bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lw-213 Synergizes with Rituximab to Inhibit Diffuse Large B-Cell Lymphomas By Upregulating CD20
Authors: Li Yuchen , Li Hui , Hui Hui , Xu Jingyan 
Published: November 2021

Abstract
Background：Downregulation of CD20, a molecular target for monoclonal antibodies (mAbs), is a clinical problem leading to decreased efficacy of anti-CD20-based therapeutic regimens.Up to one-third of diffuse large B cell lymphoma (DLBCL) patients eventually develop resistance to R-CHOP regimen, since the remaining therapeutic options are limited. LW-213, a derivative of wogonin, is reported to possess antineoplastic properties in a variety of cancers, but whether it has effects on DLBCL is n-ot known. Studies have reported that upregulation of CD20 expression b-y either HDACi or silenced SOX2 expression showed sensitizing potential in Rituximab-induced cell death in malignant B cells. Our study was to explore whether LW-213 could sensitize DLBCL to Rixutimab thus improve therapeutic efficacy.
Methods： Two DLBCL cell lines, RI-1 (ABC subtype) and Su-DHL -8 (GCB subtype), were used in our study. RI-1 and Su-DHL-8 cells were treated with LW-213 at different doses and for different times, and their proliferation and viability were detected by Cell counting kit-8 (CCK8).Flow cytometry was used to determine surface CD20 expression. Western blotting and q-PCR were applied to examine the protein and mRNA levels of CD20, SOX2, Ace-H3 and Ace-H3K27. CDC assay was used to evaluate the synergistic effects of LW-213 and Rixutimab.
Results：We showed that LW-213 inhibited the proliferation of human DLBCL cell lines (Su-DHL-8、RI-1 ) in dose-and time-dependent manners with IC 50 values at the low μmol/L levels, meanwhile it potently inhibited primary lymphoma cells derived from peripheral blood of B-cel-l lymphoma patients. Furthermore, LW-213 significantly increased CD20 surface expression and the acetylation level of histone in DLBCL cell li-nes. Inversely，the SOX2 expression level remarkably decreased. Finally,Combination with LW-213 significantly synergized Rituximab-induced cell death in vitro.
Conclusion： The results demonstrate that LW-213 sensitizes DLBCL cells to Rituximab in vitro by upregulating CD20 expression and the SOX2/ace-H3K27/ace-H3 axis may plays a critical role in CD20 upregulation processing. Even though this strategy is important in vitro models,the upregulating CD20 expression therapy against DLBCL proposed in this study warrants further study in vivo and clinical trials .
Keywords：CD20 DLBCL Rituximab SOX2 Histone Deacetylation

Disclosures
No relevant conflicts of interest to declare.
Abelacimab Reduces Thrombus Propagation in a Baboon Model of Vascular Graft Thrombosis
Authors: Wallisch Michael , Khder Yasser , Hinds Monica T. , Tucker Erik I , Bloomfield Dan , Gruber Andras 
Published: November 2021

Abstract
Background: Factor XI (FXI) inhibition demonstrated strong efficacy in preventing thrombus formation in preclinical and clinical models of arterial and venous thrombosis. However, the effect of FXI inhibition in halting the progression of a formed clot remains largely unknown.
Aims: This study aims to test whether abelacimab, a dual-acting FXI and activated FXI (FXIa) monoclonal antibody, is effective in halting clot formation and downstream growth when administered before or during active clot formation in an established baboon femoral arterio-venous (AV) shunt model.
Methods: Three baboons had a chronic femoral AV shunt put in place; platelet and fibrin deposition inside and distal to collagen- or collagen + tissue factor (TF)-coated vascular grafts were measured at baseline (control), in a therapeutic setting, where abelacimab (1 mg/kg, intravenously) was administered 30 minutes after thrombus initiation, and in a preventative setting within the first 48 h and 1 week (144 - 216 h) post-administration. Pharmacodynamic effect was measured by activated partial thromboplastin time (aPTT).
Results: Consistent with its half-life of 20 to 30 days, single iv administration of abelacimab at a dose of 1 mg/kg resulted in long-lasting (&gt; 4-week) aPTT prolongation (&gt; 2-fold). Administration of abelacimab 30 minutes after initiation of thrombosis using grafts coated either with collagen or with collagen + TF quickly halted downstream propagation of platelet and fibrin deposition compared to control. Further, downstream propagation of platelet and fibrin deposition was markedly reduced when clotting was induced by collagen, or collagen + tissue factor after abelacimab administration.
Conclusions: These data suggest that abelacimab, a dual-acting anti-FXI/FXIa monoclonal antibody with a single long-lasting iv injection has the potential to slow down the growth and reduce the size of thrombi when admistered before or after clot induction. Data indicate a potential for therapeutic benefit of targeting FXI both in therapeutic and preventive settings.
Sponsored by: Anthos Therapeutics Inc., 55 Cambridge Parkway, Suite 103, Cambridge, MA 02142
Figure 1 Figure 1.

