bluuebunny/crossref_metadata_2025 huggingface dataset:
Association of Circulating Tumor DNA from the Cerebrospinal Fluid with High-Risk CNS Involvement in Patients with Diffuse Large B-Cell Lymphoma
Authors: Wang Xiaoxiao , Gao Yan , Shan Changguo , Lai Mingyao , He Haixia , Bai Bing , Ping Li-qin , Rong Qixiang , Ai Ruyu , Wen Lei , Zhou Zhaoming , Yu Ruoying , Ou Qiuxiang , Wu Xue , Wang Xiaoxia , Shao Yang , Cai Linbo , Huang Hui-qiang
Published: November 2020

Diffuse large B-cell lymphoma (DLBCL) patients with central nervous system (CNS) involvement have dismal outcomes. The detection sensitivity of conventional diagnosis of lymphoma through cerebrospinal fluid (CSF) cytology (CC) and flow cytometry (FCM) is restricted. CSF-ctDNA has shown to be an important method of liquid biopsy in patients with CNS cancers. To assess the correlation between CSF-ctDNA and CNS involvement in DLBCL, targeted mutational profiling was performed on CSF- and plasma -derived ctDNA together with matched systemic tumor tissues in 67 DLBCL patients clinically diagnosed as high risk for CNS relapse, using a validated panel of more than 400 genes. We found that ctDNA concentration in CSF but not in plasma correlated with CNS-IPI score in DLBCL patients. In CSF-positive high-risk DLBCL patients, eighty-six gene alterations(GAs) were shared between tumor tissue and CSF while ninety-five were shared between tumor tissue and plasma. Interestingly, forty-eight GAs(CSF-CNS GAs) were identified including 24 GAs exclusively in CSF and 24GAs shared by plasma which enriched in apoptosis/cell cycle related pathway and immunity-related pathway. To better screen for CNS-related molecular features in CSF, GAs in brain tumor tissue and CSF from a PCNSL-DLBCL cohort of 10 patients were sequenced and compared to that in high-risk group. Moreover, GAs in tumor samples from an additional cohort of 40 DLBCL patients clinically diagnosed as low-risk for CNS relapse were sequenced and compared to that in high-risk group to screen out non-CNS-related features. The number of alterations in five CSF-CNS genes including BTG2, PIM1, DUSP2, ETV6, CXCR4 in CSF was found to be associated with CNS risk in DLBCL patients. Our data supported the feasibility of using CSF-ctDNA as a complementary source for early detection of CNS involvement in DLBCL. The association between GAs in five CSF-CNS genes and CNS involvement deserved further investigation to determine their relevance in patient treatment and outcome.

Disclosures

Yu: Geneseeq Technology Inc: Current Employment. Ou:Geneseeq Technology Inc: Current Employment. Wang:Nanjing Geneseeq Technology Inc: Current Employment. Shao:Nanjing Geneseeq Technology Inc: Current Employment.
Biomimetic Membrane Vesicles: Evaluation of Inflammatory Response In Vivo
Authors: Vasileva Olga , Garanina Ekaterina E , Rizvanov Albert A , Gomzikova Marina
Published: November 2020

Introduction: Biomimetic membrane vesicles are produced from live cells using cytochalasin B which disrupts the structure of the cytoskeleton and facilitate generation of membrane vesicles under subsequent vortexing. Membrane vesicles are pinched off from the cell surface, surrounded by a cytoplasmic membrane and contain the cytoplasm of parental cell. It is known that mesenchymal stem cells (MSCs) are immunoprivileged. The aim of our study was to determine whether Cytochalasin B-induced membrane vesicles (CIMVs) derived from mesenchymal stem cells retain the immunoprivileged properties of parental cells.

Method: All experiments were carried out in compliance with the procedure protocols approved by Kazan Federal University and local ethics committee (protocol #5, date 27.05.2014). To analyze the immunogenicity, murine MSCs (7.5x104 cells) either CIMVs-MSCs (15µg) were injected i.v. in 8 week old mice (Mus musculus, C57Bl/6). CIMVs were used at a concentration equivalent to 7.5×104 MSCs based on total protein concentration. Serum isolation was performed after 2 hours post-administration. Secretion of inflammatory cytokines was evaluated using multiplex analysis BioPlex Pro Mouse 23 Plex kit. (BioRad, USA).

Results: We detected all investigated cytokines in serum of control and experimental mice: Eotaxin, G-CSF, GM-CSF, IFN-g, IL-10, IL-13, IL-17A, IL-1a, IL-1b, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL12p40, KC, MCP-1, MIP-1a, MIP-1b, RANTES, TNFa. Allogenic MSCs but not CIMVs increased concentration of Eotaxin, G-CSF, IL-17A and IL-9. The level of GM-CSF, IFN-g, IL-10, IL-1a, IL-1b, IL-2, IL-3, IL-4, IL-5, IL-6, IL12p40, KC, MCP-1, MIP-1a, MIP-1b, RANTES, TNFa in mice serum were not affected by murine MSCs injection. Injection of CIMVs did not induce any statistically significant changes in cytokines level.

Conclusion: Elevated levels of Eotaxin, G-CSF, IL-17A and IL-9 cytokines after the i.v.injection of murine MSCs suggest that moderate allergy inflammation was developed after the MSCs allotransplantation. CIMVs injection did not induced increase of cytokines level in mice serum indicating absence of immunogenicity. Taken together our results demonstrate that CIMVs show less/no immunogenicity compared to parental MSCs. We believe that small diameter, better biodistribution and fusion with host cells lead to the non-immunogenicity of CIMVs. Thus, CIMVs are confirmed to be a perspective, new biomimetic vector system. This study was funded by the grant of the President of the Russian Federation for state support of the leading scientific schools of the Russian Federation НШ-2603.2020.4. Kazan Federal University was supported by the Russian Government Program of Competitive Growth.

