bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Novel 3D Extracellular Matrix Model Enriching Human Acute Myeloid Leukemia Stem Cells
Authors: Li Dandan , Lin Tara L. , Hopkins Richard , Aljitawi Omar S. 
Published: October 2019

Abstract
Background and objective: Acute myeloid leukemia (AML) develops from leukemia stem cells (LSCs), a small subset of leukemia cells possessing both self-renewal and multilineage differentiation potential similar to normal hematopoietic stem cells. The stem cell niche of the bone marrow microenvironment protects LSCs from chemotherapy, resulting in subsequent leukemia relapse. The study of AML LSCs in vitro is limited because of the lack of an ideal culture system mimicking the protective bone marrow microenvironment. The bone marrow stem cell niche is mainly composed of stromal cells, soluble cytokines and growth factors, as well as extracellular matrix (ECM). We therefore developed a 3-dimensional ECM model using decellularized Wharton's jelly from human umbilical cords to better characterize AML LSCs in vitro. Previously we have shown that leukemia cells grown in DWJM changed morphology to become spindle shaped and maintained viability but had decreased proliferation as measured by Alamar blue assay. Herein, we further characterize leukemia cells cultured in DWJM.
Methods: Wharton's jelly decellularization process included multiple osmotic shock cycles using hypertonic and hypotonic solutions, a non-ionic detergent Triton-x, an anionic detergent sodium lauroyl succinate, and an enzyme digestion with recombinant endonuclease Benzonase™. We examined three human leukemia cell lines: HL60, Kasumi I and MV411. We characterized leukemia cell proliferation by CellTrace proliferation assay and phenotype by flow cytometry for stem cell markers. Serial colony forming unit (CFU) assays were used to test the self-renewal of leukemia cells.
Results: CellTrace proliferation assay showed that, compared to cells in suspension, cells cultured in DWJM divided less frequently. To assess for LSCs properties, we measured the ALDH+ population by Aldefluor assay and found that the ALDH+ cells from Kasumi I and HL60 increased significantly in DWJM compared to suspension (p<0.05). Lastly, serial CFU showed significantly increased colony forming units in both primary and secondary plating in MV411 and HL60 cells cultured in DWJM versus suspension (P<0.05). CFU also increased in Kasumi-I cells cultured in DWJM versus suspension, though the increase was not statistically significant (P=0.08). Taken together, our findings suggest that leukemia cells cultured in DWJM demonstrated an increased self-renewal ability due to the enrichment of LSCs rather than progenitors.
Conclusion: Decellularized Wharton's jelly matrix may serve as a practical in vitro ECM model to enrich for LSCs and study the ECM-LSC interactions.

Disclosures
No relevant conflicts of interest to declare.
Analysis of the Expression of PHTF1 and Related Genes in Acute Lymphoblastic Leukemia
Authors: Huang Xin , Geng Suxia , Weng Jianyu , Lu Zesheng , Zeng Linji , Li Minming , Deng Chengxin , Wu Xiuli , Li Yangqiu , Du Xin 
Published: October 2019

Abstract
Background: In the human T-cell acute lymphoblastic leukemia (T-ALL) cell line Molt-4, siRNA-mediated suppression of BCL11B expression was shown to inhibit proliferation and induce apoptosis, which may be related to PHTF1 gene expression, and the FEM1B and Apaf-1 genes may be downstream of PHTF1. In this study, we analyzed the expression level of PHTF1 and related genes in patients with ALL to clarify the role of the PHTF1-FEM1b-Apaf-1 pathway in hematologic malignancies.
Methods: Fifteen newly diagnosed and untreated patients with AML, fourteen newly diagnosed and untreated patients with CML in chronic phase, twenty-two newly diagnosed and untreated patients with ALL, and six newly diagnosed and untreated patients with CLL were recruited. Peripheral blood mononuclear cells (PBMCs) from ten healthy individuals (HIs) served as controls. PBMCs were separated using the Ficoll-Hypaque gradient centrifugation method. All procedures were conducted in accordance with the guidelines of the Medical Ethics committees of the Health Bureau of Guangdong Province, China.Real-time PCR was used to determine the gene expression level of PHTF1 in hematologic malignancies. The PHTF1, BCL11B, FEM1B and Apaf-1 gene expression levels and correlations were analyzed in patients with primary ALL and healthy individuals (HIs).
Results: PHTF1 overexpression was found in recently diagnosed AML (p <0.001), CML-CP (p<0.001), and ALL (p= 0.016) patients in comparison with HIs. The PHTF1 expression level in CLL patients was not significantly different compared with HIs (p= 0.165). FEM1b and Apaf-1 overexpression was found in recently diagnosed ALL (p<0.005) patients compared with HIs. Positively correlated expression was found for the PHTF1, FEM1b and Apaf-1 genes in patients with ALL (p<0.005) and HIs (p<0.005), and positively correlated expression was found for the PHTF1 and BCL11B genes in HIs (p<0.005).
Conclusions: PHTF1 acts as a tumor suppressor gene, and its overexpression might be related to cell proliferation, inhibition and apoptosis. PHTF1 and BCL11B gene disorders may contribute to T-ALL pathogenesis. PHTF1 might be a therapeutic target for triggering the PHTF1-FEM1b-Apaf-1 apoptosis pathway in primary acute lymphoblastic leukemia.
Acknowledgment
The project was sponsored by grants from National Natural Science Foundation of China (No. 81100384, No. 81270648, No.91129720, and No. 30771980)

