bluuebunny/crossref_metadata_2025_split huggingface dataset:
Azacitidine Combination with Lenalidomide in Treating Higher-Risk Myelodysplastic Syndromes: A Single-Center, Phase 2 Trial
Authors: Song Dandan , Ge Zheng 
Published: December 2023

Background: Azacitidine, which works through the proteosomic destruction of DNA methyltransferase and resultant chromatin decondensation, is used primarily in patients with higher-risk myelodysplastic syndromes (MDS). Lenalidomide has been approved by the US Food and Drug Administration for the treatment of MDS patients with del(5q) cytogenetic abnormality. It has also shown activity in patients with higher-risk MDS and acute myeloid leukemia. In this trial, we assessed the efficacy and safety of combining azacytidine and lenalidomide in higher-risk MDS.
Methods: This is a single-center, phase 2 trial. Aza (75mg/m 2/d on days 1-7 of a 28-day cycle) was administered in combination with lenalidomide (10 mg/day on days 1 to 21 every 28 days) for patients. The primary endpoint was the overall response rate (ORR). Secondary endpoints were overall survival (OS), progression-free survival (PFS), and adverse events (AEs). An MDS targeted-exome-seq panel including 37 driver genes was used for screening.
Results: Between Oct 2020 to Apr 2023, a total of 41 MDS patients were enrolled. 32 patients were evaluable for efficacy. 23 (71.875%) responded to therapy: 7 achieved a complete response (CR) (21.875%), 12 a marrow CR (mCR) (37.5%), and 4 a hematologic improvement (HI) (12.5%), of whom 10 of the mCR patients also had a HI. The median follow-up was 5.6 months (1 to 27.4), the median overall survival (OS) was not reached, and the median progression-free survival was 12.2 months. The time to reach CR was 2 months (range, 1 to 4.5), and the median CR duration was 8+ months (range, 5 to 27.4) ( Table 1). 27 patients had at least one mutation, the most common being TET2 (n = 12), ASXL1 (n = 12), TP53 (n = 7), SRSF2 (n = 6), and RUNX1 (n = 6) ( Fig.1A). Compared with patients without mutations, CR/mCR rate was significantly higher for those with mutations in TP53 (100% v 48%; P =0 .025) ( Fig. 1B), ORR were numerically higher for those with TP53 (100% v 64%; P = 0.149) and TET2 (85% v 63%; P = 0.422) ( Fig. 1C). We also observed a reduction of VAF in TP53 and TET2 mutation after the combination treatment ( Fig. 1D & 1E). Common grade 3/4 nonhematologic adverse events included thrombocytopenia (18.75%), infection (12.5%), and febrile neutropenia (9.4%).
Conclusion
This trial demonstrated that the combination of azacytidine with a lenalidomide regimen is an effective therapy for higher-risk MDS patients, with high efficacy and well-tolerance. Patients with TP53 mutation appear more likely to achieve a CR/mCR.
Dasatinib and CAR-T Cell Therapy for Newly Diagnosed Ph-Positive Acute Lymphoblastic Leukemia in Adults
Authors: Zhang Mingming , Hu Yongxian , Wei Guoqing , Fu Shan , Feng Jingjing , Hong Ruimin , Chang Alex H. , Huang He 
Published: December 2023

Introduction
The use of tyrosine kinase inhibitors has improved outcomes of Ph-positive Acute lymphoblastic leukemia (ALL), but combinations with other agents are needed to achieve molecular response. CAR-T cell therapy has made a breakthrough in refractory or relapsed ALL, but has not yet been applied in newly diagnosed patients.
Methods
We conducted a phase 2 single-group trial of first-line therapy in adults with newly diagnosed Ph-positive ALL. Dasatinib in combination with a two-week vincristine and glucocorticoids regimen were administered, followed by sequential infusions of CD19 and CD22 CAR-T cells. All patients received single-agent dasatinib maintenance after CAR-T cell therapy. The primary endpoint was complete molecular response (defined as undetectable BCR/ABL1 transcript with a sensitivity of 0.001%) in the bone marrow after this treatment. Here we report initial results from this study.
Results
At the cut-off date of 31 May 2023, 18 patients have received CD19 CAR-T cell infusions, and 14 of these patients have received subsequent CD22 CAR-T cell infusions. At the end of induction therapy, the complete hematologic remission rate was 100%, and 27.8% (5/18) of the patients had a complete molecular response. The percentage of complete molecular response increased to 72.2% (13/18) after CD19 CAR-T cell therapy, and increased further to 76.9% (10/13) after CD22 CAR-T cell therapy (1 patient has not been evaluated). Notably, of the 5 patients who did not achieve complete molecular response after CD19 CAR-T cell therapy, 4 have received CD22 CAR-T cell therapy, of which 1 eventually achieved complete molecular response and 2 experienced relapses. One patient had a CD19+ CD22+ relapse accompanied by mutations in BCR/ABL1 (F317L and Y253H) and the other patient had a CD19dim CD22+ relapse. After a median follow-up of 13.5 months, 16 patients remained in complete hematologic remission, and 14 patients remained in sustained complete molecular response. No patient received allogeneic stem cell transplantation. No grade 3 or higher cytokine release syndrome or Immune effector cell-associated neurotoxicity syndrome was observed during CAR-T cell therapies.
Conclusions
Dasatinib in combination with CAR-T cell therapy has enabled chemotherapy-free treatment in newly diagnosed Ph-positive ALL. This treatment is characterized by high complete molecular response, high long-term survival, low toxicity and short treatment cycles.
IL-27 Promotes Neutrophil Extracellular Trap Generation in Sickle Cell Disease
Authors: Alagbe Adekunle Emmanuel , Novelli Enrico M , Sundd Prithu 
Published: December 2023