Disclosures

No relevant conflicts of interest to declare.
Epidemiology and Clinicopathology of Hepatosplenic T-Cell Lymphoma (HSTCL): Analysis of a Pooled Database
Authors: Haddad Philip A , Hammoud Dalia , Gallagher Kevin M.
Published: November 2020

Introduction:

HSTCL is a rare and rapidly progressive extranodal T-cell lymphoma. It accounts to for 1-2% of all T/NK cell lymphomas. Afflicting adolescents and younger adults, HSTCL commonly presents with constitutional symptoms, cytopenias, and hepatosplenomegaly. Up to a third of all the reported cases arise within a clinical context of immunosuppression. This analysis was conducted to update and expand our existing knowledge of this rare disease.

Methods:

In order to study the demographic characteristics, cytogenetic and molecular signatures, therapeutic interventions, survival, and prognostic factors, we compiled a pooled database of 360 cases. Kaplan-Meier survival curves were constructed. Cox proportional-hazards model and Log-rank tests were used to assess the influence of demographic and clinicopathology factors on overall survival (OS).

Results:

A total of 360 patients with confirmed HSTCL were identified. The median age was 30 (0.8-86) years with a peak incidence between ages 13 and 26. There is male predominance with M:F ratio of 2.6 in the whole group, 6 in the immunosuppressed, 2.2 in γδ, and 1.2 αβ. HSTCL with TCRαβ tended to be more CD5+, CD8+, and CD57+. More than 90% presented with constitutional symptoms and had bone marrow involvement. Median overall survival of the whole group was 12 months. The median duration of symptoms to diagnosis is 2 months. The median time from immunosuppression to diagnosis is 5.3 years. Sex, age, jaundice, autoimmune hemolytic anemia, hemophagocytosis, and bone marrow involvement did not affect OS. Furthermore, i7 and/or +8 and TCR type did not impact OS. Compared to no treatment, combination chemotherapy and auto and allo stem cell transplant (SCT) were statistically superior with a median OS of 1, 9, 34, and 30 months respectively. While OS of combination chemotherapy was inferior to that of SCT, there was no significant difference between auto and allo SCT. Immunosuppression adversely impacted OS in the whole group (HR:0.40, p<0.0001) and among the SCT subgroup (HR:0.38, p=0.06). There was no difference between immunosuppression with and without TNFα inhibitors. The quality of response prior to transplant also seemed to impact outcome though it did not reach statistical significance (p=0.07) with OS of 15, 30, and 44 months for none/transient, response less than CR, and CR respectively. Splenectomy improved survival in the whole group (HR:0.66; 95%CI 0.47-0.93). Further analysis revealed that splenectomy impacted survival in the context of combination chemotherapy but not SCT.

Conclusions:

This study presents an updated epidemiologic and clinicopathologic data from a pooled cohort of patients with HSTCL. It identifies the impact of immunosuppression, quality of response, treatment modalities as well as splenectomy on OS.

Disclosures

No relevant conflicts of interest to declare.
Correlation of Iron Levels with Asthma and Lung Function in Pediatric Patients with Sickle Cell Anemia
Authors: Shaikh Yusra D , Apollonsky Nataly , Bernstein Bruce
Published: November 2020

Introduction:Significant morbidity and mortality in patients with sickle cell disease (SCD) is attributed to the pulmonary sequalae of the disease. Patients with SCD often suffer airway hyper-reactivity, acute chest syndrome (ACS), chronic lung disease, pulmonary hypertension (PHTN), and obstructive sleep apnea (OSA). Recent literature has provided evidence supporting the strong association between asthma and airway hyper-reactivity in SCD. One of the factors linked to chronic inflammation and asthma is iron status. The present study examined whether iron levels are associated with pulmonary complications in pediatric patients with SCD.

Method:Through retrospective review of electronic medical records (EMR) we evaluated patients with diagnosis of asthma and SCD. All patients with available PFT (3/21/2013-3/11/2020) and iron studies were included in the analysis. Chi square and ANOVA tests were used to explore relationships of respiratory conditions with lab data and relevant medical history.

Results:The analysis reviewed information of 100 patients with SCD -- 56 males and 44 females The sample population had the following genotypes: 63% Hemoglobin (Hb) SS, 23% Hb SC, 2% Hb S Beta Zero Thalassemia, and 12% Hb S Beta Thalassemia. 38% of these patients were receiving treatment via hydroxyurea. The results generated found that patients with a large airway obstruction (LAO) had a marginally statistically significantly higher serum iron level than those with no LAO (p=0.067.) Patients with homozygous Hb S disease were four times as likely to have a history of ACS (p=0.004) than those without and were marginally significantly more likely to be SS and SB0Thal (p=0.052). Patients with history of ACS had a significantly higher mean iron saturation and lower total iron binding capacity (TIBC.) Patients with PHTN had significantly higher serum iron levels (p=0.029).

Conclusion:Our findings reveal that while iron might play a more significant role in the development of PHTN and ACS in patients with SCD, the role in asthma is borderline in our sample. These findings, although of borderline statistical significance p=0.067, are clinically noteworthy. These results may open a new window for therapy targeted at maintaining iron in normal physiologic ranges to decrease pulmonary complications in patients with sickle cell anemia. Further studies with larger samples are necessary to clarify the meaning of our marginally significant findings.

