bluuebunny/crossref_metadata_2025 huggingface dataset:
Artificial Intelligence RBC Recognition Program for Schistocyte Screening in Cytopenic Patients
Authors: Diewchim Chanida , Chalidabhongse Thanarat H. , Watanaboonyongcharoen Phandee , Kongkiatkamon Sunisa , Lawasut Panisinee
Published: December 2023

Schistocyte identification is critical for the diagnosis of thrombotic microangiopathies (TMA). It is one of the most common consultative problems for hematologists. The count of schistocytes is poorly defined by automated complete blood count (CBC) machines. Furthermore, morphological evaluation using peripheral blood smear (PBS) staining is time-consuming and exhibits high variability among inexperienced internists. Our aim is to develop and assess the effectiveness of an in-house AI program for recognizing schistocytes. In phase I of this study, we developed an AI program using deep convolutional neural networks trained from schistocyte in blood smear pictures total 1066 pictures of 1066 patients and divided into training set and validation set. In the training set, 10,377 schistocytes were identified, resulting in an accuracy of 0.76977, precision of 0.78487, recall (sensitivity) of 0.97562, and F1-score (harmonic mean of precision and recall) of 0.86991. And in the validation set, 3,564 schistocytes were identified, resulting in an accuracy of 0.68462, precision of 0.73850, recall (sensitivity) of 0.90370, and F1-score of 0.81279. Then, during phase II, PBS from 133 patients from hematology consultation service due to anemia and/or thrombocytopenia were included and scanned by an automated, cell-locating image analysis system. The program's effectiveness was validated by comparing its detection of an increased in schistocytes from patient glass slides (>1%) with the conclusion from at least two out of three hematologists who conducted conventional microscopy readings. The Cohen's kappa agreement between the AI and hematologist readings was 0.583 (p < 0.001), representing moderate agreement. There were 58 slides with schistocytes >1% as determined by AI reading, while 34 PBSs were identified as showing a significant increase in schistocytes by hematologists. The sensitivity and specificity of significantly increased schistocyte detection were 97.1% and 74.7%, respectively. Blood smears with significantly increased schistocytes detected by the program had positive predictive and negative predictive values of 43.1% and 93.3%, respectively. Poikilocytosis from thalassemic red cells were the most misinterpreted. In sum, the developed AI program for schistocyte detection showed high sensitivity and acceptable specificity, especially in high prevalence thalassemia populations, indicating its potential to assist inexperienced clinicians and reduce the need for hematologist consultations. Adding more schistocyte samples could improve the program's specificity and efficiency of TMA diagnosis in the future.
The Impact of Induction Regimens on Clinical Outcomes in Double and Triple-Hit Lymphoma: A Meta-Analysis of Comparative Studies
Authors: Haddad Philip A , Powell Frankie , Gupta Ankita , Graham Christopher
Published: December 2023

Background:

Double and triple hit lymphomas (DTHL) are aggressive high-grade lymphomas that harbor translocations of MYC and BCL2 and/or BCL6. They constitute 6-14% of all aggressive B-cell lymphoma and have been associated with poor clinical outcomes. Though there is no established standard therapy for DTHL, induction chemotherapy vacillates between R-CHOP-like regimens, R-EPOCH, or more dose-intense regimens (DI) such as R-hyperCVAD/MA and CODOX-M/IVAC with or without consolidation with stem cell transplant. Currently, there are no randomized trials establishing a standard induction regimen. Furthermore, several retrospective studies reported conflicting results regarding the comparative benefits of these various induction approaches. Therefore, we conducted this meta-analysis to evaluate the impact of induction chemotherapy regimens on the clinical outcomes of DTHL.

Methods:

A review of the medical literature was conducted using online databases. Inclusion criteria consisted of English language, diagnosis of DTHL, comparative studies of induction chemotherapy regimens, and studies that reported overall (OS) and progression-free (PFS) survival. Studies that reported the aggregate outcome of mixed samples of DTHL and other lymphomas were excluded. A meta-analysis using the fixed effects and random effects models was conducted.

Results:

Seven retrospective comparative studies with a total of 1433 patients were included. The median age was 62 years, and the median follow-up was 27.5 months. DI regimens were associated with better PFS than R-CHOP-like regimens (HR = 0.81, 95%CI 0.66-0.99, p = 0.039) but without improved OS. R-EPOCH, including R-DA-EPOCH, regimens were associated with better PFS than R-CHOP-like regimens (HR = 0.62, 95%CI 0.43-0.91, p = 0.013) but without improved OS. R-HyperCVAD/MA regimen was associated with better PFS compared to R-CHOP-like regimens (HR = 0.64, 95%CI 0.47-0.87, p = 0.005) but without improved OS.

Conclusions:

This is the first and largest meta-analysis to show that induction DI regimens, in general, and R-EPOCH and R-HyperCVAD/MA regimens, in particular, are associated with better PFS than R-CHOP-like regimens in patients with DTHL. However, this advantage did not translate into a survival benefit.
Incidence of Infections Associated with the Use of Bispecific Antibodies in B-Cell Non-Hodgkin Lymphomas
Authors: Fares Amneh A , Martinez Diana , Musleh Ud Din Saba , Wilkinson Robert , Chavez Julio C , Vargas Madueno Fernando Manuel , Sandoval-Sus Jose
Published: December 2023

Background: Despite recent advances in the treatment of B-cell non-Hodgkin lymphomas (NHL), most patients with relapsed/refractory (R/R) disease are at high risk of dying from their disease. Bispecific antibodies (BsAbs) are novel therapies with favorable response rates in R/R B-cell NHL even after cellular therapies. Mosunetuzumab, epcoritamab, glofitamab, and odronextamab are CD20 targeting BsAbs that have proven efficacy for R/R B-cell NHL. These treatments are generally well tolerated, but the infectious complications are not well described in the literature. Our study aims to quantify the relative incidence of infections associated with B-cell NHL targeted BsAbs.

