bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of Early Rasburicase on Incidence and Outcomes of Clinical Tumor Lysis Syndrome in Lymphoma
Authors: Personett Heather A , Barreto Erin F , McCullough Kristen , Dierkhising Ross , Leung Nelson , Habermann Thomas M.
Published: October 2019

Introduction: Clinical tumor lysis syndrome (TLS), which manifests most often as acute kidney injury (AKI) is a common and devastating complication of rapidly proliferating hematologic malignancies. Epidemiologic studies of attributable risk factors for AKI in this setting are small, focus on only aggressive lymphomas rather than the more prevalent intermediate risk disease subtypes, and predate contemporary risk stratification, diagnostic criteria, and supportive care. Additionally, early administration of rasburicase to enhance uric acid (UA) elimination has been adopted without robust evidence in support of its impact on clinical outcomes, specifically, prevention of AKI.

Methods: This was a retrospective cohort study of adult lymphoma patients at intermediate or high risk for TLS treated at Mayo Clinic between 2009 and 2017. Patients were excluded if they had AKI at hospital admission, received pre-admission rasburicase, or were on dialysis. Consensus definitions were used to stratify TLS risk and stage AKI. The incidence of new AKI in the setting of TLS was described along with predictors of its development, including early rasburicase use.

Results: Of the 461 patients with a new lymphoma diagnosis during the study timeframe at intermediate or high risk of TLS, 78 were excluded, primarily due to prevalent AKI at hospital admission. The 383 included patients were predominantly at intermediate risk for TLS (73%) and had a diagnosis of Diffuse Large B-cell Lymphoma (64%). Incidence of new onset AKI during hospitalization was 6% and 91% of cases were stage 1. All new onset AKI occurred within the first 5 days of hospital admission. Independent predictors of new AKI included age > 60 years, history of chronic kidney disease or cardiovascular disease, and UA >8 mg/dL at any time during hospital admission. Rasburicase was given to 48 patients, 6 (13%) of whom still went on to develop AKI. Rapid UA reduction with rasburicase did not impact the risk of developing AKI (HR 2.3; p = 0.11). Uric acid level at the time of administration did not modify the effect of rasburicase on prevention of AKI (p = 0.36 for the interaction term).

Conclusion: In conclusion, for lymphoma patients at intermediate or high risk for TLS, if not present at admission, AKI only infrequently developed during the course of a patient's hospitalization. Predictors of AKI included advanced age, comorbid conditions, and elevated UA levels. Early rasburicase administration, regardless of UA level, did not alter AKI risk.

Disclosures

No relevant conflicts of interest to declare.
Outcomes of T-Cell Depleted Haploidentical Peripheral Blood Stem Cell Transplantation for Adult Acute Leukemia: A Meta-Analysis
Authors: Durer Seren , Durer Ceren , Katsanis Emmanuel , Comba Isin Yagmur , Jamil Faiza , Aslam Shehroz , Shah Zunairah , Malik Mustafa Nadeem , Selene Insija Ilyas , Fraz Muhammad Asad , Jose Jemin Aby , Qureshi Anum , Yasir Muhammad , Amin Saher Nasir , Anwer Faiz
Published: October 2019

Background: Ex-vivo T-cell-depleted graft is used to prevent acute and chronic GVHD for haploidentical hematopoietic SCT (HSCT). We conducted a systematic review and meta-analysis to evaluate the outcomes of T-cell-depleted grafts for peripheral blood haploidentical stem cell transplantation in patients with acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML).

Method: A comprehensive literature search on PubMed, Embase, Cochrane and Web of Science was conducted up to July 25, 2018. Two independent reviewers screened the literature and extracted data. The methodological quality was assessed using modified Newcastle-Ottawa Scale. CMA software v.3 was used for the analysis. Heterogeneity among studies was assessed using the I2 test. Random-effect model was applied.

Results: The search strategy identified over 550 articles; three studies (n = 404 patients; 1 prospective; 1 multi-center) were selected for this meta-analysis. The sample size of the studies varied between 34 and 266 patients. The median follow-up ranged from 6 to 47 months. Two studies (Aversa, F. et al. 2005)(n=104 patients); (Ciceri, F. et al. 2008)(n=266 patients) used CD34+ selected cells and other study used TCR-αβ depleted PBSC graft (Kaynar, L. et al. 2016) (n=34). All studies used myeloablative (MA) conditioning. In vivo T-cell-depletion by ATG was used in two studies. Two studies using mega-dose of CD34+ did not use GVHD prophylaxis and other study with αβ T-cell depleted graft used mycophenolate mofetil (MMF). Engraftment rates ranged from 91% to 98%. Pooled rates (95%CI) of grade II-IV, III-IV aGVHD and chronic GVHD were 9% (4-20; I2=82%), 4.8% (1.7-13.1; I2=78%) and 10.7% (6.3-17.6; I2=64%), respectively. The pooled estimates (95%CI) showed a 2-year disease-free survival (DFS) rate of 30.8% (24.2-38.3; I2=54%), 2-year relapse incidence of 21.5% (17.4-26.2; I2=85%). The results are shown in figure 1.

Conclusion: T-cell-depleted peripheral blood stem cell graft for haploidentical transplantation in acute leukemia may be an alternative treatment option. Larger prospective studies are needed to evaluate the outcomes of different manipulation techniques.

Disclosures

No relevant conflicts of interest to declare.
Intraepithelial Lymphocytosis of the Resection Margin in the Monomorphic Epitheliotropic Intestinal T-Cell Lymphoma
Authors: Heo Mi Hwa , Jung Hye Ra , Do Young Rok , Ha Jung-Sook , Kim Hyera , Kim Do-Hoon , Kim Jin Young
Published: October 2019

Background: Monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL) is a disease involving the intestine and is often diagnosed in surgically resected specimens. The disease prognosis of MEITL is fatal because of rapid recurrence and leakage after excision. Recently, in microscopic finding of MEITL, intraepithelial lymphocytosis in mucosa that were grossly intact has been reported. In this study, we evaluated the intraepithelial lymphocytosis of the resection margin to determine the reason for the early recurrence or leakage in MEITL.