Disclosures

No relevant conflicts of interest to declare.
Preparation of Targeted Platelet Carrier and Its Antitumor Effect
Authors: Du Ying , Chen Baoan
Published: November 2020

Objective:Tumor targeting proteins were modified on the surface of platelets and chemotherapeutic drugs were encapsulated in platelets. Based on the fact that platelets can enter the tumor environment and interact with tumor cells, the functionalized platelets carrier has dual targeting effect to kill tumor cells.Methods:Aspirin was used to inhibit platelets aggregation and deformation in vitro to obtain complete platelets. Exogenous Tf (transferrin) was modified on platelets surface to target specific tumor cells (human multiple myeloma cell line RPMI8226). DOX (doxorubicin) was loaded into platelets. To verify the inhibitory effect of functional platelets vector on cell. To observe the distribution of targeted functional platelets vector and treatment effect on implant tumor in mice.Results:Aspirin can inhibit platelets aggregation and deformation during the preparation process effectively. Stable platelets can be modified by Tf and encapsulate DOX effectively. At the same time, the functional platelet vector can specifically aggregate in the implant tumor site in vivo and achieve effective anti-tumor effect.Conclusion:Platelets aggregation and deformation can be inhibited by aspirin effectively, which makes it a stable natural drug carrier. The antitumor effect can be achieved by the functional platelets.

Keywords:platelets, encapsulation, doxorubicin, tumor

Disclosures

No relevant conflicts of interest to declare.
Ruxolitinib for the Treatment of Polycythemia Vera: Response on Alternative Dose Versus Standard Dose
Authors: Cacciola Emma , Vecchio Veronica , Gentilini Cacciola Elio , Cacciola Rossella Rosaria
Published: November 2020

The polycythemia vera (PV) is a chronic myeloproliferative neoplasm characterized by increased hematocrit (HCT), elevated white-cell and platelet counts and splenomegaly. The most commonly used first line cytoreductive agent is hydroxyurea (HU). However, some patients have an inadequate response or have inacceptable side effects from HU. Ruxolitinib, a Janus kinase (JAK) 1 and 2 inhibitor, was shown to have a clinical benefit in patients with PV. A phase 3 study (RESPONSE) have shown that PV patients on Ruxolitinib at dose of 10 mg twice daily at week 32 had a complete hematologic remission. In this study were enrolled 40 patients with PV (30 men, 10 women; mean age 54 years, range 51-57) according to WHO criteria, assigned to Ruxolitinib at dose of 5 mg twice daily for clinical evidence of liver dysfunction. All patients were evaluated for JAK2V617F allele burden, HCT, white cells and platelets, splenomegaly and symptom burden, including pruritus, fatigue and night sweats. The mean duration of disease was 9 years. All patients received low-dose aspirin and underwent phlebotomy. Before Ruxolitinib, all patients had JAK2V617F allele burden > 50%, HCT control on phlebotomy, high white cells (12x109/L) and elevated platelets (600x109/L) and spleen volume of 450 cm3 or more as measured by magnetic resonance imaging (MRI) or computed tomography (CT). After Ruxolitinib we observed a shortened response time at week 6 marked by JAK2V617F allele burden < 50%, HCT < 45% in the absence of phlebotomy, normal white cells and platelets (6x109/L and 320x109/L, respectively) and reduced spleen size as assessed by palpation and absence of symptoms. These data suggest that the Ruxolitinib treatment at the alternative dose of 5 mg twice daily can be effective in achieving a rapid and complete hematologic remission that is still present in these patients receiving this dose of Ruxolitinib at the time of this analysis.

Disclosures

No relevant conflicts of interest to declare.
Somatic Mutations Predict Poor Prognosis in Myelodysplastic Syndrome Patients with Normal Karyotypes
Authors: Zeng Xiangzong , Dai Min , Zhang Yu , Zhou Lingling , Zhou Ya , Liu Qifa
Published: November 2020

Purpose: Somatic mutations are common in myelodysplastic syndrome (MDS), but its risk stratification is mainly based on cytogenetics. This study was to explore the prognostic significance of somatic mutations in MDS patients with normal karyotypes.

Patients and Methods: Three hundred and four patients with MDS were enrolled in this retrospective study. A genomic panel of 127 gene targets were detected by next-generation sequencing.

Results: Two hundred and Eighty-one (92.4%) patients carried at least one somatic mutation, while cytogenetics identified abnormalities in 140 (46.1%) patients. The 5 most frequently mutated genes were TET2, ASXL1, EZH2, TET1, FAT1, and TET2, TP53, TET1, EP300, SF3B1 in the patients with normal karyotypes and aberrant karyotypes, respectively. When mutations detected in >5% of the whole cohort, they were included in analysis and the results showed that the frequency of TET2, TP53, ASXL1, CD101, KDM6A, SH2B3 and IL-3RA mutations was different between two groups(all P<0.05). ASXL1, CD101, KDM6A, SH2B3, IL-3RA mutations were more common in normal karyotype group, while TET2 and TP53 were more common in aberrant karyotype group. Multivariable analysis showed that age (HR 1.02; P=0.027), IPSS-R(HR 1.80; P<0.0001), TP53(HR 2.36; P<0.0001) and DNMT3A (HR 1.83, P=0.044) were the risk factors while allo-HSCT(HR 0.50; P=0.001) was a protect factor for OS in the whole cohort. For sub-group analysis, IPSS-R(HR 1.54; P=0.005; HR 1.80; P<0.0001, respectively), TP53 mutation(HR 2.49; P=0.030; HR 2.13; P=0.005, respectively) and allo-HSCT(HR 0.52; P=0.040; HR 0.37; P<0.0001, respectively) retained the prognostic significance in both the normal karyotype and aberrant karyotype group. FAT1(HR 2.32; P=0.019), DNMT3A(HR 3.32; P=0.006) and IL-7R(HR 4.35; P=0.002) mutations were unfavorable factors for OS only in the normal karyotype group.

Conclusion: FAT1, IL-7R and DNMT3A mutations pretict poor prognosis in MDS patients with normal karyotypes.

