bluuebunny/crossref_metadata_2025_split huggingface dataset:
Erythroferrone Overexpression Mitigates Anemia and Improves Kidney Function in a Mouse Model of CKD
Authors: Czaya Brian , Nemeth Elizabeta , Ganz Tomas 
Published: December 2024

Chronic kidney disease (CKD) affects over 800 million people worldwide, most of whom develop anemia caused by relative erythropoietin deficiency, increased levels of the iron-regulatory hormone hepcidin, as well as absolute and functional iron deficiency.
Erythropoietin-stimulating agents (ESAs) and iron supplementation ameliorate anemia in CKD but patients often become hyporesponsive to ESAs. Increasing ESA doses may heighten risks of thrombosis and cardiovascular mortality, highlighting the need for alternative strategies.
Erythroferrone (ERFE) is an erythroid suppressor of hepcidin, facilitating iron mobilization for erythropoiesis. In CKD, diminished marrow erythroblasts contribute to decreased ERFE production, likely further increasing hepcidin.
To explore the potential therapeutic use of ERFE in CKD, we tested the impact of augmented ERFE on systemic iron homeostasis and kidney function. Transgenic mice with erythroid overexpression of ERFE (ERFE-Tg) had similar iron stores to WT mice at weaning but became iron overloaded as they aged. To prevent early iron overload, we placed ERFE-Tg on 4ppm diet at weaning. 8-week-old male ERFE-Tg and wild-type (WT) mice were then fed 0.2% adenine-rich diet containing 100 ppm Fe for 8 weeks to induce CKD.
At 16 weeks, both WT and ERFE-Tg mice developed CKD and anemia but ERFE-Tg had higher hemoglobin, MCV and serum iron, as well as lower liver hepcidin mRNA/liver iron content ratio, indicating effective suppression of hepcidin by ERFE. Furthermore, kidney function (BUN and serum creatinine) was less impaired in ERFE-Tg. Thus, ERFE overexpression in adenine-induced nephropathy improves CKD-associated anemia and renal dysfunction, suggesting that ERFE has therapeutic potential in CKD.
Engineering a Controllable and Reversible Switch for CAR-T Cell Immunotherapy Via a Genetic Code Expansion System
Authors: Liu Yue , An Lingna , Zhang Xi 
Published: December 2024

Background
As one of the most promising adoptive cell therapies, CAR-T therapy has achieved notable clinical effects in patients with hematological tumors. However, several obstacles remain in CAR-T therapy, such as low killing effects for some tumor targets, high-frequency recurrence, and cytokine release syndrome. These obstacles severely limit the long-term effects and the further application of CAR-T therapy. Therefore, the optimization of CAR-T cells for increased controllability and safety is urgently needed.
Methods
In this study, we engineered a genetic code expansion-based therapeutic system to achieve rapid CAR protein regulation in response to cognate unnatural amino acid at the translational level, in which the termination codon TAG was inserted into the CAR coding region. When the unnatural amino acid, N-ε-((tert-butoxy) carbonyl)-l-lysine (BOCK) is absent, the CAR protein cannot be completely translated, and CAR-T is “closed”. When BOCK is present, it recognizes and combines with the TAG, the whole CAR protein is translated, and CAR-T is “open”. Based on these findings, we subsequently investigated whether BOCK can control CAR protein expression and regulate CAR-T cell function via a series of in vitro experiments and a xenograft mouse tumor model.
Results
First, we verified that the BOCK-induced genetic code expansion system enables regulate protein expression as a controllable switch. Subsequently, we demonstrated that BOCK effectively and precisely controlled CAR protein expression and induced the activation of CAR signaling, the expression of activation markers, CD69 and CD25, and cytokine secretion. When incubated with tumor cells, BOCK regulated CAR-T cell cytotoxicity in a dose-dependent manner. Finally, a NOG mouse model analysis showed that BOCK can control CAR-T cell function and anti-tumor effect in vivo.
Conclusions
We systematically demonstrated that the BOCK-induced genetic code expansion system can effectively and precisely regulate CAR protein expression and control CAR-T cell anti-tumor effects in vitro and in vivo. We conclude that this controllable and reversible switch has a potential for more effective, secure, and clinically available CAR-based cellular immunotherapies.
Critical Roles of LILRB4 in Promoting PIM Kinase Mediated Cell Proliferation and Tumorigenesis in Multiple Myeloma
Authors: Wang Yijie , Ma Jingyuan , Sun Xiyue , Gong Lixin , Liu Lanting , An Gang , Qiu Lu-Gui , Hao Mu 
Published: December 2024