Materials and Methods: We reviewed the medical records of all patients who were diagnosed with small intestine lymphoma from January 1995 to June 2018 at our medical center. We analyzed a tissue array from 8 patients diagnosed with MEITL. The expression of CD3, CD5, and CD8 were analyzed by immunohistochemistry.

Results: A total of 8 cases of small intestine lymphoma were collected in this period. There were three men (37.5%) and five women (mean age 55 years). Two patients died without initiation of chemotherapy. Six patients received chemotherapy after resection of MEITL (4 patients received CHOP regimens, 2 patients received ProMACE-CyBOM regimens), but five patients died within 6 months (mean survival time 4.1 months, range 0.3-12.1 months). Only one patient survived for 5 years. On gross examination, the distance to the resection margin was 7.58 cm (3.8-14.0 cm) on average. Microscopic examination revealed intraepithelial lymphocytosis at least of one of the resected margins in all case. Immunohistochemical staining for CD3, CD5, and CD8 was performed on the resection margins. As a result, severe intraepithelial lymphocytosis accompanied by aberrant loss of CD5 was observed in 6 cases (75%).

Conclusion: In the MEITL, an intraepithelial lymphocytosis accompanied by aberrant CD5 loss is frequently observed in the resection margin, even if the resection margin is grossly intact, and there is no clear microscopic involvement of the lymphoma especially on low power. This fact is thought to be related to the early recurrence or leakage of MEITL at the surgical site.

Disclosures

No relevant conflicts of interest to declare.
Efficacy of Low-Dose Decitabine in the Early Relapse of AML/MDS after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Guo-Fa Xu , Ting Chen , Huanfeng Liu , Shijia Lin , Gao Lei , Zhang Cheng , Liu Yao , Zhang Xi , Zeng Defu , Peiyan Kong
Published: October 2019

Objectives: Retrospective analysis of the efficacy and adverse effects of low-dose Decitabine(DAC) in patients with early relapse of acute myeloid leukemia/myelodysplastic syndrome(AML/MDS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

Methods:Fifteen patients with AML/MDS, which were early recurrence after transplantation, received low-dose DAC treatment. The evidence of early recurrence included engraftment index(FISH detection of sex chromosomes, or STR) <95%, MRD (+), fusion gene positive or positive bone marrow blast cells (between 5%~10%). DAC was given (5-10) mg·m2-1·d-1×5 days, every 6∼8 weeks, a maximum for 8 cycles. The treatment outcome, adverse reaction and graft-versus-host disease (GVHD) were analyzed. Fifteen patients included 13 with AML,1 with MDS-RAEB-2, and 1 with chronic myelogenous leukemia (CMML).

Results: The total complete remission (CR) rate was 66.67%(10/15), the Objective response rate (ORR) was 86.67% (13/15); 5 cases of relapse in the effective patients, in which 3 cases with PR was relapse eventually, and 2 CR patients has extramedullary recurrence and bone marrow relapse, respectively, the recurrence rate was 38.46%(5/13). Grade 3 and 4 hepatic dysfunction was 26.67%, Grade 3 and 4 hematological toxicities was 60%.

Conclusion:low-dose of DAC was effective and safe in controlling early recurrence, reducing GVHD symptoms, reducing recurrence rate and prolonging patient survival time.

Disclosures

No relevant conflicts of interest to declare.
Metabolomic Profile in Patients with Autoimmune Hemolytic Anemia
Authors: Rabelo Iara Baldim , Chiba Akemi Kuroda , Moritz Elyse , Barros Melca Oliveira , Silva Ismael Dale Cotrim Guerreiro , Bordin Jose Orlando , Chauffaille Maria de Lourdes , Yamamoto Mihoko , Arrais Celso
Published: October 2019

Introduction: autoimmune hemolytic anemia (AIHA) is a rare clinical condition with hemolysis mediated by immunoglobulin binding on the surface of erythrocytes with or without complement activation. Metaboloma is the sum of the genetic configuration of an organism to the external factors. The metabolic profile involved in AIHA has not yet been described and may be the key to new findings about the mechanism of autoimmune hemolysis.

Objectives: To describe a profile of AIHA related metabolites, increased hemolysis and risk of relapse.

Patients and Method: we analyzed the plasma from 31 patients with primary AIHA compared to plasma from 150 healthy subjects using mass spectrometry (Kit AbsoluteIDQ p180, Biocrates, Austria). The statistical analysis was performed by the software available at http://www.metaboanalyst.ca/.

Results: of the 95 metabolites found in patients with AIHA, four acylcarnitines, two phosphatidylcholines (PC), asymmetric dimethylarginine (ADMA) and three sphingomyelins were increased with great statistical significance. There was an increase in PCs and polyamines (spermine and spermidine) in the AIHA group in hemolytic activity. The relationship between PCs "PC ae 34: 3 / PC ae 40: 2" seen only in the 12-months relapsed group was predictor of relapse with specificity of 81% and sensitivity of 100%.

Discussion: Increased sphingomyelin in AIHA patients may perpetuate deficient regulatory T lymphocytes in the recognition of erythrocytes as self. Accumulation of ADMA can cause downregulation of NF-kβ from macrophages by inducing one of the already known autoimmune mechanisms. Excess PCs in patients with AIHA and increased hemolysis may perpetuate effector T lymphocytes with increased susceptibility to autoimmunity and increase the production of antibodies against erythrocyte antigens. The increase of polyamines in the higher hemolysis group may induce autoimmunogenicity of DNA and interfere with the differentiation of macrophages. Finally, the presence of PC to 34: 3 / PC to 40: 2 as a predictor of relapse may help to identify cases that require follow-up more frequently or second-line therapies earlier.