Key words: Somatic mutation, Next-generation sequencing, Prognosis, Myelodysplastic syndrome

Disclosures

No relevant conflicts of interest to declare.
Diarrheal Morbidity in Patients Undergoing Hematopoietic Cell Transplantation - the Diagnostic Yield of Stool Cultures
Authors: Berger Tamar , Giladi Odil , Yahav Dafna , Ben-Zvi Haim , Pasvolsky Oren , Shargian-Alon Liat , Wolach Ofir , Rozovski Uri , Raanani Pia , Yeshurun Moshe
Published: November 2020

Diarrhea affects a significant proportion of patients undergoing hematopoietic cell transplantation (HCT). In this single center retrospective study, we explored the diagnostic yield of stool cultures for enteric pathogens among patients undergoing HCT. Between May 2007 and April 2020, a total of 1072 individuals underwent autologous (n=603) and allogeneic (n=469) HCT. Overall, 947 stool culture samples were obtained from 561 (52%) patients with diarrheal illness during hospitalization for HCT. Most (99%) samples were obtained beyond 3 days of admission and mainly (77%) during neutropenia. Overall, only four (autologous HCT, n=3; allogeneic HCT, n=1) patients were diagnosed with positive stool culture (0.42%) and in all cases, Campylobacter spp. was the pathogen identified. All isolates grew from the first stool culture obtained. The number of stool cultures needed-to-test to diagnose one case of bacterial infection was 237. The cost of diagnosing one case of bacterial infectious diarrhea was 8,770 US dollars. The median time from admission to positive stool culture sample was 12 (range, 7 to 13) days. Patients with a positive stool culture did not have discerning clinical or laboratory characteristics. In conclusion, according to our experience, the yield of stool cultures for enteropathogens in patients undergoing HCT is extremely low and thus should be avoided in most cases.

Disclosures

Wolach: Amgen: Other: Fees for lectures and Consultancy; Janssen: Other: Fees for lectures and Consultancy; Pfizer: Consultancy, Honoraria; Novartis: Consultancy, Honoraria, Other: Fees for lectures and Consultancy; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Fees for lectures and Consultancy, Research Funding; Astellas: Consultancy, Honoraria, Other: Fees for lectures and Consultancy.
Combination of Homoharringtonine with Venetoclax and Azacitidine Excerts Better Treatment Response in Relapsed /Refractory Acute Myeloid Leukemia
Authors: Yu Guopan , Xu Na , Huang Fen , Fan Zhiping , Liu Hui , Shi Pengcheng , Zhou Hongsheng , Wang Zhixiang , Zhang Yu , Liu Qifa
Published: November 2020

Overexpression of Bcl-2 is known to be an important factor for drug resistance in relapsed /refractory acute myeloid leukemia (R/R-AML). Combination of Bcl-2 inhibitor venetoclax with hypomethylating agents (HMA) has shown promising killing effect in elder patients with newly diagnosed AML, also worked in R/R-AML but with low rate of complete response (CR). High expression of Bcl-XL and/or Mcl-1 is the main reason for resistance to venetoclax. As an inhibitor of both Bcl-XL and Mcl-1, homoharringtonine (HHT) has presented a synegetic effect with venetoclax in inhibition of Mcl-1 and killing of lymphama. In this study, we analyzed the treatment response and safety of combination of HHT (1mg/m2 d1-7) with venetoclax (400mg d1-14) and azacitidine (75mg/m2 d1-7) (HVA) in R/R-AML. From 6, 2019 to 5, 2020, in Nanfang Hopsital, 22 patients with RR-AML were treated with HVA (HVA group), with a median age of 39(16-62) years, previous chemotherapy of 3(1-7) cycles. 15 (68.2%) relapsed within one year after allogenetic hematopoietic stem cell transplantation (allo-HSCT) and 12 (54.5%) previously exposed to HMA treatment. Meanwhile, 7 patients with RR-AML were treated with venetoclax combined with HMA (control group), with a median of 42(31-74) years, previous chemotherapy of 3(1-6) cycles. 2 (28.6%) relapsed after allo-HSCT and 5 (71.4%) had HMA exposure. In HVA group, totally 11/17(64.7%) patients acquired treatment response, and 7/15(46.7%) obtained CR/CRi, of whom 6 presented minimal residual disease (MRD) negative. While in control group, only 2/7(28.6%) acquired treatment response and 1 obtained CR. Subgroup analysis in HVA group showed allover response rate (ORR) was 9/13(69.2%) and CR rate was 6/11(54.5%) (all with MRD negative) in the patients with allo-HSCT. And in the patients without allo-HSCT ORR was 2/4(50.0%) and CR rate was 1/4. With a median follow-up of 6(1-7) months, one patient was bridged to allo-HSCT, 2 out of 7 patients with CR relapsed. All patients were well tolorated to both treatment regimens, showing as a median neutropenia of 9.5(5-14) days, 5 patients with Febrile neutropenia and 4 with pulmonary infection. In conclusion, combination of HHT with venetoclax and azacitidine (HVA) excerts better treatment response in R/R-AML, especially in those with allo-HSCT, and shows well tolorated. A multi-center prospective clinical trial (NCT04424147) has been initiated.

Key words: HVA regimen, relapsed /refractory acute myeloid leukemia, treatment response, safety

Disclosures

No relevant conflicts of interest to declare.
Hemorrhage Is a Major Cause of Blood Transfusion in COVID-19 Patients
Authors: Sanz Cristina , Faundez Anabel , García Carulla Anna , Rodriguez Aliberas Marta , Coromoto Anny , Pereira Arturo
Published: November 2020

INTRODUCTION

Reports from China and other countries have allowed to quickly understand the clinical and biological features of COVID-19. In contrast, little has been published on the transfusion requirements of COVID-19 patients and the clinical characteristics of transfused patients.

PATIENTS AND METHODS

For this cross-sectional study, we reviewed the blood bank and clinic records of 80 consecutive patients diagnosed with COVID 19, who required red blood cells (RBC) transfusion at the Hospital Clinic of Barcelona over a period of 60 days, from mid-March to mid-May 2020. COVID-13 was confirmed by at least two PCR test in nasopharyngeal swaps. Categorical data were described as frequencies and percentage and continuous variables were summarized as median and interquartile range (IQR)

RESULTS

Median age was 71 years (IQR: 62-76) and 59 (74 %) patients were males. The patients' main clinical characteristics and the indications for transfusion are summarized in the table. In total, the 80 patients received 2,081 RBC units in 261 independent transfusion episodes. Number of days on transfusion support ranged from 1 to 44 (median: 5).

