bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cell Type-Specific Effects of Crizotinib in Human Acute Myeloid Leukemia with TP53 Alterations
Authors: Antony Marie Lue , Noble-Orcutt Klara , Ogunsan Oluwateniayo , He Fiona , Sachs Zohar 
Published: November 2019

Introduction: In acute myeloid leukemia (AML) standard therapies induce complete remission in 50-70% of patients, but overall two-year survival is less than 20-30% because of high relapse rates. AML with TP53 alteration is largely insensitive to chemotherapy, modern targeted agents, and hematopoietic stem cell transplantation. Mutations in TP53 are seen in approximately 20% of AML and confer a particularly poor prognosis decreasing the 1-2 year survival rates to 0-10%. Leukemia stem cells (LSCs) are defined as the cells that recapitulate the leukemia and cause relapse. Recent large-scale drug-screening efforts to map treatment vulnerabilities in AML have revealed that crizotinib can reduce the in vitro viability of unselected primary human AML bone marrow samples with TP53 alterations (Tyner et al. Nature 2018). Crizotinib is a tyrosine kinase inhibitor that targets ALK, MET, and RON and is FDA approved for treatment of lung cancer. Since LSCs are largely responsible for AML mortality, we investigated whether crizotinib targets LSCs in primary, diagnostic human AML samples with TP53 alterations. We used mass cytometry (CyTOF2) to determine the cell-type specific effects of this drug on intracellular signaling states. We profiled treated samples with a panel that includes immunophenotypic markers (including LSC markers) and intracellular signaling molecules implicated in AML pathogenesis and aggressiveness.
Results: Primary human AML samples with TP53 alterations were treated with crizotinib (10 mM, n=3). There was a reduction of viability by 48 hours of in vitro treatment with crizotinib, relative to vehicle (mean 64% reduction in viability). Crizotinib abolished all colony formation in semi-solid media. Mass cytometry analysis revealed that crizotinib treatment led to a loss of cells bearing LSC-specific markers such as CD123 and TIM3. Crizotinib treatment also induced a consistent loss of phosphorylated-signaling intermediates of the STAT and p38/MAPAPKII pathways in all samples tested. Notably, crizotinib treatment also induced significant reductions in total and phosphorylated NFKB in all samples tested.
Conclusions: These data reveal cell-type specific effects of crizotinib in human AML with TP53 alterations. Crizotinib induced a specific loss of LSCs and inhibition of the STAT, p38/MAPAPKII and NFKB pathways. These data suggest that crizotinib inhibition may target LSCs in AML with TP53 alterations.
Figure

Disclosures
No relevant conflicts of interest to declare.
Prognostic Value of Diffusion-Weighted Magnetic Resonance Imaging in Newly Diagnosed Multiple Myeloma Patients Treated with up-Front Autologous Transplantation
Authors: Fernández-Poveda Elena , Cabañas Valentin , Moreno Maria José , Blanquer Blanquer Miguel , Moraleda José M 
Published: November 2019

Background and aims:
Bone disease extension in symptomatic multiple myeloma (MM) has prognostic implications. However, methods better than the traditional ones are necessary to establish the diagnosis according to the updated International Myeloma Working Group criteria (IMWG).
Magnetic resonance imaging (MRI) is a novel technique to detect bone lesions at diagnosis. The IMWG has incorporated it to the diagnostic work-up. However, due to MRI false-positive results in non-viable lesions, PET-TC is preferred during follow-up to assess prognosis.
Diffusion-weighted magnetic resonance imaging (DW-MRI) uses the diffusion of water in the bone marrow, which is greatly influenced by cellularity and tumor burden. DW-MRI has been explored for the assessment of minimal residual disease (MRD) after autologous stem cell transplantation (ASCT) in a recently published study, showing better progression free survival (PFS) in patients with negative DW-MRI.
Our aim is to compare the PFS in patients with MM with negative and positive DW-MRI at 3 months of ASCT and to assess the differences with PET-CT.
Methods:
MRI and PET/CT were performed in patients diagnosed with symptomatic MM at 3 months after ASCT. All patients received a Bortezomib based three drugs combination as induction treatment. None received maintenance.
Any ≥5 mm focal lesion seen on PET-CT with standardized uptake value (SUVmax)>2.5 for bone tissue or >3.5 for soft tissues was considered positive.
Any ≥5 mm focal lesion seen on DW-MRI that restricted the diffusion, as well as the presence of extramedullary disease, was considered positive.
Statistical analysis was performed using the Kaplan Meier test and Fisher's exact test.
Results:
The baseline characteristics of the 17 patients included are shown in Table 1.
The median PFS in the positive DW-MRI group was 28 months, whereas the median PFS in the negative DW-MRI group was not reached (p=0.029) (image 1).
Both tests were coincidently positive in 3 patients (17%) and negative in 7 patients (41%). There was no coincidence in the remaining 7 (41%) patients, 5 (29%) being DW-MRI positive and 2 (11%) being PET-TAC positive. The difference was statistically non-significant.
Conclusions:
In our population, patients with negative DW-MRI after ASCT have a significantly higher PFS. These data are favorable to the use of DW-MRI in the follow-up of MM.