Disclosures
Wallisch: Aronora Inc,: Current Employment. Khder: Anthos Therapeutics: Consultancy; Novartis: Current equity holder in publicly-traded company, Other: Retiree. Bloomfield: Anthos Therapeutics: Current Employment. Gruber: Aronora Inc.: Current Employment, Current equity holder in publicly-traded company; Oregon Health and Science University: Current Employment.
The Application of Eltrombopag to Promote Platelet Engraftment after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Li Yahan , Yujie Jiang , Liu Xin , Li Ying , Wang Xianghua , Xu Hongzhi , Wang Xin , Fang Xiaosheng 
Published: November 2021

Abstract
Background :
Eltrombopag is an agonist of the thrombopoietin receptor, and it has achieved great efficacy in the treatment of immunologic thrombocytopenic (ITP) and reduction of platelets (PLT) after hematopoietic stem cell transplantation (HSCT), but it remains unclear whether Eltrombopag can promote PLT engraftment after HSCT.
Methods :
37 patients with HSCT were administered Eltrombopag 50 mg (qd) from +1 day until PLT &gt;50x10^9/L or 1 month after HSCT. 45 patients in the same period applied TPO to promote PLT engraftment after HSCT and served as a control group. The engraftment time of PLT and WBC (white blood cell) as well as the GVHD incidence and adverse reaction were compared and analyzed in the two groups.
Results :
A total of 37 patients received Eltrombopag, 2 of the patients withdrew from the study due to intolerable gastrointestinal reactions. 35 patients eventually completed treatment with Eltrombopag. In Eltrombopag group, median time to WBC engraftment was 12 days (range, 10-17 days) and PLT engraftment was 15 days (range, 10-29 days). At the 1st month after HSCT, the median WBC was 3.93 x10 9/L (range, 0.87-40.01 x10 9/L) and the median PLT was 91 x10 9/L (range, 33-401 x10 9/L) in the Eltrombopag group. MKCs in the bone marrow were assayed at 1 month after HSCT. Median MKCs level was 40 (range, 2-345) in the Eltrombopag group. 13 (37%) patients developed grade II-IV aGVHD and 1 (2%) patient developed cGVHD in the Eltrombopag group. Of these 35 patients, 3 (8%) presented with elevated total bilirubin (TBIL), no patient had elevated alkaline phosphatase (ALP), 17（48%）developed proteinuria, 1 (2%) people with elevated blood urea nitrogen (BUN) and no patient showed elevated creatinine (CREA). A total of 45 patients were applied TPO during the same period. Among these patients, the median engraftment time for WBC and PLT was 12 days (range, 9-23 days) and 15 days (range, 9-41 days), respectively. At the 1st month after HSCT, the median number of WBC and PLT were 4.30 x10 9/L (range, 0.99-23.63 x10 9/L) and 79.5 x10 9/L (range, 5-512 x10 9/L), respectively. At the 1st month after transplantation, the median number of MKCs in the bone marrow were 44 (range, 2-788). 16 (35%) patients developed grade II-IV aGVHD and 6 (13%) patients developed cGVHD in the TPO group. Among these 45 patients, 7 (15%) presented with elevated TBIL, 5 (11%) had elevated ALP, 25 (55%) developed proteinuria, 4 (8%) people with elevated BUN and 1 (2%) patient showed elevated CREA. The engraftment time of PLT and WBC were same in the Eltrombopag group and the TPO group and the adverse reactions occurred similarly in both groups.
Conclusion :
Eltrombopag can be used to promote PLT engraftment after all-HSCT with acceptable adverse reaction.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Differential Activation of V γ4V δ1 and V γ9V δ2 Cord Blood Derived Gamma Delta T Cells upon Exposure to EBV-Lcl and K562 Cells
Authors: Ng Jeremy Wee Kiat , Lai Joey , Lim Tony Kiat Hon , Hwang William YK , Li Shang , Eaves Connie J. , Goh Yeow Tee , Cheung Alice 
Published: November 2021

