bluuebunny/crossref_metadata_2025 huggingface dataset:
Monoclonal Gammopathies and Their Significance: A Retrospective Analysis of Monoclonal Gammopathy and Renal Disease
Authors: Kumar Anupama , Drew David , Inker Leslie , Comenzo Raymond L , Varga Cindy
Published: November 2019

Background:Monoclonal gammopathy of renal significance (MGRS) describes a subtype of patients with monoclonal gammopathy of unknown significance (MGUS) who have kidney disease due to immunoglobulin deposition. Early identification of MGRS and treatment with chemotherapy can prevent progression to kidney failure. There is limited data on the characteristics of populations with concomitant MGUS and chronic kidney disease (CKD), and on the prevalence of diagnosed MGRS in this population in the real world. Methods: Through retrospective chart review, we identified 246 patients with ICD-9 or -10 codes denoting both MGUS and chronic kidney disease (CKD) between the years of 2000 and 2017. Patients with related overt malignancies such as multiple myeloma, Waldenstrom's macroglobulinemia, and amyloidosis at onset were excluded, leaving 144 evaluable patients. Results:The median age was 78, and females made up 41.0% of the population. At time of MGUS diagnosis, the median eGFR was 48 mL/min/1.73 m2, and in patients with a quantifiable gammopathy, the median M-protein was 0.54 g/dL. A total of 53/144 (36.8%) patients had a bone marrow biopsy and 19/144 (13.2%) underwent kidney biopsy. MGRS was considered as a cause of kidney dysfunction in 20/144 patients (13.9%), but only 6 were further worked up by kidney biopsy. In 3/144 (2.1%) patients, MGRS was confirmed by kidney biopsy. Ten of (6.9%) the total population and 1 (5%) of the patients with suspected or confirmed MGRS (n=20) developed malignant transformation. Conclusion: MGRS should be considered in the differential diagnosis of patients with MGUS and co-existing chronic kidney disease. Based on our findings, renal biopsy is underutilized in the work up of this treatable disease. We propose a simple algorithm for workup of suspected MGRS (see Figure 1).

Figure 1

Disclosures

Comenzo: Myself: Patents & Royalties: Patent 9593332, Pending 20170008966; Caelum: Consultancy, Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Karyopharm: Research Funding; Prothena Biosciences: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Sanofi-Aventis: Membership on an entity's Board of Directors or advisory committees; Takeda: Research Funding; Unum: Membership on an entity's Board of Directors or advisory committees, Research Funding.
Insulin-like Growth Factor-1-Induced Epithelial-Mesenchymal Transition Promotes Multiple Myeloma Progression
Authors: Peng Yue , Wang Xiaman , Shen Ying , Li Fangmei , Zhang Ru , Jia Yachun , He Aili
Published: November 2019

MM is an incurable B cell neoplasm, which is characterized by the clonal proliferation of plasma cells in the bone marrow (BM), accounts for approximately 10% of hematological malignancies. Insulin-like growth factor-1 (IGF-1) is a vital proliferation factor in myeloma cells, recent researches demonstrated that it may be involved in MM cell metastasis. In last decade, studies have demonstrated that IGF-1 induces epithelial-mesenchymal transition (EMT), which has been tightly correlated with increased motility and invasive capacity in several cancers including gastric cancer, hepatocellular carcinoma, and melanoma. However, it remains unclear whether IGF-1 induces EMT and promote myeloma progression.

In this study, we firstly observed the morphology of the myeloma cell line Karpas707 changed from tight polygon to loose spindle, a mesenchymal phenotype after treated with IGF-1 (50 ng/ml) after 72h. We supposed IGF-1 may induced EMT in myeloma cells. Furthermore, when serum-starved MM cell lines RPMI8226 and MM1.S were stimulated with 50ng/mL of IGF-1, we found that the expression of mesenchymal features was significantly upregulated in a time-dependent manner, as determined by both flow cytometry analysis and western blotting. Next we confirmed that IGF-1 can activate the PI3K/Akt signaling pathway by inducing the phosphorylation of Akt in MM cells. To investigated whether PI3K/Akt signaling pathway involved in the IGF-1 mediated EMT, we blocked PI3K/Akt signaling pathway using Akt inhibitor MK2206 in IGF-1 treated MM.1S cells. We found MK2206 reversed these changes of EMT markers. Moreover, we found the IGF-1-induced EMT promotes myeloma progression, including migration, invasion, adhesion and clone formation in MM cells, using transwell migration and invasion assay, cell adhesion assay, colony formation assay and flow cytometry. Furthermore, to further validate IGF-1-induced EMT promote MM progression, we knocked down mesenchymal marker vimentin using siRNA in MM.1S cells (VIM k/d cells). We detected that these function IGF-1 plays in myeloma was impaired in Vim k/d cells, indicating that IGF-1-induced EMT promotes MM progression. Finally, we validated that IGF1 expression is aberrantly upregulated and correlated with the expression of the EMT markers by analyzing the mRNA data of BM samples from normal donors and patients in different stages of plasma cell neoplasm from the GEO database. Taken together, these data demonstrate that the mesenchymal phenotype induced by IGF1 contributes to MM progression via PI3K/Akt pathway.

Disclosures

No relevant conflicts of interest to declare.
Efficacy and Safety of Apatinib in Relapsed or Refractory Diffuse Large B Cell Lymphoma: An Open-Label, Single-Arm, Prospective Study
Authors: Ma Xinran , Li Ling , Zhang Lei , Li Xin , Fu Xiaorui , Wang Xinhua , Wu Jingjing , Zhang Xudong , Feng Xiaoyan , Zhou Zhiyuan , Chang Yu , Li Zhaoming , Young Ken H. , Zhang Mingzhi
Published: November 2019

Background

Treatment options for relapsed or refractory diffuse large B-cell lymphoma (RR DLBCL) represent an unmet medical need. Apatinib is a new oral tyrosine kinase inhibitor mainly targeting vascular endothelial growth factor receptor 2 (VEGFR-2) to inhibit tumor angiogenesis, and has shown encouraging anti-tumor effects in multiple solid tumors. Yet clinical evidence of apatinib as a potential treatment choice for RR DLBCL remains rare. In current study, we evaluated the efficacy and safety of Apatinib for patients with RR DLBCL.