Methods: We conducted a preliminary evaluation, of an ongoing pooled analysis encompassing single agent BsAbs used in B-cell NHL with no prior use of alternate BsAbs. Exclusion criteria included studies reporting the use of BsAbs as part of combination therapy and trispecific antibodies.

Results: A total of 546 B-cell NHL patients (number of trials, n=4) were treated with BsAbs in the studied period. Pooled median follow up was 11.7 . Relevant adverse events included all grade neutropenia in 26.9% (n=147/546), grade III/IV neutropenia in 20% (n=109/546), all grade infections in 40.1% (n=219/546), grade III/IV infections in 16.8% (n=92/546), all grade pneumonia in 7.2% (n=17/235), and coronavirus 2019 (COVID-19) in 1.3% (n=3/235). Incidence of grade III/IV and all grade infections of the BsAbs in each study are shown in figure 1. In addition, all grade cytokine release syndrome was seen in 56.4% (n=308/546) and death was reported in 33 patients of which 4 (0.7%) were reported to be secondary to infections.

Conclusion: Data from this ongoing analysis better describes the various infectious risks of patients with B-cell NHL receiving BsAbs. Providers should remain vigilant regarding these risks and intervene promptly in such situations. The finalized pooled analysis with a larger sample size may corroborate these findings.
Dynamic Single-Cell Profiling Reveals Novel Immune Regulatory Mechanism of ITK Inhibitor Soquelitinib in Refractory T Cell Lymphoma
Authors: Ding Ning , Xie Yan , Guo Yong , Li Kecheng , DU Tingting , Li Zhenjun , Li Yuejiao , Cao Jiaowu , Hu Dingyao , Hsu Lih-Yun , Miller Richard , Zhu Jun , Song Yuqin
Published: December 2023

Introduction

Interleukin 2 inducible T cell kinase (ITK) plays an important role in the T cell receptor (TCR) signaling pathway. The highly selective covalent ITK inhibitor, soquelitinib (formerly known as CPI-818), is being evaluated as a monotherapy in an ongoing Phase 1/1b trial in patients with refractory T-cell lymphoma (TCL) (NCT03952078). Single cell RNA-sequencing and flow cytometry were performed on patient samples to identify potential prognostic biomarkers and to study immune regulatory mechanisms.

Methods

Paired peripheral blood and tumor tissue samples were collected from 3 responding TCL patients treated with soquelitinib. Dynamic single cell RNA-seq and flow cytometry studies were performed and changes in gene expression profiling and signature gene enrichment were analyzed at baseline and on treatment.

Results

Single cell RNA-seq of mRNA expression demonstrated that soquelitinib induced expression of cytotoxic genes (PRF1, GZMB, GZMK, GZMA, GZMH, GNLY and NKG7) and effector genes (IFNG, EOMES, CST7), and reduced exhaustion genes (PDCD1, LAG3, CTLA4, HAVCR2) in normal CD8+ cells. KEGG and GSEA enrichment analysis revealed that Interferon-Gamma, IL-2-STAT5 and TNF-alpha related signaling pathways were upregulated; the protein-protein interaction network analysis revealed that the hub gene TBX21 might play a central regulatory role. KEGG enrichment analysis also demonstrated that soquelitinib treatment significantly decreased DNA replication and cell cycle signaling in malignant T cells, and that normal CD4+ Treg cells within tumor microenvironment and peripheral blood were significantly reduced post soquelitinib treatment.

Flow cytometry analysis of the peripheral blood and intratumoral T cell subpopulations confirmed that the normal CD4+ and CD8+ T cells, especially Th1 cells and CD8+ terminally differentiated T effector memory cells were significantly increased and CD4+ Treg cells were decreased compared to baseline levels following soquelitinib treatment.

In addition, human T cell differentiation studies and EL4 syngeneic tumor bearing mice studies confirmed the immunomodulatory effect of soquelitinib in the tumor microenvironment in vitro and in vivo. Similar results were also observed in the murine syngeneic B cell lymphoma A20, breast cancer 4T1 and colon cancer CT26.WT models.

Conclusions

Our findings reveal a novel immune regulatory mechanism by which the ITK inhibitor soquelitinib induced normal CD4+ Th1 cells and CD8+ TEMRA cells and reduced CD4+ Treg cells in the tumor microenvironment in responding patients. These findings demonstrate the potential of soquelitinib as a novel immunotherapy for the treatment of T-cell lymphomas and solid tumors.
Antinuclear and Specific Autoantibodies Predict Autoimmune Disease in Sickle Cell Patients: A Longitudinal Cohort Study
Authors: Truffinet Frederic , Compain Caroline , Manea Maria Elena , Habibi Anoosha , Fois Elena , Audureau Etienne , Hue Sophie , Michel Marc , Bartolucci Pablo
Published: December 2023

Introduction: Sickle cell disease (SCD), the most frequent genetic disorder, can be associated with systemic autoimmune diseases (ADs). High levels of antinuclear antibodies (ANA) are commonly described in SCD, as alloimmunization after blood transfusion. However, the specificity and the prognostic value of ANA have not been fully determined.

