bluuebunny/crossref_metadata_2025_split huggingface dataset:
HT-6184 Inhibits the Formation of the NLRP3 Inflammasome in Human Monocytic THP-1 Cells
Authors: Bearss Benjamin , Bursey Devan , Avei Taylor , Mollard Alexis , Bearss Jared , Janat-Amsbury Margit Maria , Bearss David J. 
Published: December 2023

Background
The NLRP3 inflammasome is a multiprotein complex composed of several proteins, including NLRP3, ASC, Nek-7, and pro-caspase-1. Many immune cell populations express the NLRP3 inflammasome. In the presence of various exogenous stimuli such as lipopolysaccharide (LPS) or extracellular ATP, the NLRP3 inflammasome components assemble and become activated, releasing pro-inflammatory cytokines IL-1β and IL-18. One of the critical steps in inflammasome assembly is the recruitment of ASC and the formation of ASC protein aggregates called specks. These specks are large enough to be measured through microscopy, and the ASC speck count indicates NLRP3 activation. The release of IL-1β and IL-18 and the upregulation of NLRP3 inflammasome component genes have been observed in many inflammatory and hematological disorders, suggesting the NLRP3 pathway is involved in the disease pathogenesis of many diseases.
Purpose
HT-6184 is a small molecule that blocks the assembly of the NLRP3 inflammasome through allosteric binding to the protein Nek7. Here we demonstrate the ability of HT-6184 to modulate NLRP3 inflammasome formation by measuring ASC specks using a human monocytic THP-1 cell line engineered to express a GFP-labeled ASC protein.
Methods
THP1-ASC-GFP cells were cultured in RPMI 1640 with 10% heat-inactivated FBS, 100 μg/mL normocin, and 100 μg/mL Pen-Strep. Cells were seeded at 400,000 cells per well in a 96-well plate and left overnight. Cells were then treated with vehicle or LPS for 3 hours. Following LPS stimulation, cells were treated with vehicle or HT-6184 for 2 hours. After incubation, cells were treated with 5 μM Nigericin and imaged in 3-minute intervals for 1 hour. The images were analyzed for specks.
Results
By treating THP1-ASC-GFP with LPS and nigericin, we induced the formation of ASC specks and visualized them using fluorescent microscopy. Cells treated with 200 nM HT-6184 had significantly fewer ASC specks than the LPS and nigericin-treated control. ASC speck inhibition by HT-6184 was dose-dependent, as inhibition of speck formation decreased with lower doses of HT-6184.
Conclusions
HT-6184 effectively prevents the formation of ASC specks in THP-1-ASC-GFP cells. Since THP-1 cells are a monocytic cell line, this data suggests our ability to inhibit the NLRP3 pathway in hematological disorders effectively. By preventing inflammasome formation and activation, we expect to be able to reduce disease burden in hematological diseases.
Ex-Vivo Human Whole Blood Assay for Evaluating the Pharmacodynamics of HT-6184: Reduction in NLRP3 Inflammasome Mediated Cytokine Production
Authors: Avei Taylor , Bursey Devan , Bearss Benjamin , Mollard Alexis , Bearss Jared , Janat-Amsbury Margit Maria , Bearss David J. 
Published: December 2023