Disclosures
No relevant conflicts of interest to declare.
Multicenter Study of Ruxolitinibcombined DEP Regimen As a Salvage Therapy for Refractory/Relapsed Hemophagocytic Lymphohistiocytosis
Authors: Wang Jingshi , Wang Zhao 
Published: November 2019

Hemophagocytic lymphohistiocytosis (HLH) is a refractory immune disorder with a significant risk of death.Current therapies have improved the survival of HLH patients; however, approximately 30% patients do not respond to current therapies. At present, there is nounanimously recommended salvage HLH treatment regimen, and there are a few case reports or clinical reports with small sample sizes concerning salvage therapy after first line treatment failure. Therefore, alternative therapies for relapsed/refractory HLH is needed. Previous studies have found that ruxolitinib can improve the inflammatory status well, but the remission depth is not sufficient.
This prospective study aimed to investigate the efficacy of ruxolitinib combined with our previous DEP (doxorubicin-etoposide-methylprednisolone) regimen(DEP-Ru) as a salvage therapy for refractory/relapsed(R/R)HLH.All patients who did not achieve at least partial response 2 weeks after initial standard HLH therapy or relapsed after remission with HLH-94 regimen were enrolled in this study between June 2018 and June 2019. Response to salvage therapy was assessed every 2 weeks after initiation of DEP-Ru therapy and patients were followed until death or until July 2019.
Fifty-four R/R HLH patients were enrolled, including 28 cases of Epstein-Barr virus-associated HLH(EBV-HLH), 5 cases of macrophage activation syndrome (MAS), 3 cases of lymphoma-associated HLH, and 6 cases of familial HLH, 1case of pregnancy, 1 case of drug, and 2 cases of infection. There were 8 cases with unknown underlying diseases. One case was lost to follow-up, Eight cases (15.1%) achieved complete response and 31 cases (58.5%) achieved partial response. The overall response was 73.6% (39/53).Thirty two of the 54 patients survived to subsequent chemotherapy or allogenic hematopoietic stem cell transplantation.
Our study suggests that DEP-Ru regimen compared with DEP regimen had similar efficacy, and no serious complications such as gastrointestinal bleeding occurred in DEP regimen. DEP-Ru has higher efficacy for MAS and HLH with unknown reason , while EBV-HLH and lymphoma-associated HLH have lower efficacy. DEP-Ru regimen is an effective salvage regimen for R/R HLH, which can prolong patient survival and bridge the gap between HLH and its underlying diseases. This study was registered in the Clinical Trail (ClinicalTrails.gov Identifier: NCT03533790).

Disclosures
No relevant conflicts of interest to declare.
Cold Plasma with Immunomodulatory Properties Has Significant Anti-Lymphoma Activities in Vitro and In Vivo
Authors: Cheng Fengdong , Yan Dayun , Chen Jie , Keidar Michael , Sotomayor Eduardo 
Published: November 2019