Conclusion: Our data demonstrate a distinct profile of metabolites in AIHA patients, as well as in individuals with higher hemolytic activity. There was a relationship of phosphatidylcholines strongly predictive of AIHA relapse.

Disclosures

No relevant conflicts of interest to declare.
Analysis of Differential Circrnas Expression Profile Identifies Novel Biomarkers for Acute Monocytic Leukemia
Authors: Wu Balu , Pan Yunbao , Jin Yanxia , Zhao Mingfeng , Zhou Fuling
Published: October 2019

Malignant acute monocytic leukemia (AML-M5) is a subtype of AML that is associated with hyperleukocytosis, extramedullary disease, and abnormal coagulation. Accumulating evidence has revealed that aberrant circular RNAs (circRNAs) expression play important roles in carcinogenesis and tumor progression. However, the roles of circRNAs in AML-M5 remains unclear. In this study, next-generation deep sequencing was performed to identify the circRNAs profiles in bone marrow from 6 AML-M5 patients and 3 healthy donors. A total of 78 circRNAs were identified to be associated with AML-M5 carcinogenesis. Two circRNAs (hsa_circ_0100160 and hsa_circ_0012152) were selected for further validation by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) in 30 AML patients and healthy control. Both of them were significantly upregulated in AML patients (P < 0.05). The AUC value of ROC curve was 0.934 with sensitivity of 90.9% and specificity of 81.8%, suggesting that hsa_circ_0012152 could distinguish the AML-M5 patients from healthy donors. Bioinformatics analysis predicted that hsa_circ_0012152 interact with hsa-miR-4763-3p and its corresponding mRNAs, which will be our further study. In summary, Our study provides an overview of the circRNA profiles in AML-M5, which will facilitate future investigations into the function of hsa_circ_0012152.

Disclosures

No relevant conflicts of interest to declare.
Disruption of SOCS3 Promotes the Anti-Cancer Efficacy of Primary NK Cells
Authors: Naeimi Kararoudi Meisam , Elmas Ezgi , Lamb Margaret , Chakravarti Nitin , Trikha Prashant , Lee Dean Anthony
Published: October 2019

Genetic modification of NK cells to enhance cancer immunotherapy has potential application to treat a wide range of cancers. Recently, we developed a new strategy in which CRISPR/Cas9 elements are introduced into NK cells as ribonucleoproteins (RNPs) via electroporation, followed by expansion on feeder cells expressing 4-1BBL and membrane-bound IL-21 to generate large numbers of genetically modified NK cells. This method was used to genetically modify several genes in primary and expanded NK cells including suppressor of cytokine signaling 3 (SOCS3). SOCS3 negatively regulates cytokine signaling through the JAK/STAT pathway. We hypothesized that disruption of SOCS3 in primary NK cells using Cas9/RNPs could potentially maintain STAT3 signaling levels and subsequently increase their proliferation and cytotoxic function.

We designed gRNAs to target exon 2 of the SOCS3 gene and electroporated them along with Cas9 protein as Cas9/RNPs into primary NK cells using the Lonza 4D electroporator. We tested 6 different conditions of gRNAs alone or in combination. The NK cells in the control group were electroporated with no Cas9/RNPs. After electroporation, the cells were rested in culture media supplemented with 100 IU of human IL-2 for 48 hours and then expanded using irradiated feeder cells. At day 7, an equal number of cells were restimulated with irradiated feeder cells to test the effect of SOCS3 KO on proliferation. Western blot was used to assay the knock out efficacy at the protein level. Calcein assay and IncuCyte Zoom (Essen) were performed to measure cytotoxicity against two cancer cell lines, K562 and Daoy.

Our results showed a significant reduction in SOCS3 protein levels in 3 conditions (gRNA1, gRNA3 and gRNA1+ gRNA3) in comparison to the control group. The calcein assay and IncuCyte zoom showed the modified SOCS3 KO NK cells could kill the tumors more efficiently in comparison to the control. Proliferation data showed the SOCS3 KO cells can grow faster than the control group.

In conclusion, our data demonstrate the role of SOCS3 and JAK/STAT pathway in NK cell function and suggests that SOCS3 may be a good target for genetic modification to improve cancer immunotherapy using NK cells.

Disclosures

Lee: CytoSen Therapeutics: Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding; Merck, Sharp, and Dohme: Consultancy; Courier Therapeutics: Equity Ownership, Membership on an entity's Board of Directors or advisory committees.
A Highly Efficient and GMP Compliant Protocol to Manufacture CCR5 Edited Cells to Treat HIV Infection
Authors: Romito Marianna , Meyer Emily , Poddar Sushmita , Heinz Helena , Rositzka Julia , Hildenbeutel Markus , Gautron Anne-Sophie , Busser Brian W , Kok Yik Lim , Turchiano Giandomenico , Haas Simone , Schäfer Richard , Smith Julianne , Metzner Karin J , Mussolino Claudio , Duchateau Philippe , Cathomen Toni , Cornu Tatjana I
Published: October 2019

Targeted genome editing in blood and immune cells enable new therapeutic applications, especially for infectious diseases. We present a GMP-compliant protocol to manufacture CCR5-edited CD34+ hematopoietic stem and precursor cells (HSPCs) with the goal to cure patients suffering from chronic infection with human immunodeficiency virus type 1 (HIV1). We hypothesize that genetic disruption of the CCR5 gene, which encodes the major HIV1 co-receptor, in HSPCs will give rise to an HIV-resistant immune system after transplantation. We have developed engineered nucleases based on transcription activator-like effector nucleases (TALENs) targeting CCR5. Electroporation of CD4+ T-cells and CD34+ HSPCs with mRNAs encoding TALENs revealed disruption of up to 80% of CCR5 alleles in CD4+ T-cells and over 90% of alleles in HSPCs. The high gene editing frequencies in T-cells and HSPCs were confirmed by deep sequencing, and no cleavage activity above background levels were detected at the top 20 predicted off-target sites. CCR5-edited CD4+ cells preserved their proliferation capacity and their biological function. Importantly, these cells showed significantly reduced CCR5 expression and became resistant to infection with the R5-tropic HIV-1JR-FL virus. The CCR5-edited HSPCs maintained their proliferation potential and their capacity to differentiate into the various blood lineages in vitro and in vivo, and clonal analysis revealed bi-allelic CCR5 disruption in more than 75% of cells. In summary, our developed protocol enables highly efficient and GMP-compliant knockout of the CCR5 locus in clinically relevant cells, so forming the foundation for a planned phase I/II clinical study.