Bleeding was the indication for transfusion in 55 patients (69%), and included either large hematomas in 22 and external hemorrhage in 31. Anemia of critical illness was the reason for transfusion in 22 (27%) patients. Most patients were on anticoagulants at the time of transfusion or the two days before (table). In total, 138 of the 261 transfusion episodes (59%) were related to spontaneous (94) or procedure-related (44) bleeding. Spontaneous bleeding was more frequent in the retroperitoneal space and the gastrointestinal apparatus. Tracheostomy with endotracheal intubation, surgical interventions, and cannulation of femoral vessels were the main procedures behind non-spontaneous bleeding.

Hemorrhage was protracted and difficult to control and patients often required transfusion support for several days. Not rarely, hemorrhage recurred when the patient was restarted on anticoagulants. It was not unusual for bleeding from an anatomical location to be followed by, and overlapped with bleeding from a different location.

Seventeen patients died during the period on study. None of the deaths was ascribed to hemorrhage or the blood transfusion.

DISCUSSION

Bleeding, mostly related to the use of anticoagulants, was the main indication for RBC transfusion in patients with COVID-19.

Figure 1

Disclosures

No relevant conflicts of interest to declare.
Dynamic BH3 Profiling As Pharmacodynamic Biomarker for the Activity of BH3 Mimetics
Authors: Pan Rongqing , Wang Youzhen , Qiu Shumei , Ryan Jeremy , Halilovic Ensar
Published: November 2020

Pharmacodynamic (PD) biomarkers provide information about the pharmacologic effects of a drug on its target, i.e. to assess whether a given agent is engaging its target in the expected manner. There is currently no convenient PD marker for BH3 mimetics. BH3 profiling (BP) is a functional assay developed to measure cells susceptibility to apoptosis and cell's dependence on certain Bcl-2 anti-apoptotic proteins for survival. Dynamic BH3 profiling (DBP) measures the changes of BP signals after drug perturbations. In this study, we studied whether DBP with lymphoid cells can serve as a PD biomarker for the activity of BH3 mimetics.

Using DBP and cell lines with defined dependency, we first showed that BH3 mimetics BCL201 (S55746) and S63845 are highly selective for Bcl-2 or Mcl-1, respectively. Next, we treated primary human lymphoid cells with BCL201 and then conducted DBP. We asked if pretreatment with BCL201 altered mitochondrial sensitivity to different BH3 peptides. After treatment with 1 μM BCL201, an amount consistent with achievable levels in vivo, T cell sensitivity to the Mcl-1 selective MS1 peptide, but not Bad peptide or Bfl-1 selective FS1 peptide, increased significantly, from 20% to 51%. These results suggest that BCL201 treatment increases T cell dependency on Mcl-1 and that DBP of T cells with MS1 peptide may serve as a PD marker for BCL201 activity. Similar results were also observed in B cells.

Next, we conducted DBP on healthy T cells treated with S63845. T cell mitochondrial sensitivity to MS1 peptide or Bcl-xL selective HRK peptide did not change. However, the treatment increased mitochondrial sensitivity to the Bad and FS1 peptides significantly, boosting the signal from 10.2% to 57.2% and 46.8% respectively, suggesting DBP of T cells with Bad or FS1 peptides can be robust PD markers for S63845 activity. This results also indicate that S63845 treatment augmented T cell dependency on Bcl-2 and Bfl-1 for survival.

Currently, we are investigating whether DBP can be used as a PD biomarker for in vivo studies. Our preliminary data suggest that DBP works well across different species (rat, mouse, and human). More importantly, DBP can detect changes in mitochondrial apoptotic signaling of ex vivo rat cells after in vivo treatment with BH3 mimetics.

Conclusions: Exposure to BH3 mimetics causes changes in mitochondrial apoptotic signaling of T/B cells which are readily detectable by DBP. DBP with lymphoid cells may provide a robust PD biomarker for clinical trials testing BH3 mimetics, either as monotherapy or in combination with other agents.

Disclosures

Wang: Novartis Institutes for Biomedical Research: Current Employment. Qiu:Novartis Institutes for Biomedical Research: Current Employment. Halilovic:Novartis Institutes for Biomedical Research: Current Employment, Current equity holder in publicly-traded company.
Levels of N-Terminal Pro-B-Type Natriuretic Peptide Relate to Risk of Infection in Multiple Myeloma Patients
Authors: Huang Weimin , Wei Xiaolei , Wei Qi , Wei Yongqiang , Feng Ru
Published: November 2020

Background

Infections are a major complication in multiple myeloma (MM) patients and are associated with inferior outcomes. Thus there is a pressing need to predict risk of infection in MM patients.

Methods

Data from 110 MM patients were retrospectively analyzed. Patients were divided into high and low N-amino terminal fragment of the N-terminal pro-B-type natriuretic peptide (NT-proBNP) groups using the cut-off value 300ng/L at diagnosis.

Results

This study showed patients with high NT-proBNP had significantly higher beta-2-micorglobin (B2M), creatinine, lactate dehydrogenase (LDH), Eastern Cooperative Oncology Group performance status (ECOG-PS) score, international staging system (ISS) score and lower estimated glomerular filtration rate (eGFR), hemoglobin, albumin than patients with low NT-proBNP. During the follow-up period, 46% presented at least one grade ≥3 infection. Patients with high NT-proBNP had high cumulative incidences of infections at the first 4 months and at the end of follow-up. Multivariate Cox regression analysis proved NT-proBNP to be an independent significant prognostic factor for grade ≥3 infections (relative ratio [RR] 2.190, 95% confidence interval [CI]: 1.126-4.261, P = 0.021).