Abstract
Gamma-delta (γδ) T cells have emerged as a promising candidate for adoptive cellular immunotherapy. To harness and maximize the anti-leukemia properties of these cells, we sort to comprehensively profile the transcriptomic signatures and immune repertoire of in vitro expanded γδ T cell products. Given the reported diverse TCR γδ repertoire and naïve nature of γδ T cells found in human cord blood (CB γδ), we serially track the molecular and cellular changes in these cells upon activation in expansion cultures. Based on the established viral reactivities of γδ T cell as well as prior studies showing their cross reactivities against leukemia and cancer cells, we had previously shown that stimulating CB γδ with an irradiated EBV-LCL feeder cell-based rapid expansion protocol (REP) is capable of generating cell products with potent and specific cytotoxicity against human AML cells. In the present study, using single cell RNA sequencing (scRNA-seq) coupled with single cell TCR γδ repertoire analysis, we compared the transcription signatures between our REP expanded γδ T cell (REP γδ) and non-manipulated γδ T cells reported in literatures, showing the progressive acquisition of an adult PB derived γδ T cell (PB γδ)-like cell states. Time course analysis demonstrated complex T cell activation and maturation trajectories correlating with variable level of clonal induction throughout the course of in vitro expansion. At the end of expansion, the harvested REP γδ are predominantly of the V γ4V δ1 subtype. Nevertheless, upon exposing REP γδ to target leukemia cell line K562, outgrowth of other non-V γ4V δ1 as well as the semi-invariant V γ9V δ2 cells were observed. Taken together, our data shows that as CB γδ expand and differentiate in culture, they adopt an adult PB γδ-like program. More importantly, our data highlights the rich clonal composition of in vitro expanded CB γδ, with different clonotypes being variably activated upon exposure to different stimuli. Such characteristics can potentially overcome the challenges of cancer heterogeneity and cell persistence, with the potential of improving outcomes in cell immunotherapy.

Disclosures
No relevant conflicts of interest to declare.
Fanci-FANCD2 Promotes Genome Stability and DNA Repair By Down-Regulating BLM Helicase Activity
Authors: Liang Fengshan , Nagarajan Arvindhan , Pillai Manoj M , Sung Patrick , Kupfer Gary M. 
Published: November 2021

Abstract
Background: Fanconi anemia (FA) is a genetic disease characterized by bone marrow failure, developmental defects, and higher risk of cancer. Mutations in FA genes have been detected commonly in a large swath of cancers. In the FA DNA repair pathway, DNA damage induces the mono-ubiquitination of the FANCI-FANCD2 (ID2) heterodimer and this regulation licenses the execution of downstream DNA damage signaling and repair steps. In response to replication stress, FANCD2 also prevents replication fork collapse during S phase. Bloom syndrome (BS) is also a genomic instability disease, characterized by growth abnormalities and cancer predisposition. The single BS protein, BLM helicase, participates in DNA repair by promoting DNA end resection and double Holliday junction dissolution. It has been shown that BLM is involved in restart of stalled replication fork. FA and BS have functional interactions. In tumor DNA sequencing of the Yale Precision Tumor board, we identified a somatic 6 amino acid deletion in FANCD2 in a head and neck tumor, while a germline point mutation was found on the other allele. We have identified a FANCD2-L822A mutant with defective BLM binding, which was used to further investigate the role of FANCD2-BLM interaction in genome stability and DNA repair.
Methods: Highly purified proteins were used to investigate how ID2 affects helicase and DNA end resection activity of the BLM complex. HeLa, FANCD2-deficient, and FANCD2 corrected fibroblast cell lines were used to examine pRPA2 and RAD51 foci formation. We also used DNA fiber assay to detect end resection and isolation of proteins on nascent DNA (iPOND) assay to examine the RAD51 recruitment on replication fork.
Results: A somatic 6 amino acid deletion (p819-824) in FANCD2 was identified in a head and neck tumor. FA-D2 mutant cells expressing the mutant cDNA demonstrated defects in FANCD2 mono-ubiquitination and DNA damage hypersensitivity. A FANCD2-L822A mutant with defective BLM binding was identified (Figure A, B). We found that Bloom helicase and its DNA end resection activity within BLM-DNA2-RPA were negatively regulated by the heterodimer ID2 (Figure C, D). Both DNA and BLM binding of the ID2 are required for the inhibitory function. The premature DNA end resection and HU sensitivity in FANCD2 deficient and mutant cells are rescued by BLM knockdown. By iPOND assay, we discovered that FANCD2 antagonizes BLM to promote RAD51 recruitment on HU-stalled replication fork.
Conclusions: Our study suggests that the DNA end resection activity of BLM-DNA2 is tightly regulated by FANCD2 to ensure that the nuclease DNA2 normally resects the DNA intermediate needed for efficient DNA repair and RAD51 recruitment to protect replication forks. Our findings highlight that ID2-BLM interaction functions in DNA damage repair to maintain genome stability.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Single-Cell Sequencing Technology Reveals the Heterogeneity of the Immune Microenvironment and Key Molecules of Drug Resistance in Angioimmunoblastic T-Cell Lymphoma
Authors: Jin Hui , Fan Lei , Lu Xueying , Cao Lei , Wu Zijuan , Wang Luqiao , Zhu Huayuan , Li Jianyong 
Published: November 2021