Methods

In this open-label, single-arm, prospective study, we enrolled patients aged 14-70 years with treatment failure of at least two chemotherapeutic regimens using Simon's two stage design. All patients took Apatinib at an initial dose of 500mg on a 4 weeks cycle at home and visited outpatient clinic every 2 cycles to evaluate the efficacy and to record the adverse events. Dose interruptions and reductions were only allowed when there were grade 3 haematological adverse events or grade 2 non-haematological adverse events. We considered objective response rate (ORR) as the primary end point, and progression-free survival (PFS), overall survival (OS) plus duration of response (DoR) as the secondary end point. (This trial was registered at ClinicalTrials.gov, identifier: NCT03376958.)

Results

From January 2017 to February 2019, we screened 35 and enrolled 32 eligible patients. At the cutoff point (April 2019), we noted 2 (6.3%) complete responses, 12 (37.5%) partial responses and 9 (28.1%) stable diseases, attributing to an ORR of 43.8% and a disease control rate (DCR) of 71.9%. The median PFS was 6.9 months (95%CI 5.8-7.9) and the median OS was 7.9 months (95%CI 7.0-8.7). Kaplan-Meier estimates of PFS and OS are shown in Figure 1. For patients achieved PR, The median DoR was 5.0 months (95%CI 3.5-6.5). At data analysis time, the median duration of follow-up was 8.6 months (95%CI 7.3-9.4), and 16 (50.0%) patients remained on treatment. 21 patients showed tumor shrinkage comparing with their baseline data and the best percentage changes from baseline are shown in Figure 2. The most common grade 3-4 adverse events (AE) were hypertension (12.6%), hand-foot syndrome (9.4%) and leukopenia (6.3%). No Apatinib-related deaths were noted. AEs arose in at least one patient are shown in Table 1.

Conclusion

With the potential superiority of home administration without frequent hospitalization, apatinib can be an arrow in the bow for RR DLBCL patients considering its favorable efficacy, manageable AEs, improved patient adherence and economic effectiveness. Further investigations into this topic will be of profound significance.

Disclosures

No relevant conflicts of interest to declare.
Genomic Landscape and Risk-Stratification for De Novo Acute Myeloid Leukemia with Normal Cytogenetics and No NPM1 or FLT3-itd Mutation
Authors: Zhou Ya-Lan , Wu Li-Xin , Gale Robert Peter , Wang Zi-Long , Li Jin-Lan , Jiang Hao , Jiang Qian , Jiang Bin , Cao Shan-Bo , Lou Feng , Sun Ying , Wang Chengcheng , Li Ting , Liu Yan-Rong , Wang Yu , Chang Ying-Jun , Xu Lan-Ping , Zhang Xiaohui , Liu Kaiyan , Ruan Guorui , Huang Xiaojun
Published: November 2019

Introduction-About 25% of persons with new-diagnosed acute myeloid leukemia (AML) have normal cytogenetics and no NPM1 or FLT3-ITD mutation. The prognosis and best therapy of these persons is controversial.

Methods-We evaluated 809 consecutive newly diagnosed adult with normal cytogenetics and 231 of whom had no NPM1 or FLT3-ITD mutation identified by targeted regional sequencing. 158 achieved a complete remission within 2 cycles of induction therapy and were assigned to 2 different post-remission strategies: (1) 6 courses of consolidation chemotherapy (N=95); or (2) 2-4 courses of consolidation chemotherapy and an allotransplant (N=63).

Results-In multi-variable analyses a WBC ≥13·6×10E+9/L, mutated IDH2, not having a bi-allelic CEBPA mutation at diagnosis, a positive measurable residual disease (MRD)-test during consolidation and not receiving an allotransplant were independently associated with a higher cumulative incidence of relapse (CIR) and worse event-free survival (EFS). Amongst subjects with IDH2 mutations, non-bi-allelic CEBPA mutations or a positive MRD-test, subjects receiving an allotransplant had a lower 5-year CIR (16% [95% confidence interval, 6, 26%]; vs. 83% [72, 95%]; hazard ratio, HR=8·77 [4·05, 13·49]; P < 0·001) and better 5-year EFS (74% [60, 88%] vs. 15% [5, 25%]; HR=0·16 [0·09, 0·29]; P < 0·001). In contrast, in subjects with none of these adverse predictive variables there was no difference in CIR and EFS between those receiving an allotransplant and those who did not.

Conclusions-Our data suggest a strategy to identify which persons with AML with normal cytogenetics and no NPM1 or FLT3-ITD mutation benefit from an allotransplant.

Trial Registration: Registered in the www.clinicaltrials.gov, NCT01455272 and NCT02185261.

Keywords: Acute myeloid leukemia, mutations, prognosis, targeted regional sequencing, measurable residual disease, risk stratification.