Introduction
Multiple myeloma (MM) is the second most common hematologic malignancy. Although multiple targets on MM cells are discovered and applied in clinical treatment, relapse is almost inevitable in MM patients. Our previous study found a high-risk cell cluster from MM patients with overall survival less than 2 years by single-cell RNA sequencing (scRNA seq). In this specific cluster, leukocyte immunoglobulin-like receptor B4 (LILRB4) was highly expressed, indicating the critical role of LILRB4 in myelomagenesis and drug resistance. However, the mechanisms of LILRB4 in MM development has not been fully understood. Here, we investigated the role of LILRB4 in tumorigenesis and MM cell proliferation.
Methods and Results
Our clinical data showed that MM patients with LILRB4 overexpression had poor prognosis and decreased overall survival. Compared with newly-diagnosed MM patients, expression of LILRB4 was higher in the MM patients relapsed after treatment, indicating the significance of LILRB4 in MM progression and drug resistance. In vitro experiment showed that MM cells with LILRB4 overexpression enhanced the cell colony-forming ability and promoted cell proliferation. Conversely, knocking-out LILRB4 induced cell apoptosis. We used MM cells with LILRB4 overexpression to establish myeloma xenograft model and results showed that compared with LILRB4neg MM cells, mice injected with LILRB4high MM cells displayed higher tumor formation rate and lower survival rate. Then, we analyzed RNA-seq data to investigate the mechanisms of LILRB4 in MM proliferation. Transcriptome analysis indicated that overexpression of LILRB4 in MM cell lines activated NF-kB, glycolysis, hypoxia and MTORC1 signaling pathways. Among these, NF-kB pathway has been proved in previous studies. More important, RNA-seq data showed that PIM kinase, correlating with cancer cell proliferation and apoptosis, was significantly upregulated in MM cells with LILRB4 overexpression, which has been confirmed in our in vitro experiments. We also found that LILRB4 upregulated the expression of PIM through activating Src-homology 2-containing protein tyrosine phosphatase (SHP) and signal transducer and activator of transcription (STAT) protein, indicating the underlying mechanism of MM cell proliferation. Interestingly, transcription factors promoting B cell program were upregulated, and the immunophenotypic marker related to B cell differentiation was downregulated after LILRB4 overexpression. These findings indicated that LILRB4 may induce myeloma cell dedifferentiation and promote tumorigenesis.
Conclusion
In conclusion, LILRB4 is highly associated with poor prognosis of MM patients and has the ability of tumorigenesis. LILRB4 promotes MM cell proliferation by activating SHP and STAT to upregulate the expression of PIM kinase, which plays critical roles in MM pathogenesis.
Deciphering Inflammasome Activation in Gaucher Macrophages and Its Role in the Development of Multiple Myeloma
Authors: Reinecke Vivien , Hofbauer Daniel , Bittenbring Jörg-Thomas , Lischer Christopher , Berges Johannes , Nusser Gina , Bitterer Katrin , Voelkl Simon , Mougiakakos Dimitrios , Mackensen Andreas , Bruns Heiko 
Published: December 2024

Gaucher disease (GD) is the most common lysosomal storage disease and corresponds to the accumulation of glucocerebroside in macrophages due to defective synthesis of the β-glucocerebrosidase. This lysosomal storage leads to the characteristic engorged macrophages, known as Gaucher macrophages (GMs) and contributes to the secretion of inflammatory cytokines, bone disease, splenomegaly and cytopenia typically encountered in patients with GD. Interestingly, patients with GD have an increased incidence of both polyclonal and monoclonal gammopathies with an increased risk of multiple myeloma (MM).
We have recently been shown in MM that the NLRP3 inflammasome in TAMs is triggered by “frustrated phagocytosis” and represents a critical and early step in the initiation and progression of MM (Hofbauer et al., Immunity, 2021). Therefore, we hypothesize that the inflammasome is activated in GMs, and excessive production of interleukin (IL)-1β and IL-18 by macrophages promotes the development of MM.
We identified subsets of pro-inflammatory macrophages in GD patients with increased transcription of inflammasome associated-markers (AIM2, CASP1, IL-1B, IL-18, and NLRP3) by single-cell RNA sequencing. We verified this inflammasome activation in GMs by immunohistochemistry and flow cytometry through detection of active caspase-1 and ASC aggregation. Mechanistically, we demonstrated in macrophages from healthy donors, that excessive addition of glucocerebroside or inhibition of β-glucocerebrosidase leads to lysosomal rupture, caspase-1 activation and secretion of IL-1β and IL-18. Furthermore, the coculture of primary isolated MM cells with GMs resulted in a significantly higher proliferation and metabolic activity of the tumour cells. Notably, blockade of inflammasome activation by MCC950 (a specific inhibitor of the NLRP3 inflammasome) results in reduced IL-1β and IL-18 secretion from ex vivo cultured GMs.
These data suggest that the inflammasome is activated in GMs by disruption of the lysosomal integrity and that therapeutic inhibition of the NLRP3 inflammasome in GD patients may reduce the risk of clonal evolution of plasma cells.
Characterizing and Targeting Follicular Lymphoma's Common Progenitor Cells
Authors: Atkins Oscar , Zhang Lingling , Hung Miu Shing , Reinhardt Christian , Calado Dinis 
Published: December 2024