Conclusions

This study revealed the potential of NT-proBNP for predicting risk of infection in MM patients. NT-proBNP may be applied as a criterion to identify patients with high risk of infection, who can benefit from prophylactic measures.

Disclosures

No relevant conflicts of interest to declare.
The Prognostic Significance of Minimal Residual Disease in Multiple Myeloma in the Molecular Genetic Groups Msmart 3.0
Authors: Garifullin Andrey , Voloshin Sergei , Linnikov Sergey , Martynkevich Irina , Kuvshinov Alexey , Kuzyaeva Anastasiya , Chubukina Zhanna , Shuvaev Vasily , Kleina Elizaveta , Bakai Marina , Sсhmidt Alexander
Published: November 2020

Assessment of the role of genetic abnormalities and minimal residual disease (MRD) is an active developing area in hematology. The use of genetic methods makes it possible to predict the course of the disease and apply an individualized approach to antimyeloma therapy. At the same time, the identification of MRD after therapy determines possibility of relapse.

Aim. To identify the prognostic potential of MRD in patients in the standard and high molecular risk groups.

Materials and methods. We analyzed 72 patients with MM (median age was 59 years (range 37-80), male/female - 1.3:1). All patients received initial therapy with proteasome inhibitors and / or immunomodulators. High dose therapy (MEL200) and autologous stem cell transplantation (ASCT) was carried out 50 (69%) patients. Standard cytogenetic and FISH methods were used to stratify patients in risk groups of mSMART 3.0. The standard risk (SR) was established in 52 (72%) patients, the high risk (HR) - in 20 (28%) patients. The MFC MRD status of bone marrow was evaluated after 4-6 cycles of induction therapy or after ASCT with use of 5-colors flow cytometry. MRD-negative status (MRD-) was based on level of clonal plasma cells <10-4 in bone marrow sample.

Results. The MRD- was reached in 36% (26/72) patients. The median of OS in MRD+ group was 104 months, in MRD- was 146 months (p=.01). The median of PFS in MRD+ group was 26 months, in MRD- was 70 months (p=.00021). 2-years PFS in MRD+ group was 56%, in MRD- group was 100% (p=.00021). We divided patients into the following groups for evaluation the effect of MRD on survival in risk groups: SR МRD+ 34/72 (47%), SR МRD- 18/72 (25%), HR МRD+ 12/72 (17%) and HR MFC МRD- 8/72 (11%). The median of OS in HR MRD+ group was 72 months, in SR MRD+ - 104 months, in HR MRD- - 146 months, in SR MRD- was not achieved (p=.02). The median of PFS in HR MRD+ group was 24 months, in SR MRD+ - 26 months, in HR MRD- - 68 months, in SR MRD- - 70 months (p=.003). The 2-years PFS in HR MRD+ group was 44%, in SR MRD+ group was 50% and in SR MRD- and HR MRD- groups were 100% (p=.003).

Conclusion. The absence of MRD is the most important prognostic factor. The leveling of negative effect of genetic abnormalities become possible when the MRD-negative response status is achieved. Presumably, this is due to the elimination of clonal plasma cells owing to the use of optimal antimyeloma therapy which is based on the risk stratification.

Disclosures

Martynkevich: Pfizer: Honoraria, Speakers Bureau; BMS: Honoraria, Speakers Bureau; Novartis: Honoraria, Speakers Bureau. Shuvaev:Novartis: Honoraria, Speakers Bureau; BMS: Honoraria, Speakers Bureau; Pfizer: Honoraria, Speakers Bureau.
Factors Associated with Survival and Severe Cytokine Release Syndrome of Patients with Relapsed/Refractory B-ALL Receiving CD19 CAR-T Cell Therapy
Authors: Li Xi , Luo Chengxin , Xu Shuangnian , Ma Yanni , Chen Jieping
Published: November 2020

Background : Chimeric antigen receptor-modified T (CAR-T) cells against CD19 have achieved great therapeutic effect in patients with relapsed/refractory B-cell acute lymphoblastic leukemia (B-ALL). However, factors associated with the duration of survival and the occurrence of severe cytokine release syndrome (CRS) after CD19 CAR-T cell therapy have not been fully defined.

Patients and Methods: We analyzed 24 patients with relapsed/refractory B-ALL who received CD19 CAR-T cell therapy in a phase Ⅰ/Ⅱ clinical trial (ClinicalTrials.gov, NCT02349698). Infused CAR-T cell characteristics, its expansion kinetics in vivo, patient characteristics and laboratory parameters were analyzed for their correlation with overall survival (OS), progression-free survival (PFS), and the incidence of severe CRS.

Results: Sixteen patients achieved complete remission (CR), among which 13 achieved minimal residual disease-negative CR. Patients achieved CR had significantly prolonged OS and PFS compared with those who did not. Multivariable cox regression showed that lower peak of CAR-T cell expansion and the occurrence of severe CRS were significantly associated with both inferior OS and inferior PFS, and higher proportion of effector memory T cells in the infused cells was only significantly associated with inferior PFS. Nineteen patients developed CRS, among which 6 developed severe CRS. Compared patients with mild CRS, patients with severe CRS experienced worse vital sign instability and pancytopenia, higher incidences of coagulation and organ dysfunctions, and higher concentrations of procalcitonin and cytokines. Multivariable logistic regression showed that higher pretreatment disease burden was independent predictor of severe CRS.

Conclusions: Clinical and biological factors correlated with survival and severe CRS of refractory/relapsed B-ALL patients receiving CD19 CAR-T cell therapy were identified. These data may be helpful for improving the efficacy of CAR-T cell therapy, facilitating the recognition and early intervention of severe CRS.