*Correspondence

Profs. Guo-Rui Ruan and Xiao-Jun Huang

Peking University Peoples Hospital and Institute of Hematology

No.11 Xi-Zhi-Men South Street, Beijing 100044, China

T 86-10-88324672

F 86-10-88324672

Disclosures

No relevant conflicts of interest to declare.
The Role of Direct Oral Anticoagulants in Antiphospholipid Syndrome
Authors: Alsheef Mohammed Abdullah , Alomar Mukhtar , Alarfaj Ohoud , AlHamidi Hussam A , Zaidi Abdul Rehman Z.
Published: November 2019

Background

Antiphospholipid Syndrome (APS) is defined by thrombosis and/or pregnancy morbidity in patients with antiphospholipid antibodies (aPL). There remains a pressing need for identification of clinical and laboratory parameters that define patients at most considerable risk for APS-related events on Direct oral anticoagulants (DOACs).

Aims

To demonstrate the efficacy and safety of DOAC (Rivaroxaban) compared with Warfarin (Vitamin K Antagonist) in patients with thrombotic APS.

Methods

A retrospective cohort study of thrombotic APS patients who were started on anticoagulant therapy, either VKA or DOAC, from 2010 -2016.

Results

We investigated 73 patients diagnosed with APS. 83.5% of the patients were female. The age range at diagnosis was between 26-40 (52%). 67% of the cases were diagnosed in the form of DVT while the rest had arterial or unusual site venous thrombosis. 43% of the patients had systemic lupus disease. 28% of the cases had a single-positive aPL profile, 23% had double-positive, and 49% had triple-positive aPL. 68.5% of the cases were treated with warfarin, while 31.5% of the cases were switched from warfarin to Rivaroxaban. 15% of patients had major and clinically relevant non-major bleeding (2 patients had major bleeding in the warfarin group, and four patients in the rivaroxaban group). Recurrence of thrombosis (arterial and venous) was 43% with Rivaroxaban and 30% with warfarin. The average duration from starting Rivaroxaban to development of complications such as bleeding and thromboembolic events was mostly after 1-2 years (70%).

Conclusions

Rivaroxaban should be used with extreme caution in APS patients, especially patients with a full positive aPL profile and arterial thrombosis.

Disclosures

No relevant conflicts of interest to declare.
CRISPR Dropout Screens Identify DHODH,PIK3C3, and Crkl as Potential Therapeutic Targets in Acute Myeloid Leukemia
Authors: Takacs Gregory , Zhou Yujia , King David J. , Meacham Amy , Alex Loguinov , Tagmount Abderrahmane , Sobh Amin , Russo Max , Vulpe Chris D , Cogle Christopher R.
Published: November 2019

Relapsed disease is still a major challenge in AML despite recently approved targeted therapies. New therapeutic targets are needed. To identify vulnerabilities in AML, screening data from a lentiviral-based genome-wide pooled CRISPR-Cas9 library (DepMap) was analyzed by examining gene knockout (KO) dependency scores in 15 AML cell lines (HEL, MV411, OCIAML2, THP1, NOMO1, EOL1, KASUMI1, NB4, OCIAML3, MOLM13, TF1, U937, F36P, AML193, P31FUJ). 438 gene KOs were identified as essential to AML cell survival in 14/15 AML cell lines. 247/438 (56%) genes had no active compound or approved drug interacting with its gene-products (DGIdb, canSAR Black) but were tractable targets based on 3D modeling or ligand-based druggability scoring (canSAR Black). This refined list of targets made up a new screening platform termed disKO. For clinical relevance, a primary AML specimen was treated with a custom-made lentiviral-based pooled CRISPR-Cas9 library including disKO targets using guide RNA's constructed against a parsed version of the druggable genome. Following transduction, PCR and oligo-sequencing were performed on Days 0, 7, and 14. Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout (MAGeCK) was used to identify gene KOs associating with AML dropout over time. Four gene KOs from disKO were identified as essential in primary AML: NUP93, DHODH, CRKL, and PIK3C3. NUP93 loss of function mutations have been associated with steroid-resistant nephrotic syndrome in patients, suggesting inhibition may be toxic. DHODH inhibitors are currently being tested in AML. PIK3C3 codes for lipid kinases involved in autophagocytosis, and PIK3C3 inhibitors are in preclinical stage of development. CRKL is a proto-oncogene encoding src homology domains (SH2, SH3), which activate RAS and JNK signaling pathways. Although CRKL is a substrate of BCR-ABL CML, its essentiality in our study was found in AML cell lines and primary AML lacking the BCR-ABL fusion. Differential gene expression analysis showed CRKL to have lower expression in AML when compared to normal hematopoietic cells. Its druggability score based on six 3D structures and 1.3 ligand z-score indicates promising pharmacologic inhibition. In conclusion, a new CRISPR-Cas9 screening method termed disKO led to the identification of three viable targets for AML: PIK3C3, DHODH, and CRKL. Current efforts are focused on target validation followed by compound screening to find inhibitors with therapeutic potential.

Disclosures

No relevant conflicts of interest to declare.
Calr Mutational Status in Egyptian Patients with Myeloproliferative Neoplams
Authors: Soliman Eman A. , El-Ghlban Samah I. , Abdelaleem Abdelaleem H. , Abdel-Aziz Sherin , Shamaa Sameh , Abdel-ghaffar Hasan A.
Published: November 2019

It has been known that the insertion/deletion mutation of CALR gene is the second deriver mutation in myeloproliferative neoplasm (MPN) of essential thrompocythemia (ET) and primary myelofibrosis (PMF). As the molecular workup has been incorporated for the prospective screening and diagnosis of MPN in our Oncology Center. An Egyptian 87 cohort of patients with non-mutated JAK2 (58 ET and 29 MF) were investigated using polymerase chain reaction (PCR) as a pilot study. We found that 37 out of 87 patients (42%) were carrying CALR mutations (30 out of ET (52%) and 7 out of MF (24%)). Sanger sequencing was used to determine the type of CALR mutations in all positive patients and we found that 13 out of 37 (35%) had type 1/type 1-like and 36 out of 37 (97%) with type 2/type 2-like. This CALR mutation profile in Egyptian patients appear different from the western status as type2/type 2-like is the highest in our patients (97%) versus 35-45% and type1/type 1-like was 35% versus 55-65% compared to western results. Meanwhile, the clinical course and phenotype of our cohort of patients is not similar to that in western as there is no significant difference of overall survival between type1/type1-like and type2/type2-like. This finding might be due to the different environmental and genetic backgrounds of Egyptian population. A part of it might be related to the HCV infection as 12 out of 37 (32%) had HCV infection. Further study is in progress on a large number of patients to correlate that with the clinicopathological status, response to therapy and the mechanistic pathway of oncogenic transformation.

