiency may explain the tissue iron excess, the cause of hepcidin deficiency is unclear. Based on expression profiling of hypoxia-regulated genes, we propose that hepcidin deficiency reflects aberrant activation of hypoxia signaling by manganese excess in the absence of true hypoxia. Overall, this data suggests novel interactions between iron and manganese with implications for our understanding of the mechanisms by which aberrant metal homeostasis leads to human disease.

Disclosures

No relevant conflicts of interest to declare.
Spliceosome: Physiology and Disease Pathogenesis
Authors: Krainer Adrian
Published: October 2019

Pre-mRNA splicing is an essential step for the expression of intron-containing genes, i.e., the majority of genes. Splicing is a high-fidelity process, as required for the correct expression of proteins. However, there is flexibility in the selection of competing splice sites, which gives rise to alternative splicing, a regulated process that greatly increases the diversity of the proteome. Splicing is catalyzed in a stepwise manner by the spliceosome, a macromolecular machine that consists of 5 small RNAs and more than 100 proteins. Key insights about the structure and dynamics of spliceosomal complexes have recently been obtained through cryo-electron microscopy studies. Dysregulated splicing can arise from mutations in the splice sites or regulatory elements of individual genes, from alterations in the levels of regulatory splicing factors (activators and repressors), or from mutations in splicing-factor genes. All of these scenarios can give rise to various cancers, depending on the affected gene and the cellular context. Interestingly, recurrent somatic heterozygous mutations in particular splicing factors involved in branchpoint-sequence and 3'-splice-site recognition have emerged as key drivers of certain myeloid neoplasias. This presentation will review relevant features of the spliceosome machinery, the functional implications for normal and pathological conditions, and the potential for novel therapies.

Disclosures

Krainer: Ionis Pharmaceuticals: Consultancy, Honoraria, Patents & Royalties, Research Funding; Stoke Therapeutics: Consultancy, Equity Ownership, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Co-founder, Patents & Royalties; Cold Spring Harbor Laboratory: Employment, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties; H3 Biomedicine: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees.
Next Generation Sequencing for Blood Group Antigen Profiling
Authors: Lane William
Published: October 2019

There are over 300 serologically defined red blood cell (RBC) antigens and 33 serologically defined platelet antigens, most of which have known genetic changes in 45 RBC or 6 platelet genes that correlate with antigen expression. Exposure to non-self RBC antigens during transfusion or pregnancy can lead to the development of alloantibodies, which on re-exposure can lead to clinically significant and even fatal complications. Therefore, it is vital to know which antigens are present on RBCs. However, traditional serologic phenotyping methods are labor intensive, costly, sometimes unreliable, and reagents are not always available. As such, routine antigen typing is only done for ABO and D antigens.

A large percentage of blood is given for hematologic malignancies that will soon get routine whole genome sequencing (WGS). For a minor added cost this data could be used for RBC antigen prediction. However, there are no published reports of using WGS data to predict RBC antigens. This is likely for several reasons: (1) none of the existing WGS data sets have paired serologic RBC phenotypes, (2) there are no fully annotated and complete databases of genotypes to phenotypes, (3) all of the known alleles are defined using cDNAs numbered relative to the start codon without human genome coordinates, and (4) lack of software capable of automated RBC antigen prediction.

We have created a fully interactive web site of all known blood group genotype to phenotype correlations, fully annotated with relevant information, and mapped to and visually overlaid to their corresponding human reference genome gene sequences, with algorithms to predict antigen phenotypes from inputted sequences. These predictions are part of the General Genome Reports for the 100 patients getting WGS as part of The MedSeq Project. In addition, each patient is undergoing an extensive antigen phenotypic workup using traditional blood bank serology and currently available SNP PCR based assays, which are being used to validate and improve our prediction strategies.

As clinical WGS becomes pervasive we hope that comprehensive blood group prediction will be done on everyone, allowing for easy identification of rare donors and the prevention of alloantibody formation using extended upfront matching of antigens from sequenced recipients and donor.