Disclosures
No relevant conflicts of interest to declare.
B Cell Lymphoma Unclassifiable, with Features Intermediate Between Diffuse Large B Cell Lymphoma and Burkitt Lymphoma and Diffuse Large B Cell Lymphoma NOS with Doble/Triple Translocations: Immunophenotypic Analysis
Authors: González de Villambrosia Sonia , Colorado Mercedes , Insunza Andres , Batlle Ana , López-Pereira Brenda , Martin Guillermo , Ibarrondo Paloma , Montes-Moreno Santiago , Conde Eulogio 
Published: October 2019

Abstract
Background: Diffuse large B-cell lymphoma (DLBCL) is a clinically and molecularly heterogeneous disease. We can identify two subgroups with aggressive clinical course and higher risk of treatment failure after standard therapy: B cell lymphoma unclassifiable (BCLU) with features intermediate between DLBCL and Burkitt lymphoma (BL) and B-cell lymphomas with double/triple translocation (DHL/THL). Previous reports suggest that these types of lymphomas may show a common immunophenotype, providing another tool in the challenge of their diagnosis.
Objetives: To analyze the immnunophenotype (IF) of BCLU and DH/TH lymphomas by multiparametric flow cytometry and compare it with the IF of a series of cases with DLBCL and BL.
Methods: we analyzed the inmunophenotype (four-color flow cytometry on a FACSCalibur flow cytometer) and cytogenetic studies (FISH to detect MYC, BCL2 and/or BCL6 rearrangement) of cases diagnosed of BCLU and DH/TH lymphomas. Control cases of DLBCL and BL were consecutively collected from our database. Fisher`s Exact test was used to compare proportions between two groups. P-values <0.05 were considered statistically significant.
Results: We analyzed 23 controls (14 DLBCL and 9 BL) and 17 cases: 9 DHL (8 BCL2/MYC+ and 1 BCL6/MYC+), 3 THL (BCL2/BCL6/MYC+) and 5 BCLU (1 IGH-MYC+, 3 MYC+ and 1 unknown). Six of the 17 cases (35.3%) had decreased expression of CD19 while this was exceptional in DLBCL (1/10 P=0.073) and BL (0/9 P=0.054). All of the cases were positive for CD20 but with different intensities: only 5.9% expressed high CD20 compared to 42.9% of DLBCL (p=0.021) and 55.6% of BL (P=0.010). Most of the cases (12/17) had intermediate expression of CD20 and only 4/17 had weak expression. CD10 expression was typical in BL (100%) and frequent in the cases (82.4%), while it was only present in 20% of DLBCL (P<0.0001). CD38 expression was high in 100% of BL, 6.3% of the cases and none of DLBCL. It was intermediate in 64.7% of the cases and in 35.7% of DLBCL (P=0.015). BCL2 overexpression was detected in 57.1% of the cases; neither DLBCL nor BL (P= 0.009) had BCL2 overexpression.
Conclusions: We identify a common immunophenotype in DH/TH lymphomas and BCLU: decreased expression of CD19 and CD20, CD10 expression, overexpression of BCL2 and intermediate expression of CD38. This immuphenotype may be useful for identifying cases for confirmatory cytogenetic studies. Larger studies are needed to validate these results.