Disclosures

No relevant conflicts of interest to declare.
Human PAR4 Is a More Potent Receptor for Activating Platelets Than Mouse PAR4
Authors: Renna Stephanie , Edelstein Leonard C. , McKenzie Steven Edward
Published: November 2019

PAR4 is a protease-activated receptor with major roles in both platelet aggregation and platelet procoagulant function, contributing to both hemostasis and thrombosis in vivo. It is a major target for anti-thrombotic agents in current development. There are notable differences in the amino acid sequences between human (hu) PAR4 and mouse (mu) PAR4 in domains associated with the mechanisms of receptor action. These include the second transmembrane domain, which has a Valine at position 120 in muPAR4 while it is Alanine or Threonine in huPAR4 (Edelstein, Nature Med 2014). Other differences include 4 non-conservative amino acid changes in extracellular loop 2 and a major non-conservative change (mu = Cysteine, hu = Glutamine) in helix 8 in the cytosolic carboxy terminal (Ramachandran, Mol Pharm 2017).

We generated a unique set of mice which enable us to compare for the first time the potential differences in platelet activation between huPAR4 and muPAR4 in the platelet context, rather than in heterologous cells. We generated and characterized 5 independent lines of mice transgenic for human PAR4, using an approach with a large genomic clone which we have implemented successfully in the past. Each of these huPAR4 transgenic lines has been bred to the mouse PAR4 knockout mice generated by Coughlin and colleagues (generously provided by S. Kunapuli, Temple University). The mice are referred to as PAR4 tgKO mice; 3 express the hu Thr120 allele and 2 the Ala120 allele. The level of huPAR4 expression in the tgKO platelets is equivalent to that of muPAR4 in wild-type mouse platelets.

Washed platelets from wild-type mice, PAR4 tgKO mice, and muPAR4 KO mice were stimulated with a range of concentrations of PAR4 activating peptide (PAR4-AP, AYPGKF) and the activation of αIIbβ3 and expression of P-selectin on the surface were determined with flow cytometry. As expected, muPAR4 knockout mice showed no response to the treatment, but reacted normally to other agonists. While we observed small differences between the hu Ala120 and Thr120 tgKO mice, consistent with prior reports by Bray, Edelstein and colleagues for human platelets, we observed large and statistically significant differences between all tgKO mouse platelets tested and wild-type mouse platelets.

In summary, all other things being equal (i.e. the same platelet context), human PAR4 is a more potent receptor for platelet activation than mouse PAR4. Studies are in progress to elucidate the contribution of the different functional domains and the roles of heterotrimeric G proteins, calcium and other signaling intermediates.

Figure

Disclosures

No relevant conflicts of interest to declare.
Fludarabine and Cyclophosphamide Based Conditioning Is Associated with Good Outcomes in Patients Undergoing Matched Sibling Donor Transplants for Aplastic Anaemia
Authors: George Biju , Abraham Aby , Korula Anu , Devasia Anup , Lakshmi Kavitha M , Kulkarni Uday Prakash , Abu Backer Fouzia N , Srivastava Alok , Mathews Vikram
Published: November 2019

Background: Fludarabine based conditioning protocols are increasingly being used for transplant (SCT) in patients with aplastic anaemia (AA) especially in those who have received multiple transfusions.

Patients and Methods: Between January 2004 and June 2019, 229 patients with AA and HLA identical sibling donors underwent SCT at CMC Vellore India using Fludarabine [180 mg/m2 over 6 days] and Cyclophosphamide [120 mg/kg over 2 days]. Few patients received low dose ATG [ATGAM 40 mg/kg]. Data on HSCT and outcomes were collected from the institutional database and individual medical records.

Results: The median age was 23.9 years (range: 1.5 - 58) including 151 males and 78 females including 78 children (33.4%). Donors were matched sibling (n = 215) or family donors (n = 14). The graft source was Bone marrow in 11 and peripheral blood stem cells in 218. GVHD prophylaxis consisted mainly of cyclosporine and methotrexate mainly with few receiving post-transplant cyclophosphamide.

Engraftment occurred in 90% with graft failure in 2.6% and early death in 7.4%. Regimen related toxicity (RRT) was seen in 4.7% and included veno-occlusive disease of liver and hemorrhagic cystitis. Acute GVHD (grade 2-4) occurred in 26.4% while chronic GVHD was seen in 40.3%. The 5 year overall survival (OS) for the entire group is 75.9 + 4.9%. The 5 yr OS was 81.4 + 3.9% for ages 0 - 20, 76.3 + 4.4% for ages 21-40 and 55.3 + 9.3% for ages > 40 years. Age > 40 years (p = 0.000), presence of fever requiring hospital admission within 4 weeks prior to SCT (p = 0.001) and acute GVHD (p = 0.051) were identified as risk factors associated with a poor outcome on a univariate analysis while on a multivariate analysis, older age and fever continued to remain significant.

Conclusion: This is the largest series of patients with AA undergoing SCT using a fludarabine based conditioning and is associated with improved survival following sibling donor HSCT for AA. Presence of fever requiring a hospital admission immediately prior to SCT is associated with poor outcomes.