Follicular lymphoma (FL) is the most prevalent low-grade germinal centre (GC) derived non-Hodgkin lymphoma. Despite being considered an indolent lymphoma its clinical course is heterogenous with some patients experiencing frequent relapses and eventual transformation to a high-grade disease associated with an especially poor prognosis.
Through phylogenetic analysis of successive human FL tumors, evidence has emerged regarding the existence of common progenitor cells (CPCs). These precursor cells serve as a reservoir of cells that are able to evade killing with standard treatments for FL and give rise to relapse and transformed FL. They carry the t(14:18) translocation and are enriched for somatic mutations in chromatin-modifying genes including CREBBP and KMT2D. Developing therapies to specifically target and eliminate these common progenitor cells will be crucial in finding a cure for FL.
To understand this population of therapy-resistant precursor cells, we engineered mouse models that faithfully replicate the early and recurrent gene mutations observed in FL, including BCL2 translocation, CREBBP and KMT2D loss-of-function (LoF) mutations (BCK mice). We then subjected mice to a standard human-like treatment regimen of R-CHOP (Rituximab, Cyclophosphamide, Doxorubicin, Vincristine, Prednisone). Administration of the mouse-adapted R-CHOP resulted in the complete depletion of B cells from control mice. However, treatment of BCK mice revealed a resistant B-cell population. Notably, these resistant cells predominantly displayed a memory B-cell phenotype.
Single cell RNA-seq analysis of these therapy-resistant cells identifies unique transcriptional signatures and enrichment of pathways that can be targeted therapeutically. Comparison with human FL scRNAseq datasets shows that these therapy-resistant precursor cells are enriched in FL tumors of patients with early relapse.
Finally, we have used an innovative in vitro platform that harnesses a GC B-cell culture system to amplify CPCs, facilitating subsequent high-throughput drug screening. This approach has allowed us to identify specific vulnerabilities within the CPC population, paving the way for the development of therapies tailored to overcome FL resistance mechanisms.
Through these concerted efforts, we edge closer to the long-awaited goal of discovering a definitive cure for Follicular Lymphoma.
Venous Thromboembolism in Individuals with Sickle Cell Disease- a Retrospective Review of This Understudied Problem
Authors: Ajayi Ayotola , Zhang Li , Agarwal Shreya , Bhasin Neha 
Published: December 2024

Individuals with sickle cell disease (SCD) are proposed to be at a higher risk for venous thromboembolism (VTE) compared to the general population. Observational studies show a 25% increased risk of first VTE in SCD with a mean age of incidence of 30 years, which is much lower than the general population of 65 years. The mechanisms for this increased VTE risk have been proposed to be increased hemolysis causing increased venous stasis and vasculopathy, increased endothelial activation with increased activation of the coagulation pathway and chronic inflammation.
In this study, we reviewed the medical records of 783 individuals with SCD who were followed in our lifelong sickle cell center between May 2013 to May 2023 to determine the prevalence of thrombosis and associations of these events with laboratory and clinical risk factors of thrombosis. We collected retrospective data including patient demographics, clinical history, and laboratory data at the time of VTE events and an evaluation of the presence of a central venous catheter (CVC) in our population. VTE was defined as occurrence of deep vein thrombosis (DVT), pulmonary embolism (PE), cerebral sinus venous thrombosis (CSVT), or right atrial thrombosis.
The prevalence of VTE in our population was 8.2% (n = 64 out of 783), with a median age of 33 years. Out of 64 total patients who developed a VTE during our study period, 21 developed a PE; this was 33% of the VTE events noted. A high body mass index (BMI) (&gt;30 kg/m2) was noted among 22% of the patients in the VTE group compared to 7.6% of the non-VTE group (p&lt;0.001). Smoking was noted in 41% of the VTE group patients compared to 10% in the control group (p&lt;0.001). CVCs were present in 55.6% of the patients in the VTE group compared to those in the non-VTE group (10%) with a p &lt;0.001. Twenty-two patients (34.4%) of the patients in the VTE group had a history of a VTE event prior to our study period and were having a recurrent VTE episode during our study period between 2013 and 2023. In addition, out of those patients who developed a VTE during our study period, 20 patients (31%) developed a recurrent VTE event during our study period.
Our results show that there is a higher rate of obesity and smoke exposure among patients with SCD who develop a VTE event. PE's are a substantial number of VTE events in the SCD population. We also noted a significant correlation between CVCs and VTEs in SCD. There is high recurrence rate of VTEs among SCD patients. Based on these findings, future prospective data are needed to determine if these associations are clear risk factors for VTE in SCD so these risk factors can be minimized in patients at risk for VTE events.
The Addition of Lenalidomide to R-CHOP Is Effective for Prevention of CNS Relapse in High-Risk Patients with Diffuse Large B-Cell Lymphoma
Authors: Wei Xiaolei , Guo Xutao , Wei Qi , Xiong Dan , Zhang Peng , Zhang Zewen , Wei Yongqiang , Feng Ru 
Published: December 2024