The NLRP3 inflammasome is an intracellular multiprotein complex that plays a crucial role in the innate immune response by mediating the maturation and release of pro-inflammatory cytokines IL-1 beta, and IL-18. The release of these cytokines leads to the downstream release of TNF-alpha, IL-6, which propagate a robust inflammatory response. Dysregulation of the NLRP3 inflammasome pathway has been linked to various inflammatory diseases, suggesting the possibility for novel therapeutic strategies targeting NLRP3 activation. NLRP3 has been linked to regulating hematopoietic stem progenitor cell (HSPCs) development, expansion, release, and mobility. In addition, various hematological disorders such as myelodysplastic syndrome, myeloproliferative neoplasms, acute and chronic leukemias, and graft-versus-host disease (GvHD) have all been shown to involve NLRP3 inflammasome activation. Given its significant role in these conditions, inhibiting the NLRP3 inflammasome may be a novel therapeutic strategy in hematology and hemato-oncology
This study utilized an ex-vivo human whole blood assay to investigate the pharmacodynamics of HT-6184, an allosteric Nek7 inhibitor. We used LPS and ATP to prime and activate the NLRP3 inflammasome and examined the effects of pre-treatment with various concentrations of HT-6184 (starting at 1 mM) on the production of critical pro-inflammatory cytokines, such as IL-1b, IL-18, IL-6 and TNF-a. Notably, preincubation with HT-6184 at concentrations as low as 4.1 nM led to a significant reduction in cytokine production (p&lt;0.0001). Specifically, TNF-α and IL-6 were reduced by 38% and 58%, respectively, while IL-1β and IL-18 were decreased by 44% and 53% respectively.
These results provide further evidence of the ability of HT-6184 to modulate the activity of the NLRP3 inflammasome in a complex biological milieu. Our study suggests that HT-6184 has significant therapeutic potential in treating a broad spectrum of inflammasome-mediated diseases. In addition, this assay has the potential to be used to measure the pharmacodynamic properties and the efficacy of HT-6184 and other inflammasome inhibitors in future healthy volunteer clinical trials. In conclusion, this study highlights the importance of innovative and physiologically relevant assay systems in identifying and evaluating potential therapeutic agents for inflammatory diseases.
SGN-35C: A Novel CD30-Directed Antibody-Drug Conjugate for the Treatment of Lymphomas
Authors: Hamblett Kevin J. , Cochran Julia , Snead Katie , Jin Steven , Yumul Roma , Simmons Jessica , Stone Ivan , Lyski Ryan , Schrum Jason P. , Lim Andrea R. , Clarke Astrid 
Published: December 2023

SGN-35C is an antibody drug conjugate composed of an anti-CD30 antibody conjugated to a camptothecin-derived topoisomerase 1 (TOP1) inhibitor payload. SGN-35C was designed to leverage the antibody backbone from brentuximab vedotin (BV) and the novel mechanism of action of camptothecin-derived ADCs. Here, we evaluated the mechanism of action, cytotoxicity, and safety in in vitro and in vivo models.
To characterize the mechanism of action of SGN-35C in vitro and in vivo experiments were performed in CD30-positive anaplastic large cell lymphoma (ALCL), Hodgkin lymphoma (HL) cell lines, and versions of these cell lines that are resistant to BV. SGN-35C binding, internalization, induction of DNA damage, cell cycle analysis, and in vitro cytotoxicity were evaluated. To assess if payload released in CD30-positive cells could kill adjacent CD30-negative cells, an in vitro bystander assay of CD30-positive and CD30-negative co-cultures was developed. The in vivo antitumor activity of SGN-35C was evaluated in animals bearing subcutaneous lymphoma models. SGN-35C antitumor activity was evaluated in a xenograft model of heterogeneous CD30 expression consisting of a mixture of a BV-resistant Karpas-299 cell line mixed with parental Karpas-299 cells to assess bystander activity in vivo. The tolerability of SGN-35C was assessed in a non-human primate model.
SGN-35C binding to CD30-positive cells was similar to the unconjugated anti-CD30 antibody and demonstrated internalization into CD30-expressing cells. Treatment of cells with SGN-35C for 24 hours led to an increase of the DNA damage marker phospho-γH2A.X, which induced an increase in the fraction of cells in S-phase, consistent with the mechanism of camptothecin-based TOP1 inhibitors. Cytotoxicity was observed in ALCL and HL cell lines treated with SGN-35C, including an ALCL cell line resistant to BV. In a mixture of CD30-positive and CD30-negative cells, SGN-35C treatment induced cytotoxicity of the CD30-negative cells, demonstrating that SGN-35C can elicit bystander activity. A single dose of SGN-35C led to significant tumor growth inhibition in animals bearing lymphoma xenografts, including a BV-resistant model and a model of heterogeneous CD30 expression, demonstrating that SGN-35C can elicit bystander activity in vivo. SGN-35C demonstrates a tolerability profile in non-human primates that is consistent with camptothecins. A Phase 1, first-in-human study is planned to evaluate the safety and anti-tumor activity of SGN-35C in lymphoid malignancies.
Efficacy of Direct Oral Anticoagulants Vs Warfarin Vs Low Molecular Weight Heparin in the Treatment of Acute Splanchnic Vein Thrombosis in Patients with Underlying Myeloproliferative Disorder
Authors: Jamil Maria , Nasser Zeinab , Jacob Brigid , Mangal Ruchi , Donthireddy Vijaya 
Published: December 2023