Disclosures

No relevant conflicts of interest to declare.
Targeting Polo-like Kinase 4 for Therapeutic Benefit in the Patients with Diffuse Large B-Cell Lymphoma
Authors: Zhao Yi , Han Yang , Yang Juan , Hu Shunfeng , Wang Xin
Published: November 2019

Abstract:

Polo-like kinase 4 (PLK4) is a serine/threonine kinase that plays a critical role in regulating centriole duplication. PLK4 overexpression causes centriole amplification and genomic instability, which is the hallmark of cancer. Thus, PLK4 has been suggested as a tumor promoter role in solid malignancies. CFI-400945 is a highly selective small-molecule inhibitor of PLK4. CFI-400945 treatment results in centriole duplication defects, which leads to mitotic catastrophe and cell death. The antineoplastic effects of CFI-400945 have been reported in many cancers and this inhibitor has entered Phase I clinical trials for solid tumors. However, the role of PLK4 and cellular effects of CFI-40045 in hematological malignancies remains elusive.

In the present study, we found that PLK4 expression is up-regulated at both transcriptional and translational levels in diffuse large B-cell lymphoma (DLBCL) cell lines (Fig. 1a, Fig. 1b). DLBCL patients displayed high protein levels of PLK4 as visualized by immunohistochemistry (Fig. 1c). In vitro, treatment of DLBCL cell lines with CFI-400945 induced a decrease in cell proliferation and an increase in cell apoptosis and a block at the G2-M transition in a dose-dependent manner (Fig. 1d, Fig. 1e, Fig. 1f). Furthermore, we found that the antineoplastic effects of CFI-400945 in DLBCL cell lines were due to the increased DNA damage, as observed by increased accumulation of γH2AX through phosphorylation of CHK1/CHK2 (Fig. 1g). Since PLK4 inhibition affects DNA damage, we next aimed to explore the effects of PLK4 expression on sensitivity of DNA damage-inducing drugs (e.g., doxorubicin). We found that silencing of PLK4 via shRNA improve the anti-tumor effects of doxorubicin in DLBCL cell lines (Fig. 1h, Fig. 1i). PLK4 inhibition in combination with doxorubicin significantly increased phosphorylation of H2AX compared to cells treated with doxorubicin alone (Fig. 1j).

In summary, our data indicates that PLK4 is aberrantly expressed in DLBCL cell lines and tissues. Targeting PLK4 with the selective inhibitor CFI-400945 suppresses cell proliferation and induces apoptotic deaths and causes DNA damage in vitro. Our findings established PLK4 as a potential therapeutic target in DLBCL. Silencing of PLK4 expression induces accumulation of DNA damage and enhances the cytotoxic effects of doxorubicin. Thus, the levels of PLK4 may serve as a determinant of doxorubicin sensitivity and a predictive biomarker of the patients with DLBCL treated with R-CHOP based therapy.

Figure 1

Disclosures

No relevant conflicts of interest to declare.
Anemia Is Negatively Correlated with Cardiac Index in Hereditary Hemorrhagic Telangiectasia(HHT) Patients with Liver Arteriovascular Malformations (AVMs)
Authors: Farooq Hafsa , Lee Alfred I , Young Lawrence , Henderson Katharine
Published: November 2019

OBJECTIVE:Patients with hereditary hemorrhagic telangiectasia (HHT) and liver arteriovascular malformations (AVMs) may develop high output cardiac failure. Iron deficiency anemia is common in HHT patients and is believed to adversely impact cardiac index (CI) but has not been systemically studied in this population. We performed a retrospective study to assess the relationships among hemoglobin, iron, and CI.

METHOD:Chart records were reviewed of HHT patients with liver AVMs who were seen at Yale HHT Center from 2003-2019 and who had undergone cardiac catheterization with calculation of CI. For each patient, clinical history, laboratory studies, and CI were recorded. All variables were categorized according to median of interquartile range and analyzed using Spearman's correlation coefficients.

RESULT:Seventeen patients with HHT and liver AVMs were identified who had undergone cardiac catheterization. Median age was 68 years (range, 36-85); 15 were female and 2 males. Median values of hemoglobin, ferritin, and CI were 10.6 g/ld., 35 ng/mL, and 4.77L/m/m2, respectively. A statistically significant, inverse correlation between hemoglobin and CI was observed (R = -0.45, P = 0.03). Serum iron and CI were also negatively correlated although the association was not significant (R = -0.3, P = 0.17). No significant correlation of serum ferritin with CI was seen (R = 0.05, P = 0.8).

CONCLUSION:Anemia is negatively correlated with CI in HHT patients with liver AVMs. The impact of iron indices on CI is uncertain and partly limited by our small sample size. Larger studies incorporating multivariable analyses are warranted.

Figure

Disclosures

No relevant conflicts of interest to declare.
Cytokine Release Syndrome Is an Independent Risk Factor Associated with Platelet Transfusion Refractoriness after CAR-T Treatment for Relapsed/Refractory Acute Lymphoblast Leukemia
Authors: Liu Yadan , Hu Yongxian , Huang He
Published: November 2019

