bluuebunny/crossref_metadata_2025_split huggingface dataset:
Disease Risk Comorbidity Index for Patients Receiving Haploidentical Allogeneic Hematopoietic Transplantation
Authors: Mo Xiaodong , Zhang Xiaohui , Xu Lanping , Wang Yu , Yan Chenhua , Chen Huan , Chen Yuhong , Han Wei , Wang Fengrong , Wang Jingzhi , Liu Sining , Liu Kaiyan , Huang Xiaojun 
Published: November 2019

Purpose or Background:
We aimed to develop a disease risk comorbidity index (DRCI) based on Disease Risk Index (DRI) and Hematopoietic Cell Transplantation-Specific Comorbidity Index (HCT-CI) in patients receiving haploidentical hematopoietic stem cell transplantation (haplo-HSCT).
Method or Case:
We examined 889 patients undergoing haplo-HSCT from 2015 to 2016. We used a Cox multivariable model to identify factors prognostic of disease-free survival (DFS) in a training subset (n = 593). A weighted score using these factors was assigned to the remaining patients (validation cohort; n = 296). This work was supported by the Capital's Funds for Health Improvement and Research (grant number 2018-4-4089).
Results or Progress:
The multivariable model identified two independent predictors of DFS: DRI and HCT-CI before HSCT. A weighted score of 2 was assigned to very high risk DRI, and a weighted score of 1 was assigned to high-risk DRI and intermediate- and high-risk HCT-CI in the scoring system (i.e., haplo-HSCT). In the validation cohort, the 3-year DFS was 65.2% (95% CI, 58.2-72.2%), 55.8% (95% CI, 44.9-66.7%), and 32.0% (95% CI, 5.8-58.2%) for the low-, intermediate- and high-risk group, respectively (P=0.005). Haplo-DRCI can predict relapse (P<0.001), non-relapse mortality (NRM, P<0.001), DFS (P<0.001), and overall survival (OS, P<0.001) in total population and in disease-specific subgroups, particularly in acute leukemia patients. Increasing score was also significantly predictive of increased relapse (P<0.001), increased NRM (P=0.001), decreased DFS (P<0.001), and decreased OS (P<0.001) in an independent historical cohort (n=526).
Conclusion or Discussion:
These data confirmed that haplo-DRCI can effectively risk stratifies haplo-HSCT recipients and provide the tool to better predict who will best benefit from haplo-HSCT.

Disclosures
No relevant conflicts of interest to declare.
Gfi1 Upregulates c-Myc Expression and Promotes c-Myc-Driven Cell Proliferation
Authors: Zhang Yangyang , Dong Fan 
Published: November 2019

Gfi1 is a zinc-finger transcriptional repressor that plays an important role in hematopoiesis. When aberrantly activated, Gfi1 may function as a weak oncoprotein in the lymphoid system, but collaborate strongly with c-Myc in lymphomagenesis. c-Myc is a transcription factor that is frequently activated in human cancers including leukemia and lymphoma mainly due to its overexpression as a result of gene amplifications and chromosomal translocations. c-Myc overexpression may also result from stabilization of c-Myc protein, which is highly unstable and rapidly degraded through the ubiquitin-proteasome pathway. The mechanism by which Gfi1 collaborates with c-Myc in lymphomagenesis is incompletely understood. c-Myc activates gene expression by forming a heterodimeric complex with the partner protein Max, but may also repress target genes through interaction with transcription factor Miz-1. We previously showed that Gfi1 indirectly interacts with c-Myc through Miz-1 and collaborates with c-Myc to repress CDK inhibitors p21Cip1 and p15Ink4B. In this study, we show that Gfi1 augmented the level of c-Myc protein transiently expressed in Hela cells and the levels of MycER fusion protein stably expressed in the mouse pro-B Ba/F3 and myeloid 32D cells. The C-terminal ZF domains of Gfi1, but not its transcriptional repression and DNA binding activities, were required for c-Myc upregulation. Notably, although Miz-1 has been shown to stabilize c-Myc protein, the expression of c-Myc V394D mutant, which is defective in Miz-1 interaction, was still upregulated by Gfi1, suggesting that Gfi1-mediated c-Myc upregulation was independent of Miz-1 interaction. We further show that Gfi1 overexpression led to reduced polyubiquitination and increased stability of c-Myc protein. Interestingly, the levels of endogenous c-Myc mRNA and protein were augmented upon induction of Gfi1 expression in Ba/F3 and Burkitt lymphoma Ramos cells transduced with the doxycycline-inducible Gfi1 lentiviral construct, but reduced in Gfi1-knocked down human leukemic HL60 and U937 cells. Additionally, targeted deletion of Gfi1 resulted in reduced c-Myc expression in mouse lineage negative bone marrow cells, which was associated with a decline in the expression of c-Myc-activated target genes. The oncogenic potential of Myc derives from its ability to stimulate cell proliferation. Our results demonstrate that inducible expression of Gfi1 in Ba/F3 cells expressing MycER promoted Myc-driven cell cycle progression and proliferation. Thus, in addition to its role in c-Myc-mediated transcriptional repression, Gfi1 upregulates c-Myc expression at both mRNA and protein levels, leading to enhanced expression of c-Myc-activated genes and augmented cell proliferation driven by c-Myc. Together, these data may reveal a novel mechanism by which Gfi1 collaborates with c-Myc in lymphomagenesis.