We aimed to evaluate the association of ANA with AD development during long-term follow-up in SCD patients. Our secondary goal was to assess mortality and serious SCD-related events in this population.

Patients and methods: We conducted a single-center longitudinal cohort study on 66 adult SCD patients with ANA titers ≥1/320 at baseline followed over a 13 year-period. Patients known to have an AD (n=9) were excluded from the risk analysis.

Results: During follow-up, 13 out of 57 patients (23%) developed AD. Patients with ANA titers ≥1/1280 had a 3-fold increased risk of developing AD (RR 3.4[1.4;8.6]) and a 9-fold increased risk in the presence of specific autoantibodies (anti-DNA, anti-ENA, RF or ACPA) (RR 9.0[3.7;23]) with a positive (PPV) and negative predictive value (NPV) of 90%. When these two immunological features were associated, specificity and PPV reached 100%, with a NPV of 84%. There was no significant difference in the occurrence of serious SCD-related events or mortality rate according to AD.

Conclusions: Sickle cell patients with high level of antinuclear antibodies and specific autoantibodies are at high risk of developing AD. The occurrence of AD does not seem to have a significant impact on the course and severity of SCD.
A Phase 1b Study of ELA026, a Monoclonal Antibody Targeting Signal Regulatory Protein-α/β1/γ, in Patients with Newly Diagnosed and Previously Treated Secondary Hemophagocytic Lymphohistiocytosis
Authors: Maiti Abhishek , Daver Naval , Iyer Swami P. , McCall David , Gohil Satyen H , Lopez Jimenez Javier , Lane Hayley , Kim Benjamin , Dao Kim-Hien , Patou Gary , Panicker Sandip , Covert Kelly , Parry Graham Carl , Allen Carl E
Published: December 2023

Introduction: ELA026 is under investigation in secondary hemophagocytic lymphohistiocytosis (sHLH), a life-threatening, hyperinflammatory condition that is most often triggered by malignancies, autoimmune disorders, and infectious diseases. There are no approved therapies for sHLH, and current treatment approaches, particularly for malignancy-associated HLH (mHLH), are suboptimal. For example, in adult patients with mHLH, median overall survival (mOS) is approximately 2 months (Lee et al. 2023). ELA026 is a monoclonal antibody directed against signal regulatory protein-α/β1/γ and is intended to target the activated myeloid and T cells that drive sHLH.

Methods: This is an open-label, single-arm, Phase 1b global multicenter study (NCT05416307) to evaluate the safety, efficacy, pharmacokinetics, and pharmacodynamics of ELA026 in newly diagnosed and previously treated sHLH patients, aged 12 years or older (6 years or older in select countries including the US). In Cohort 1, an intra-patient dose escalation of ELA026 occurs in the absence of a dose limiting toxicity (DLT) and optimal biomarker response. Response is evaluated as described in Locatelli et al. 2020.

Results: Six mHLH patients enrolled in Cohort 1 with median age of 50 years (range 21-67). Three patients were refractory to etoposide-based therapies and/or anti-cytokine therapies. No DLTs or study drug-related serious adverse events occurred. Five patients were deemed evaluable for efficacy based on >1 week of ELA026 exposure (efficacy assessment is pending). All 5 patients received concurrent chemotherapy for their malignancies. As of July 31, 2023, 3 patients are alive and 2 patients have died from complications of malignancy (mOS has not been reached but has exceeded 3 months). Three patients who had periods of ELA026 monotherapy displayed early, progressive monocyte and lymphocyte reductions with associated improvements in C-reactive protein and other sHLH-associated biomarkers. Updated study results and progress will be provided at the meeting.

Conclusion: Preliminary data suggest ELA026 is well tolerated in sHLH patients and induces early biomarker changes that warrant further clinical investigation.
TET2-Mediated Dysregulation of Heterochromatin in Age-Related Clonal Hematopoiesis
Authors: Hong Tingting , Li Jia , Guo Lei , Cavalier Maryn , Wang Tianlu , Fang Shaohai , Guzman Anna , Wohlan Katharina , Kapadia Chiraag , Yang Yaling , Yin C. Cameron , Li Shaoying , You M. James , Cheng Xiaodong , Zhou Yubin , Goodell Margaret , Huang Yun Nancy
Published: December 2023

Aging is one of the major negative factors of normal hematopoiesis, characterized by a decline in the self-renewal capabilities of hematopoietic stem and progenitor cells (HSPCs), alongside a myeloid lineage bias, thereby sabotaging the immune system and increasing the risk of malignant transformation. Epigenetic deregulation is a hallmark feature representative of aging and significantly contributes to age-related dysfunction in multiple systems. DNA methylation drifting is commonly observed during aging, especially within the partially methylated domains (PMDs). PMDs are intriguing heterochromatin regions featured by their late-replicating, lamina-associated, and B-compartment localized properties in HiC analysis. While heterochromatin is generally considered devoid of genes, it nonetheless plays a pivotal role in maintaining genomic stability. However, the link between age-related DNA methylation loss in PMDs and heterochromatin dysfunction remains elusive.