Disclosures

Gautron: Cellectis SA: Employment. Busser:Cellectis: Employment, Patents & Royalties: Cellectis. Smith:Cellectis. Inc: Employment, Patents & Royalties. Duchateau:Cellectis: Employment, Patents & Royalties: Cellectis. Cathomen:TRACR Hematology: Consultancy; Cellectis: Research Funding; Miltenyi Biotec: Research Funding. Cornu:Cellectis: Research Funding; Miltenyi Biotec: Research Funding.
In Elderly Patients with Newly Diagnosed Aggressive B-Cell Lymphoma, the Rate of Change in Body Weight after the First Course R-CHOP (-like) Chemo Therapy Has Effect on Their Survival
Authors: Ayako Muramatsu , Hiroaki Nagata , Kodai Kuriyama , Yoshiko Hirakawa , Muneo Oshiro , Toshiki Iwai , Hitoji Uchiyama
Published: October 2019

Background

The incidence of B-cell lymphoma increases with age, and over 40% occurs in patients at age of 70 years old or more. Aggressive B-cell lymphoma was often treated with R-CHOP (-like) regimen. However, in elderly B-cell lymphoma patients, treatment intensification often must be lowered due to the risks of comorbidities and organ function deterioration, and treatment outcomes are worse compared to younger patients. The optimal dose of R-CHOP (-like) therapy is necessary to improve the outcome of the elderly patients with B-cell lymphoma. We conducted a retrospective cohort study examining the influence of the rate of change in body weight after the first chemo therapy on their outcomes and survival.

Methods

Clinical records of 111 patients who had received R-CHOP (-like) regimen were retrospectively analyzed.

They were all over 73 years old, and newly diagnosed with aggressive B-cell lymphoma by WHO 2008 criteria.

They were treated at Japanese Red Cross Kyoto Daiichi Hospital between January 1st 2008 and December 31st 2017.

Data on clinical characteristics and treatment modalities were obtained through the review of medical charts.

Progression-free survival (PFS) and overall survival (OS) were estimated using the Kaplan-Meier method.

The impact of variables on PFS and OS was evaluated by univariate log-rank tests and by multivariate analysis using the Cox proportional hazards model.

Results

Median age at diagnosis was 78 years old (73-94).

The 2-year OS rate was 77.5% (95%Cl: 68.3-84.3%), PFS rate was 77.4% (95%Cl: 68.2-84.3%) in all patients.

The 5-year OS rate was 62.3% (95%Cl: 50.2-72.2%), PFS rate was 55.5% (95%Cl: 43.4-66%) in all patients.

The average rate of change in body weight after the first therapy was 4.59%.

Large changes in body weight ( >9.3%) after the first therapy had worse clinical outcomes with shorter median OS (1.43 years vs. NA, P <0.001 HR 4.39, 95% CI 2.14 to 8.99, see figure 1) and median PFS (1.43 years vs. 6.9 years, P<0.001, HR 3.65, 95%CI 1.82to 7.29, see figure 2).

Conclusion

Large changes in body weight ( >9.3%) after the first therapy were associated with poor outcomes in elderly people with newly diagnosed aggressive B-cell lymphoma.

This suggests that adjusting drug dosage on and after the second therapy in those patients has possible to be improve their survival.

Disclosures

No relevant conflicts of interest to declare.
Efficacy and Safety of Ruxolitinib in Treatment of Steroid Refractory/Refractory Acute Graft-Versus-Host Disease: A Prospective Multicenter Research
Authors: Fan Zhiping , Deng Lan , Li Xudong , Zhang Qing , Xuan Li , Huang Fen , Lin Ren , Jin Hua , Xu Na , Shi Pengcheng , Wang Zhixiang , Ye Jieyu , Jiang Ling , Sun Jing , Liu Qifa
Published: October 2019

Objectives To explore the efficacy and safety of ruxolitinib in the treatment of steroid refractory/refractory acute graft versus host disease (aGVHD).

Methods A prospective analysis was conducted in the 31 steroid refractory/refractory aGVHD cases who treated with ruxolitinib from October 2017 to June 2018. The first dose of ruxolitinib was 5-10mg bid, and the dose was adjusted to 5mg qd for maintenance treatment when complete remission (CR) status lasted for 2 weeks after steroid refractory/refractory aGVHD turned into CR.

Results Of all the 29 steroid refractory/refractory aGVHD patients, 15 patients were treated with ruxolitinib in case of disease progression after receiving at least one second-line treatment, and 14 cases directly with ruxolitinib after the occurrence of corticosteroid refractoriness. The time of starting ruxolitinib treatment after the occurrence of aGVHD was 10 (4-81) d, the median time of onset was 8 (3-18) d. The overall response rate was 89.6% (26/29) including 22 complete responses (75.9%), and 4 partial responses (13.8%). 12 patients with thrombocytopenia need to reduce the dose of ruxolitinib, and 1 patient withdrew treatment for obvious bleeding tendency. With the median follow-up 3 (2-8) months after treatment with ruxolitinib, 28 patients survived and 1 died of severe pneumonia related to aGVHD, and the overall survival was 96.6% (28/29).

