bluuebunny/crossref_metadata_2025 huggingface dataset:
The PP2A-B56α Heterocomplex Regulates Response to Venetoclax Plus Azacitidine Treatment in AML
Authors: Romero-Murillo Silvia , Peris Irene , Marcotegui Nerea , Vicente Carmen , Odero Maria D. , Narla Goutham
Published: December 2023

Venetoclax (VEN), a selective inhibitor of BCL2, combined with azacitidine (Aza) has emerged as the new standard of care for patients with acute myeloid leukemia (AML) ineligible for intensive chemotherapy. However, the long-term medical benefit is modest, and relapse appears to be unavoidable. We have recently reported that PP2A activators enhance the efficacy of VEN and VEN+Aza in primary samples and pre-clinical models of AML, and that the PP2A-B56α complex drives the synergistic pro-apoptotic activity observed. Our aim here is to further decipher the role of PP2A-B56α in VEN+Aza treatment response in AML. We used CRISPR-Cas9 to generate HL-60 sublines lacking the B56α or B55α PP2A regulatory subunits. Interestingly, while there were no significant differences in their baseline rate of proliferation, HL-60 B56α knockdown (KD) clones were significantly more resistant to VEN and VEN+Aza than wild type (WT) cells. In contrast, HL-60 B55α KD clones were sensitive to VEN+Aza, pointing out a specific role for specific PP2A heterotrimers, in this case the PP2A-B56α complex in the combined treatment response. Protein expression analysis of known substrates of PP2A-B56α demonstrated that pSer62-MYC and total MYC decreased after VEN+Aza treatment in WT cells but not in KD cells. It is known that the PP2A-B56α complex directly dephosphorylates MYC-Ser62, inducing MYC ubiquitin-mediated proteasomal degradation. Moreover, p21/CDKN1A, that is transcriptionally activated by p53 and inhibited by MYC, was up-regulated at both the mRNA and protein level after VEN+Aza treatment in WT cells but not in KD cells. HL-60 is a p53 null cell line; therefore, p21 up-regulation upon the combined treatment might be MYC-dependent. Of note, treatment of HL-60 B56α KD cells with the MYC inhibitor MYCi975 rescued the VEN+Aza synergy seen in these cells, confirming the relevance of MYC degradation to VEN+Aza response. Interestingly, VEN+MYCi975 had synergistic anti-leukemic effects in these same cell lines. Furthermore, CDKN1A expression was also rescued upon the triple therapy consisting of VEN+Aza+MYCi975. Combined, our results suggest that the PP2A-B56α complex is involved in the regulation of VEN+Aza response through changes in MYC expression. Interestingly, a novel small molecular glue (PMG) developed by our group, which specifically stabilizes the PP2A-B56α complex, enhanced VEN plus Aza response in vitro; however, this triple therapy did not work in HL-60 B56α KD cells and we observed a clearly decrease in MYC protein expression upon the triple therapy only in HL-60 B56α WT cells, supporting the rationale to translate these novel PMGs into the clinic. Altogether, our data suggest that PP2A-B56α might have an important role in VEN plus Aza treatment response through the regulation of MYC degradation.
Cooperative Super-Enhancer Inactivation through Loss of Crebbp and KMT2D Skews B Cell Fate Decisions and Yields T Cell-Depleted Lymphomas
Authors: Li Jie , Chin Christopher , Ying Hsia-Yuan , Meydan Cem , Teater Matthew , Xia Min , Farinha Pedro , Takata Katsuyoshi , Chu Chi-Shuen , Rivas Martin A , Chadburn Amy , Steidl Christian , Scott David W. , Roeder Robert G. , Mason Christopher , BÃ©guelin Wendy , Melnick Ari M
Published: December 2023

Mutations in chromatin modifiers are a hallmark of many tumors, especially lymphomas arising from germinal center (GC) B cells. Given that most of these lymphoma mutations induce aberrant gene repression, it is surprising that they often co-occur in individual patients. The most common pairing are mutations affecting CREBBP and KMT2D, even though regulating overlapping gene enhancers and pathways. Hence their co-occurrence is especially puzzling. Herein, we report that combined haploinsufficiency of Crebbp and Kmt2d (C+K) do indeed induce a more severe lymphoma phenotype (vs either allele alone) and unexpectedly conferred an immune evasive phenotype manifesting as reduced CD8+ T cell infiltration. This was linked to profound repression of immune synapse genes that provide costimulatory effects to T-cells and others. Repression of immune synapse genes was reflected in impaired response to T-cell directed cell fate decisions and weakening of costimulatory signals. From the epigenetic perspective we observed interaction and mutually dependent cooperative binding of C+K to chromatin. Notably loss of C+K cooperativity was selectively severe at superenhancers (vs enhancers), especially those driving expression of immune synapse signaling genes and pointing to a particular dependency for both co-activators at these specialized regulatory elements. Therefore, we show that critical features of the lymphoma microenvironment are shaped by specific combinations of chromatin modifier somatic mutations, induced by the dependency of immune synapse superenhancers on cooperative actions of CREBBP and KMT2D.
The Potential Utility of Cerebral Fluid sLR11 and sIL-2R As a Marker for CNS Involvement of Malignant Lymphoma
Authors: Shimizu Naomi , Nakao Sanshiro , Nakaseko Chiaki , Shimizu Tomo , Ebinuma Hiroyuki , Bujo Hideaki
Published: December 2023

Introduction: LR11 is a type I membrane protein that releases serum-soluble LR11 (sLR11) by proteolytic shedding. In the past, we reported that patients with acute leukemia and malignant lymphoma showed significantly increased serum sLR11 levels compared to those in normal controls. The diagnosis of central nervous system involvement of malignant lymphoma (CNS ML) is often difficult to diagnose by cytology. This study analyzed the potential utility of cerebral fluids (CFs) sLR11 and sIL-2R as diagnostic markers for CNS ML.