In this study, we juxtapose the influence of aging on both wild-type (WT) and Tet2 knockout (KO) HSPCs. Tet2 depletion was found to counteract the functional deterioration typically associated with aging by preserving robust self-renewal and repopulation capacities in aging HSPCs. This resistance to functional decay may account for the age-related clonal hematopoiesis observed in Tet2KO HSPCs. At a molecular level, we identified distinct epigenetic regulatory mechanisms mediated by Dnmt3a and Tet2 at heterochromatin. These mechanisms are associated with alterations in 3D genome architecture during HSPC aging. Age-related dysregulation of heterochromatin leads to the upregulation of endogenous retroviruses (ERVs), which subsequently activate intracellular innate immune response and contribute to the functional decline of aging HSPCs. The application of reverse transcriptase inhibitors was shown to suppress ERV production and interferon-stimulated genes (ISGs) expression, thereby ameliorating age-related defects in aged HSPCs. Our findings provide compelling evidence supporting the intricate interplay between DNMT and TET in regulating DNA (de)methylation equilibrium in heterochromatin and euchromatin during stem cell aging. Moreover, our study offers critical insights into the mechanisms underlying age-related heterochromatin dysfunction, which contributes to the functional deterioration of HSPCs during aging.
Safety and Efficacy of GPRC5D CAR T Cell Therapy in Relapsed/Refractory Multiple Myeloma Patients
Authors: Li Shiqi , Yuan Zhongtao , Liu Lin , Li Yu , Luo Le , Chen Yingnian , Zhang Lihua , Li Guangchao , Luo Min , Zeng Kaikai , Ma Donghui , Wang Sanbin
Published: December 2023

B-cell maturation antigen (BCMA)-directed chimeric antigen receptor (CAR) T-cell therapies have shown breakthrough therapeutic effects in treating relapsed/refractory (R/R) multiple myeloma patients. However, relapses, including some that are BCMA negative, remain a challenge.

Here we have developed a second-generation GPRC5D CAR-T, composed of a GPRC5D-binding scFv (derived from a humanized rabbit monoclonal antibody), a 4-1BB costimulatory domain, and a CD3ζ signaling domain. Following pre-clinical studies, we initiated a phase 1 dose-escalation clinical trial. Lymphodepletion with fludarabine and cyclophosphamide was performed before administering 3E6, 6E6, or 1E7/kg anti-GPRC5D CAR T cells.

To date, we have enrolled a total of 7 R/R MM patients, including 3 patients with prior BCMA CAR-T treatment. All patients received autologous CAR T-cell infusions and were clinically evaluated between February 7th, 2023, and June 26th, 2023. No dose-limiting toxicities (DLT) were observed, even at the highest dose of 1E7/kg. Grade 3 or higher adverse events (AEs) were mainly hematological toxicities, such as neutropenia (6 [85.7%]), anemia (3 [43%]), and thrombocytopenia (3 [43%]). Cytokine release syndrome occurred in 6 (85.7%) of 7 patients, with all cases classified as grade 1 or 2. Notably, no neuro-toxicities were observed in any of the seven patients.

The overall response rate was 85.7% (6/7 patients), comprising 3 complete responses or better, 3 partial responses, and one no-response (NR). The NR patient had previously shown no response to BCMA CAR-T therapy, with FACS analysis confirming the absence of both BCMA and GPRC5D on their tumor cells. Interestingly, the other two patients previously treated with BCMA CAR T-cell therapy, one reached complete response (CR), and the other reached partial response (PR). Remarkably, one patient diagnosed as COVID-19 positive two days before CAR-T infusion still received the CAR-T cell product, which led to a dramatic in vivo expansion of CAR-T cells, resulting in CR within one month after cell infusion.

In conclusion, GPRC5D CAR T-cell therapy demonstrated promising clinical efficacy and tolerable safety in patients with R/R MM. For those who relapsed after anti-BCMA CAR T-cell therapy, GPRC5D CAR T-cell therapy offered an effective option (NCT05739188).“
A Phase 1 Study to Assess the Effect of Supratherapeutic Doses of Iptacopan on Cardiac Conduction Parameters and Safety
Authors: Schmouder Robert , Kaetterer Noemi , Kulmatycki Kenneth , Nidamarthy Prasanna Kumar
Published: December 2023

Introduction: Iptacopan is a first-in-class, oral, selective inhibitor of factor B, a key component of the complement system alternative pathway. Inhibition of the complement system represents a potential mechanism for treatment of several diseases including paroxysmal nocturnal hemoglobinuria and atypical hemolytic uremic syndrome. The purpose of this randomized, participant-blinded, placebo-controlled phase 1 study was to assess the effect of supratherapeutic doses on the exposure-response relationship of iptacopan concentration and cardiac conduction, and to assess the cardiac safety and tolerability of ascending, supra-therapeutic single doses of iptacopan.

Methods: Supratherapeutic doses of iptacopan 400, 800, and 1200 mg were included in this study, representing ≤6-fold higher dose than the planned clinical dose of 200 mg. The primary endpoints were change from baseline in Fridericia-corrected QT interval (QTcF) and safety endpoints. The analyses included data from this study pooled with part 1 data (single dose) from the first-in-human (CLNP023X2101) study (n=86). The study population included healthy men and women aged 18-55 years. The relationship between iptacopan plasma concentration and change from baseline in QTcF was estimated using a linear mixed effects model.

Results: Thirty-two patients were randomized and received placebo (n=8) or iptacopan (n=24). The median age of participants in the 400-mg, 800-mg, 1200-mg, and placebo groups was 51.5, 35, 36, and 37 years, respectively. The upper bound of the 2-sided 90% CI for linear mixed effects model-derived placebo-adjusted change from baseline in QTcF at the geometric mean C max was 2.82 ms for 400-mg, 2.94 ms for 800-mg, and 3.22 ms for 1200-mg doses. Hence, there was no evidence of clinically meaningful QTc prolongation ( Table). The exposure-response analysis of PR and QRS intervals showed no iptacopan concentration-related effects. While the pre-specified and more simple exposure-response analysis of heart rate using change from day 1 baseline (0 hour) revealed an increase of 3 to 5 beats per minute, in a post hoc analysis using 24-hour, time-matched comparison for each participant at each time point before and after study treatment (the more common and gold-standard analysis), iptacopan had no effect on heart rate. No deaths, no moderate or severe treatment-emergent adverse events, and no treatment cohort discontinuations due to adverse events were reported.