Abstract
Objective:
Angioimmunoblastic T-cell lymphoma (AITL) is a common subtype of peripheral T-cell lymphoma (PTCL). AITL is an aggressive malignancy with a poor prognosis, and its clinical manifestations vary greatly among individuals. The current chemotherapy regimens based on anthracycline show limited efficacy, and there is no best rescue treatment for patients with relapsed and refractory (RR) AITL. In addition, the lack of optimal AITL models in vitro greatly limits the basic research on the mechanism of disease occurrence and progression, and also hinders the development of new drugs and preclinical trials. Our study aims to deeply analyze the tumor heterogeneity and clonal evolution of AITL, discovering key molecules of drug resistance and potential theraputic targets.
Methods :
We detected fresh lymph node samples from newly diagnosed and relapsed/ refractory AITL patients using single-cell RNA sequencing, combined with imaging mass cytometry (IMC) and whole exome sequencing. IMC was performed to analyze the spatial position relationship and protein expression characteristics of different subgroups in the tumor microenvironment of AITL. In addition, AITL patient-derived organoid model was established to study the regulatory role of YY1 and its inhibitors in relapsed and refractory AITL.
Results :
ScRNA-seq revealed the significant differences in the tumor microenvironment of newly diagnosed and RR-AITL patients (Fig A,B). B cells and myeloid subgroups may play important roles in the development of AITL (Fig C). Transcription factor YY1, highly expressed in follicular helper T cell (Tfh) of RR-AITL patients, promoted the proliferation and drug resistance of AITL cells (Fig E,F). The proportion of CD8+ T cells in the RR-AITL sample was reduced, while the proportion of Treg was increased, as well as the depletion of T cells (Fig G,H). Furthermore, the stemness of B cells in RR-AITL was enhanced and exhibits significant malignant characteristics (Fig C,I-K). We also found decreased interaction in RR-AITL samples (Fig L,M). Moreover, for the first time, we established AITL patient-derived organoid models that can be stablely cultured in vitro (Fig N). On this basis, we could further clarify the important roles of transcription factor YY1 in the drug resistance of AITL, evaluate the cytotoxic effect of YY1 inhibitor NP-001 on AITL tumor cells.
Conclusion :
In conclusion, our study revealed the differences between newly diagnosed and relapsed /refractory AITL in terms of immune microenvironment, single-cell transcriptomes, and signal pathway activation. YY1 may serve as an novel target for drug resistance for RR-AITL patients. These findings may provide a theoretical foundation for improving the clinical treatment of AITL.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Comparison of Prophylactic and Preemptive Donor Lymphocyte Infusion in Patients with Very High Risk Acute Leukemia after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Yang Luxin , Luo Yi , Huang He , Tan Yamin , Shi Jimin , Zhao Yanmin , Yu Jian , Lai Xiaoyu 
Published: November 2021