Conclusion Ruxolitinib is safe and effective in the treatment of steroid refractory/refractory aGVHD, and can be used as a second-line treatment for refractory aGVHD.

Disclosures

No relevant conflicts of interest to declare.
Abnormal Microenvironment of Bone Marrow B Lymphocytes in Patients with Primary Immune Thrombocytopenia
Authors: Yu Tianshu , Hou Ming , Liu Xinguang , Han Panpan
Published: October 2019

BACKGROUNDS:

Primary immune thrombocytopenia (ITP) is an autoimmune disorder characterized by peripheral blood platelet count<100x10^9/L and increased risk of bleeding, but its exact pathogenesis remains unclear. Previous researches mostly emphasized on periphral blood, in order to elucidate the pathogenesis and provide new strategies for treatment, a series of studies about bone marrow B cells were carried out in our investigation.

METHODS:

5ml bone marrow blood samples from 11 newly diagnosed ITP patients and 7 allo-HSCT healthy donors were collected into heparin anticoagulant tubes , bone marrow mononuclear cells (BMMCs) were separated with Ficoll density gradient-centrifugation in two hours. Different subsets of B lymphocytes were determined by multicolor flow cytometry including naive B cells, total memory B cells , plasma cells and regulatory B cells (Bregs) ,as well as some chemokine receptors of B cells (CXCR5, BAFFR, BCMA, TACI) . RNA were extracted from BMMCs using Trizol reagent, transcription factors related to development and differentiation of B cells (PRDM1, Pax5, IRF-4, XBP1) were detected by real-time PCR.

RESULTS:

The percentage of B cells (CD19+) in bone marrow lymphocytes in ITP patients was significantly lower than that in healthy donors (6.16±0.74% vs. 17.28±2.43%, P < 0.0001). The proportion of naive B (CD19+CD27-) in B cells was also lower compared with normal controls (59.11±7.60% vs. 81.58±4.00%, P=0.041), and the proportion of memory B (CD19+CD27+) was higher(40.17±7.67% vs. 18.01±3.89%, P=0.045),but there was no significant difference in plasma cells (CD19-CD138+). Besides,there was a decrease of Breg (CD19+CD24highCD38high) in ITP patients compared with healthy donors (20.33±5.05% vs. 57.98±9.76% , P = 0.008), and its percentage of total lymphocytes was also significantly lower unsurprisingly (1.38±0.38% vs. 12.23±2.88%, P < 0.001). The level of BAFFR in mature B cells was elevated in ITP patients (80.72±4.53% vs. 45.81±8.49%, P = 0.002). However, no significant difference was observed in other three chemokine receptors. All four Transcription factors related to B cell development and differentiation was not found to be significantly different between ITP patients and healthy controls.

CONCLUSIONS:

Our results showed that memory B cells which represent the active form were increased, and Bregs which mediate immune tolerance were much more decreased in ITP patients. As BAFFR is the only specific receptor of B cell activating factor(BAFF), its elevated expression suggested the BAFF-BAFFR system enhanced chemotactic function of B cells in ITP patients. All those results indicated that the bone marrow B cells in patients with ITP were in a state of immune overstimulation, this may potentially constitute a novel therapeutic target.

Figure. Figure.

Disclosures

No relevant conflicts of interest to declare.
Profile of Capillary-Leak Syndrome in Patients Received Chimeric Antigen Receptor T Cell Therapy
Authors: Hu Yongxian , Feng Jingjing , Shao Mi , Huang He
Published: October 2019

Background: Autologous T cells modified to express a chimeric antigen receptor (CAR-T) has demonstrated exciting efficacy in treating leukemia and there has been some reports about the toxicities recently. However, the spectrum of capillary-leak syndrome (CLS) associated with CAR-T cell therapy has not been systematically evaluated, which can be a life threatening complication as results of the cytokine release syndrome (CRS). Therefore, as the use of CAR-T therapy continues to expand to broader applications, it is prudent to characterize the profile of CLS to help providers guide safe management.

Method: We reviewed all acute lymphoblastic leukemia (ALL) patients who had participated in the clinical trial from our center to receive CAR-T therapy between 2016-2018. Patients analyzed in the study received either CD19 CAR-T cells or CD19 plus CD22 CAR-T cells. The diagnosis of CLS includes edema, acute hypotension and hemoconcentration with hypoproteinemia or hypoalbuminemia. CRS grading was evaluated with Lee's criteria for CRS.

Result: 42 ALL patients were included in this study with the mean age of 27(8-52) years old. 11(11/42, 26.2%) patients were diagnosed as CLS and 31 were not. It was observed that CLS was more common in patients who developed severe CRS. Patients with CLS was found to have high rate of hypotension and use of gamma globulin.(Table 1) Top level concentration of serum IL-6 in CLS patients was much higher than that in non-CLS patients (16438.7 vs 3292.7 pg/mL, p=0.0016), which is consistent with the well recognized concept of IL-6 as an indicator of CRS.(Figure 1) It is important to notice that CLS patients had lower levels of serum total protein (TP, 43.7 vs 52.8 g/L, p=0.0005) and serum albumin (ALB, 27.4 vs 33.8 g/L, p=0.0011), while the hemoglobin (HGB) concentration showed no difference, suggesting that TP and ALB might be better indicators for CLS than HGB, although hemoconcentration, hypoproteinemia and hypoalbuminemia are both important in diagnosis.(Figure 2) Moreover, there was no significant difference in age, gender, Ph type of ALL, type of CAR-T cells infused and death ratio.(Table 1) Although CRS has been reported to be related with disease burden before the therapy, our data showed no difference of it between the patients with and without CLS.

Conclusion: In conclusion, we have evaluated a basic profile of CLS among CAR-T patients in our center and the study indicates that CLS warrants extra attention for patients who receive CAR-T therapy. Further investigations are required to elucidate best practices for prevention and management of CLS in CAR-T therapy.