Conclusions: Three supratherapeutic doses were well tolerated and were not associated with any relevant change in QTcF. Iptacopan did not affect PR or QRS intervals and a time-matched post hoc analysis showed no effect on heart rate. These results indicate that the anticipated clinical dose of iptacopan (200 mg twice daily) has no adverse clinical effects on cardiac function.
Characterizing the Immature Immunophenotype of Sickle Cell Disease Monocytes during Vaso-Occlusive Crisis
Authors: Gingell Luke , Hrinczenko Borys
Published: December 2023

Sickle cell disease (SCD) is marked by episodic vaso-occlusive crisis (VOC). Painful VOC is the leading cause of emergency room visits and hospitalization in SCD, contributing heavily to the morbidity and mortality of this disease. Recurrent VOC creates a pro-inflammatory state that induces phenotypic alterations in the innate immune system. This inflammation-induced immune profile remodeling results in chronically active monocytes and neutrophils that antagonizethe vascular endothelium and further predispose SCD patients to VOC. While the mechanism of neutrophil involvement in VOC has been elucidated, less is known about the pathologic role of monocytes. Monocytes are of particular interest to VOC pathophysiology because they are especially malleable to inflammatory signaling. Indeed, inflammatory disease states such as COPD, obesity and atherosclerosis are known to influence monocyte development and alter monocyte subpopulations. Recently, nonclassical monocytes (CD14 dimCD16 +) with high heme oxygenase-1 expression have been shown to have a critical role in preventing VOC, generating renewed interest in monocytes as a therapeutic focus in SCD. In this study, we describe the monocyte subsets found in SCD patients during VOC by performing immunophenotypic flow cytometric, enzymatic, and morphologic analysis on peripheral blood obtained during hospitalization for VOC. Herein, we add to the growing body of evidence suggesting aberrant monocyte populations underpin VOC pathophysiology. We found that SCD monocytes during VOC possess an immature phenotype as demonstrated by 1) decreased CD4 positivity (p < .01), 2) low α-naphthyl butyrate esterase (ANBE) expression, and 3) naïve morphologic features (high nuclear/cytoplasmic ratio, indented or less lobular nuclei, and decreased cytoplasmic vacuolization). We additionally found an increase in CD14 +CD16 -CD4 - monocytes (p < .01), a subset associated with the impaired immune response of post-trauma patients. Interestingly, we also found a large proportion of CD14 +CD4 -HLA-DR - monocytes which, under normal circumstances, are exclusively found in neonates (p < .01). Finally, we report an increase in nonclassical monocytes (CD14 dimCD16 +), providing further evidence for the connection between this subset and the molecular mechanisms driving VOC. Taken together, these findings suggest that SCD patients in VOC deploy relatively immature monocytes that possess a lower capacity for antigen presentation, likely contributing to the immune dysfunction observed in SCD.
Delineating the Functions of Cohesin Stag1 during Terminal Erythroid Differentiation
Authors: Adams Sarah , Dhara Anita , White Rachael , Prabahar Archana , Jiang Peng , Gnanapragasam Merlin Nithya , Ramamoorthy Mahesh
Published: December 2023

Erythroid terminal differentiation is defined by a series of precise, orchestrated and dynamic changes in the genomic architecture that is consistent with the transcriptional programing towards producing mature blood cells. In recent times, these architectural shifts are under scrutiny, not only because they shed light into the mechanisms underlying erythroid differentiation, but also because corruption of the organization principle can result in compromised hematopoietic function and tumor formation. Functionally, genome organization relies upon the concerted action of proteins such as the cohesion complex and CTCF. In this study, we identify Stag1, a cohesin protein, as a factor that contributes to the erythroid transcriptional machinery during terminal differentiation. Stag1 levels increase during terminal differentiation and depletion of Stag1 either using short hairpin RNA or using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR-Cas) in Hudep2 or CD34+ cells, results in a drastic reduction of both adult globins, α- and β-globin. This is not the case when Stag1's paralogue, Stag2, is knocked-down. Further, we observe faster proliferation in erythroid cells after Stag1 depletion and these cells also demonstrate an undercondensed nucleus and a larger cell-size. This could reflect the delayed differentiation, as the cells proceeded to undergo slower differentiation as compared to their WT counterparts. Metaphase spreads of the Stag1 depleted cells suggests an altered cohesion phenotype and further provides clues into a possible epigenetic defect underlying the observed phenotypes in erythroid cells, which is being further investigated.
Prior Daratumumab Exposure and It's Correlation to Parameters Related to Apheresis Prior to Hematopoietic Stem Cell Transplantation in Multiple Myeloma
Authors: Unis Graham D , Gudiel Cesia , Philon Ernest , Finn Laura E.
Published: December 2023