Materials and Methods: We enrolled patients suspected of CNS ML and administrated intrathecal anticancer drugs from 2017 to 2023. We analyzed CFs levels of sLR11 and sIL-2R, and cytological malignant grades. Then, we analyzed the relationship between changes in these CFs markers and clinical outcomes.

Results: We studied 26 patients (16 men and 10 women; mean age: 72 years old). The 20 patients were diagnosed as CNS ML in clinical conditions, and the other 6 patients were treated with prevention because of ML of the testis and adrenal glands. The sLR11 of CFs levels significantly and positively correlated with the sIL-2R of CFs levels (r = 0.6618, p = 0.0002) (Figure 1A). As shown in Figure 1B, in CNS invasion groups, only four patients showed class V in the cytology of CFs, the other patients showed negative results. There were no relations between the number of cell numbers and sLR11 and sIL-2R of CFs. As shown in Figure 2, the levels of sLR11 of CFs in diagnosis tended to be higher (p = 0.1831) in the CNS invasion group (36.5±30.0 ng/ml) than in the no-invasive group (23.9±5.8 ng/ml). The levels of sIL-2R of CFs in diagnosis were significantly higher (p = 0.0035) in the CNS invasion group (986.0±1274.3 U/ml) than in the no-invasive group (74.2±78.2 U/ml). The LR11 and sIL-2R of CFs post-treatment were lower in the remission group (n=9; 13.46±6.37 ng/ml, <50.0 U/ml) than non-remission group (n=5; 22.40±14.32 ng/ml, 78.4±41.1 U/ml), p = 0.2861, 0.0629 respectively.

Conclusions: Patients diagnosed with CNS ML showed high levels of sLR11 and sIL-2R of CFs. These two may be good markers for evaluating diagnosis and treatment effects. Further examination of a large number of cases is required to elucidate these mechanisms.
Pathophysiology and Treatment Opportunities of CX3CL1 in Liver Acute Gvhd
Authors: Xu Ziwei , Zhao Xiaoyan , Li Ziying , Hu Yu , Wang Huafang
Published: December 2023

Acute graft-versus-host disease (aGVHD) is one of the most familiar and severe complications of hematopoietic stem cell transplantation (HSCT). Liver aGVHD, which has limited response to treatment and high mortality, is a critical concern among clinicians in the field. Chemokine (C-X3-C motif) ligand 1 (CX3CL1), also called fractalkine, as a chemokine and adhesion molecule, plays an important role in the pathogenesis of rheumatoid arthritis, primary biliary cirrhosis, and multiple sclerosis. Whether CX3CL1 is involved in liver aGVHD has not been investigated. We collected peripheral blood samples from 10 patients with liver aGVHD during the onset phase and 15 patients without aGVHD after HSCT. The results of the enzyme-linked immunosorbent assay (ELISA) indicated a significant increase in serum CX3CL1 levels among patients with liver aGVHD. Similarly, we observed elevated levels of CX3CL1 in the serum of a mouse model with aGVHD, and found a positive correlation between elevated bilirubin levels and higher serum CX3CL1 levels. The expression of CX3CL1 was markedly elevated in the liver of aGVHD mice by real-time quantitative polymerase chain reaction (RT-qPCR), western blotting (WB). The results of immunohistochemistry showed that the expression of CX3CL1 in the bile duct epitheliums of aGVHD mice was significantly increased compared with those of control mice, but not in hepatocytes. To further analyze CX3CL1 expression, we isolated hepatocytes and bile duct epitheliums from mice and measured the levels using RT-qPCR and WB. Similar to the results of immunohistochemistry, the expression level of CX3CL1 in bile duct epitheliums from aGVHD mice was increased significantly, while the expression level in hepatocytes was unchanged. The prominent bile duct damage observed in liver aGVHD may be attributed to the high CX3CL1 expression in bile duct epitheliums instead of hepatocytes. Anti-CX3CL1 monoclonal antibody (mAb) significantly improved the clinical symptoms and survival rates of liver aGVHD mice, and the liver enzymes and bilirubin levels were decreased remarkably after treatment. Additionally, HE staining showed that the application of anti-CX3CL1 mAb obviously reduced the infiltration of inflammatory cells in bile duct epithelium of liver aGVHD mice. In conclusion, these results provide evidence for the close relationship between CX3CL1 and the pathogenesis of liver aGVHD, suggesting anti-CX3CL1 mAb as a promising strategy for the management of liver aGVHD.

Keywords: Hematopoietic stem cell transplantation, Liver aGVHD, CX3CL1, Bile duct
Emicizumab Versus Immunosuppression for Acquired Hemophilia Α (AHA)
Authors: Hart Christina , Klamroth Robert , Sachs Ulrich J. , Greil Richard , Knoebl Paul N. , Oldenburg Johannes , Miesbach Wolfgang , Pfrepper Christian , Trautmann-Grill Karolin , Pekrul Isabell , Holstein Katharina , Eichler Hermann , Weigt Carmen , Schipp Dorothea , Werwitzke Sonja , Tiede Andreas
Published: December 2023

Introduction: AHA is a serious autoimmune bleeding disorder caused by neutralizing antibodies against coagulation factor VIII (FVIII). Standard of care involves immunosuppressive therapy (IST) to suppress formation of anti-FVIII autoantibodies. Recently, prophylaxis with emicizumab (EMI) has been used instead of IST to manage patients with AHA.

Aim: This analysis aimed to compare outcomes of treatment with EMI or IST in patients with AHA. It was prospectively planned as an exploratory analysis of the GTH-AHA-EMI trial (NCT04188639, registered at www.clinicaltrials.gov).