Central nervous system (CNS) relapse is an uncommon but devastating event in patients in diffuse large B-cell lymphoma (DLBCL). Although several prophylaxis attempts including intrathecal methotrexate and intravenous high-dose methotrexate were conducted to reduce the CNS relapse rate, the optimal management remained uncertain. Herein, we performed an open-label, multicenter trial to explore the efficacy and safety of the combination of lenalidomide and R-CHOP (R2-CHOP) for preventing the CNS relapse in the high-risk diffuse large B cell lymphoma.
Eligible patients were aged 18-80 years, newly diagnosed DLBCL and high risk for CNS recurrence: CNS-IPI:4-6 or involvement of testicular, kidneys or adrenal glands. Lenalidomide orally 25 mg per day was administered on days 1 through 10 of each cycle and delivered concomitantly with standard dose R-CHOP-21 regimen. All patients received aspirin 100mg per day prophylaxis throughout. The primary endpoint was 2-year central nervous system relapse rates. The secondary endpoint were 2-year overall survival, progression-free survival, and treatment-emergent adverse events. This trial is registered with ClinicalTrials.gov, number NCT04544059.
A total of 64 DLBCL patients with high-risk for CNS relapse were included in this study and 56 of them was evaluated. The median age of the patients in this study was 57 years old (range 26-77) and 48.2% patients were female. 91.1% patients had a CNS-IPI score ≥4 and the left patients may be involved with testicular, kidneys or adrenal glands. With a median follow up of 20 months, only 2 (3.6%) patient developed CNS relapse. The 2-year progression-free survival and overall survival were 77.8% and 79.7%, respectively.
In summary, lenalidomide with R-CHOP is effective for prevention of CNS relapse in high-risk patients with diffuse large B-cell lymphoma.
Keywords:
Diffuse large B-cell lymphoma, central nervous system relapse, lenalidomide.
Modulating mRNA Translation Fidelity As a Novel Mechanism for AML Chemoresistance
Authors: Mayerhofer Christina , Li Dan , Kristiansen Trine A. , Mayerhofer Ernst , Sharda Azeem , He Lingli , Mazzola Michael , Schiroli Giulia , Gustafsson Karin , Keyes Sam , Kiem Anna , Crompton Eve , Xu Yanxin , Dou Zhixun , Vidoudez Charles , Miller Peter G. , Van Gastel Nick , Scadden David 
Published: December 2024