Disclosures
No relevant conflicts of interest to declare.
The Effect of Chronic Viral Hepatitis on Serum Ferritin Level in Thalassemia Patients
Authors: Rujeerapaiboon Natthapat , Tantiworawit Adisak , Piriyakhuntorn Pokpong , Rattanathammethee Thanawat , Hantrakool Sasinee , Chai-Adisaksopha Chatree , Rattarittamrong Ekarat , Norasetthada Lalita , Charoenkwan Pimlak 
Published: November 2019

Background: Serum ferritin is widely used as a marker of iron overload in thalassemia patients. However, the ferritin level is affected by active infections or inflammation. The association between viral hepatitis and serum ferritin level in thalassemia patients is still unclear. This study aimed to determine the effect of chronic viral hepatitis on serum ferritin level in thalassemia patients.
Methods: This was a cross-sectional study in thalassemia patients aged ≥15 years-old at Chiang Mai University hospital. We expected that thalassemic patients in our clinic have a mean serum ferritin of 767 ng/mL with a standard deviation of 210 ng/mL. As a result, we have to enroll a total of 28 patients to demonstrate 30% difference of mean serum ferritin when the power was set at 80% with alpha level of 0.05. Information on chronic viral hepatitis, mean serum ferritin and liver iron concentration (LIC) as measured by T2* MRI were collected. Chronic viral hepatitis status was confirmed by either HBV DNA or HCV RNA testing. Patients were categorized to hepatitis and non-hepatitis group. Serum ferritin levels were compared between two groups. LIC measurement was used as a gold standard for iron overload. Subgroup analysis was performed according to iron overload and transfusion requirement status. Categorical and continuous variables were compared using the Chi-squared test and T-test, respectively. The correlation between viral loads and mean serum ferritin levels was analyzed by Pearson's correlation.
Result: Of 32 thalassemia patients (25 non-transfusion dependent [NTDT] and 7 transfusion dependents [TDT]), 13 patients had chronic viral hepatitis (7 with hepatitis B and 6 with hepatitis C infections). The LIC between hepatitis and non-hepatitis groups were not significantly different (7.28 [SD 4.7] vs 9.08 [SD 5.2] mg Fe/g, p=0.19). In the higher LIC group (≥ 5 mg Fe/g), the mean serum ferritin level was higher in the hepatitis group than non-hepatitis group (1,776 [SD 488] vs 967 [SD 860] ng/mL, p=0.03). For the lower LIC group (<5 mg Fe/g), the mean ferritin levels were not significantly different between the hepatitis and non-hepatitis groups (646 [SD 224] vs 459 [SD 205] ng/mL, p=0.22). The correlation between the viral load and mean ferritin level in NTDT group showed a significant linear correlation with R=0.7 (p=0.04).
Conclusions: We observe a higher serum ferritin level among thalassemia patients who concurrently have chronic viral hepatitis. Chronic viral hepatitis is a possible cause of a falsely high ferritin level in these patient population. Furthermore, the viral load is positively correlated with serum ferritin level.