Methods: Individual patient data were retrieved from the GTH-AHA-EMI (n=47, treatment with EMI) and the GTH-AH 01/2010 (n=101, treatment with IST). Propensity score (PS) matching was used accounting for covariates that were previously established to influence bleeding risk and overall survival. Standardized mean differences were used to compare baseline characteristics before and after matching.

Results: Baseline characteristics of the study populations were very similar and further improved by PS matching. IST treated patients had a high risk of bleeding in the first 3 weeks (0.25 to 0.30 clinically relevant new bleeds [CRNB] per patient-week), whereas EMI treated patients were largely protected from bleeding throughout the entire observation period (<0.1 CRNB per patient-week). For the first 12 weeks of observation, the negative-binomial model-based mean bleeding rate of EMI treated patients was 68% lower as compared to IST treated patients (incident rate ratio 0.325, 95% confidence interval [CI] 0.182-0.581, p<0.001). During the first 12 weeks, infections occurred with similar frequency but were less often fatal (IST: 36 events in 29 patients [29% of patients], 11 fatal events; EMI: 11 events in 10 patients [21%], no fatal events). Thromboembolic events were less frequent with EMI (1 event (2%], no fatal event) as compared with IST (7 events in 7 patients [7%], 4 fatal events). Overall survival after 24 weeks was 91% and 76% (hazard ratio 0.44, 95% CI 0.24-0.81, p=0.008, figure).

Conclusions: This PS-matched individual patient data analysis showed better bleed protection and improved survival in patients treated with EMI as compared to patients treated with IST. These observations suggest a change of clinical practice. Patients with newly diagnosed AHA should be offered prophylaxis with EMI to reduce the risk bleeding, and provided sufficient time to allow for clinical stabilization and improvement of their general health status before IST is considered.
Preclinical Studies Support the Clinical Development of LP-284 in Relapsed and Refractory B-Cell Lymphomas
Authors: Zhou Jianli , Sturtevant Drew , Kulkarni Aditya , Bhatia Kishor , Ewesuedo Reginald
Published: December 2023

Genomic instability, a hallmark of many cancers, often results from corrupted DNA damage response/repair (DDR) pathways. Targeting DDR deficiencies in cancer therapy has been successful in solid tumors with dysfunctional homologous recombination repair (HR). However, while less recognized, lymphomas also carry DDR defects such as HR deficiency (HRD) and can account for approximately 18% of all lymphomas. LP-284 is a fully synthetic DNA-damaging agent that induces double-stranded DNA breaks and displays elevated potency in cancer cells with HRD or transcription-coupled nucleotide excision repair defects, including multiple advanced B-cell lymphomas. In preclinical pharmacology studies, the mantle cell lymphoma (MCL) xenograft tumor mouse model derived from JeKo-1, which has mutated TP53 and checkpoint kinase 2 (CHEK2), an HRD score of 46, and was refractory to ibrutinib and bortezomib, showed near complete response after LP-284 treatment. It is noteworthy that 40~50% of MCL patients may be HRD due to the presence of mutations in the ataxia-telangiectasia mutated (ATM) gene. HRD is also likely to be prevalent in high-grade lymphomas, as IGH/MYC translocation or BCL2 expression has been reported to be associated with HR inhibition. Studies of LP-284 in the MYC/BCL2 dual-translocated OCILY1 tumor xenograft model have shown a robust efficacy profile with an average of 99% tumor growth inhibition.

Key pharmacokinetic studies demonstrated that LP-284 is neither a substrate nor modulator of major CYP-450 enzymes, multi-drug resistance transporters, renal drug transporters, or hepatic drug transporters. Safety pharmacology studies indicated that LP-284 does not inhibit hERG potassium channel and has a low potential to affect cardiovascular functions. Toxicology repeat dose studies of LP-284 in Sprague Dawley rats and Beagle dogs, using intravenous infusion on Days 1, 8, and 15 in a 28-day cycle, were generally well tolerated with notable adverse effects in Beagle dogs consisting mainly of reversible hematologic toxicity and testicular toxicity. The highest non-severely toxic dose (HNSTD) in dogs was considered to be 0.4 mg/kg/dose. Overall, based on its impressive preclinical anti-tumor efficacy profile, low drug-drug interaction liability, and findings from toxicological studies, a Phase 1 study is currently planned with LP-284 to assess its safety, tolerability, pharmacokinetics, and clinical activity in patients with relapsed or refractory B-cell lymphoma. Clinical development will be informed by an emphasis on patient selection strategy based on DDR/HR biomarkers.
Interactive Case-Based Education on Pyruvate Kinase Deficiency Improves Hematologists' Competence in Implementing the Latest Diagnostic and Management Approaches
Authors: Rohani-Montez S Christy , Koekkoek Sanneke , Al-Samkari Hanny
Published: December 2023

INTRODUCTION

The diagnosis of pyruvate kinase deficiency (PKD) is challenging, resulting in long diagnostic delays, sometimes into late adulthood. Management has previously only been supportive, but is evolving, therefore, physicians need to understand the latest evidence-based approaches. This study was conducted to determine whether online interactive case-based independent medical education could improve clinicians' competence in diagnosing and managing PKD according to the latest evidence.

METHODS

174 hematologists participated in a text, case-based activity, and completed pre- and post-questions (Al Samkari H. Investigating Anemia and the Possibility of Pyruvate Kinase Deficiency: What Is Your Next Step? www.medscape.org/viewarticle/989782) Educational effect was assessed using a 3-question repeated-pair design with pre-/post-assessment. A paired samples t-test was conducted for significance testing on overall average number of correct responses and for confidence rating, and a McNemar's test was conducted at the learning objective level (5% significance level, P <.05). Cohen's d with correction for paired samples estimated the effect size of the education on number of correct responses. Data were collected from 3/21/2023-6/14/2023.