Disclosures

No relevant conflicts of interest to declare.
Impact of Body Mass Index (BMI) on Survival Endpoints in Multiple Myeloma
Authors: Kocoglu Mehmet Hakan , Patel Ameet , Rapoport Aaron P. , Ma Ning , Ding Jennifer , Elder Simran , Hossain Aneesha , Badros Ashraf Z.
Published: October 2019

Background: The role of metabolism in outcomes of cancer patients is under investigation. A higher body mass index (BMI) substantially increases the risk of cancer and impacts the treatment. High BMI is also associated with increased risk of MM (Euro J Cancer 2011). However no data is available in the effect of BMI on myeloma survival after autologous stem cell transplantation (ASCT)

Methods: We retrospectively reviewed patient charts who had ASCT between Jan 2010 - November 2011. Patients were selected if they have adequate data, two lines of induction treatment prior to transplantation. Most of the patients received maintenance treatment. IRB granted exemption for anonymous data collection. We collected demographic data, cytogenetic risk, response with induction and after transplant, progression free and overall survival. BMI was estimated at the time of ASCT.

Results: A total of 223 post ASCT myeloma patients met inclusion criteria. Median age of the cohort was 58 years, 48.8% were African-American, 51% Caucasian, 1% Asian, and 0.5% Hispanic. Median BMI was 29.7 and 30 in female and males respectively. Median BMI was 29.4 in Caucasians and 29.6 in African-American cohorts. 24% of patients had high risk cytogenetics. 72% of patients experienced very good partial response (VGPR) or better post-transplant.

On multivariate analysis, PFS and OS showed no association at any of the four BMI levels (p=0.573), age, gender, race, or depth of response. High risk cytogenetics was associated with PFS (p= 0.022). There was no association with high risk cytogenetic features at any BMI level, age, gender, race (p=0.245). For patients with BMI > 40, PFS differed significantly when compared to patients with normal BMI: 37 months versus 50, p=0.52. Unadjusted hazard ratio (HR) 2.10 (95% CI: 0.62-7.18) with respect to patients with normal BMI. Subgroup multivariate analysis with patients with morbid obesity did not show any association with depth of response (partial response or VGPR or better) or high-risk cytogenetic features.

Conclusion: BMI as a surrogate marker does not significantly impact long term survival outcomes in patients with multiple myeloma in the post-transplant setting and thus should not factor in transplant decision-making. Patients with morbid obesity appear to have a trend towards shortened progression free survival and may warrant further investigation.

Figure. Figure.

Disclosures

Badros: Karyopharm: Research Funding; Celgene: Consultancy, Research Funding; GSK: Research Funding.
Coagulation Factor XII Plays an Important Role in Procoagulant Activity of Apoptotic Cells
Authors: Yang Aizhen , Wu Yi
Published: October 2019

Apoptosis can be induced in a variety of pathological disorders, including inflammation, autoimmune diseases, and chemotherapy. When cells undergo apoptosis, they express phosphatidylserine (PS) on cell membrane surface and thus become procoagulant. Although it has been known that the procoagulant activity of apoptotic cells are tightly associated with thrombotic disorders, such as atherothrombosis and Trousseau syndrome, the mechanisms by which apoptotic cells activate the coagulation system and enhance blood clotting are largely unknown. In this study we investigated which coagulation factor(s) is involved in this process. Using western blotting and chromogenic substrate assay, we found that incubation with apoptotic cells induced by Dexamethasone (DXMS), but not with viable cells, resulted in rapid cleavage and activation of FXII. Moreover, apoptotic cells-mediated FXII activation was significantly increased in the presence of prekallikrein (PK) and high molecular weight kininogen (HK), other two components of plasma contact system. However, incubation of apoptotic cells did not cause dramatic changes of other coagulation factors, suggesting a selective association of FXII activation with apoptotic cells. Activation of FXII by apoptotic cells was markedly inhibited by a specific anti-kallikrein antibody, indicating the activation of the contact system by apoprotic cells. Flow cytometric measurement showed that FXII bound to apoptotic cells in a concentration-dependent manner, which was inhibited by annexin V and PS liposome. A surface plasmon resonance assay showed a direct binding of FXII to PS (KD=3.9E-9 M). When challenged by apoptotic cells, clotting time of plasma from FXII-knockout mice was significantly prolonged, which was reversed by replenishment with human FXII. Moreover, an inhibitory anti-FXII antibody completely prevented apoptotic cells-induced intrinsic tenase complex formation. Consistently, apoptotic cells significantly increased thrombin production in normal plasma, which were attenuated by PS blocker annexin V, an inhibitory anti-FXII antibody, and the deficiency of FXII, respectively. Addition of human FXII to XII-deficient plasma recovered thrombin generation. As evaluated by ELISA, the levels of thrombin-antithrombin complex in circulation were significantly increased when apoptotic cells were intravenously injected into wild-type mice, but not in FXII-knockout mice. In conclusion, FXII plays an important role in apoptotic cells-mediated procoagulant activity.

Disclosures

No relevant conflicts of interest to declare.
Combined Single-Cell and Spatial Transcriptomics to Deconvolute the Hematopoietic Stem Cell Niche
Authors: Haas Simon , Baccin Chiara , Al-Sabah Jude , Velten Lars , Lars Steinmetz , Trumpp Andreas
Published: October 2019

Coordinated interaction of many cell types is required to facilitate hematopoietic and mesenchymal stem cell maintenance and differentiation in the bone marrow. However, the molecular factors and cell types involved in this complex interplay remain poorly understood. Here we developed a combined single cell and spatial transcriptomics approach to address this problem.