Disclosures
No relevant conflicts of interest to declare.
C-Myc-Driven and PRMT5-Dependent Regulation of Multiple Myeloma Cell Proliferation through TNRC6B Gene
Authors: Xu Peipei , Dan Guo , Chen Bing , Zhao Quan , Zhou Rong-Fu , Ouyang Jian 
Published: November 2019

Objective: The protein arginine methyltransferase 5 (PRMT5) catalyzes the symmetrical bimethylation of arginine residues, which plays an important regulatory role in the life process. Transcription factor c-Myc is widely proved involved in the alteration and development of tumors, while the regulatory mechanism of c-Myc-mediated gene expression has not been fully understood. In this article we try to investigate the functional role of PRMT5 and c-Myc in regulating multiple myeloma cell proliferation.
Methods: The protein and mRNA expression levels of PRMT5 in myeloma cells and normal cells were detected by Western Blot and qPCR. Lentivirally transduced shRNAs targeting PRMT5 (sh-PRMT5) was constructed using lentivirus mediated RNAi technology, and was packaged to infect MM cells to select positive colonies. The effect of PRMT5 on the proliferation of MM cells was detected by cck-8 assay. Annexin V/7-AAD double staining flow cytometry was used to detect cell survival and apoptosis rate. The expression of apoptotic suppressor gene c-Myc in sh-PRMT5 cell lines was detected by Western Blot and qPCR. Finally, double-knockdown of PRMT5 and c-myc was performed and gene expression differentiation were identified by RNA sequencing (RNA-seq).
Results: PRMT5 expression in MM cell lines (RPMI8226 and U266) was relatively high compared with peripheral blood mononuclear cells (PBMCs), which was correlated with progression-free survival (PFS) and overall survival (OS) in MM patients. After sh-PRMT5 infection to MM cell lines, puromycin selection was performed and efficient gene knockdown was evaluated by Western Blot and qPCR. After targeted silencing of PRMT5 expression, the proliferation of MM cells was decreased and apoptosis was significantly increased (P < 0.05). C-Myc protein and mRNA levels were significantly downregulated after PRMT5 gene silencing. The results showed that PRMT5 promoted the proliferation of MM cells and inhibited the apoptosis of MM cells by regulating the expression of apoptotic suppressor gene c-Myc. Subsequently, RNA-seq confirmed SMCHD1 as the common candidate target gene of c-myc and PRMT5. Therefore, we speculated that PRMT5 and c-Myc may jointly regulate TNRC6B gene to promote the proliferation of MM cells.
Conclusion: Altogether, our study not only proposed the mechanism PRMT5 and c-myc in regulating multiple myeloma related gene expression, but also provided a new strategy and theoretical basis for clinical treatment of multiple myeloma.

Disclosures
No relevant conflicts of interest to declare.
Combined Targeting of Distinct c-Myc and JunB Transcriptional Programs for Multiple Myelioma Therapy
Authors: Lind Judith , Czernilofsky Felix , Vallet Sonia , Fan Fengjuan , Sattler Martin , Bakiri Latifa , Wagner Erwin F , Percherstorfer Martin , Podar Klaus 
Published: November 2019