Disclosures
No relevant conflicts of interest to declare.
The Neutrophil-Lymphocyte Ratio Is a Prognostic Marker in Patients with EBV-Positive Diffuse Large B-Cell Lymphoma
Authors: Beltran Brady E , Malaga Jose M , Chavez Julio C , Sotomayor Eduardo M. , Castillo Jorge J 
Published: October 2019

Abstract
Introduction: Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) is a rare hematologic malignancy with a poor prognosis when treated with current therapies. Clinical factors have been developed to prognosticate survival in EBV-positive DLBCL patients; however, more refined, easy to use and reliable prognostic tools are needed. The neutrophil to lymphocyte ratio (NLR) has been reported prognostic in patients with DLBCL (Troppan et al. BJC 2014). We have investigated the prognostic value of the NLR in the overall survival (OS) of patients with untreated EBV-positive DLBCL.
Methods: We included patients with a pathological diagnosis of EBV-positive DLBCL who were diagnosed and treated at our institution between 2001-2014. We excluded cases with primary cutaneous CNS involvement, and patients with >50% incomplete data. IRB approval was obtained prior to research. Pathological samples were reviewed by hematopathologists to confirm the diagnosis. Pertinent clinicopathological data such as age, sex, performance status, LDH levels, stage, extranodal sites of disease, absolute neutrophil and lymphocyte counts were collected through chart review, and are presented using descriptive statistics. The NLR was calculated by dividing the absolute neutrophil by the lymphocyte count, and dichotomized in NLR>=5 and NLR<5. Survival curves were estimated using the Kaplan-Meier method and compared using the log-rank test. Univariate Cox models were fitted to evaluate hazard ratios (HR) for OS.
Results: A total of 46 patients were included in our analysis. The median age was 73 years (range 25-95 years) with male predominance (70%). Poor performance status (ECOG >1) was seen in 25 (58%), elevated LDH levels in 19 (46%), 1+ extranodal site in 24 (57%), and advanced stage (stage 3 and 4) in 23 (53%) of patients. Based on the NLR, 13 patients (39%) had NLR>=5. Patients with NLR >=5 were more likely to present with poor performance (ECOG >1; 85% vs. 15%; p=0.04), elevated LDH levels (69% vs. 31%; p=0.04), advanced stage (III and IV, 69% vs. 31%, p=0.05), and IPI score 3-5 (77% vs.23%; p=0.02). There were no differences in age, sex and number of extranodal sites. NLR>=5 was associated with a worse OS (HR 2.67, 95% CI 1.01-7.01; p=0.047).
Conclusion: The NLR appears as a novel and easy to use prognostic factor for OS in patients with untreated EBV-positive DLBCL. Our findings support the need for validation of the NLR in larger retrospective or prospective studies in patients with EBV-positive DLBCL.
Figure 1. Figure 1.

Disclosures
No relevant conflicts of interest to declare.
MYD88 L265P Mutation Is a Possible Unfavorable Prognostic Factor in Patients with Diffuse B-Cell Lymphoma
Authors: Sidorova Anna A. , Zvonkov Eugene E. , Sudarikov Andrey B. , Severina Nataliya A. , Kovrigina Alla M. , Troitskaya Vera V. , Savchenko Valeri G. 
Published: October 2019