RESULTS

48 hematologists completed all pre- and post-assessment questions. Overall significant improvements were seen after participation: 56% average correct response rate at pre-assessment vs 81% at post-assessment; P<.001, Cohen's d= 0.70. In addition, 65% of hematologists improved their competence, selecting correct responses in the post assessment after having selected incorrect responses in the pre-assessment. Significant improvements were achieved with regards to competence in diagnosing PKD and selecting an appropriate treatment approach (both P <.01). After participating, 42% had measurable improved confidence (P<.001) in their ability to implement new treatment approaches for patients with PKD.

CONCLUSIONS

This study demonstrates the success of online, interactive case-based education in improving clinicians' competence in diagnosing and managing PKD according to the latest evidence. Both improvement and reinforcement in the context of a linked learning assessment have been shown to positively correlate with increases in confidence as well as intention to make clinical practice changes (Lucero KS, Chen P. J Eur CME. 2020 Oct 12;9(1):1834759), suggesting that most clinicians who participated in this activity are likely to make improvements in their practice. This could lead to earlier appropriate treatment and improved overall outcomes for these patients.
Thrombocytopenia - Is It an Impactful Prognostic Indicator in Patients with Liver Cirrhosis?
Authors: Araji Ghada , Arafa Fatema , Mustafa Ahmad , Rajavel Anisha , Chehab Hamed , Niazi Muhammad , El-Sayegh Suzanne , Deeb Liliane
Published: December 2023

Background: Several prognostic scores have been developed to guide management strategies in cirrhotic patients. Child-Pugh and Model of End-stage Liver Disease (MELD) are the typical scores currently used to predict outcomes in cirrhosis, however both do not include platelet count. Although platelet count may provide clues regarding severity of the disease, there is scarcity of data supporting its utility as a prognostic factor in patients with cirrhosis. Hence our study aimed to investigate the association between thrombocytopenia and development of complications as well as mortality in cirrhotic patients.

Methods: TheNational Inpatient Sample (NIS) Database was used to sample individuals with liver cirrhosis between 2016 and 2018. Baseline demographics and comorbidities were collected using ICD-10-DM codes. Patients younger than 18, those with missing data, and with history of thromboembolism were excluded. Study patients were then stratified into those with and without thrombocytopenia. Greedy propensity matching using R was performed to match the two cohorts in 1:1 ratio on age, gender, race, use of anti-thrombotic agents, smoking, and nine other comorbidities including diabetes, dyslipidemia, hypertension, morbid obesity, chronic kidney disease, end stage renal disease, chronic heart failure, history of cancer, and history of transient ischemic attack or stroke. Multivariate logistic regression model was used to analyze outcomes. P-value of <0.05 was considered statistically significant.

Results: Out of a total of 402,104 cirrhotic patients, 129,461 had thrombocytopenia. Thrombocytopenic patients were younger, male pre-dominant, and healthier with lower prevalence of all comorbidities and anti-thrombotic use. Thrombocytopenic patients had statistically significant higher mortality (6.4% vs 5.6%), venous thromboembolism (1.6% vs 1.5%), variceal bleeding (6.9% vs 4.3%), spontaneous bacterial peritonitis (3.9% vs 3%), portal hypertension (35.8% vs 23.9%), hepatorenal syndrome (5.5% vs 3.8%), hyponatremia (23.7% vs 19%), jaundice (0.6% vs 0.4%) and abnormal coagulation profile (2.2% vs 1.6%). Multivariate logistic regression model after matching showed similar outcomes with thrombocytopenic patients having higher mortality (OR: 1.13 [1.09 - 1.18];p<0.001) and statistically significant higher rates of all other complications.

Conclusion: Thrombocytopenia was significantly associated with higher mortality and complications in liver cirrhosis. These findings highlight the importance of considering thrombocytopenia as a poor prognostic factor when evaluating patients with liver cirrhosis. Further randomized controlled trials are needed to corroborate these findings and explore the benefit of addition of platelet count to the current prognostic scores used in clinical practice to estimate outcomes in liver cirrhosis.
PPIL2 Is a Target of the JAK2-STAT5 Pathway and Mediates p53 Polyubiquitination and Degradation
Authors: Wang Pan , Han Xu , Ren Kehan , Li Ermin , Bi Honghao , Aydemir Inci , Ji Peng
Published: December 2023

The activated JAK2-STAT pathway is characteristic of myeloproliferative neoplasms (MPNs). We previously revealed that Plek2, a downstream target of the JAK2/STAT5 pathway, functions as a scaffold protein to activate Akt and plays an important role in the pathogenesis of JAK2 V617F-induced MPNs. To further explore the mechanisms of the Plek2 signalosome, we performed a comprehensive proteomic analysis of Plek2 interacting proteins and found peptidylprolyl isomerase-like 2 (PPIL2) as a potential novel effector of the signalosome. As a U-box E3 ubiquitin ligase, PPIL2 was reported to be involved in cancer metastasis. However, its role in normal and malignant hematopoiesis remains unknown. We found that PPIL2 is required for human and murine erythropoiesis in vitro. Knockout of PPIL2 through CRISPR/Cas9 significantly reduced cell proliferation and differentiation and led to marked apoptosis. Mechanistically, PPIL2 interacts with and catalyzes p53 polyubiquitination at lysine 24 to promote proteosome-mediated p53 degradation, thereby preventing p53-dependent cell cycle inhibition and apoptosis. We further revealed that PPIL2 is a downstream target of the JAK2/STAT5 pathway and is upregulated in a JAK2 V617F-positive MPN mouse model and in patients with MPNs. Loss of Ppil2 ameliorated JAK2 V617F-induced myeloproliferative phenotypes including erythrocytosis, neutrophilia, thrombocytosis, and splenomegaly in JAK2 V617F-knockin mice. The same findings were also observed when the JAK2 V617F-knockin mice were treated with cyclosporin A, an inhibitor of PPIL2. Our findings reveal PPIL2 as an effector of the JAK2/STAT5 pathway and Plek2 signalosome. PPIL2 plays an important role in normal erythropoiesis and the pathogenesis of JAK2 V617F-induced MPNs by degrading p53, pointing to PPIL2 as a potential therapeutic target for the treatment of MPNs.
Metabolic Tumor Volume from Baseline PET/CT Improves Stratification in the High IPI Risk Patients with Diffuse Large B-Cell Lymphoma
Authors: Hanzhen Zhang , Wei Xiaolei , Zhou Wenlan , Xu Zihan , Chen Junjie , Wei Yongqiang , Wu Hubing , Feng Ru
Published: December 2023