c-Myc plays a pivotal role in multiple myeloma (MM) pathogenesis; and the BET protein BRD4 is a key regulator of c-Myc transcription. Recently, a pathophysiologic role in MM has also been attributed to the AP-1 family member JunB. Approaches to target transcription factors (TFs), such as c-Myc and JunB, currently emerge among the most promising novel anti-MM strategies, with a potentially high therapeutic index. However, redundancy phenomena represent a major challenge for targeting c-Myc- or JunB-mediated tumor proliferation programs.
MZ-1 is a novel proteolysis-targeting chimera combining the recognition sequence for the E3-ligase Von-Hippel-Lindau with JQ1, a moiety that targets BRD4. Indeed, beyond direct inhibition by JQ1, MZ-1 significantly decreased BRD4 as well as c-Myc protein levels in MM cell lines and primary cells. Consequently, MZ-1 inhibited MM cell growth through G0/G1 arrest; and induced tumor cell death. Moreover, patient-derived BMSC- or exogenous IL-6- induced BRD4/c-Myc upregulation in MM cells was inhibited by MZ-1 treatment, indicating that targeting BRD4, at least in part, overcomes the protective effect of the microenvironment. Of note, MZ-1 did not have an impact on BMSCs/IL-6- induced upregulation of JunB RNA or protein levels. Conversely, dox-induced knockdown of BMSC/IL-6-triggered JunB upregulation in TetshJunB/MM.1S cells did not decrease BRD4/c-Myc RNA or protein levels. Similar data were obtained in other MM cell lines and primary cells. RNAseq, unbiased GSEA, and luciferase reporter assays of representative target genes further supported the co-existence of c-Myc- and JunB-mediated proliferative programs, which are initiated by the same stimuli. Importantly, MZ-1 in combination with dox-induced knockdown of BMSC/IL-6-triggered JunB upregulation in TetshJunB/MM.1S cells significantly decreased both BRD4/c-Myc as well as JunB protein levels; synergistically inhibited tumor cell proliferation and induced cell death. Similar results were obtained in other MM cell line and primary cells upon treatment with MZ-1 and transient siJunB. Finally, our preliminary results show the synergistic in vivo activity of MZ-1 in BMSC:TetshJunB/MM.1S versus BMSC:TetshSCR/MM.1S-carrying doxycyclin- treated NSG mice. Taken together, these data for the first time delineate the existence of distinct c-Myc and JunB transcriptional programs that contribute to MM proliferation. They thereby provide the preclinical rationale for the development of therapeutic strategies that combine targeting of distinct transcriptional programs to improve outcome in MM.

Disclosures
Vallet: Roche Pharmaceuticals: Consultancy; Pfizer: Honoraria; MSD: Honoraria. Podar:Celgene: Consultancy, Honoraria; Amgen Inc.: Honoraria; Roche Pharmaceuticals: Research Funding; Takeda: Consultancy; Janssen Pharmaceuticals: Consultancy, Honoraria.
A Novel Role for Malate Dehydrogenase 1 in Acute Myeloid Leukemia Progression
Authors: Jin Jie , Huang Shujuan , Pan Jiajia 
Published: November 2019

The tumor suppressor role of the metabolic genes is rarely reported. Here, we report that malate dehydrogenase 1 (MDH1) acts as an anti-oncogene in acute myeloid leukemia (AML) progression. In a screen of various leukemia cell lines, as well as peripheral blood mononuclear cells from patients with AML and healthy individuals, we observed that patients with AML had lower mRNA expression of MDH1 than healthy individuals. In addition, we found that overexpression of MDH1 in the AML cell lines MV4-11 and THP-1 in vitro promoted their differentiation, reduced their growth, induced apoptosis, and increased the proportion of cells in S phase, which were more sensitive to the chemotherapeutic drugs cytarabine and daunorubicin. Using an in vivo xenograft mouse model of AML, we found that disruption of MDH1 expression influenced tumor burden and mouse survival. These findings were consistent with an inverse relationship between MDH1 expression and overall survival in two independent cohorts of AML patients. We also identified changes in the expression of other genes, concomitant with reduced MDH1 expression in patients with AML, that largely explain the anti-oncogenic role of MDH1 in AML progression. Notably, downregulation of P53 and upregulation of the MAPK pathway in low MDH1-expressing MV4-11 cells were reversed upon overexpression of MDH1. In conclusion, we demonstrated that MDH1 plays a tumor-suppressing role in AML and can be used as a novel mimic drug therapy in the future.

Disclosures
No relevant conflicts of interest to declare.
Immunosenescent Characteristics of T Cells in Young Patients Following Haploidentical Stem Cell Transplantation from Parental Donors
Authors: Lee Ga Hye , Hong Kyung Taek , Choi Jung Yoon , Shin Hee Young , Lee Won-Woo , Kang Hyoung Jin 
Published: November 2019