Platelet transfusion refractoriness (PTR) has been widely observed in hematopoietic stem cell transplantation (HSCT) and chemotherapy in some researches. However, PTR not only has unclear mechanism but no recognized standard treatment guidelines, even has a life-threatening effect on hematological patients. Our study is aim to evaluate the rate of PTR and related factors of PTR in patients with refractory/relapsed acute lymphocytic leukemia (R/R ALL) who received chimeric antigen receptor T cell therapy (CAR-T). We retrospectively collected the data of 49 R/R ALL patients between July 2011 and June 2019 in our single center, and divided them into chemotherapy group and CAR-T group, including 26 patients only receiving chemotherapy and 23 undergoing CAR-T therapy. All R/R ALL patients underwent multiple first lines or second lines of chemotherapies. As for CAR-T group, the fludarabine and cyclophosphamide were administrated three days to lower the tumor burden before CAR-T cell infusion. Age, sex, height, weight, the platelet count before and after transfusion, transfusion unit are all included in our data. In terms of the CAR-T group ,we also collect the peak body temperature , the peak c-reactive protein ,CRS grade , IL6 , IL10 , IFN-γ. The pre-transfusion and post-transfusion platelet count were obtained after each transfusion, the 12h CCI<5000 (corrected count increment) is used to evaluate if the patient has PTR. By chi-square test, we found 13 patients became PTR in all patients, of 2 patients (15.4%) is in chemotherapy group while 11 (84.6%) is in CAR-T group. Moreover, 7.7% (2/26) PTR in chemotherapy group is markedly lower than 47.8% (11/23) in CAR-T group (P=0.001). The associated factors with PTR in this study is cytokine release syndrome (CRS) grade. Then we found the significant elevated cytokine levels in CAR-T group. By Mann-Whitney U test, the mean levels of IL-6 in PTR group versus non-PTR group were 6.58 vs 17.91 (p<0.001). Serum mean levels of IL-10 in PTR group versus non-PTR group were 7.58 vs 16.82 respectively (p=0.001) while serum levels of IFN-γ were 6.58 versus 17.91 respectively (P<0.001). With the decline of cytokines, the 12h CCI of partial patients with PTR will be increased than before, some patients even recover to normal platelet transfusion increment. The rate of PTR in patients undergoing CAR-T therapy was much higher than those who received chemotherapy only. CRS was the only independent risk factor associated with PTR after CAR-T treatment. High serum levels of cytokines (IL-6, IL-10 and IFN-γ) during CRS might contribute to is PTR.

Disclosures

No relevant conflicts of interest to declare.
Safety and Utilization of Midline Catheters in the Management of Acute Myeloid Leukemia: A Single Center Experience
Authors: Torregrosa Diaz Jose Miguel , Gardeney Helene , Gallego Hernanz Maria Pilar , Desmier Deborah , Leleu Xavier , Rammaert Blandine
Published: November 2019

BACKGROUND: Tunneled central venous catheters (TCVCs) are used in cancer patients for irritant or vesicant infusion such as chemotherapy or antibiotics. However, they can be associated which significant complications. Midline catheters (MCs) are peripheral intravenous access devices that ends in a large peripheral vein, easier to install and which may reduce the need for TCVCs. Therefore, the aim of this study is to assess the rate of adverse outcomes with both kind of catheter, in hospitalized patients for the treatment of acute myeloid leukemia.

METHODS: We retrospectively compared the use and outcomes of TCVCs and MCs at our institution between December 2017 and December 2018. Data were collected using electronic medical records. Both devices were used indistinctly among our hospitalized patients for chemotherapy, antibiotics and parenteral nutrition. The following complications were recorded: catheter related bloodstream infection (defined as documented bacteriemia and differential time to positivity pointing to TCVC as main source), local signs of infection (such as erythema or inflammation) at insertion site, thrombosis which required an anticoagulant treatment, extravasation and local bleeding at insertion site.

RESULTS: Through this period, 39 TCVCs and 50 MCs were used among 56 different patients. As expected duration of the device was shorter for MCs (mean 24days) as they were ablated on discharge, compared to TCVCs (mean 32.5 days). Median aplasia duration (defined as polymorphonuclear neutrofils count in peripheral blood <500/mm3) of each chemotherapic episode was 13 days [0 - 55 days] for patients harboring a TCVCs and 17 [0 - 46] for patients with MCs, with no statistically-significant difference. Main complications were infection related, TCVCs being more prone to have positive bloodstream cultures [20 (52.6%) vs 15 (29.4%), p= 0.027] and local signs of infection [7 (18.4%) vs 1 (2%); p=0.019] than MCs (and probably related to length of stay).

No extravasations were found in any group. As expected local bleeding was more frequent inTCVCs [6 (15.8%) vs 1 (2%); p=0.039].

Whole complications (infection, thrombosis and bleeding) were more frequent among TCVCs (n=35) than among MCs (n=18) even when reported to the length of the catheter (1.08 complications/day in TCVCs and 0.75 complications/day in MCs).

SUMMARY/CONCLUSION: MCs insertion -quick and easy- makes them user-friendly for hematologists and can be safely used among acute leukemia patients.

Disclosures

Leleu: Oncopeptide: Honoraria; Carsgen: Honoraria; Takeda: Honoraria; Amgen: Honoraria; Sanofi: Honoraria; Incyte: Honoraria; Novartis: Honoraria; Celgene: Honoraria; Janssen: Honoraria; BMS: Honoraria; Merck: Honoraria; Karyopharm: Honoraria.
A Hydrogel Based 3D Culture System for Hematopoietic Differentiation of Induced Pluripotent Stem Cells
Authors: Sidhu Ishnoor , Barwe Sonali , Kolb E. Anders , Kiick Kristi Lynn , Gopalakrishnapillai Anilkumar
Published: November 2019

The generation of hematopoietic stem and progenitor cells (HSPCs) from induced pluripotent cells (iPSCs) holds a great potential in development of cell therapies, modeling hematological malignancies, and testing new drugs. Hematopoiesis is regulated by bone marrow (BM) microenvironmental factors such as BM architecture, and cell-cell and cell-matrix interactions. The current methods for iPSC differentiation into HSPCs rely on cell-cell interaction (embryoid body formation and feeder cell co-cultures) or cell-matrix interaction (ECM coated dishes). Generation of a 3-dimensional (3D) culture environment attempts to capture both cell-cell and cell matrix interactions in studying the differentiation of the iPSCs. This study reports development of a 3D culture system for hematopoietic differentiation of iPSCs.