Abstract
INTRODUCTION
Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma. Activated B-cell like (ABC) molecular subtype DLBCL is characterized by more aggressive behavior and poor clinical outcomes with standard R-CHOP chemotherapy. Approximately 30% of all patients (pts) with ABC DLBCL have recurrent mutation in the MYD88 gene (a change from leucine to proline at position 256 - L256P). We report the impact of the MYD88 mutation status on the clinical course and outcome of pts with ABC DLBCL.
METHODS
In present study MYD88 mutation status was investigated in untreated pts with ABC DLBCL.
The diagnosis of ABC DLBCL was established according to the WHO 2008 classification. DNA from 41 cryopreserved and 12 formalin-fixed paraffin-embedded tumor samples were tested in our study. Sanger sequencing and real-time allele-specific PCR was used to assess MYD88 L265P mutation status.
RESULTS
We report data for 53 pts (34 male, 19 female) with a median age of 53 (18-74) years. MYD88 L265P mutation was detected in 26,4% (14/53) of pts. The international prognostic index (IPI) score was 4-5 in 7/14 (50%) of pts with MYD88 mutated DLBCL and 12/39 (30,8%) pts with MYD88 wild type DLBCL, whereas nearly half of all pts presented with Ann Arbor stage IV disease. The majority of pts 13/14 (93%) with MYD88-positive DLBCL had elevated lactate dehydrogenase levels versus 26/39 (66%) pts with MYD88 unmutated DLBCL. Extranodal was more common in pts with MYD88 mutation than in pts with MYD88 unmutated DLBCL (78,5% vs 54%). All pts received high-dose chemotherapy and were followed for 50,5 (11-95) month. Pts with MYD88 mutation had similar complete response rate comparing to pts without MYD88 mutation (78,6% vs 84,6%). The disease relapsed in 45,4% of pts with MYD88 mutation and in 24,2% of pts without MYD88 mutation. Mortality was in 45,4% of pts with MYD88 mutation and in 24,2% of pts without MYD88 mutation. Overall survival (OS) was better in pts with MYD88 unmutated DLBCL than in pts with MYD88 positive DLBCL (5-year OS, 73% vs 31%).
CONCLUSION
IPI high risk and extranodal involvement is common in ABC DLBCL pts with MYD88 L265P mutation. MYD88 L265P mutation can be a predisposing factor for poor OS probability in pts with ABC DLBCL.

Disclosures
No relevant conflicts of interest to declare.
Consolidative Treatment after Salvage Chemotherapy Improved Prognosis in Patients with Relapsed Extranodal Natural Killer/T-Cell Lymphoma
Authors: Bi Xi-Wen , Zhang Wen-Wen , Huang Jia-Jia , Xia Zhongjun , Li Zhi-Ming , Jiang Wenqi 
Published: October 2019

Abstract
The optimal treatment strategy and prognosis for patients with relapsed natural killer/T-cell lymphoma (NKTCL) remain largely unknown. The treatment modalities and prognosis of fifty-six patients with relapsed NKTCL were retrospectively reviewed. Twenty-three (41.1%) patients had locoregional relapse alone, while 33 (58.9%) had distant with or without locoregional relapse. Chemotherapy was the initial salvage treatment, followed by radiotherapy (RT) or autologous hematopoietic stem cell transplantation (AHSCT) as consolidative therapy, depending on the status of remission and the pattern of relapse. The 5-year overall survival (OS) after relapse was 41.4% for the entire cohort. Complete remission (CR) after salvage treatment was associated with a substantially better survival (5-year OS after relapse: 74.7 vs. 7.8%, P < 0.001). For patients with locoregional relapse alone, consolidative RT after response to salvage chemotherapy significantly improved prognosis compared with follow-up (5-year OS: 83.3 vs. 41.7%, P = 0.047). For patients with distant relapse, the addition of consolidative AHSCT after response to chemotherapy significantly prolonged survival than follow-up (2-year OS: 100.0 vs. 20.0%, P = 0.004). Patients without consolidative treatment after response to salvage chemotherapy had a comparable survival to those who experienced stable or progressive disease after chemotherapy, regardless of the relapse pattern. Asparaginase (ASP)-containing salvage chemotherapy failed to convey a survival advantage over ASP-absent chemotherapy (5-year OS: 44.2 vs. 39.3%, P = 0.369). Patients who received ASP-containing chemotherapy in the first-line treatment had a poorer response to salvage chemotherapy and worse prognosis after relapse, compared with those receiving ASP-absent first-line chemotherapy. Consolidative RT or AHSCT improved prognosis in patients with relapsed NKTCL who responded to initial salvage chemotherapy. The role of ASP in salvage chemotherapy after relapse required further exploration in prospective studies.

Disclosures
No relevant conflicts of interest to declare.
The Frequency of Occurrence of Minimal Residual Disease into Different Prognostic Groups of Patients with Chronic Lymphocytic Leukemia
Authors: Kuvshinov Aleksei , Voloshin Sergei , Martynkevich Irina , Martynenko Ludmila , Garifullin Andrei , Kleina Elizaveta , Abdulkadyrov Kudrat 
Published: October 2019