In recent years, cancer immunotherapy has revolutionized cancer care. Remarkable clinical efficacy and durable responses to checkpoint blockade antibodies or to genetically engineered T-cells (CAR T cell) have been observed in patients with multiple cancers. However, not all cancer patients benefit from these therapies and as such novel immunotherapeutic approaches are needed. The cold atmospheric plasma (CAP) is a form of near room temperature ionized gas, which has shown a promising application in cancer therapy given its antitumor effects in vitro as well as in vivo. For the first time, we have shown that upon CAP treatment, the viability of the immune cells remained unaffected. Strikingly, we observed a significantly stronger immune activation of those CAP treated cells when compare with helium gas control. First, we have demonstrated that even without LPS stimulation, in vitro exposure of peritoneal elicited macrophages (PEM) to CAP for 15 or 30 seconds was sufficient to trigger the production of the pro-inflammatory cytokines IL-12 and IL-6. In addition, decreased production of anti-inflammatory cytokine IL-10 and diminished the expression of PD-L1 were observed in CAP-treated PEM as well. It indicated that CAP treatment may potentially facilitate PEMs as better activator of T cells. Second, in lieu of the stimulatory effects of CAP upon PEM, we asked whether CAP could enhance T-cell activation. So we isolated T cells from spleens of C57BL/6 mice and exposed these T cells to CAP followed by anti-CD3/CD28 stimulation. Our study shown that the production of IL-2 and IFN-g were significantly increased by T-cells treated with CAP as compared with helium gas control. Furthermore, to gain insights into effects of CAP upon immune responses in vivo, we isolated lymph nodes from OTII mice and directly exposed these LNs with CAP, helium gas control or left untreated. Then CD4+ T cells were further isolated from the LNs and cultured with macrophages in the presence or absence of OVA peptide for 48 hours, respectively. Surprisingly, CD4+ T cells isolated from CAP-treated LNs displayed enhanced function indicated by its increased production of IL-2 and IFN-g. More importantly, strong in vivo anti-tumor effects were observed when adoptively transfer CAP exposed T cells to lymphoma bearing animals.
Taken together, we have shown for the first time an elevated immune-stimulatory effect of CAP upon both APCs and T cells in vitro and in vivo. Our finding may potentially shed light of a novel therapeutic approach for future cancer immunotherapy.

Disclosures
No relevant conflicts of interest to declare.
Characterisation of Citrullinated TFPI and Truncated TFPI Constructs By PAD4 in Model and Plasma Systems
Authors: Thomassen M.G.L. Christella D. , Suylen Dennis P.L. , Hackeng Tilman M. , Koenen Rory R. 
Published: November 2019

Background: The Kunitz type tissue factor pathway inhibitor (TFPI) is an important regulator in hemostasis. Decreased concentrations are a risk factor for thrombosis and complete TFPI deficiency is associated with lethality. TFPI is also reported to be an important link between inflammation and thrombosis. Neutrophil extracellular traps (NETs) formed by NETosis can bind TFPI which then can be cleaved and inactivated by neutrophil elastase (NE) during thrombotic events. In neutrophils, the enzyme peptide arginine deiminase 4 (PAD4) is central in the citrullination of histones prior to the externalization of DNA during NETosis. In this study, we provide evidence that PAD4 also might regulate the activity of TFPI by posttranslational modification of its functional arginines into citrulline.
Aim: To study the effect of citrullination of TFPI and various TFPI constructs on their functional activity on FXa or thrombin generation.
Methods: Citrullination of TFPI by the neutrophil protein PAD4 was studied in a model system (FXa inhibition) and in plasma system (thrombin generation). Various TFPI constructs, Kunitz (K) domains K1K2, K2, and TFPI1-161 were used to study the effects of citrullination on inhibition of FXa. LC-MS was used to locate the specific sites of citrullination.
Results: This study shows that PAD4 very efficiently citrullinates full lenght TFPI. Very low concentrations of PAD4 were sufficient (Ki 0.4 nM) to abolish FXa inhibition by TFPI. Citrullination is calcium-ion, time- and concentration-dependent. The truncated variants K1K2 and TFPI 1-161 and the isolated K2 domain were citrullinated less efficiently by PAD4 than TFPI, implying the presence of specific binding sites for PAD4 at the C-terminus of TFPI. The presence of phospholipids inhibited the citrullination reaction, an effect only seen for TFPI and not for all the other TFPI variants. Thus, the presence of the C-terminus in TFPI appears to be favourable for citrullination by PAD4.
Thrombin generation in TFPI-deficient plasma triggered with TF or Russell's viper venom (RVV)-X showed almost complete absence of anticoagulant activity of citrullinated TFPI
Conclusions: To conclude, only TFPI is very sensitive to citrullination by PAD4. Citrullinated TFPI has lost its ability to inhibit FXa. This process might play a role in the increased thrombosis risk with inflammation. Further experiments are needed to determine the physiologic or pathologic relevance of this process.