Abstract
Relapse remains to be the biggest problem for patients with very high risk acute leukemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Donor lymphocyte infusion (DLI) has been widely applied to the prevention and preemptive intervention of relapse post-transplantation. However, the optimal time to implement DLI in patients with a high risk of relapse remains controversial. During August 1, 2014 and August 30, 2020, four hundred and thirty-one consecutive acute leukemia patients with very high-risk features underwent allo-HSCT in our center. Ninety-five patients received prophylactic DLI (pro-DLI) while one hundred and fifty-five patients received preemptive DLI (pre-DLI). Patients were stratified into five risk groups by the disease-risk stratification system (DRSS) which recently introduced by European Society for Blood and Marrow Transplantation: low risk (N=44), intermediate-1 (N=67), intermediate-2 (N=60), high (N=36), and very high (N=43). In the very high group, patients who received pro-DLI achieved higher 3-year overall survival (OS) (50.8% versus 31.8%, P=0.05), leukemia-free survival (LFS) (52.5% versus 31.6%, P=0.03) and graft-versus-host disease (GVHD)-free and relapse-free survival (GRFS) (47.9% versus 31.6%, P=0.04) compared with pre-DLI. Pro-DLI cohort was associated with a trend of lower 3-year cumulative rates of relapse (42.8% versus 57.9%, P=0.12) and non-relapse mortality (NRM) (4.6% versus 10.5%, P=0.43). In the subgroup of intermediate-2 and high (N=96), pro-DLI cohort was associated with superior survival and relapse rate, though without statistical significance. Further analysis was conducted separately between patients with positive minimal residual disease (MRD) and negative MRD pre-transplant in intermediate-2/high subgroup. In the patients with positive MRD before all-HSCT, pro-DLI achieved significant higher 3-year OS (66.7 % versus 26.2%, P=0.028), PFS (68.6 % versus 24.7%, P=0.015) and GRFS (68.6 % versus 14.4%, P=0.007), along with lower cumulative of relapse (31.4 % versus 59.3%, P=0.1) and NRM (0 versus 16%, P=0.1) compared with pre-DLI. However, pro-DLI patients was associated with inferior OS (52.7 % versus 77.8%, P=0.046), PFS(54.8 % versus 75.3%, P=0.247) and NRM (25% versus 3%, P=0.02) in patients with negative MRD. In addition, no difference was found in patients in the group of low and intermediate-1 risk. Taken together, we recommend early pro-DLI in patients of very-high risk group and positive MRD of intermediate-2/high group. Pre-DLI given timely after MRD turning positive achieved better outcome in the remaining patients.

Disclosures
No relevant conflicts of interest to declare.
Histopathologic Characterization of Vexas Syndrome
Authors: Barouqa Mohammad I. , Ravindran Aishwarya , Chen Dong , Oliveira Jennifer L , Olteanu Horatiu , Howard Matthew T , Zheng Gang , Patnaik Mrinal M. , Begna Kebede H. , Bock Allison M. , Go Ronald , Koster Matthew , Kourelis Taxiarchis , Mangaonkar Abhishek A. , Ryu Jay , Samec Matthew , Sartori Valinotti Julio C. , Warrington Kenneth , Reichard Kaaren K. 
Published: November 2021

Abstract
Introduction:
VEXAS syndrome (vacuoles, E1 ubiquitin ligase, X-linked, autoinflammatory, somatic) is a newly recognized inflammatory disorder caused by somatic mutations in the UBA1 gene. Bone marrows from these patients reveal a range of morphological changes in hematopoietic precursor cells. In this study, we aim to assess the laboratory indices and morphologic spectrum of bone marrow pathology in VEXAS syndrome.
Methods:
We identified 16 cases of VEXAS syndrome. All cases had confirmed UBA1 mutation. We reviewed bone marrow biopsies corresponding to the date of diagnosis. This study was approved by the Mayo Clinic Institutional Review Board.
Results:
All patients were male with a median age of 73 years - associated autoimmune disorders included Sweet syndrome, inflammatory arthritis, relapsing polychondritis and granulomatosis with polyangiitis.
14/16 patients had anemia with median hemoglobin of 10.4 (Range: 6.7- 14.1 g/dL). 15/16 had macrocytosis with median MCV 110.4 (Range: 94.8- 123.1 /fL). 5/16 had thrombocytopenia with median platelet count 174 (Range: 20- 500 x10^9/L). 7/16 had leukopenia with median WBC 3.65 (Range: 2.4- 11.6 x10^9/ L). The ESR and CRP medians were 61.0 mm/hr and 81.5 mg/L, respectively.
Karyotype was performed in 12 patients of which 11 were normal and the remaining case showed a complex karyotype. An NGS panel targeting the most frequent myeloid disorder associated gene mutations was negative in 10/15 cases. GS for myeloid mutations revealed pathogenic mutations in 5 patients, involving genes TET2 (2/5), DNMT3A (2/5), and TP53 (1/5).
Conclusions:
Bone marrow findings in VEXAS syndrome, in this series of 16 patients, are individually non-specific, yet when taken altogether in the overtly abnormal cases, are very suggestive when the clinical index of suspicion is high. In such scenarios, the combined clinical and bone marrow findings should prompt discussion and consideration for UBA1 mutation testing given the significant clinical implications for patient management and prognosis.

Disclosures
Patnaik: StemLine: Research Funding; Kura Oncology: Research Funding. Warrington: Eli Lilly: Research Funding; Kiniksa: Research Funding.