A number of recent studies have brought to light the potential detrimental effect the biologic monoclonal antibody daratumumab, directed towards CD38, may have on the collection of stem cell collection for hematopoietic stem cell transplantation (HSCT). In our evaluation of this claim, we retrospectively reviewed multiple myeloma patients who underwent autologous HSCT at a major tertiary transplant center between 2017 and 2022. A total of 240 patients were identified which included 62 patients who received daratumumab prior to stem cell collection and 178 patients who had no prior daratumumab exposure. Basic demographic data was collected as well as radiation, chemotherapy, and smoking history. No significant differences were identified between the groups based on demographics, smoking, or radiation exposure. Parameters related to peripheral stem cell collection were identified and recorded. We found that in patients exposed to daratumumab prior to apheresis, the mean number of days of apheresis required to reach the target collection goal was statistically increased compared to control (1.42 days ± 0.07 vs 1.26 ± 0.03; p<0.05). Furthermore, medications used to increase peripheral stem cell collection yield were also increased. It was demonstrated that the mean number of doses of plerixafor and G-CSF analogs were increased in the daratumumab group vs the control (1.08 ± 0.12 vs 0.6 ± 0.05; p<0.001 and 5.98 ± 0.31 vs 5.17 ± 0.06; p <0.001 respectively). Total number of stem cells collected both as an absolute number and controlled for the number of days of apheresis were lower in the daratumumab group compared to the control (5.31x10^6 ± 0.31 vs 6.68x10^6 ± 0.20; p<0.05 and 4.16x10^6 ± 0.32 vs 5.84x10^6 ± 0.21; p < 0.001 respectively). These results suggest that pre-apheresis exposure to daratumumab has a detrimental effect on peripheral stem cell collection. These results have a significant implications for both long term patient care and healthcare system resource utilization. The clinical outcome implications of daratumumab are currently not well known and further study is needed to determine if alterations in stem cell collection protocols would result in improved stem cell collection yields for patients with pre-collection daratumumab exposure.

All data is expressed as mean ± SEM
Bendamustine Bridging Versus Standard of Care Chemotherapy on Toxicity Profile for Multiple Myeloma Patients Undergoing CAR T Cell Therapy
Authors: Leyfman Yan , Rajeeve Sridevi , Sheng Tianxiang , Diniz Marcio , Richard Shambavi , Jagannath Sundar , Rossi Adriana C
Published: December 2023

Introduction

Multiple myeloma (MM) remains an incurable malignancy arising from plasma cells, however great promise has been seen with the use of Chimeric Antigen Receptor (CAR) T cells. Autologous CAR-T products require bridging therapy, however little is known about different agents used in this manner. Some studies indicate decrease in disease burden is crucial, while others note an association with higher rates of toxicity and negative effect on survival. As commercial CAR T cell therapy is indicated after failure of at least 4 prior treatments, careful consideration of bridging therapy is crucial, particularly in patients with aggressive and high-risk disease. This study aims to compare the impact of bendamustine to standard of care (SOC) bridging therapy in MM patients undergoing CAR T cell therapy.

Methods

A single-center retrospective chart review of 19 adult patients with relapsed/refractory multiple myeloma treated with CAR T cells, who also received bridging therapy with either bendamustine (n=9) or SOC/Other (n=10) was conducted at Mount Sinai Hospital. Patient demographic information (gender, race, MM subtype, history of prior transplant) was collected and toxicity profiles (infection, pancytopenia, cytokine release syndrome (CRS), and immune effector cell-associated neurotoxicity syndrome (ICANS)) were compared. Descriptive statistics were used to analyze demographic characteristics and toxicity profiles between the two cohorts.

Results

Comparative analysis of demographics showed that both cohorts were predominantly males with IgG MM subtype, and varied in terms of race ( Figure 1a). The Other cohort had a higher proportion of patients with a history of autologous stem cell transplant (ASCT) (70%) compared to the bendamustine group. In the SOC group, two patients developed infections, whereas none were observed in the bendamustine cohort ( Figure 1b). Pancytopenia was observed in both groups, but interestingly, 100% of patients experienced it in the SOC cohort. CRS and ICANS were comparable between the two groups ( Figure 1b). Comparative statistics demonstrated no significant difference between the two groups ( Figure 1c).

Conclusion

Our study findings indicate that there was no significant difference in toxicity profiles between the two pre-conditioning therapy groups. Given our limited sample size, future studies with a larger are indicated to validate these results.
Screening Cerebrospinal Fluid for Aggressive B-Cell Lymphoma: Significance of Clonal Populations Found with 8-Color Flow Cytometry
Authors: Ho Carrie I , Lopedote Paolo , Lin Olivia , Fromm Jonathan R , Gopal Ajay K , Milano Filippo , Smith Stephen D
Published: December 2023

Background: Secondary central nervous system (CNS) lymphoma confers a poor prognosis, and prompt identification is critical for guiding management. Over the last decade, we have employed cerebrospinal fluid (CSF) analysis with 8-color flow cytometry (FCM) in screening patients deemed at risk of CNS relapse. However, there is little data to guide interpretation and management of positive results, and concurrent clinical and radiographic features of FCM positive (FCM+) patients, remain areas of uncertainty.

Methods: We analyzed newly diagnosed (ND) and relapsed/refractory (R/R) diffuse large B cell lymphoma (DLBCL) and high grade B cell lymphoma (HGBCL) patients who underwent a screening lumbar puncture (LP) from 2011-2020. Clinical, imaging, and CSF findings (differential, cytology, FCM) were collected. Patients with prior CNS involvement or FCM showing CLL/low-grade B-NHL were excluded. “FCM-only” patients were defined as FCM+ but without any other evidence of CNS spread (negative cell count/cytology, MRI, and no symptoms), whereas others were deemed to have overt CNS disease.