Disclosures
No relevant conflicts of interest to declare.
Glucose Metabolism in Sickle Cell Disease
Authors: Hassan Taif , Shah Binal N. , Zhang Xu , Gordeuk Victor R 
Published: November 2019

Background. Whether glucose metabolism differs in sickle cell anemia (SCA) patients versus controls is not clear. Transfusional iron overload and Inflammation might enhance glucose intolerance, while lower BMI and increased hypoxic responses might be protective.
Methods: We prospectively determined indicators of glucose metabolism in 46 African-American hemoglobin SS or S/β0-thalassemia subjects >35 years of age who did not have the diagnosis of DM, and examined the relationship of these parameters to BMI and serum ferritin concentration. We compared the SCA subjects to 46 African-American controls from the National Health and Nutrition Examination Study (NHANES) who did not have the diagnosis of DM and were matched by age, sex, BMI and serum ferritin to the SCA subjects.
Results: The BMI was >25 kg/m2 in 23 SCA subjects (48%) and the serum ferritin was >300 ng/ml in 30 (65%) consistent with increased body iron stores. The fasting blood glucose was >125 mg/dL in 3 of 46 SCA subjects (6.5%) suggesting unsuspected DM. The fasting glucose concentration independently correlated with serum ferritin concentration (r=0.336, P=0.024) and the fasting insulin level correlated independently with both BMI (r=0.583, P=0.00002) and serum ferritin (r=0.300, P=0.016). Measures of glucose metabolism did not differ significantly between the SCA subjects and NHANES controls, even though the SCA subjects had lower blood pressures, WBC counts and blood lipid levels.
Conclusions: Measures of glucose metabolism in older SCA patients are similar to controls matched for BMI and iron status despite marked differences in blood pressure, hemoglobin, WBCs and blood lipid levels.

Disclosures
Gordeuk: Inctye: Research Funding; CSL Behring: Consultancy, Honoraria, Research Funding; Ironwood: Research Funding; CSL Behring: Consultancy, Honoraria, Research Funding; Imara: Research Funding; Emmaus: Consultancy, Honoraria; Novartis: Consultancy, Honoraria, Research Funding; Inctye: Research Funding; Modus Therapeutics: Consultancy, Honoraria; Pfizer: Research Funding; Pfizer: Research Funding; Global Blood Therapeutics: Consultancy, Honoraria, Research Funding; Ironwood: Research Funding; Imara: Research Funding.
Immunophenotype Drift of Residual Plasma Cells Indicates Therapeutic Response and Prognosis in Multiple Myeloma
Authors: Du Chenxing , Gang An , Xu Yan , Mao Xuehan , Yan Yuting , Liu Jiahui , Fan Huishou , Sui Weiwei , Deng Shuhui , Zengjun Li , Zhao Jiawei , Yi Shuhua , Zou Dehui , Qiu Lugui 
Published: November 2019

Background
Chemotherapy resistance remains a significant hurdle in the treatment of multiple myeloma (MM). However, it is difficult to discriminate the potential refractory patients from the very early stage. Flow cytometry is a convenient tool to detect the residual myeloma cell tiding, indicating therapeutic response sensitively.
Methods
From June, 2014 to December, 2016, 172 sequential patients with newly diagnosed multiple myeloma were enrolled in the BDH2008/02 clinical trial. Patient informed consent was obtained in accordance with the Declaration of Helsinki. 144 patients with at least two flow cytometry detections were analyzed. Bone marrow samples were detected by an eight-color EuroFlow panel. CD20 negative and CD81 positive is defined as normal phenotype.
Results
We conducted a median of 3-time (2-8) flow cytometry detection on each patient. When newly diagnosed and achieved best response, CD20, CD81 expression rates were 29.9%, 9.7% and 14.9%, 64.4% (P=0.0091, P<0.0001), respectively. According to the status variation of CD20 and CD81, all patients were divided into three groups: both markers were always normal (Group A), either CD20 or CD81 was abnormal at diagnosed and turned normal during therapy (Group B) and markers stayed abnormal (Group C). Patients with undetectable residual tumor cells were also classified as Group A. The overall response rate of the patients in Group C was inferior to Group B (>PR rate: 54.3% vs. 71.4%, P=0.021). And the OS of Group C was significantly worse than Group A and B (47.9 months vs. not reached vs. not reached, P=0.036).
Conclusion
CD20/CD81 switching to normal phenotype during therapy indicates therapeutic response and an improved outcome than that staying abnormal. The expression tiding of CD20 and CD81 may be a reasonable combination to dynamically stratify MM patients, directing the choice of maintenance therapy.
Figure

Disclosures
No relevant conflicts of interest to declare.
Cost per Progression Free Survival (PFS) of Daratumumab, Elotuzumab, Carfilzomib and Ixazomib Combinations with Lenalidomide/Dexamethasone (DRd, ERd, KRd, IRd) in Patients with Relapsed/Refractory Multiple Myeloma (RRMM)
Authors: Santana Pamela , Saad Ricardo , Kolanian Adrielle , Fioratti Cyntia , Junqueira Marcela , Decimoni Tassia 
Published: November 2019