Background: In the rituximab era, the international prognostic index (IPI) risk stratification was not enough to predict outcome in patients with diffuse large B cell lymphoma (DLBCL). Previous studies showed the Metabolic tumor volume（MTV） from baseline [ 18F]FDG PET/CT could be used to predict outcome. In this study, we evaluated the prognostic value of IPI and MTV in DLBCL patients.

Patients and methods: A total of 262 de novo DLBCL patients who were treated with the R-like chemotherapy were retrospectively analyzed in this study. The IPI and MTV were compared to the Overall Survival (OS) and Progression Free Survival (PFS).

Results: The MTV is better than TLG and SUVmax for predicting disease progression. The cutoff value of MTV was 444 cm 3. Patients with high MTV showed inferior OS and PFS ( p < 0.001, respectively). Although the IPI remains predictive it distinguishes only 2 risk groups, rather than the 4 groups originally described. Further analysis showed high MTV could identify a subgroup of patients with extremely inferior OS and PFS in high IPI risk patients ( p = 0.0089 and p= 0.024, respectively). Multivariate analysis revealed that high MTV is a significant poor prognostic factor for both OS (HR 3.168[1.81-5.56], p <0.001) and PFS (HR 2.637 [1.51-3.71], p <0.001).

Conclusions: Our study suggests that MTV from baseline PET/CT is an important prognostic factor in DLBCL patients, allowing the identification of an inferior outcome subgroup in high-risk patients, which may help to guide treatment in clinical trial.
Investigation of the Impact of in Utero Thirdhand E-Cigarette Exposure on Platelet Function
Authors: Umphres Shelby , Alshbool Fatima , Khasawneh Fadi
Published: December 2023

Cardiovascular disease (CVD) is the leading cause of death worldwide, with many deaths being due to heart attacks and stroke. To this end, platelets are key players in maintaining and regulating “hemostasis” in the cardiovascular system. However, a disruption in the platelet function (hyperactivity) can promote the genesis of occlusive (thrombosis-based) CVD. Though there are many known risk factors for CVD, the number one preventable risk factor is exposure to tobacco or tobacco related products. Interestingly, the use of emerging tobacco products, specifically e-cigarettes (e-cigs) has been on the rise, achieving unprecedented levels, due to the misconception that they are a safe(r) alternative to traditional cigarette smoking, especially amongst pregnant women or women of childbearing age. Therefore, there has been efforts- including by our laboratory- to characterize the negative effects of smoking on the CV system. To this end, we have previously shown that direct exposure to e-cigs increases the risk of occlusive CVD. However, the effects of indirect exposure to e-cigs, namely thirdhand exposure, which is formed from toxicants “produced” by e-cig vapor that accumulate on surfaces such as upholstery, curtains, car seats, carpet etc., are still unknown, including under in utero settings. To this end, the present studies aim to characterize the impact that in utero thirdhand e-cig (IUTHEC) exposure exerts on platelet function and the CV system and examine the mechanism of these effects. Utilizing a custom e-cig vape system mice were housed with clean air or e-cig-exposed material for one week prior to mating and for the duration of their pregnancy. Using this approach to mimic a real life exposure scenario, our preliminary results indicate that IUTHEC exposed pups exhibited a shortened bleeding and thrombus occlusion times, in two widely used hemostasis and thrombosis models. In terms of the mechanism of this prothrombotic phenotype, it was found that platelet aggregation, as well as dense and alpha granule secretion were potentiated in the IUTHEC exposed mice in response to agonist stimulation. Furthermore, integrin activation and phosphatidylserine exposure, additional markers of platelet function, were also found to be enhanced. These data together demonstrate for the first time that IUTHEC increases the risk of thrombotic CVD in part via modulation of platelet activation. Moreover, our findings highlight the negative health effects of exposure to IUTHEC, which is an underappreciated threat to human health.
A Prospective, Multinational Study of Clinical and Biological Factors Associated with Short Overall Survival in Multiple Myeloma
Authors: Tang Andy , Kim Kihyun , Chen Fiona , McQuilten Zoe K , Mollee Peter , Rajagopal Rajeev , Quach Hang , Ho Phoebe Joy , Harrison Simon J. , Spencer Andrew , Chng Wee-Joo
Published: December 2023

Introduction: Multiple myeloma (MM) is an incurable cancer, but a subset of patients can achieve long-term remission and overall survival (OS). We aimed to study the clinical and biological factors associated with short OS defined as less than 3 years.

Methods: A prospective, observational, multinational study was conducted by Australian and New Zealand (ANZ) and Asia-Pacific (APAC) Myeloma and Related Disease Registry (MRDR) from January 2013 to January 2023, across 5 countries (Australia, New Zealand, Korea, Singapore, Malaysia). Eligible patients were ≥18 years old, diagnosed with MM per International Myeloma Working Group (IMWG) criteria, received a bortezomib-containing triplet, and had at least 3 years of follow-up or had deceased within 3 years. Descriptive analysis was completed using chi-squared test for categorical variables and Wilcoxon rank-sum test for continuous variables. The Kaplan-Meier method was used for analysing OS and follow-up data. The ‘stepwise’ function in Stata was used for the forward stepwise logistic regression, with the p-value set to 0.1. All analyses were completed using Stata 16.