Disclosures

No relevant conflicts of interest to declare.
Regulation of Hematopoietic Stem Cell Mitochondrial Metabolism
Authors: Ghaffari Saghi
Published: October 2019

Hematopoietic stem cells (HSCs) like most, if not all, adult stem cells are primarily quiescent but have the potential to become highly active on demand. HSC quiescence is maintained by glycolytic metabolism and low levels of reactive oxygen species (ROS), which indicate that mitochondria are relatively inactive in quiescent HSC. However, HSC cycling - and exit of quiescence state - require a swift metabolic switch from glycolysis to mitochondrial oxidative phosphorylation. To improve our understanding of mechanisms that integrate energy metabolism with HSC homeostasis, my laboratory has been focused on the transcription factor FOXO3, which is critical for the maintenance of HSC quiescence and redox state and is implicated in HSC aging. We showed recently that FOXO3 is key to HSC mitochondrial metabolism, independent of its inhibition of ROS or mTOR signaling. Mitochondria divide and fuse constantly in part to segregate and dispose of their damaged counterparts. These processes are influenced by and highly linked to mitochondrial metabolism. We have recently developed imaging approaches to study HSC mitochondrial divisions. Mechanisms by which FOXO3 regulates HSC mitochondria and the impact of impaired FOXO3 on the HSC health and activity, and mitochondrial network will be discussed. Detailed understanding of the mitochondrial metabolism and divisions in HSC and their relationship to nuclear transcription are likely to have broad implications for the state of HSC fitness, regenerative capacity and aging.

Disclosures

No relevant conflicts of interest to declare.
My D88-Dependent Interplay Between Myeloid and Endothelial Cells in the Initiation and Progression of Obesity-Associated Inflammatory Diseases
Authors: Li Xiaoxia
Published: October 2019

Low-grade systemic inflammation is often associated with metabolic syndrome, which plays a critical role in the development of the obesity-associated inflammatory diseases, including insulin resistance and atherosclerosis. Here, we investigate how Toll-like receptor-MyD88 signaling in myeloid and endothelial cells coordinately participates in the initiation and progression of high fat diet-induced systemic inflammation and metabolic inflammatory diseases. MyD88 deficiency in myeloid cells inhibits macrophage recruitment to adipose tissue and their switch to an M1-like phenotype. This is accompanied by substantially reduced diet-induced systemic inflammation, insulin resistance, and atherosclerosis. MyD88 deficiency in endothelial cells results in a moderate reduction in diet-induced adipose macrophage infiltration and M1 polarization, selective insulin sensitivity in adipose tissue, and amelioration of spontaneous atherosclerosis. Both in vivo and ex vivo studies suggest that MyD88-dependent GM-CSF production from the endothelial cells might play a critical role in the initiation of obesity-associated inflammation and development of atherosclerosis by priming the monocytes in the adipose and arterial tissues to differentiate into M1-like inflammatory macrophages. Collectively, these results implicate a critical MyD88-dependent interplay between myeloid and endothelial cells in the initiation and progression of obesity-associated inflammatory diseases.

Disclosures

No relevant conflicts of interest to declare.
Studies on the Etiology of the Elevated Serum Isomerase in Chronic Myelocytic Leukemia
Authors: ISRAELS L. G. , DELORY G. E. , HNATIUK L. , FRIESEN E.
Published: October 2019

1. The phosphohexose isomerase activity of the serum of patients with chronic myelocytic leukemia closely parallels the rise and fall in total granulocyte count.

2. Its separated leukocytes the isomerase activity of the granulocytes was found to be approximately three times that of the lymphocytes.

3. The serum isomerase activity was estimated in dogs with induced leukocytosis and leukopenia. The enzyme activity bore a closer relationship to leukocyte destruction than to leukocyte production.