iPSC colonies were encapsulated in 3D Matrigel or polyethylene glycol (PEG) based hydrogels containing synthetic integrin binding peptide (GRGDSPC) and enzymatically degradable peptide (GGPQGIWGQGKG) and cultured in feeder-free cell culture and maintenance medium (mTeSR1™, Stem Cell Technology). There were notable morphological differences between the 3D encapsulated and 2D cultured iPSC colonies (Fig 1A). The 3D encapsulated colonies were more compact with a spheroid morphology in PEG compared to Matrigel, whereas colonies in 2D were more diffused. Viability of the encapsulated colonies was evaluated in situ using cytotoxicity kit. The 3D encapsulation did not have an adverse effect on the viability and growth of iPSCs (Fig 1B). The encapsulated iPSCs maintained the pluripotent phenotype in 3D as assessed by TRA-1-60 staining (Fig 1C).

To test the efficiency of these 3D model systems to generate HSPCs, the encapsulated iPSCs were subjected to hematopoietic differentiation using STEMdiff™ Hematopoietic Kit (Fig 1D). Following differentiation, the single cells were collected and immunophenotype analysis was performed by flow cytometry. Above 50% population stained positive for HSPC surface marker CD34 both in PEG hydrogel and Matrigel (Fig 1E).

To test the robustness of the 3D system in inducing hematopoietic differentiation, we encapsulated the isogenic trisomic 21 (Tri21) iPSCs and subjected to hematopoietic differentiation and flow cytometry analysis as described above. As reported previously in 2D system (Banno et al., 2016, Cell Reports), Tri21 iPSCs showed higher percentage of hematopoietic commitment into erythroid and megakaryocytic lineages compared to disomic 21 (Di21), as characterized by the presence of respective surface markers (Fig 1F).

In conclusion, 3D culture system was suitable for inducing hematopoietic differentiation of iPSCs in isogenic disomic and trisomic lines. The 3D model can be used to generate patient-specific HSPCs.

Figure 1

Disclosures

No relevant conflicts of interest to declare.
Thrombopoietin Has Protective Effect in Neonatal Hypoxia-Ischemia Rat Model
Authors: Li Liang , Yang Liuming , Xin Hongwu , Chong Beng H , Yang Mo
Published: November 2019

Thrombopoietin (TPO) is a growth factor for the megakaryocytic lineage. The expression of TPO and TPO receptor (c-mpl) in the central nervous system (CNS) and the role of TPO in neural cells and brain damage models were investigated. Our results showed the expression of TPO in human cerebral hemisphere, cerebellum, cerebrospinal fluid and blood plasma. We found that TPO had a protective effect in hypoxic-ischemic rat model, as indicated by the increased ipsilateral brain weight and neuron density in a neonatal rat model of hypoxic-ischemic brain damage. Recoveries of sensorimotor functions and histopathology were observed in these animals that received TPO. In addition, TPO could promote C17.2 cells proliferation by activating PI3K/Akt signaling pathway, and the proliferation could be reduced to nearly basal level by the pre-treatment with LY 294002. The phosphorylation of AKT, which is a hallmark of activation of each molecule was significantly enhanced after the treatment with TPO in the cells, peaking at 30 min after stimulation with TPO. TPO was also found to have an anti-apoptotic effect which mediated via Bcl-2/BAX and suppressing the mitochondrial membrane potential. Results showed the increased level of Bcl-2 and decreased level of BAX were in the time-dependence manner (0, 5, 15, 30 and 60 mins) in these cells. In addition, the mitochondrial membrane potential was significantly decreased by adding 100 ng/ml TPO. Our results indicated that TPO have neural protective effects.

Disclosures

No relevant conflicts of interest to declare.
CUDC-101 Overcomes Arsenic Trioxide Resistance Via Caspase-Dependent PML-Rarα Degradation in Acute Promyelocytic Leukemia
Authors: Zhang Tianzhuo , Wang Jishi
Published: November 2019

While arsenic trioxide (ATO) treatment has transformed acute promyelocytic leukemia (APL) from the most fatal to the most curable hematological cancer, many high-risk APL patients who fail to achieve a complete molecular remission or relapse become resistant to ATO. Studies have suggested that HDAC inhibitors (HDACi) have been targeted as promising agents for the treatment of several cancers. In this study, a novel pan-HDACi 7-(4-(3-Ethynylphenylamino)-7- methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide (CUDC-101) was used to tentatively to treat hematologic disease for the first time. The CCK-8 assay demonstrate that CUDC-101 exerts much stronger anti-proliferative effects than ATO in APL and ATO-resistant APL cell lines. Previous experiments on cell proliferation inhibition prompted investigation of whether CUDC-101 could induce apoptosis in APL and ATO-resistant APL cells. Then the results of apoptosis assay strongly support CUDC-101 as an effective apoptosis inducer that induced apoptotic cell death in both APL and ATO-resistant APL cell lines, more effectively than ATO. Our data also demonstrate that CUDC-101 is a novel drug that has a negligible cytotoxic effect on non-cancerous cell lines including normal CD34+ cells and BMSCs from APL patients. Mechanistically, we conclude that CUDC-101 directly induce hyperacetylation of histone 3, which subsequently lead to the activation of caspase 3, the activated-caspase 3 cleaves the PML-RARα fusion protein subsequently facilitates APL and ATO-resistant APL cells undergoing apoptosis. The precise mechanism of CUDC-101-induced degradation of PML-RARα remains unclear. Yet the apoptosis inducing effect of CUDC-101 offers a potent therapeutic advantage of CUDC-101 and its role in overcoming APL warrants further validation in patient-derived samples in future studies. Finally, using a NB4 xenograft mouse model, we demonstrated that CUDC-101 significantly represses leukemia development in vivo compared with ATO. These findings suggest that the CUDC-101 is efficient in inhibiting tumor growth in vivo and performs better than ATO. To the best of our knowledge, this is the first report that CUDC-101 functions as HDACi and has potential therapeutic value for the treatment of both APL and relapsed APL with ATO resistance in vitro and in vivo. In conclusion, these results suggested that CUDC-101 can serve as a potential candidate drug for APL, particularly for ATO-resistant APL.