Results: Of the 1539 patients enrolled, majority were from ANZ (1071 and 359, respectively), with male preponderance (61.0%). Half were above 65 years old. High risk cytogenetic abnormalities, a criterion of the Revised International Staging System (R-ISS), was unknown for half of the patients (49.4%). 45.3% patients presented as ISS stage 1, 21.9% stage II, and 32.8% stage III, respectively at study entry . The most frequent front-line agent used in combination with bortezomib-based triplet was cyclophosphamide [VCd, 1380 (89.7%)], followed by thalidomide [VTd, 92(6.0%)], lenalidomide [VRd, 63 (4.1%)] and daratumumab [DVd, 4 (0.3%)]. The median follow-up was 29.9 months (95%CI: 28.3-31.8) and the median OS was 87.7 months (95%CI: 80.9-93.2). Thirty percent patients had OS<3 years. Clinical and biological factors associated with long OS (≥3 years) were compared with those <3 years (Table 1). Baseline factors associated with higher odds of short OS included age≥65, Eastern Cooperative Oncology Group performance status (ECOG PS)≥2, ISS III, lower creatinine clearance and platelet count, high LDH and poor cytogenetic abnormalities (Table 1). In a multivariate analysis, age≥65 (OR:2.50; p<0.001), ECOG≥2 (OR:2.08; p<0.001), ISS III (OR:2.48; p<0.001), t(4;14)[OR:2.04; p=0.004], t(14;16)[OR:6.33; p=0.004] and 17p deletion (OR:2.77; p =0.001) were associated with a significantly shorter OS.

Conclusion: From clinician perspective, identifying patients with an increased likelihood of shorter OS through the determination of patient- and disease-specific risk factors might be clinically relevant to facilitate better treatment decisions.
Acute Abdominal and Pelvic Complications in Patients with Non-Promyelocytic Acute Myeloid Leukaemia Undergoing Intensive Induction Chemotherapy Are Associated with Favorable Prognosis
Authors: Murphy Philip T , Alsaleh Mohammed , Cheema Harris , John Quinn , Glavey Siobhan , Sargent Jeremy , Swan Dawn Jennifer
Published: December 2023

Acute abdominal abnormalities are relatively uncommon during the treatment of acute leukaemia but, historically, have been associated with high mortality rates (Hawkins JA, Mower WR, Nelson EW. Acute abdominal conditions in patients with leukaemia. Am J Surgery 1985;150:739-742). Given the improvement in diagnostic and supportive measures for patients with acute myeloid leukaemia (AML) over time, we aimed to determine the rate and outcomes of acute abdominal and pelvic complications in a cohort of AML patients at our hospital.

We retrospectively reviewed the medical records, radiology reports and laboratory test results of 70 consecutive newly diagnosed, non-promyelocytic AML patients undergoing intensive induction chemotherapy between January 2012 and December 2019. Seventeen patients developed 22 episodes of acute abdominal or pelvic pathology and were investigated by radiology (abdominal X ray, CT scan and/or MRI). Table 1 summarises the underlying causes for these complications, as well as neutrophil count and platelet count at time of presentation. Colitis was the most common complication, occurring 6 times. The neutrophil count was <1x10 9/L in 15 of the 22 episodes. Platelet count <20x10 9/L occurred in 8 of the episodes, including the 2 cases of ureteric obstruction by clot. All complications resolved completely with conservative management, except for 1 patient who required surgical drainage of an intraperitoneal and psoas muscle abscess in addition to intravenous broad spectrum antibiotics.

In conclusion, acute abdominal and pelvic complications were relatively rare in our cohort of patients. All but 1 of these episodes resolved with conservative management alone. These results suggest that, with current diagnostic and therapeutic measures, the prognosis for non-promyelocytic AML patients who develop such complications is favorable.
DNA Vaccines Against GPRC5D in Myeloma
Authors: Li Yong
Published: December 2023

Introduction:

Myeloma is an incurable disease despite improved survival from the ongoing immunotherapy revolution. Virtually all myeloma cases are preceded by MGUS, a condition affecting over one million Americans. Smoldering myeloma is an asymptomatic clonal plasma cell disorder between MGUS and myeloma. Smoldering myeloma is distinguished from MGUS primarily for clinical reasons because the risk of progression to malignancy in the first five years after diagnosis is different: 10% per year in smoldering myeloma vs. 1% per year in MGUS. Yet the risk of progression is not constant, and in a subset of MGUS patients, the risk evolves from a low chance of progression to a high degree of probability of progression to MM. Therefore, patients with smoldering myeloma and MGUS are potential subjects for myeloma prevention. GPRC5D is an emerging novel immunotherapeutic target for myeloma, with several ongoing clinical trials using CAR T cells and antibodies directed against GPRC5D.

Methods:

Nanoplasmid is a next-generation plasmid technology that bypasses the antibiotic selection when amplified in E. coli by expressing a 150 bp RNA-OUT antisense RNA and uses the highly productive heat-inducible R6K origins for DNA replication. With a <500 bp backbone, Nanoplasmids achieve significantly higher expression levels than pVAX1, the most widely used plasmid in DNA vaccine development. We used Nanoplasmid to express human GPRC5D and introduced it into mice via electroporation xenografted with mouse cancer cells that express the human GPRC5D gene. We examined the animals and determined whether the prophylactic vaccines prevented myeloma development and progression.