Disclosures

No relevant conflicts of interest to declare.
Clinical Analysis of Hemorrhagic Cystitis after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Shen Biao , Pang Aiming , Jiang Erlie , Feng Sizhou , Han Ming-Zhe
Published: November 2020

Objective: To analyze the risk factors and survival of the patients with hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation (HSCT).Methods: Propensity score Matching was conducted for HC patients among 523 patients who received HSCT in Hematopoietic Stem Cell Transplantation Center, Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences from August 2016 to August 2019. Non-HC patients were selected with a ratio of 1:2 (HC: non-HC) and then statistically analyzed with SPSS 22.0. Results:130 HC patients and 233 non-HC patients were included in the study. There were HC of grade I-II in 114/130 cases(87.69%) and of grade III-IV in 16/130 (12.31%), The median time of onset was 33 days after HSCT(range 3-339 days).Univariate analysis indicated that HC occurrence were associated with the source of stem cells, infusion CD34+ cells count, morphologic remission before transplantation, HLA matching, urinary BK virus load and the drug of Bu in conditioning regimens. Multiple regression analysis showed that morphologic remission before transplantation, HLA matching and infusion CD34+ cells count were associated with HC. K-M survival analysis showed that the 1-year survival rate of HC and non-HC was 91.8% vs 86.3%, and the 2-year survival rate was 52.5% vs 50.6%, respectively, showing no statistical difference. ROC curve showed that when the level of BKV is higher than 5×108 copies /ml, the risk of HC would increase.

Conclusion: HC was associated with morphologic remission before transplantation, HLA matching and infusion CD34+ cells count. There was no significant difference in survival between HC and Non-HC groups. the level of urinary BKV load higher than 5×108 copies /ml may cause hemorrhagic cystitis symptoms.

Key words: hemorrhagic cystitis; Allogeneic hematopoietic stem cell transplantation; Risk factors; Prognosis

Disclosures

No relevant conflicts of interest to declare.
Outcomes of Red Cell Exchange in Pregnant Patients with Sickle Cell Disease: A Case Series
Authors: Khan Suheb Mahammed , Abughanimeh Omar , Ebers Steven , Eclov Julie , Bobr Aleh , Koepsell Scott A , Nester Alex , Harper James , Gundabolu Krishna , Buddharaju Laxmi Narayana
Published: November 2020

Background:

Pregnant patients with sickle cell disease (SCD) have higher morbidity compared to patients without SCD. SCD during pregnancy can increase the risk of fetal and obstetrical complications including preterm birth, preeclampsia, and others. Moreover, during pregnancy SCD can become more severe resulting in more sickle cell vasoocclusive crisis. Prophylactic transfusion during pregnancy has been used in practice to reduce sickle cell pain crisis since hydroxyurea is contraindicated during pregnancy. However, using prophylactic red blood cell exchange (RBCX) has been a controversial topic. In this study, we aim to evaluate the outcomes of red blood cell exchange in pregnant patients with SCD

Methods:

This is a retrospective study. We evaluated the charts of three pregnant patients who were enrolled in the chronic RBCX program at the University of Nebraska Medical Center at the time of their pregnancy. Data was collected to assess the sickle cell disease related complications during pregnancy, outcomes of pregnancy, and safety of the red blood cell exchange.

Results:

A total of 19 exchange procedures were performed for three pregnant patients while being enrolled in the chronic red blood cell exchange program. Patients demographic is summarized in table 1. The indication for enrollment in the red cell exchange program were recurrent vasooclussive crisis in patient 1 and 3, avascular necrosis in patient 2. The pregnancies were uncomplicated except for preeclampsia in the third patient resulting on early delivery. Overall, the three patients had less frequent visits to the emergency room for sickle cell related complications after starting apheresis and during pregnancy (Figure 1). We believe that the modest improvement that was noticed for the third patient was due to late enrollment in the exchange transfusion program and her older age compared to the two other patients

Conclusion:

Our study shows that red blood cell exchange for SCD patients during pregnancy can be safe, feasible, and can reduce the visits to the emergency room due to SCD related complication. Further larger studies are warranted to confirm this.

Disclosures

Gundabolu: BioMarin:Consultancy;Bristol Myers Squibb pharmaceuticals:Consultancy.
Association of Pulmonary MALT Lymphoma and Sarcoidosis
Authors: Baig Nabil , Rao Ravi Dama , Daniel Vijai , Kandaswamy Chitra , Vedire Apurva
Published: November 2020

Introduction

Sarcoidosis is a systemic granulomatous disease of unknown etiology characterized by the chronic inflammation. Mucosa associated lymphoid tissue (MALT) lymphoma is a low-grade marginal zone B-cell lymphoma of MALT often etiologically associated with chronic inflammation in the underlying organ. A rare association between sarcoidosis and MALT lymphoma has been reported.

Case Description

We present the case of a 75-year-old woman who presented with chronic cough and dyspnea of several years' duration. She was found to have fluctuating lung infiltrates on serial imaging studies. A bronchial biopsy revealed chronic inflammation, and no evidence of infection. She was then treated with oral steroids with a transient benefit. However, the infiltrate and symptoms recurred, therefore 8 months later, she underwent repeat imaging. This showed a relapse of the infiltrate and enlarged mediastinal lymphadenopathy. The lung infiltrate was biopsied (percutaneously). This demonstrated a MALT lymphoma. Imaging studies done for staging showed a large retroperitoneal lymph node. A biopsy of this node revealed non-caseating granulomas, consistent with sarcoidosis. Chemotherapy with bendamustine and rituximab was initiated for therapy of the MALT lymphoma.

Discussion

A rare association between pulmonary MALT lymphoma and underlying inflammation caused by sarcoidosis has been previously described. In our patient, sarcoidosis was diagnosed in a biopsy of a retroperitoneal lymph node, but we suspect that there is likely involvement of the mediastinal nodes.