Introduction:
Myeloproliferative neoplasms (MPNs) are hematopoietic stem cell disorders characterized by clonal proliferation of myeloid-lineage cells. Venous thrombosis poses a significant morbidity risk for MPN patients. The current recommended treatments for acute splanchnic vein thrombosis (SVT) are warfarin and low molecular weight heparin (LMWH). Direct oral anticoagulants (DOACs) are potential alternatives due to their predictable dose response and oral administration, but limited data exists for their use in acute SVTs. This retrospective study aims to assess the efficacy and outcomes of DOACs, LMWH, and warfarin in treating acute SVTs in patients with underlying MPN.
Methods:
We included patients of all ages with underlying myeloproliferative neoplasms and splanchnic vein thrombosis, with no underlying liver diseases, from Henry Ford Hospital Clinics in metropolitan Detroit, Michigan between 2013 and 2023. Patient data, which included age, gender, race, and BMI were recorded. The primary outcome was complete radiographic resolution (CRR) of the SVT, and secondary outcomes included recanalization, thrombosis progression, recurrent thrombosis, major bleeding, thrombocytopenia, and skin necrosis. The outcomes were compared between the three anticoagulant groups and patients using aspirin.
Results:
Among the 34 MPN patients with SVT, warfarin was prescribed most frequently (n=19), followed by enoxaparin (n=5) and DOACs (n=5). Although no significant differences were observed, warfarin showed the highest CRR rates (41.2%), while enoxaparin and DOACs had equal CRR rates (25%). DOACs showed recanalization rates similar to warfarin and higher than enoxaparin (44.4% and 47.1% vs. 20.0%, respectively). LMWH was associated with increased recurrent thrombosis rates (50.0%), while DOACs had a higher risk of major bleeding. All three anticoagulants had similar effects on thrombocytopenia and SVT progression. Aspirin use was linked to a higher risk of major bleeding (42.9%), but not using aspirin correlated with complete SVT resolution (30.8%), SVT progression (20.8%), recurrent thrombosis (29.2%), and thrombocytopenia (28.6%).
Conclusions:
Although limited by a small sample size, our study found no significant differences among the three anticoagulant groups. This contributes to establishing the role of DOACs in treating SVTs in patients with underlying MPN. Further studies with larger sample sizes are needed to explore the efficacy of DOACs compared to warfarin. Additionally, investigating the potential benefits of early thrombolysis in conjunction with anticoagulation for SVT patients with underlying MPN should be pursued, as the rates of complete resolution in these patients were low.
Application of Comprehensive Nursing Program Based on Evidence-Based Concept in Patients with HIV-Related Lymphoma
Authors: Lv Jing , Jiang Cuiting , Wang Xia , Deng Yuhong , Luo Qin , Liu Tingting , Liu Yao 
Published: December 2023

Objective: To explore the application and effect of comprehensive nursing program based on evidence-based concept in patients with HIV-related lymphoma.
Methods: 148 patients with HIV-related lymphoma were randomly divided into control group and observation group, with 74 cases in each group （Table 1). The control group was given routine nursing, while the observation group was given comprehensive nursing on the basis of routine nursing. The HIV Self-management Scale, Herth Hope Index and WHO Quality of Life Questionnaire for HIV Brief Version were used to evaluate the score of self-management, hope and quality of life of patients in the two groups before and after intervention. The level of chemotherapy compliance and the incidence of complications were also compared between the two groups.
Results: Compared with the control group, the scores of self-management, hope and quality of life in the observation group were significantly higher (P&lt;0.05) , the compliance of chemotherapy and the incidence of complications in the observation group were significantly lower than those in the control group (P &lt;0.05) (Table 2).
Conclusion: The comprehensive nursing program based on evidence-based concept can effectively improve the self-management ability, the level of hope and quality of life of the patients with HIV-related lymphoma after chemotherapy, it is worth popularizing in clinic to improve the compliance of patients and reduce the incidence of complications.
Radiotherapy Combined Chimeric Antigen Receptor T-Cells Therapy in Relapsed or Refractory Diffuse Large B-Cell Lymphoma（R/R DLBCL）
Authors: Zhou Lili , Liu Yifan , Ye Shiguang , Li Ping , Li Shaoguang , Liang Aibin 
Published: December 2023