Results:

We found that mice vaccinated with the human GPRC5D Nanoplasmids had both humoral and cellular responses against the GPRC5D protein. The DNA vaccines were active in reducing tumor development from human GPRC5D-expressing tumors.

Conclusions:

GPRC5D-targeting DNA vaccines are a cost-effective prevention agent for MGUS and smoldering myeloma.
Survival Trends and Prognostic Factors of Patients with Newly Diagnosed Multiple Myeloma Accompanied with Extramedullary Disease
Authors: Wang Jing , Shen Na , Shen Xuxing , Zhang Run , Jin Yuan , Li Jianyong , Chen Lijuan
Published: December 2023

Extramedullary disease (EMD) is an unusual event in patients with MM. This study aimed to assess the prognostic impact of EMD and develop an EMD-based risk model to estimate the survival of patients with newly diagnosed multiple myeloma (NDMM). A total of 518 patients were enrolled in this study, of which 121 presented with EMD at the initial diagnosis. Patients were divided into non-EMD, extramedullary-bone-related (EM-B), and extramedullary-extraosseous (EM-E) groups. Clinical characteristics were compared using the chi-squared test or Fisher's exact test. Survival curves were plotted using the Kaplan-Meier method, and a nomogram was constructed based on the Cox proportional hazards model. Compared to patients without EMDs, patients with EM-E were younger ( P=0.028), and those with EM-B had less renal damage ( P<0.001). The EM-E group had the worst progression-free survival (PFS) and overall survival (OS). In addition, patients with multiple sites of EMD invasion or high Ki67 expression had poor OS. Lenalidomide-based treatment showed the worst outcome, and autologous stem cell transplantation (ASCT) remarkably improved the survival of patients with EMD. A prognostic model (MM prognostic index, MM-PI) comprising lactate dehydrogenase (LDH), circulating plasma cells (CPC), del(17p), and type of extramedullary involvement was developed, and a 4-factor nomogram. In conclusion, we established a risk model incorporating extramedullary disease that provides accurate and individualized survival estimates for patients with NDMM.
The Clinical Complexity of Langherans Cell Histiocytosis in Adult Patients: Experience of the Institute of Hematology of Bologna
Authors: Broccoli Alessandro , Bagnato Gianmarco , Argnani Lisa , Cantelli Martina , Casadei Beatrice , Pellegrini Cinzia , Zoli Sabrina , Sutto Emanuele , Coppola Paolo Elia , Gentilini Marianna , Morigi Alice , Carella Matteo , Gugliotta Gabriele , Nanni Laura , Stefoni Vittorio , Zinzani Pier Luigi
Published: December 2023

Langerhans cell histiocytosis (LCH) in adult patients is a rare disease with protean manifestations, including single system unifocal (SSU) or multifocal (SSM) presentation or multisystemic (MS) involvement. Clinical care of adult patients is based on case series and reports. Regimens applied in pediatric patients show unacceptable toxicity in the adult setting.

Our single-center experience with LCH is presented. Data on 30 patients diagnosed between 1997 and 2022 have been reviewed. Patients with isolated pulmonary histiocytosis were excluded. Responses to any treatment were classified according to the acknowledged criteria. Progression-free survival (PFS) after frontline treatment was calculated on all treated patients. Overall survival (OS) was determined on the whole cohort irrespective of any treatment received.

Eighteen male and 12 female patients received a diagnosis of LCH at a median age of 46 years (range 20-75). Sixteen patients had single system (SS) disease, with SSU involvement in 12 cases and SSM involvement in 4 cases. The skeleton was the mostly involved system (Figure), while SSU extra-skeletal disease was detected in 25% of cases. Fourteen patients had MS disease: axial skeleton, lungs, hypophysis, skull, skin and appendicular skeleton were the predominantly involved sites, in this order. Ten patients with SS disease received chemotherapy because of symptoms (mostly bone pain) or due to multifocal dissemination. Two patients underwent radiotherapy (RT), 2 had their disease completely excised and 1 received systemic steroids. One patient was observed. A complete response (CR) was obtained in 60% of treated cases, while a partial response (PR) in 27%. Four patients relapsed after frontline therapy and were rescued with RT or autologous transplant. Among patients with MS disease, chemotherapy (MACOP/VNCOP-B, cladribine, vinblastine) was applied in 6 cases; 2 patients received RT and 1 systemic steroids. One patient underwent surgery and 4 were observed because of asymptomatic disease. A CR was obtained in 70% of treated patients and a PR in 10%. Five patients relapsed, with symptomatic disease requiring treatment (chemotherapy and RT) in 3 cases. Median PFS among all treated patients was 177 months. OS was 91% at 20 years.

Appropriate treatment based on disease extension, site involvement and clinical manifestations yields high remission rates and long term PFS and OS. The optimal approach to relapsed patients remains undefined.
PIEZO1-TMEM16F Interplay in Hereditary Xerocytosis
Authors: Liang Pengfei , Zhang Yang , Wan Yui Chun Serena , Ma Shang , Dong Ping , Lowry Augustus , Francis Samuel , Khandelwal Sanjay , Delahunty Martha , Telen Marilyn J. , Strouse John J , Arepally Gowthami , Yang Huanghe
Published: December 2023