4. These findings suggest that the elevated serum phosphohexose isomerase activity found in patients with chronic myelocytic leukemia probably originates from disintegrating granulocytes.
Paroxysmal Nocturnal Hemoglobinuria: a Case Report with a Negative Ham Presumptive Test Associated with Serum Properdin Deficiency
Authors: BLAISDELL RICHARD K. , PRIEST ROBERT E. , BEUTLER ERNEST
Published: October 2019

1. A case of paroxysmal nocturnal hemoglobinuria (PNH) has been presented in which the Ham presumptive test for PNH was negative.

2. The basis for this false negative test and the atypical results of the Ham complete acid hemolysis test for PNH appeared to be due to a loss of the in vitro PNH hemolytic activity of the serum associated with a deficiency of properdin.

3. The acidosis accompanying the renal failure in this patient did not appear to accelerate the hemolytic process, for there was no apparent reduction of hemolytic activity in vivo when the acidosis was corrected by the administration of sodium bicarbonate.

4. The autopsy observations suggest that the protracted deposition of massive quantities of iron in the kidneys may play a robe in the not uncommon occurrence of renal disease in PNH.
On the Presence of Hemoglobin in Erythroleukemia Cells
Authors: COWLES JACK , SAIKKONEN JOUKO , THORELL BO
Published: October 2019

The neoplastic cells in acute erythroleukemia of the fowl appear to contain microspectrographically detectable amounts of hemoglobin in their cytoplasms. The acute myeloleukemia cells show no measureable specific absorption at all in the Soret band spectral region. Similar findings were obtained on cells from one case of human erythroleukemia and one of acute lymphatic leukemia.
The Effects of Acute Protein Depletion on the Distribution of Radioiron in Rat Tissues
Authors: BETHARD W. F. , WISSLER R. W. , THOMPSON J. S. , SCHROEDER M. A. , ROBSON M. J.
Published: October 2019

1. The distribution of Fe59 in plasma, bone marrow, erythrocytes, liver, and spleen was determined at specified intervals after single intravenous injections of the radioisotope into normal and protein-deficient rats.

2. In normal rats, injected radioiron followed the usual metabolic pathways; i.e., through bone marrow to erythrocytes, with small, consistent amounts to liver and spleen.

3. In protein-deficient animals normal Fe59 time-concentration relationships were disrupted completely.

4. Less iron was recovered from the tissues of protein-deficient rats than from normal animals.

5. Storage iron in liver and spleen was not conjugated with polysaccharide, even though tissue protein was seriously depleted.
The Pathogenesis of Transferred Chloroleukemia in the Albino Rat
Authors: HARRIS CHARLES , BURKE WILLIAM T. , GRUENSTEIN MARGOT , SHAY HARRY
Published: October 2019

The pathogenesis of transferred chloroleukemia in the rat was studied and the following results were derived. This type of leukemia colonized in an orderly manner, after intraperitoneal inoculation of leukemic cells, first in the retroperitoneal fat, then almost simultaneously in liver, spleen, lungs, and bone marrow. Only after the leucocyte pool was saturated with leukemic cells did the peripheral blood show an increase in both abnormal and normal components.
Studies on Glutathione Stability in Erythrocytes of Cases with Past History of Favism or Sulfa-Drug-Induced Hemolysis
Authors: SZEINBERG A. , ASHER Y. , SHEBA CHAIM
Published: October 2019

1. The glutathione stability test of Beutler has been performed on blood samples from cases with past history of hemolytic anemia due to Vicia faba or sulfa drugs and some members of their families.

2. In all the affected cases the level of reduced glutathione (GSH) in erythrocytes fell below 30 mg./100 cc. RBC after two hours incubation with acetylphenylhydrazine (APH). Four out of 13 relatives examined showed the same results.

3. Out of 117 control cases only five showed similar results. All the five cases were Jews of Sefardic origin.

4. Indications have been found that by addition of glucose to the blood samples tested by the above method more clear-cut results could be obtained.

5. The decrease of the reduced glutathione level in the abnormal cells incubated with acetylphenylhydrazine could not be prevented by addition of glucose.