Large-scale single-cell transcriptional profiling in conjunction with a multi-layered sorting approach allowed us to generate a complete and evenly sampled transcriptional map of all major bone and bone marrow populations. Our dataset covers all cell types or differentiation trajectories involved in mesenchymal and hematopoietic stem cell differentiation, osteogenesis, adipogenesis, myelopoiesis, erythropoiesis, lymphopoiesis, memory T cell formation as well as bone marrow neural innervation and vascularization at the single cell level. Using this data, we derive fundamental properties of the described cell types, clarify the cellular source of signals affecting stem cell differentiation processes and provide a systems view on putative intercellular interactions. Systematic spatial transcriptomics, using laser-capture microdissection of selected bone marrow niches followed by transcriptional profiling and bioinformatic cellular deconvolution, allowed us to confirm predicted interactions and map the cellular composition of distinct bone marrow niches.

Our analyses highlight the importance of pre-adipogenic CXCL12 abundant reticular cells as key niche cells for stem cell maintenance, provides a holistic systems view of the hematopoietic stem cell niche and offers a novel approach to systematically deconvolute the molecular, cellular and spatial composition of complex tissues.

Disclosures

No relevant conflicts of interest to declare.
Adding Slow Release Tetracosactide Acetate to Eltrombopag Improves Haematological Response in ITP Patients with Platelet-Count Fluctuation
Authors: Iuliano Francesco , Iuliano Eleonora , Rossi Marco
Published: October 2019

Background and aim

Tetracosactide Acetate (SR-TA) stimulates the release of corticosteroids such as cortisol from the adrenal gland. It is absorbed on to a zinc phosphate complex which ensures the sustained release of the active substance from the intramuscular injection site. SR-TA is currently used in the management of ulcerative colitis and Crohn's disease, juvenile/adult rheumatoid arthritis and osteoarthrosis. However, its properties indicate that this compound may be particularly useful in patients unable to tolerate oral glucocorticoid therapy or where normal therapeutic doses of glucocorticoids turned to be ineffective. Eltrombopag is an oral, non-peptide thrombopoietin receptor agonist that has been shown efficacy and safety in chronic immune thrombocytopenia (ITP) patients not responding to previous therapy. Based on these premises, we treated with a combination of Eltrombopag and SR-TA selected ITP patients undergoing a platelet count fluctuation despite a standard eltrombopag dose was given .

Material and methods

A total of 6 adult patients, female (60%),median (range) age 37 (28 - 54) years, median (range) baseline platelet count 31 (28 - 46) × 109/L, median of 3 (2 - 4) prior ITP therapies, received a combo of Eltrombopag 50 mg by mouth daily and SR-TA 1 mg intramuscularly daily until the target platelet count of 50× 109/L was reached, then a dose of 1mg of SR-TA was given every 3 days; in responding patients , i.e. without platelet count fluctuation, the dose was further reduced to 1mg per week.

Results

5 out of six patients responded to treatment reaching and maintaining the platelet count between 50× 109/L and 100× 109/L. After a median follow-up of 6 months with combo treatment, all patients maintained their response. Self-administration was started by 100% of patients (6/6). No adverse events have been observed.

Our experience has shown that eltrompopag/SR-TA combo is effective and safe in maintaining the platelet count above 50x109/L in those patients not reaching this target with the TPO mimetic alone.

Disclosures

No relevant conflicts of interest to declare.
Loss-of-Function Mutation in tet2 in Zebrafish Leads to Early MDS like Phenotype
Authors: Rajan Vinothkumar , Woodside Rachel , Prykhozhij Sergey V , Berman Jason
Published: October 2019

Myelodysplastic syndrome (MDS) is a pre-leukemic state characterized by the failure of the bone marrow to produce mature and functional blood cells. Nearly one-third of MDS patients progress to acute myeloid leukemia (AML). AML is the most common form of acute leukemia in adults and accounts for a high level of mortality in pediatric leukemia.

Loss-of-function mutations in the Ten-11 Translocation 2 (TET2) gene are implicated in MDS and AML, but evidence that TET2 mutations are also found in healthy individuals as a result of clonal hematopoiesis calls into question the contribution of TET2 to leukemogenesis. Many studies have tried to mutate the DNA binding domain in TET2 both in mouse and zebrafish, but the blood phenotypes observed have been inconsistent.

We created a tet2 zebrafish mutant using CRISPR-Cas9 technology, where we deleted 2.1 kb from exon 2, confirmed by cDNA analysis. A recent publication suggests the importance of Ser99 in the stability of the TET2 protein (Wu et al., 2018) and this conserved serine residue is contained in the region of deletion in our zebrafish tet2 mutant.

These tet2 mutant fish were incrossed to create a maternal-zygotic mutant with no tet2 expression. Importantly, quantitative PCR demonstrated that there was no compensatory effect from tet1 and tet3 in the context of tet2 loss.

Whole mount in situ hybridization performed on 24-48h tet2 mutant zebrafish embryos provided evidence of a significant reduction of early and mature myeloid and erythroid cells through staining for spi1/pu.1 (myeloid), gata1 (erythroid) myeloperoxidase (mpx; neutrophils), carboxypeptidase A5 (cpa5; mast cells) and l-plastin (lcp1; macrophages). The stem cell compartment was not impacted, as shown by wild-type runx1/c-myb expression. Further, o-dianisidine staining revealed low hemoglobin content in tet2 mutants compared with control embryos. Overall, the early embryos elucidate a refractory anemia like phenotype.

Flow cytometry analysis of kidney marrow from 3-month-old adult tet2 mutant fish similarly demonstrated a decrease in the myeloid and erythroid compartments, in keeping with characteristics typical of low stage MDS. We are currently employing RNAseq and MeDIP analysis on blood cell populations from tet2 mutants to determine the genetic and epigenetic signatures associated with this mutation.

This zebrafish model of tet2 loss uniquely provides with a dynamic phenotype that starts with refractory anemia with progression through low stage MDS. Thus this model will be beneficial in elucidating the mechanisms underlying how tet2 contributes to MDS and AML and provide a preclinical platform to screen for therapeutic interventions.