Cell surface exposure of phosphatidylserine (PS), an anionic phospholipid that is usually confined to the inner leaflet of the plasma membrane, triggers a plethora of cellular responses. PS exposure in RBCs contributes to blood coagulation, promotes RBC aggregation and adhesion to endothelial cells, and accelerates the removal of aging RBCs from circulation. The existence of CaPLSase, a passive phospholipid transporter that catalyzes rapid PS exposure in response to intracellular Ca 2+ elevation, was first observed in RBCs in the 1990s. However, the molecular identity of RBC CaPLSase, its activation mechanism and its role in red cell disorders remain elusive. Here we demonstrate that TMEM16F, the long-sought-after RBC CaPLSase, is activated by calcium influx through the mechanosensitive channel PIEZO1 in RBCs. PIEZO1-TMEM16F functional coupling is enhanced in RBCs from individuals with hereditary xerocytosis (HX), a RBC disorder caused by PIEZO1 gain-of-function (GOF) channelopathy. Enhanced PIEZO1-TMEM16F coupling leads to an increased propensity to expose PS, which may contribute to HX clinical manifestations including anemia, splenomegaly and post-splenectomy thrombosis. Pharmacological inhibition of PIEZO1 prevents calcium-induced dehydration, hemolysis and PS exposure in HX RBCs. Our study reveals an activation mechanism of TMEM16F and its pathophysiological function in HX, providing insights into potential treatment.
Bone Marrow Microenvironment-Responsive Nanomedicine Promotes Anti-Leukemia Activity In Vivo
Authors: Wang Dongyao , Chen Haoting , Zhu Xiaoyu
Published: December 2023

Background: Leukemia escape and relapse is the leading cause of death after allogeneic hematopoietic stem cell transplantation (allo-HSCT) for acute myeloid leukemia (AML). Work over the past decades has demonstrated that the CD47- signal-regulatory protein α (SIRPa) interaction mediated cancer cell immune escape. Although CD47-SIRPα interaction blockade contributes to cancer cell clearance by macrophages, how to deliver the anti-CD47 to bone marrow niche through an efficient drug delivery strategy and improve the cytotoxic activity of macrophages is an important issue that needs to be addressed. Methods: Here, we performed immunohistochemical (IHC) analysis and flow cytometry analysis of TIM-3 expression using samples of bone marrow from patients with or without relapse. In addition, according to the natural “myeloid niche homing” characteristic of AML cells and the low-oxygen microenvironment in bone marrow, we constructed β-cyclodextrin azobenzene linker (AZO-β-CD)-modified fusion protein and amantadine-modified HSA protein (HSA-ADA), followed by the addition of anti-CD47. Furthermore, the nanogel TIM3-targeting antibodies were modified on the surface of the nanogel to finally construct the supramolecular nanogel delivery system. Moreover, we detected the effect of target nanomedicine in a leukemia xenograft mouse model. Results: We demonstrated that in the bone marrow microenvironment of patients with relapse, the expression of TIM-3 significantly increased, especially in the CD34 +CD38 + leukemic progenitors, but not in the HSCs. Furthermore, we verified using nanomedicine could significantly increase the number of macrophages in the bone marrow. Importantly, the blockade of CD47-SIRPα signaling using nanomedicine markedly improved the anti-tumor activity in a leukemia xenograft mouse model. Conclusion: Thus, our findings reveal that nanomedicine could inhibit CD47-SIRPα signaling and contribute to anti-leukemia function effectively, which may be a new strategy for preventing and treating the recurrence of AML after transplantation.
A Prognostic Nomogram Survival Model for Newly Diagnosed Patients with AIDS-Related Diffuse Large B-Cell Lymphoma: A Multicenter Cohort Study in China
Authors: Yang Tao , Liang Yang , Wang Chaoyu , Liu Jun , Wu Yan , Min Haiyan , Huang Yunhong , Wei Guo , Zhang Wei , Wang Min , Tang Xiaoqiong , Yang Zailin , Jun Li , Zhou Hui , Liu Yao
Published: December 2023

BACKGROUND

The prognosis of patients with AIDS-related diffuse large B-cell lymphoma(AR-DLBCL) becomes very poor when the lymphoma relapses or is refractory to first line immunochemotherapy. We aimed to develop a novel ARDPI nomogram prognostic model for risk stratification so as to guide individualized treatment to achieve sustained remission and improve the overall prognosis for newly-diagnosed AR-DLBCL patients.

METHODS

We interrogated data from 306 patients with newly-diagnosed AR-DLBCL. We filtered variables using LASSO regression and Cox regression to identify prognostic co-variates and develop a survival model, we termed AR-DLBCL Prognostic Index (ARDPI). We evaluated model discrimination, calibration and clinical benefit by Area Under the Receiver-Operator Characteristic (AUROC), calibration plots and decision curve analysis (DCA). Next, we compared the ARDPI model discrimination, calibration and clinical benefit with the IPI and NCCN-IPI models using the same methods. Lastly, we stratified patients into three survival risk cohorts based on ARDPI model by X-tile selecting cutoff point.

RESULTS

7 co-variates were independently correlated with survival and were used to develop the ARDPI model including age, lymphocyte monocyte ratio (LMR), CD5 expression on lymphoma cells, blood EBV-DNA copy number, CD4/CD8 ratio, central nervous system (CNS) involvement and anti-HIV therapy (ART). AUROCs of ARDPI model for 1-, 3-, and 5-year were 0.80 (95% Confidence Interval [CI], 0.72, 0.88), 0.78 (0.69, 0.87) and 0.77 (0.63, 0.91). Predicted and calibrated values were concordant. The DCA curve had higher net benefit using the ARDPI model. Prediction accuracy of the ARDPI model was better compared with the IPI and NCCN-IPI models. For example, 3-year survival AUROC in the ARDPI model was 0.78 (0.69, 0.87) compared with the IPI (0.53 [0.43, 0.63] P< 0.001) and the NCCN-IPI (0.52 [0.42, 0.62] P< 0.001). Using the ARDPI model, we identified 3 survival risk cohorts with 3-year survivals of 0.80 , 0.38 and 0.09 ( P<0.001).

CONCLUSION

The ARDPI has good survival prediction accuracy in newly-diagnosed persons with AIDS-related DLBCL and using it has clinical benefit.Accuracy is better than the IPI and NCCN-IPI models. Validation of our conclusions is needed.