6. The decrease of reduced glutathione level in normal and abnormal cells incubated at 37 C. for 24 hours without acetylphenylhydrazine was prevented by addition of glucose. No such protective action of glucose on GSH in hemolysates had been found.
The Effect of a Riboflavin Antagonist Upon Leukocytes of Normal and Shay Myeloid Chloroleukemic Rats
Authors: MUSSER ELIZABETH A. , HEINLE ROBERT W.
Published: October 2019

1. Administration of a riboflavin antagonist to nonleukemic Wistar rats resulted in granulocytosis without lymphopenia.

2. The analog did not have any beneficial effect on the course of myeloid chloroleukemia in Wistar rats. There was a tendency for the leukemic rats receiving the analog to have larger numbers of abnormal granulocytes than blank-injected controls, although the differences were not statistically significant.

3. Administration of TEPA resulted in rapid and prolonged remission of the myeloid chloroleukemia, as noted by others.
Myelomatosis with the "a2-Type" Serum Protein Pattern
Authors: OWEN J. A. , RIDER W. D.
Published: October 2019

1. A case of myelomatosis with "a2-type" myeloma protein in the serum is described.

2. Previously reported cases of myelomatosis with this rare type of serum protein pattern have been reviewed.

3. It is concluded from the limited data available that the clinical features of cases with the "a2-type" serum protein pattern and the prognosis are, in general, not different from those of patients with other protein abnormalities.
Observations on the Anemia of Cryptogenetic Splenomegaly II. Expansion of the Plasma Volume
Authors: MCFADZEAN A. J. S. , TODD DAVID , TSANG K. C.
Published: October 2019

In cryptogenetic splenomegaly, although there is consistently a significant reduction in hemoglobin concentration and in the red cell count, the total red cell volume may be reduced, normal or increased.

Following splenectomy the total red cell volume is either within normal limits or is increased, yet significant anemia persists.

Before splenectomy a significant increase in plasma volume is consistently present. Following splenectomy the plasma volume falls, but it remains significantly greater than normal. It would appear that the greater the plasma volume before splenectomy the greater is the fall following splenectomy. This fall in plasma volume is not a prompt response to splenectomy, for the fall which occurs one month after splenectomy is equalled or exceeded by that occurring in the succeeding four months.

It is concluded that an expanded plasma volume before splenectomy contributes with the hemolysis previously reported to the development of the anemia encountered in cryptogenetic splenomegaly. Expansion of the plasma volume at least contributes to the anemia which persists following splenectomy.
Starch Block Electrophoretic Studies of Human Hemoglobin Solutions II. Results in Cord Blood, Thalassemia and other Hematologic Disorders: Comparison with Tiselius Electrophoresis
Authors: JOSEPHSON AARON M. , MASRI M. S. , SINGER LILY , DWORKIN DAVID , SINGER KARL
Published: October 2019

The results of starch block electrophoresis of normal adult hemoglobin in the various disease states has been reported. The slow-moving fraction, A2, is elevated in all patients so far studied with thalassemia. Some patients with Addisonian pernicious anemia also show an increase in this fraction.

Cord blood contains very little A2.

The A2 fraction has no correlation with the severity of the disease.

The A2 fraction found on starch seems to be identical with the small, fast-moving component noted in the Tiselius apparatus at acid pH.

Iron deficiency anemia of the acquired type tends to have a lower amount of A2 than normal, with an increase to average normal values after therapy.
Sickle Cell Disease in India
Authors: SHUKLA R. N. , SOLANKI B. R. , PARANDE A. S.
Published: October 2019

Five cases of sickle cell disease are reported with complete clinical, hematologic and genetic studies. Four were cases of sickle cell anemia and one had sickle cell thalassemia. The importance of the presence of a focus of sickling in this area has been discussed.
Osmotic Fragility of Human Blood Platelets: Phase Contrast and Electron Microscopic Studies
Authors: GUREVITCH J. , NELKEN D. , DANON D.
Published: October 2019

Phase contrast and electron microscopic studies on the morphologic changes of platelets in various hypotonic salt solutions are presented.