Results: There were 415 LPs among 361 patients (243 ND, 172 R/R). 21 (5%) patients were FCM+, 10 ND and 11 R/R patients (Table 1). Four had FCM-only involvement (2 ND, 2 R/R). Among these, the clonal population was 1.63% of WBC (range, 0.3-2.7%), and 3 had concurrent bone marrow (BM) or peripheral blood (PB) involvement. All 4 received intrathecal (IT) therapy. At median follow up of 23 months, 3 (75%) have died (2 from CNS disease progression, 1 unknown due to loss in follow up), and 1 remains alive in remission 44 months later following IT methotrexate (MTX) and CAR T-cell therapy.

17 had overt CNS disease (8 ND, 9 R/R). The clonal population was 9% (range, 0.2-98%). Eight (47%) had concurrent BM or PB involvement. Twelve (70%) received IV high dose MTX, and the remainder received only IT therapy. At median follow up of 42 months, 9 (53%) have relapsed (2 CNS only, 1 systemic, 6 both), and 9 have died. Median time to CNS relapse was 5 months.

Conclusions: CSF 8 color FCM screening of DLBCL/HGBCL patients yields a low proportion of positive results, mostly in the setting of overt CNS disease. However, even FCM-only patients had poor outcomes (2/4 with CNS progression), suggesting that blood contamination does not explain CSF findings, and that these patients should be approached as secondary CNS involvement. Better pt selection for CSF screening is needed to reduce unnecessary LPs. Intensified or novel approaches for all FCM+ patients are needed.
Flumatinib for Newly Diagnosed Chronic Phase Chronic Myeloid Leukemia: An Open-Label, Multi-Center Study
Authors: Wen Ziyu , Zhao Ying , Li Guowei , Su YongZhong , Chen FeiHeng , Tao HongFang , Zheng ZhengJin , Lin Wuqiang , Chen XiuLi , Meng Meili , Yu Lian , Chen LongTian , Huang JianQing , Wu Guocai , Zhao Mengxia , Liu Zhenfang , Zhu Hongqian , Jiang Yirong , Liu Qifa , Liu Xiaoli , Xu Na
Published: December 2023

Background Tyrosine kinase inhibitors (TKIs) ushered in the era of targeted therapy for chronic myeloid leukemia(CML), which greatly improved the prognosis of patients with CML. Flumatinib is a second-generation tyrosine kinase inhibitor (TKI), which is currently approved for patients with Philadelphia chromosome-positive chronic myeloid leukemia (Ph+ CML) in China. And some current clinical trials revealed that flumatinib has been shown to be a more potent inhibitor of BCR::ABL than imatinib.

Methods In this open label, single-arm, multicenter study, 127 patients with newly diagnosed chronic-phase chronic myeloid leukemia (CP-CML) treated with flumatinib were enrolled. The primary endpoint of this study was early molecular response (EMR) rates at 3 months. The secondary were the rates of complete cytogenic response (CCyR), major molecular response (MMR) at 3, 6, 12 months and treatment-related adverse events (AEs). The trial is registered with ClinicalTrials.gov NCT04591197.

Results From October 2020 to December 2022, a total of 158 patients with CP-CML were assessed for eligibility, of whom 127 were followed up for more than 3 months. 79(62%) patients were men and 48 (38%) were women. The median age was 47 years (range, 19-82 years), The median time from diagnosis to treatment was 6 days (0-176 days) and the median follow-up was 9.7 months (range 3.1-26.6 months). Of evaluable 127 patients, EMR was achieved by 85% (95% confidence interval [CI], 77.6%-90.7%) of patients at 3 months. After the initiation of flumatinib treatment, the incidence of MMR at 3, 6, and 12 months was 24%, 59%, and 73%, respectively. Patients receiving flumatinib had higher rates of CCyR at 6 months and MMR 12 months than the data from DASISION (dasatinib) and ENESTnd (nilotinib) therapy (85% vs 73% vs 67% and 73% vs 46% vs 44%). Progression-free survival (PFS) rate at 1-year was 97.2%. Diarrhea (34%) were the most commonly reported adverse events. Most treatment-related adverse events were grade 1/2. None of the patients discontinued or died due to AEs.

Conclusion Flumatinib as first-line therapy in patients with newly diagnosed CP-CML was not inferior to nilotinib and dasatinib and well tolerated.
Provisional Prognostic Score for Plasmablastic Lymphoma
Authors: Haddad Philip A , Lerner Jaren
Published: December 2023

Background

Plasmablastic Lymphoma (PBL) is a rare and aggressive subtype of non-Hodgkin's lymphoma. It tends to be associated with immunosuppressive clinical contexts, such as HIV infection and immunosuppressive therapies for autoimmune disorders and organ transplants. However, due to the rarity of the disease, which needs to be better understood, varied treatment approaches have been implemented, and no optimal data or guidelines for managing PBL are available. Therefore, we conducted this study to develop a provisional PBL prognostic score (PBLPS) for this rare disease.

Methods

We used our constructed PBL database, which contains retrospective data on 300 cases. Such data included demographics such as sex, age, and race. It also included disease presentation symptoms, duration of symptoms before diagnosis, site(s) of the disease, blood counts, coexisting comorbidities, disease immunohistochemical and molecular phenotype, types of treatment, and survival outcomes. Of the 300 cases, 291 had complete survival and outcomes data, the sample chosen for this study. Cox proportional-hazards model and Log-rank tests were used to assess the influence of clinicopathologic factors on overall survival (OS). We included factors that statistically impacted OS and scored them by the impact of their hazard ratios.