Introduction: Pediatric and adolescent patients in need of allogeneic hematopoietic stem cell transplantation generally receive stem cells from older, unrelated or parental donors when a sibling donor is not available. Despite encouraging clinical outcomes, it has been suggested that immune reconstitution accompanied by increased replicative stress and a large difference between donor and recipient age may worsen immunosenescence in pediatric recipients. Therefore, in this study paired samples were collected at the same time from donors and recipients of haploidentical hematopoietic stem cell transplantation (HaploSCT).
Methods: We conducted flow cytometry-based phenotypic and functional analyses and telomere length measurements of 21 paired T-cell sets from parental donors and children who received T cell-replete HaploSCT with post-transplant cyclophosphamide (PTCy) at Seoul National University Children's Hospital between February 2014 and January 2017. The conditioning regimen was comprised of targeted busulfan (total target area under the curve, 75,000 mg•h/L) with intensive pharmacokinetic monitoring, fludarabine and cyclophosphamide.
Results: Fourteen pediatric, adolescent, and young adult patients with malignant disease and seven with nonmalignant disease were included with a median post-transplantation period of 16.9 months (range, 12.4-38.8). Senescent T cells, CD28- or CD57+ subsets of both CD4+ and CD8+ T cells, were significantly expanded in patients compared with parental donors. Further, not only CD4+CD28- T cells, but also CD4+CD28+ T cells showed reduced cytokine production capacity and impaired polyfunctionality compared with parental donors, whereas their TCR mediated proliferation capacity was comparable. Of note, the telomere length in patient T cells was preserved, or even slightly longer, in senescent T cells compared with donor cells. We also found that the patients had a higher level of γ-H2AX-expressing CD28- senescent T cells compared with the donors, which is used as a DNA damage marker. Regression analysis showed that senescent features of CD4+ and CD8+ T cells in patients were influenced by donor age and the frequency of CD28- cells, respectively.
Conclusions: Our data suggest that T cells undergo premature immunosenescent changes and exhibit functional defects in pediatric HaploSCT recipients. Further, there is an increased level of DNA damage in patient CD4+ T cells compared to those of parental donors. Therefore, long-term, comprehensive immune monitoring of these patients is necessary.

Disclosures
No relevant conflicts of interest to declare.
Cytomegalovirus Infection in Bone Marrow-Derived Mesenchymal Stem Cells: Implications for Functions and Exploration of Relevant Mechanisms
Authors: Lv Weiran , Fan Zhiping , Huang Fen , Xu Na , Xuan Li , Lin Ren , Zhao Ke , Liu Qifa 
Published: November 2019

Human cytomegalovirus (HCMV) infection could lead to life-threatening complications in immunocompromised and immunosuppressed populations, especially after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Reactivated CMV infections can contribute to poor graft function (PGF) characterized by multilineage hematopoietic failure after allo-HSCT. Bone marrow derived mesenchymal stem cells (BM-MSCs) play an important role in supporting hematopoiesis and modulating differentiation of hematopoietic primitive stem cells. Nevertheless, little is known about the dysfunctions and underlying mechanism closely associated with hematopoiesis of BM-MSCs after HCMV infection and no previous studies have concentrated on the dysfunctions of primary BM-MSCs from patients with CMV viremia following allo-HSCT. In the current study, we explored whether BM-MSCs could be infected by HCMV and evaluated the function of BM-MSCs with HCMV infection and the relationship between the dysfunction and mitogen activated protein kinases (MAPKs). Moreover, we retrospectively investigated the relation between CMV infection and PGF among 801 patients who received allo-HSCT in Nanfang Hospital. Our results showed that HCMV infection was detected both in BM-MSCs with HCMV AD169 infection in vitro and in BM-MSCs derived from patients with CMV viremia following allo-HSCT. Dysfunctional BM-MSCs with HCMV infection, which were characterized by impaired proliferation, osteogenesis potential, mRNA level of stem cell growth factor (SCF), colony-forming unit (CFU) plating efficiency and higher levels of apoptosis, adipogenesis potential, mRNA level of interleukin 6 (IL-6) and transforming growth factor β1 (TGF-β1) were revealed. Activation of p38 was detected in BM-MSCs with HCMV infection and function of infected BM-MSCs were enhanced by SB203580 treatment in vitro. Retrospective clinical data analysis showed that haploidentical donor (HRD) transplant and CMV infection (in 30 days) were risk factors of PGF. In summary, dysfunctional BM-MSCs were observed after HCMV infection which might induce hematopoiesis suppression and SB203580 treatment in vitro functionally improved BM-MSCs through downregulation of the p38MAPK pathway and our clinical analysis showed that CMV infection is a risk factor of PGF which further demonstrated hematopoietic inhibition of CMV.

Disclosures
No relevant conflicts of interest to declare.