Disclosures

No relevant conflicts of interest to declare.
Efficacy and Tolerance of Interferon-α in Hypereosinophilic Syndromes. a Retrospective Multicenter Study on 29 Patients
Authors: De Malglaive Pauline , Groh Matthieu , Lefèvre Guillaume , Morati Hafsaoui Chafika , Alexis Magda , Hunault Mathilde , Seneschal Julien , Charlanne Hilaire , Preudhomme Claude , Bonnotte Bernard , Kahn Jean Emmanuel
Published: November 2019

Background: Corticosteroids and imatinib are the first line treatment in hypereosinophilic syndromes (HES) according FIP1L1-PDGFRA (F/P) positivity. In HES unrelated to F/P, second line available treatment are historically interferon alpha (IFN-α) and hydroxyurea, while anti-interleukin-5 antibodies are currently investigated. Data concerning efficacy of IFN-αin HES are limited to small case series and one large international study with a short-term follow-up.

Methods: We performed a multicenter, retrospective, descriptive study to evaluate the efficacy and safety of IFN-αin F/P-negative HES patients. Complete hematologic responses (CHR : eosinophils < 0,5 G/L) and partial hematologic responses (PHR: eosinophils between 0,5 to 1,5 G/L or a decrease of more than 50%), as well as tolerance and corticosteroid sparing-effect were analyzed at 1, 3, 6 and 12 months.

Results: Twenty-nine patients (9 women, with a mean [+/-SD]age at diagnosis of 38 [+/-13]years were included. Fifteen patients (52%) had idiopathic HES, 10 (34%) had a lymphocytic HES, and 4 (14%) had a clonal HES. The most common clinical manifestations were cutaneous (69%) and gastrointestinal involvement (31%). IFN-αwas used in pegylated form in 16 patients at an average dose of 113 [+/-45 μg]/week and in non-pegylated form in 13 patients at an average dose of 12 [+/-9]MIU / week. At one month, a CHR or PHR was observed in 24 patients (83%), including 16 in CHR. At 1 year, 16 patients were still treated with IFN-α, (9 in CHR and 7 in PHR). Treatment with IFN-αwas associated with a significant decrease in the mean daily prednisone dose from 20 [+/-16]mg/day at inclusion to 11 [+/-6]mg/day at 6 months and at 9 [+/-9]mg/j at 1 year. After a median of 15 months (range 1 to 161 months), 21 patients (72%) discontinued IFN-αmainly due to adverse events (27%) or loss of efficacy (31%). At the end of the follow-up, 8 patients were still on IFN-αwith a median duration of treatment of 21.5 months (3 to 161 months), including 6 in CHR and 2 in RHP. In these 8 patients, pegylated IFN-αwas used at the median maintenance dose of 50 μg/week with maintenance of a corticosteroid sparing-effect (median daily prednisone dose at 5.5 mg/day).

Conclusions: IFN-αappears to be an effective treatment for corticosteroid-resistant or corticosteroid-dependent F/P-negative HES. Its effectiveness can be maintained over long term and at low dose in nearly a quarter of patients, with good tolerance. However, a loss of efficacy or adverse effects justify discontinuation of IFN-αin the majority of cases, underlying the need of alternative therapies

Disclosures

No relevant conflicts of interest to declare.

OffLabel Disclosure:

Interferon-Alpha is an off-label drug used in hypereosinophilic syndromes since more than 20 years.
Diagnostic Performance of Heparin-Platelet Factor 4 Antibodies in the Identification of Heparin-Induced Thrombocytopenia in Cancer Population
Authors: Oo Thein H. , Rojas Hernandez Cristhiam Mauricio
Published: November 2019

Introduction: Cancer patients appear to have a higher risk of heparin induced thrombocytopenia (HIT) related complications than non-cancer patients; yet data on the performance of conventional diagnostic tools for HIT in cancer is limited.

Our aim was to determine among cancer patients with a 4T score ≥ 4, the performance of the conventional cut-off for HIT antibody testing (IgG anti PF4) to discriminate between serotonin release assay (SRA) positive and negative cases.

Methods: Retrospective and prospective analysis of cases (2002-2019) was performed of the electronic medical records of adult cancer patients at MD Anderson Cancer Center with suspected HIT. Cases were included in the analysis if the 4T score was ≥ 4 and investigated with IgG anti-PF4 optical density (HIT OD) and SRA.

Logistic regression model and the receiver operating characteristic curves were conducted to identify the sensitivity and specificity of different cut-off points for the HIT OD to discriminate HIT cases based on the SRA status.

Results: Among 50 cases, 18 were SRA positive. Median HIT OD was 1.03. At a cut-off point of 0.4, the HIT OD performed with a sensitivity of 0.89 and a specificity of 0.50 to discriminate the cases of SRA positive HIT. When the cut-off HIT OD was 1.0, the sensitivity was 0.78 with a specificity of 0.66.

Conclusions: Our findings suggest that in cancer patients the performance of IgG anti-PF4 is similar to that of non-cancer patients for the identification of HIT cases.

Disclosures

Oo: Janssen and Janssen: Other: Research: site co-investigator ; Medical Education Speakers Network: Honoraria.