Background:
CAR (chimeric antigen receptor) T-cell therapy has transformed the treatment of patients with recurrent or refractory hematological malignancies. However, some patients cannot benefit from CAR-T cell therapy. Radiation therapy (RT) can induce immune effects and has synergistic effects in combination with other treatments. With the help of RT's immunomodulatory effects, a successful bridging strategy can be developed. This study aimed to investigate the efficacy and safety of CAR-T combined radiotherapy in patients with relapsed or refractory diffuse large B-cell lymphoma (R/R DLBCL).
Methods:
We retrospectively analyzed the clinical data of 57 R/R DLBCL patients who received radiotherapy combined CAR-T therapy in Tongji Hospital of Tongji University between January 2018 and February 2023 (NCT02537977). 18 Patients (31.58%) received radiation therapy within 1 year before CAR-T therapy and 1 patient (1.75%) received radiation therapy 1 month later after CAR-T therapy. Patients were divided into two groups: CART group and CAR-T+RT group. Clinical features, treatment response and incidence of adverse events of 57 patients were collected and analyzed to demonstrate the efficacy of CAR-T combined radiotherapy in patients with R/R DLBCL.
Results:
Of the 57 patients with R/R DLBCL treated with CAR-T therapy, 19 patients (33.33%) received CAR-T combined radiotherapy. The overall response rate (ORR) is 63.00% and 35.80% for patients in CAR-T+RT group and CAR-T group, respectively. The response rate is better for patients in CAR-T+RT group than that of CAR-T group P=0.05). One-year overall survival (OS) is 72.40% (95%CI, 61.90-82.90% ) for patients in CAR-T+RT group, while 1-year OS is 47.40% (95%CI, 39.30-55.50%) for patients in CAR-T group. The median progression-free survival (PFS) is 91 days (95%CI, 35.12-146.88) for patients in CAR-T group and the median PFS is not reached for patients in CAR-T+RT group. There is significantly different in OS and PFS for patients in CAR-T+RT group and CAR-T group ( P=0.02 and P=0.01). Only one patient with CNS lymphoma developed grade 4 CRES. 9 patients (47.37%) occurred grade 1-2 CRS, no patient occurred ≥grade 3 CRS for patients in CAR-T+RT group; 2 patients (3.51%) occurred grade 3 CRS, 25 patients (65.80%) occurred grade 1-2 CRS, no patient occurred grade 4-5 CRS for patients in CAR-T+RT group.
Conclusion ：
Our study shows that CAR-T combined radiation therapy can obtained better remission rate for R/R DLBCL patients. It is also can effectively prolong PFS and OS, improving the survival of patients. Radiotherapy combined with CAR-T therapy showed efficacy and manageable tolerability, making it a promising treatment option for patients with R/R DLBCL.
Dihydroartemisinin Synergistically Promotes the Antileukemic Activity of Venetoclax By Downregulating Mcl-1 in Acute Myeloid Leukemia Cells
Authors: Zhuang Haihui , Chen Yao , Li Fenglin , Jin Jie , Lu Ying 
Published: December 2023

Background: Venetoclax is a BH3 mimetic, which selectively inhibits Bcl-2. For older persons with newly diagnosed AML, the FDA has approved the use of venetoclax in conjunction with hypomethylating drugs or low-dose cytarabine. However, the selective inhibition of Bcl-2 in AML often results in the overexpression of Mcl-1, another dominant anti-apoptotic Bcl-2 family protein conferring venetoclax resistance. Dihydroartemisinin (DHA) is a herbal monomer that is taken orally to cure malaria. Studies have shown that DHA downregulates MCL-1 protein expression in AML cells. In the present study, we investigated whether the combination of venetoclax and DHA efficiently promotes apoptosis in AML cells.
Methods: We assessed the combined effect of venetoclax and DHA on cell viability, apoptosis, combination index, and cell cycle in AML cells from AML cell lines and deno AML patients. Western blot analysis was used to validate the mechanisms of venetoclax and DHA.
Results: Venetoclax efficiently impaired cell viability and dose-dependently promoted apoptosis when combined with DHA; their synergism was aptly represented by the combination index. Venetoclax and DHA had significant synergistic inhibition of cell viability both in cells from AML cell lines and in deno AML patients. The combination of venetoclax and DHA impaired cell cycle progression by arresting the cell cycle in S-phase and inducing apoptosis. Mechanistically, DHA mitigated venetoclax-induced upregulation of Mcl-1 and BCL-XL. Moreover, the combination inhibited the expression of C-MYC.
Conclusions: Our findings demonstrate the synergistic, preferential antileukemic effects of venetoclax and DHA on AML cells, providing a rationale for preclinical and clinical trials by combining these agents already being used in clinical practice to treat AML.
Ruxolitinib or Interferon-α Treatment As a Protective Strategy for Patients with Philadelphia-Negative Myeloproliferative Neoplasms (MPN) during the COVID-19 Pandemic
Authors: Gao Zicheng , He Zherou , Zhou Xuan , Wen Ziyu , Chen Hong , Wang Qiang , Liang Hanyin , Luo Dongmei , Liu Xiaoli , Liu Qifa , Xu Na 
Published: December 2023