Disclosures

No relevant conflicts of interest to declare.
Efficacy and Safety of Low-Dose Decitabine in Low- or Intermediate 1-Risk MDS
Authors: Cao Zhijian , Ma Jun
Published: October 2019

OBJECTIVES: Decitabine has been approved in China for treatment with high-risk Myelodysplastic syndromes (MDS) patients (pts) in 2008. However data of decitabine on efficacy and safety of low-dose decitabine in low- or intermediate 1-risk MDS was absence. Herein,3 years datas in our centre of low- or intermediate 1-risk MDS treated with decitabine were analysed to evaluate the efficacy and safety in two subgroups.

METHODS: Inclusion Criteria:Patients of age > 18; Patients with low-or intermediate 1-risk MDS. The IPSS is used to classify patients with MDS across disease subgroups. ECOG performance status ≤ 2. And normal hepatic,renal and cardiac function. Study Design: Decitabine 20mg/m2 intravenously (IV) over the course of one hour daily for 3 days. Courses were repeated every 28 days when possible. Dose reductions for grade 3 and 4 toxicities were allowed for decitabine (to 10 mg/m2 or 15 mg/m2). The primary endpoint was overall response rate (ORR).

RESULTS: In total, 37 patients were enrolled and treated from September 2014 to October 2017 in our center. The median follow-up was 26 months (range, 4-49 months). The median time from diagnosis to therapy with decitabine was 3 weeks (range, 1-52 weeks). Their F/M ratio was 16/21 and the median age of the enrolled population was 58 years (range, 40-71 years). More than 70% of patients had 2 to 3 cytopenias, and 43% were transfusion dependent at enrollment. Most patients had intermediate 1-risk MDS by IPSS (n=29). Next-generation sequencing was performed in 35 patients (90%). The most frequently detected mutations included TET2 (n=15; 41%), SF3B1 (n=7; 19%), ASXL1 (n=5, 13%), RUNX1 (n= 5, 13%), and SRSF2 (n=3, 8%). The ORR after 2 cycles of decitabine was 65% (24/37),with 35% of patients (13/35) achieving a CR. Hematologic improvement was seen in 19%(7/37). The median time to best response was 3 months (range, 1-12 months). The median number of cycles received was 5 (range, 1-13 cycles). 19% of patients (7/37) became transfusion independent. The EFS and OS were 68% and 78% respectively in 1 year. The treatment was well-tolerated, with most adverse events being of grade 1 to 2, Nausea (24%), Fatigue (14%), Infection/Neutropenic fever (19%), Constipation (6%), Diarrhea (6%). Grade 3 and higher nonhematologic adverse events were rare. Grade 3 adverse events were Infection/Neutropenic fever (6%), but rare, and no grade 4 adverse events were observed.

CONCLUSIONS: These data confirm that decitabine is a feasible and effective agent for low- or intermediate 1-risk MDS pts, and its need longer time to response (more than 2 cycles). Safety profile was acceptable and the most adverse events were Nausea, Fatigue, and Infection/Neutropenic fever.

Disclosures

No relevant conflicts of interest to declare.
Co-Transplantation of Mesenchymal Stem Cells Can Ameliorates Acute Gvhd and Viremia after Allogeneic Hematopoietic Stem Cell Transplantation for Severe Aplastic Anemia: A Multi-Center Retrospective Study of 119 Patients
Authors: Chen Minmin , Zheng Zhizhong , He Guangsheng , Lin Shengyun , Zhang Donghua , Hu Huijing , Liu Jia , Chen Zhizhe , Hu Jianda , Yang Ting
Published: October 2019

BACKGROUD:Mesenchymal stem cells (MSCs) are known to exhibit immunomodulatory, anti-inflammatory, and pro-angiogenic properties, and therefore have the potential to improve the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for severe aplastic anemia (AA).

OBJECTIVE: To explore the efficacy and safety of allo-HSCT by co-transplantation with MSCs in AA patients.

METHODS: We conducted a multi-center retrospective study to comparing the incidence and severity of acute GvHD, transplant-related complication and overall survival (OS) of allo-HSCT with or without co-injection of MSCs in AA patients. A total of 119 consecutive severe AA patients (64 males/55 females) undergoing allo-HSCT at 4 hospitals from January 2012 to February 2018 were analyzed. The median age of the patients is 23 (range 1-56 years). Patients received conditioning regimens based on cyclophosphamide or busulfan, among whom 50 (42%) were MSD-HSCT, 21(17.6%) MUD-HSCT and 48 (40.4%) haplo-HSCT. All patients received cyclosporine A with short course methotrexate for prevention of GVHD. 89 patients transplanted with donor HSCs only, and 30 patients transplanted with HSCs and MSCs .MSCs infusion dose was 0.5-3.0×106/kg, at -1~+1 days following HSCs infusion. None of these patients had severe infection or organ failure before HSCT.

RESULTS: The two groups were well matched demographically. All patients achieved successful engraftment within one month post transplant. There was no difference in time to leukocyte and platelet engraftment in the two groups. The incidence of aGVHD grade II-IV (7.9 % versus 6.7 %, P =1.000) was similar in the HSC versus MSC groups, but there was a trend to a lower incidence of aGVHD grade III-IV in the MSC group (3.4% versus 0%, P=1.0). Moreover, a lower incidence of viremia in the MSC group was observed (CMV: 26.7% vs 62.9%, P < 0.001; EBV: 50% vs 80.9%, P = 0.002).With a median follow-up time of 811 days (range:28-2348days),the 2-yr OS was similar in both groups (82.8% versus 71.1%, P =0.498). A combination of bone marrow and peripheral blood as the sources of stem cells co-transplanted with MSCs (n=9) demonstrated an improved survival benefit (2-yr OS=100%).

CONCLUSION: Co-transplantation with MSCs could ameliorate the challenges of aGVHD and viremia and facilitate survival in allo-HSCT for AA patients.

Disclosures

No relevant conflicts of interest to declare.