Abstract
Background. It is known, that genetic factors and the absence of minimal residual disease (MRD) are strongly affecting prognosis of chronic lymphocytic leukemia (CLL).
Aim. To determine the influence of genetic abnormalities (GA) on achieving MRD-negative remissions in patients with CLL.
Methods. Twenty-four adult pts (median age 57 years, range 35-67; male 14, female 10) with newly diagnosed CLL were included. The CLL was diagnosed according to the standard basic examination (complete blood count with differential, multicolor flow cytometry (MFC) of blood and bone marrow (BM), lymph node and BM immunohistochemistry (IHC), computered tomography). Cytogenetic studies were performed on blood samples using standard GTG-method. Interphase FISH analyses were performed according to the manufacturer's protocol using DNA probes: LSI 13(RB1)13q14, LSI ATM (11q22), CEP12, LSI TP53 (17p13.1) (Abbott). Immunophenotype (IFT) of CLL cells assessed with combinations: CD3/CD19, CD19/CD5, CD19/CD11c, CD19/CD20, CD19/CD22, CD19/CD23, CD19/CD25, CD19/CD38, CD19/CD43, CD19/CD81, CD19/HLA-DR, and CD19/CD5/CD23. We have used NCI revised guidelines (Hallek M, et al., 2008) for treatment initiation and assessment of response after completion of primary therapy with rituximab-based regimens (FCR or RB). All patients treated subsequently with rituximab maintenance. MRD was detected by MFC.
Results. The frequency of GA in CLL was 50.0% (12/24): 15.0% (3/20) - by conventional karyotyping, 47.8% (11/23) - by FISH analyses and 9.5% (2/21) - using both methods. Stratification of patients into prognostic groups based on identified GA. Favorable prognosis (Group 1) - patients with del(13q) (n = 5); neutral prognosis (Group 2) - normal karyotype (n = 12) or trisomy of chromosome 12 (n = 3); unfavorable prognosis (Group 3) - del(11q) (n = 3) or the complex karyotype (n = 1).
Statistically significant differences in the frequency of achieving MRD-negative remissions between FCR (5/11) vs. RB (5/13) were not detected (p<0.05).
Complete remissions (CR) were reached in 37.5% (9/24) pts, partial remissions (PR) - 62.5% (15/24). The MRD-negative remissions were reached in 10 patients: in Group 1 - 2/5 (40%; CR - 1), in Group 2 - 5/15 (33.3%, CR - 6), in Group 3 - 75.0% (3/4; CR - 2).
Statistically significant differences in PFS were detected between MRD-negative vs. MRD-positive groups (p=0.03). Median PFS in MRD-negative has not been reached. Median PFS MRD-positive was 33.1 month.
Conclusions. Further researches aimed at examining the relationship between the presence or absence of MRD and genetic prognostic groups, will help to understand the most important factors affecting the overall and progression-free survival.

Disclosures
No relevant conflicts of interest to declare.
Neutrophil-Secreted Proteinase 3 Mediates Metastasis of Prostate Cancer Cells Expressing RAGE to the Bone Marrow
Authors: Kolonin Mikhail , Sergeeva Anna , Staquicini Daniela , Molldrem Jeffrey J. , Pasqualini Renata , Arap Wadih 
Published: October 2019

Abstract
Human prostate cancer non-randomly metastasizes to the bone marrow, but the biological basis for such site-specific tropism remains largely unresolved. Differential expression of molecules in the bone marrow microenvironment "niche" has recently been proposed to play a role. In previous work, combinatorial selection of random peptide libraries in end-of-life cancer patients has revealed an unexpected interaction between the receptor for advanced glycation end products (RAGE), a molecule expressed on malignant cell surfaces in advanced prostate cancer, and proteinase 3 (PR3), a serine protease abundantly present on neutrophils and promyelocytes within the bone marrow microenvironment. Because RAGE is selectively overexpressed in prostate cancer bone metastases, we hypothesize here that its specific binding to PR3 might mediate homing of prostate cancer cells to the bone marrow. We demonstrate that PR3 non-proteolytically binds to RAGE on prostate cancer cell surfaces and thereby promotes tumor cell activation and motility. We also show that the downstream signal transduction cascade triggered by RAGE/PR3 binding involves p44/42 (Erk1/2) and JNK1 phosphorylation. Finally, we use a mouse model of experimental metastasis to demonstrate that RAGE protein expression on human prostate cancer cells promotes their homing to bone marrow within a short time frame. These results validate a functional protein interaction between RAGE and PR3 and uncover a mechanism through which neutrophils mediate prostate cancer cell metastasis to the skeleton.

Disclosures
Sergeeva: Astellas: Patents & Royalties. Molldrem:Astellas Pharma: Patents & Royalties.