Results

The median OS of the cohort was 25 months. The following dichotomous variables were identified as impactful prognostic factors in this cohort: LDH>500 (p=0.04), CD4<100 (p=0.006), bone marrow (BM) involvement (p<0.0001), EBER-negative (p=0.01), HHV8+ (p=0.02), and involvement of liver (p=0.003), lungs/pleura (p<0.0001), and upper GI tract (p=0.001). A prognostic model was devised using these variables to identify different levels of risk. Involvement of BM and involvement of any of the worse prognosis organs (liver, lung/pleura, and/or upper GI tract) were each assigned a score of 2 when present. CD4<100, EBER-negative, and LDH>500 were assigned 1 point each as they had the lowest hazard ratios of all the other variables. In this exploratory cohort, we assigned low risk a score of 0, intermediate risk a score of 1-2, and high risk a score of 3-7. This prognostic score system led to our cohort's most optimal risk discriminatory model, where low, intermediate, and high risk had a median OS of NR, 15, and 4 months, respectively (p<0.0001).

Conclusion

This PBLPS is a promising new tool for risk-stratifying patients with PBL. However, it still requires prospective validation.
Inhibition of CDC20 Suppressed the Development and Progression of Mantle Cell Lymphoma
Authors: Chen Yingtong , Yang Ping , Wang Jing , Gao Shuang , Ke Xiaoyan , Jing Hongmei
Published: December 2023

Background: Cell division cycle 20 homologue (CDC20) is an important factor regulating the cell mitosis phase. CDC20 is highly expressed in several types of cancers, which is related to the clinicopathological characteristics and prognosis of cancer patients. This study aimed to explore whether CDC20 could affect the development and progression of mantle cell lymphoma (MCL) and the underlying mechanism.

Methods: The expression level of CDC20 was detected in peripheral blood mononuclear cells (PBMCs), bone marrow mononuclear cells (BMNCs) and pathological tissues of MCL patients and MCL cell lines (Z138, Mino, Rec1). The CDC20 inhibitor apcin was used to evaluate the effect of CDC20 on the cell phenotypes of MCL cells. The changes in cell proliferation, apoptosis, cell cycle, cell migration and invasion were determined using CCK-8, flow cytometry, and Transwell assays, respectively. The anti-tumor effect of apcin were investigated in vivo in the Z138-driven xenograft tumor model. RNA-seq was performed to explore the pathways that changed in MCL cells after apcin treatment, and then these possible changes were verified in vitro and in vivo by western blot.

Results: CDC20 was overexpressed in PBMCs, BMNCs, and pathological tissues of MCL patients and MCL cells compared with their respective control group. In Z138, Mino, and Rec1 cells, apcin could inhibit cell proliferation, migration, and invasion, promote cell apoptosis, and induce cell cycle arrest. Moreover, apcin confirmed its safety and anti-tumor activity in vivo. In apcin-treated MCL cells and mice, the PI3K/AKT pathway was significantly inhibited (Figure 1).

Conclusions: This study confirms the essential role of CDC20 in MCL tumorigenesis. Anti-tumor therapy targeting CDC20 is expected to be a powerful treatment strategy for MCL patients.
Decitabine in Combination with Aclacinomycin, Cytarabine and Granulocyte Colony-Simulating Factor (CAG) Had a High Response Rate in Children with Advanced Myelodysplastic Syndrome and Acute Myeloid Leukemia with Myelodysplasia-Related Changes
Authors: Ren Yuanyuan , Chen Yumei , Wang Shuchun , Liu Tianfeng , Wan Yang , Zhang Jingliao , Yang Wenyu , Guo Ye , Zhu Xiaofan
Published: December 2023

Advanced myelodysplastic syndrome (MDS) is a rare disease in children with poor prognosis. Allogeneic transplantation is the only curative option. However, there has been no established treatment prior to transplant. We conducted a retrospective study exploring the response rates and safety of decitabine combined with aclacinomycin, cytarabine and granulocyte colony-stimulating factor (CAG) for induction treatment of advanced MDS and acute myeloid leukemia with myelodysplasia-related changes (MDR-AML) in children. Patients from 1 to 14 years administrated in Pediatric Blood Disease Center of CAMS Blood Disease Hospital between January 1the 2019 and December 31th 2022 were screening. All diagnosed with MDS-EB or MDR-AML according to WHO 2016 criteria and treated with decitabine combined with CAG were identified. The efficacy of one-cycle treatment of MDS-ED was evaluated based on The International Working Group Response Criteria 2006. The response for MDR-AML was evaluated according to the criteria for AML published by ELN. The response of MDR-AML was defined as complete remission (CR) or CR with incomplete count recovery (CRi). The response of MDS-EB was defined as CR or bone marrow CR with hematological improvement (HI). A total of 13 patients, including MDS-EB (n=7) and MDR-AML (n=6), were enrolled for analysis. 5-day decitabine was administrated, combined with modified CAG regimen for 7 days. Eventually 12 patients of those were available for evaluation. For patients with MDS-EB, four achieved CR and three reached marrow CR with HI. Four patients with MDR-AML, three achieved CR and two reached CRi. The overall response rate was 100%(12/12). The most frequent adverse events were myelosuppression and suppression-induced infections, but no infection-related mortality was recorded. There had been no cardiotoxic event happened, either. Only one patient experienced fatal bleeding in early stage after treatment. The median follow-up time was 16 months (range 1-48). At last follow-up, 10 patients who had underwent allo-HCT were alive without disease. The remaining 2 patients died of disease progression. The 2-year estimated overall survival was 62.9%. Decitabine combined with modified CAG had a high response rate for children with advanced MDS and MDR-AMl, and it can bridge patients to curative allogeneic transplantation.