OBJECTIVES: Multiple myeloma (MM) is the second most common hematological malignancy with several available therapies and an extensive pipeline. A metanalysis comparing pivotal relapsed/refractory (RRMM) studies that assessed DRd, ERd, KRd and IRd showed that DRd had the best PFS results. The goal of this analysis was to compare the cost per progression-free survival (PFS) for each of the comparators from the perspective of the Brazilian private healthcare system.
METHODS: We calculated the cost per PFS individually using the most recent available data from follow-up (FUP) studies of the therapies assessed. PFS values were based on POLLUX 3-year FUP (DRd), ENDEAVOR 4-year FUP (ERd), ASPIRE 3-year FUP (KRd) and TOURMALINE-MM1 (IRd). Only drug acquisition costs (with wastage) were considered in the analysis (according to drug label dosage) and were retrieved from the Brazilian official price list (Jul/19). Cost per PFS was calculated by dividing the total cost in the PFS period by time in PFS (in months). Cost in 18 months was assessed because it is the maximum time that carfilzomib is recommended to be used.
RESULTS: DRd showed the best PFS result compared with ERd, KRd and IRd (42.0 vs. 19.4, 26.1, 20.6, respectively). Total costs in the period of PFS for each of the comparators were BRL 2.02 million (DRd), BRL 1.29 million (ERd), BRL 1.14 million (KRd) and BRL 0.98 million (IRd). Costs per PFS- were BRL 48,060, BRL 66,343, BRL 43,822, BRL 47,760 and costs in the 18 first months were BRL 1.01 million, BRL 1,13 million, BRL 0.95 million and BRL 0.84 million for DRd, ERd, KRd and IRd, respectively.
CONCLUSIONS: DRd showed the best PFS result, however its cost per PFS is the 3rd lowest among the comparators. As costs are directly dependent on PFS, the longer the PFS the higher are the costs. Additionally, daratumumab has continuous usage meanwhile carfilzomib has label recommended usage for just 18 cycles due to toxicity and tolerability-related events.

Disclosures
Santana: Janssen pharmaceuticals: Employment. Saad:Janssen pharmaceuticals: Employment. Kolanian:Janssen pharmaceuticals: Employment. Fioratti:Janssen pharmaceuticals: Employment. Junqueira:Janssen pharmaceuticals: Employment. Decimoni:Janssen Pharmaceuticals: Employment.
Outcomes of Haploidentical Haematopoietic Stem Cell Transplantation for Paroxysmal Nocturnal Haemoglobinuria
Authors: Liu Limin , Zhou Huifen , Miao Miao , Wu Depei , Tang Xiaowen , Qiu Huiying , Han Yue , Jin Zhengming , Chen Suning , Sun Aining , Wang Qingyuan 
Published: November 2019

Background: Haematopoietic stem cell transplantation (HSCT) is the only curative therapy for paroxysmal nocturnal haemoglobinuria (PNH). Haploidentical HSCT (haplo-HSCT) is an alternative treatment method for PNH patients lacking suitable identical donors. The current study evaluated the feasibility of haplo-HSCT in PNH patients.
Methods: We retrospectively analysed the outcomes of 28 PNH patients who received haplo-HSCT between October 2010 and December 2018.
Results: The median age was 28 (range 6-54) years. One patient (underwent HSCT in December 2018) was not evaluable for engraftment or haematopoietic recovery because of early death on day +14 due to septicaemia. The 27 evaluable patients all achieved myeloid engraftment and complete chimerism, and one patient experienced secondary graft failure. Three patients demonstrated delayed platelet recovery, and one patient demonstrated platelet graft failure. The median time to neutrophil recovery was 12 (range, 9-26) days, and the median time to platelet recovery was 16 (range, 11-75) days. The cumulative incidences were 29.63% for grades I to IV acute graft-versus-host disease (aGVHD) and 14.82% for grades II to IV aGVHD. No patient experienced grades III to IV aGVHD. With a median follow-up time of 37 (range 4-78) months, the cumulative incidence of chronic graft-versus-host disease (cGVHD) was 28.81%, and the cumulative incidence of moderate-severe cGVHD was 11.73%. No patient relapsed during the follow-up period. The 12-month transplantation-related mortality (TRM) rate was 15.25%. The probabilities of 3-year overall survival (OS) and graft-versus-host disease-free, failure-free survival (GFFS) were 84.8 ± 7.1% and 77.0 ± 8.3%, respectively.
Conclusions: The preliminary results indicated that haplo-HSCT is a feasible option for PNH patients lacking suitable identical donors.

Disclosures
No relevant conflicts of interest to declare.