Background: Both myeloproliferative neoplasms (MPNs) and coronavirus disease 2019 (COVID-19) are characterized by systemic inflammation and with intrinsic thrombotic risk. Series of studies found that ruxolitinib treatment was efficacy for COVID-19 in patients with hyperinflammation, and interferon-α as the innate immune system rapidly to combat viral infections, has been considered a potential therapeutic strategy to treat COVID-19 disease. However, ruxolitinib or interferon-α treatment with response outcomes for MPN patients were still unidentified.
Patients and methods : We prospectively analyzed the outcome of ruxolitinib (n=175) or interferon-α (n=165) treatment group versus other available therapy (n=478) group for MPN patients during the COVID-19 Pandemic (from May, 2020, to December, 2022). Outcomes were the rate of thrombosis, bleeding, acute respiratory distress syndrome (ARDS) and death.
Results: There were no difference between ruxolitinib and interferon-α treatment group in thrombosis rate, bleeding, the rate of ARDS and death. Thrombosis occurred lower frequently in patients treated with ruxolitinib or interferon-α than other available therapy group (1.1% vs 1.8% vs 5.6%, p = 0.004), rates of bleeding (1.7% vs 1.8% vs 2.0%, p = 0.93), rates of ARDS (1.1% vs 1.8% vs 4.2%, p = 0.048), and death (6.8% vs 3.0% vs 6.9%, p = 0.22) of COVID-19 disease were similar.
Conclusion: Ruxolitinib or interferon-α therapy was associated with lower thrombosis event and lower rate of ARDS for MPN patients in COVID-19 Pandemic.
Keywords: MPN, Ruxolitinib, thrombosis event, interferon-α
Internal and External Validation of the Predictive Scoring Systems from Essential Thrombocythemia to Post-ET Myelofibrosis
Authors: Xiang Danhong , Yang Xiudi , Zhu Jingjing , Huang Jian 
Published: December 2023

Essential thrombocythemia (ET) is one of the myeloproliferative neoplasms with a high risk of progression to post-ET myelofibrosis (post-ET MF). However, there still lacked the prediction model of post-ET MF. Here, we conducted a study of 708 ET patients identified by 2016 WHO diagnostic criteria in 13 hematology centers between 01 January 2010 to 01 July 2023, aimed to identify risk factors progression to MF and develop a prediction nomogram for post-ET MF(ChiCTR2200061208).
The training cohort included 543 ET patients with ET from 8 hematology centers. The external validation cohort included 165 subjects from another 5 hematology centers. Univariable and multivariable analysis identified the red blood cell distribution width (RDW) level (P=0.002), lactic dehydrogenase (LDH) level (P=0.003), smoking(P=0.003) and splenomegaly (P=0.000) were independent risk factors of post-ET MF(Table A). These independent risk factors were included to develop a prediction model of nomogram (Figure B). The bias-corrected AUC were both high in the training cohort (AUC=0.899) and external validation cohort (AUC=0.892). Analysis of calibration curves showed that the apparent and bias-corrected lines were overlapped with the ideal line in both training and external validation cohorts, and the Hosmer−Leeshawn tests showed that the model were a good fit (P = 0.73 in training cohort, P=0.43 in external validation cohort) (Figure C, D). Decision curve analysis(DCA) was conducted to assess the clinical usefulness of nomogram by quantifying the net benefits at defferent threshold probability of post-ET MF. Threshold probabilities for the standardized net benefit nomogram in predicting the probability of the occurrence of post-ET MF ranged from 0.02 to 0.87 in the training cohort, 0.01 to 1 in the external validation cohort.
In conclusion, we developed a nomogram that can predict the risk of progression to MF in ET patients, which can help clinicians adjust therapy timely to prevent disease progression to a great degree.
Acknowledgement: This research was funded by the Key R&D Program of Zhejiang, No. 2022C03137; Public Technology Application Research Program of Zhejiang, China, No. LGF21H080003; Zhejiang Medical Association Clinical Medical Research special fund project, No. 2022ZYC-D09.
Correspondence to: Dr Jian Huang, Department of Hematology, The First Affiliated Hospital of Zhejiang University School of Medicine. No. 79 Qingchun Road. Hangzhou, Zhejiang, PR China.