Disclosures

Rao: Bayer & Daiichi Sankyo:Consultancy.Daniel:GSK:Other: Paid speaker for COPD medications.
T(4; 14) Is Not a Poor Prognostic Factor for Newly Diagnosed Multiple Myeloma Patients Undergoing Autologous Stem Cell Transplantation in New Drug Era
Authors: Geng Chuanying , Yang Guangzhong , Gao Wen , Wang Huijuan , Li Yanchen , WU Yin , Zhou Huixing , Zhang Zhiyao , Jian Yuan , Chen Wenming
Published: November 2020

Background

The prognostic value of t(4;14) in newly diagnosed multiple myeloma (MM) patients undergoing autologous stem cell transplantation (ASCT) remains controversial. We aimed to evaluate the prognostic value of t(4; 14) for newly diagnosed MM patients undergoing ASCT in new drug era.

Methods

We retrospectively analyzed 419 newly diagnosed MM patients under 66 years of age in Beijing Chaoyang hospital, Capital Medical University. The propensity score matching technique was used to reduce the bias between groups.

Results

Among 419 patients, 243 (58.0%) patients received ASCT after induction therapy including new drugs with 12 months. Patients with ASCT had significantly longer median overall survival (OS) (69.8 vs. 59.3 months, p=0.034) and progression-free survival (PFS) (35.0 vs. 23.9 months, p=0.001) than non-ASCT patients. Univariate Cox proportional hazards regression analyses showed that ASCT was correlated with longer OS (HR=0.720, 95%CI: 0.531-0.977, p=0.035) and PFS (HR=0.618, 95%CI: 0.470-0.814, p=0.001). The favorable effect of ASCT on PFS was confirmed in multivariate (HR=0.376, 95%CI: 0.195-0.723, p=0.003), but it had no impact on OS in multivariate analysis (p=0.289). In the propensity score matching analysis, 116 patients, 58 in each group, were identified. Among 116 matched patients, patients with ASCT had longer PFS (52.1 vs.23.3 months, p=0.002) and OS (89.2 vs.50.2 months, p=0.010). Among 121 patients received ASCT and t(4; 14) measurement by fluorescence in situ hybridization, 30 (24.8%) patients presented translocation t(4; 14). Univariate Cox analyses showed that t(4; 14) was not correlated with PFS (p=0.347) and OS (p=0.454). Multivariate Cox analyses also showed that t(4; 14) was not correlated with PFS (p=0.466) and OS (p=0.349). There was no significant difference in PFS (36.5 vs.37.0 months, p=0.344) or OS (54.0 vs.81.7 months, p=0.452) in ASCT patients with or without t(4;14).

Conclusion

These results demonstrated that ASCT was a favorable prognostic factor for newly diagnosed MM patients under 66 years of age, however, t(4; 14) had no prognostic value for newly diagnosed MM patients received ASCT in new drug era.

Disclosures

No relevant conflicts of interest to declare.
Coagulation Dysfunction and Hematological Changes in 633 Patients with COVID-19
Authors: Yang Mo , Deng Huixia , Yang Liuming , Li Liang , Ye Jieyu , Cheng Yucai , Tan Yafang , Chong Beng H , Li Qiang
Published: November 2020

Background: A previously unknown beta-coronavirus was discovered through the use of unbiased sequencing in samples from patients with pneumonia. The virus was named as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by the international committee for the classification of viruses (ICTV). The disease caused by this virus was named as coronavirus disease 2019 (COVID-19). In addition to pulmonary manifestations, hematological changes such as lymphocytopenia, thrombocytopenia, and coagulation dysfunction can also be found in COVID-19 patients, and the mechanism is still unclear.

Case data and methods: A total of 633 COVID-19 patients from Wuhan hospital of China were retrospectively analyzed. Clinical case data of all patients were collected, including gender, age, chronic underlying diseases, outcome, and blood laboratory test results. The hematological features of COVID-19 patients and the factors affecting their outcome were analyzed.

Results: Of 633 patients with COVID-19, the median age was 62 years (interquartile range, IQR, 51.0-70.0) and 330 (52%) were men. Lymphocytopenia (lymphocyte count, 1.0 ×109 / L [IQR, 0.7-1.4]) occurred in 317/607 patients (52%), thrombocytopenia (platelet count <100 × 109/ L) occurred in 14/62 death patients (23%), prolonged prothrombin time (13.8 seconds [IQR, 13.1-15.1]) in 289/486 patients (59%), increased D-Dimer level (0.7 mg/L[IQR, 0.2-2.9]) in 230/411 patients (57%) and increased C-reactive protein levels (10.7 mg/L [IQR, 2.2-49.7]) in 217/426 patients (51%) . Compared with the survival patients, death patients have higher white blood cell count (11.7 × 109/L [IQR, 8.4 to 15.6]), neutrophil count (10.8 × 109/L [IQR, 7.8 to 13.9]), neutrophil count/lymphocyte count (20.5 [IQR, 12.4-34.2]), activated partial thromboplastin time (36.8 seconds [IQR, 31.3-42.3]), prothrombin time (17.1 seconds [IQR, 14.7 to 19.7]), D-Dimer level (4.6 mg/L [IQR, 1.0 to 7.8]), C-reactive protein level (111.8 mg/L (IQR, 53.1 to 196.6), and low lymphocyte count (0.5 × 109/L [IQR, 0.3 to 0.7]). The results of logistic multivariate regression analysis showed that age, neutrophil count, prothrombin time, and C-reactive protein were risk factors for patients with COVID-19.

Conclusion: Hematological changes are common in patients with COVID-19. The early stage of the disease is mainly characterized by lymphocytopenia, thrombocytopenia, and the late stage may be characterized by more severe lymphocytopenia, even neutrophils elevation, elevated C-reactive protein, and severe coagulation disorder. The pathogenesis may be mediated by a direct viral infection and/or indirect immunopathology.

Disclosures

No relevant conflicts of interest to declare.