Acute Myeloid Leukemia (AML) is a highly aggressive blood cancer that initially responds to high-dose chemotherapy, but the majority of patients relapse. To understand the mechanisms underlying chemoresistance of AML, we investigated the metabolic adaptations in AML cells that occur following chemotherapy but before overt disease relapse. In AML patient-derived xenografts (PDX), we applied untargeted metabolomics and showed that chemoresistant AML cells have significantly elevated levels of branched-chain amino acids (BCAAs) compared to untreated cells. Elevated BCAA levels and their catabolism have been implicated in AML but their upregulation to drive chemoresistance is not well studied. Transcriptome analysis and stable isotope tracing supported the upregulation of BCAA pathways during chemotherapy in vivo. Our results showed that AML cells, compared to normal granulocyte-monocyte progenitor cells, had significantly higher valine uptake, suggesting a dependency on BCAA metabolism for survival during chemotherapy. When BCAA restriction was combined with chemotherapy, survival was significantly improved with an accompanying clearance of residual AML cells. However, relapse still occurred.
Unexpectedly, the surviving cells had significantly higher levels of protein synthesis during maximum response to chemotherapy and BCAA restriction. We anticipated that BCAA restriction after chemotherapy would lower mTOR activity and thus protein synthesis rates. Unexpectedly, we found that ribosomal gene transcripts were enriched, and OPP incorporation in newly synthesized peptides increased in AML cells restricted for BCAAs combined with chemotherapy in vivo. Using low-input Ribo-seq to analyze paired transcriptome and translatome data, we found activated ribosomal protein translation and elevated production of the dioxygenase, 2-oxoglutarate-and-iron-dependent oxygenase 1 (OGFOD1). OGFOD1 modulates the fidelity of ribosomal codon-anticodon pairing through ribosome hydroxylation in proximity to the decoding center where this pairing takes place. Accordingly, AML cells with increased OGFOD1 in vivo had reduced ribosome stalling by ribosome profiling. Conversely, inhibiting OGFOD1 led to translation arrest, reduced protein production and improved animal survival following chemotherapy. Our data therefore suggest that OGFOD1 may enable cells to bypass BCAA shortage by permitting less stringent translation accuracy and targeting OGFOD1 and restricting amino acids can counteract this chemoresistance mechanism. Modifying translation fidelity is a previously unrecognized strategy in AML and has important potential implications across other oncologic contexts.
CD47 Upregulation Inhibits Phagocytic Clearance of Alloreactive Donor T Cells after Allo-HCT
Authors: Shaikh Haroon , Flamann Cindy , Matos Carina , Ali Hla , Kern Michael AG , Kreutz Marina , Buettner-Herold Maike , Lischer Christopher , Voelkl Simon , Kellner Christian , Bitterer Katrin , Saul Domenica , Holler Ernst , Zernecke Alma , Mougiakakos Dimitrios , Beilhack Andreas , Bruns Heiko 
Published: December 2024

Acute graft-versus-Host disease (aGvHD) is a life-threatening complication that frequently occurs after allogeneic hematopoietic cell transplantation (allo-HCT). Resolving inflammation is a key therapeutic goal in aGvHD. Macrophage-mediated phagocytosis contributes substantially to resolve inflammatory processes. However, the role of phagocytosis after allo-HCT still remains unexplored.
Therefore, we analyzed the expression of the “don´t eat me” signal CD47 on allogeneic T cells from aGvHD patients and in two MHC major mismatch aGvHD mouse models (FVB à C57BL/6, 9 Gy TBI and C57BL/6 à Balb/C, 8 Gy TBI). We evaluated the effect of in vitro and in vivo administration of an anti-CD47 antibody on antibody-dependent cellular phagocytosis (ADCP) of alloreactive T cells using flow cytometry and confocal microscopy, bioluminescence imaging, cytokine measurements and histopathology.
Initial analysis of a global dataset revealed increased CD47 expression on intestinal T cells in patients with chronic gastrointestinal inflammation. Subsequently, we examined CD47 levels on T cells in the allo-HCT setting. Notably, intestinal T cells in patients with severe GvHD (Lerner grade III) significantly upregulated CD47 (p = 0.0079). There were no significant changes in CD24 (another “don´t eat-me”-signal) or in the “eat-me”-signals calreticulin and ATP10A. This upregulation of CD47 correlated with reduced macrophage-mediated phagocytosis. Similarly, intestinal alloreactive donor T cells highly upregulated CD47 in mice developing aGvHD after allo-HCT (p = 0.0317). In vitro studies revealed that CD47 expression is regulated by NF- kB in T cells after TCR activation and primed T cells inhibit ADCP in a CD47-dependent fashion. Transfer of CD47 knock-out T cells and bone marrow resulted in swift clearance of donor T cells after allo-HCT resulting in significantly improved clinical scores and survival compared to controls (p = &lt;0.0001). Crucially, application of an anti-CD47 antibody significantly antagonized the impaired ADCP of alloreactive T cells. In anti-CD47 treated mice, we observed increased phagocytosis of T cells in the gastrointestinal tract (GIT) and induction of immunosuppressive responses in vivo. Besides an increase of the myeloid-derived suppressor cell population, systemic pro-inflammatory cytokine levels such as TNF, IL-1β and IFN-β decreased in the serum.
In conclusion, our data suggest that high CD47 expression impairs phagocytic clearance of alloreactive T cells after allo-HCT, potentially contributing to exaggerated inflammatory responses in the GIT of GvHD patients. Thus, CD47 emerges as potential therapeutic target to eliminate alloreactive T cells, with anti-CD47 treatment aiding the resolution of inflammation in GvHD.