Introduction: Neutrophil Extracellular Traps (NETs) are neutrophil-derived damage-associated molecular patterns (DAMPs) composed of decondensed chromatin decorated with citrullinated histones and neutrophil proteases. Recently, we have shown that caspase-4-dependent activation (cleavage) of the pore-forming protein, Gasdermin-D (GSDMD) in neutrophils promotes NETs generation leading to lung injury in SCD. IL-27, an anti-inflammatory cytokine with some pro-inflammatory properties is significantly elevated in the plasma of SCD patients, however, it remains debatable whether IL-27 signaling in neutrophils serves to promote or inhibit NETs generation in SCD.
Methods: Citrated blood samples from SCD or race matched control humans were incubated with saline, hemin (20 mM), or hemin (20 mM) + recombinant human IL-27 (200 ng/mL) for 15 minutes at room temperature. Platelet-poor-plasma from the three treatment groups were incubated with sytox green and fluorescent Abs against neutrophil elastase (NE) and histones for in situ staining of extracellular DNA, NE and histones, respectively. Imaging flow cytometry was used to detect circulating NETs (cNETs) as 1-2 mm fragments triple-positive for extracellular DNA, NE, and histones.
Results: Incubation with hemin led to significant increase in cNETs concentration in SCD but not control human blood, which was further increased following incubation of SCD human blood with hemin in presence of IL-27.
Conclusion: Our current findings suggest that IL-27 signaling promotes NETs generation by neutrophils in SCD patient blood. Currently, studies are underway to identify the molecular mechanism underlying IL-27-dependent NETs generation in SCD.
RAD51 Is Required for the Occupancy of Replication Licensing Factors on the Myeloma Genome
Authors: Zhao Jiangning , Talluri Srikanth , Kumar Subodh , Morelli Eugenio , Potluri Lakshmi B. , Shammas Masood A. , Munshi Nikhil C 
Published: December 2023

Genomic instability and extended replicative potential arise early during oncogenic transformation and contribute to the development and progression of cancer. Although chromosomal instability and replication are distinct pathways, they share certain proteins and activities. DNA replication starts by sequential loading of replication licensing factors at replication origins. We have observed that RAD51 - a keystone protein in stabilizing the genome through homologous recombination (HR) and a dependency in multiple myeloma (MM) - interacts with the MCM complex. In fact, RAD51 knockdown inhibits DNA replication in MM cells. Because RAD51 organizes the initial steps of homologous recombination, we investigated whether RAD51 plays a similar setup role in replication - namely, that RAD51 is required for the occupancy or proper assembly of replication licensing factors (ORC1-ORC6, CDC6, CDT1, MCM2-7) on the MM genome.
To investigate this, we either inhibited (by knockdown or inhibitor) or overexpressed RAD51 in MM cells and monitored the occupancy of the replication complex on genomic DNA using ChIP with an antibody against MCM4. Subsequently, the proteins in the complex were released from DNA and identified by Western blotting. Although RAD51-KD (or inhibition) did not reduce the expression or stability of licensing factors, the occupancy of all these factors (MCM4, MCM2, MCM5, MCM6, MCM7, CDC6) on the MM genome was inhibited. Consistently, RAD51-overexpression led to an increase in the genomic occupancy of most of the licensing factors (ORC1, ORC2, MCM4, MCM2, MCM5, MCM6, MCM7, CDC6) tested. This was validated using antibodies against other members of the complex i.e., ORC1 and CDC6.
Overall, we demonstrate that RAD51 in MM cells interacts with components of the replicative protein complexes (ORC1-ORC6, CDC6, CDT1, MCM2-7) that license and ensure DNA replication once per cell cycle. Our data also demonstrate that RAD51 is required for the occupancy of most of these factors on genomic DNA in MM cells, which may explain its ability to impact DNA replication in MM cells. We are currently trying to understand if RAD51 brings one or more of these factors to the origin of replication and if some of its activities (such as ATPase, DNA binding) are involved in loading these factors onto DNA. Based on its impact both on replication and HR, RAD51 is an attractive target in myeloma. Our inhibitor studies provide further rationale to study therapeutic strategy to target RAD51 in MM.
Phenotypic and Proteomic Characterization of the Human Erythroid Progenitor Continuum Reveal Dynamic Changes in Cell Cycle and in Metabolic Pathways
Authors: Papoin Julien , Yan Hongxia , Leduc Marjorie , le-Gall Morgane , Narla Anupama , Steiner Laurie A. , Gallagher Patrick G , Hillyer Christopher D. , Gauthier Emilie-Fleur , Narla Mohandas , Blanc Lionel 
Published: December 2023

Human erythropoiesis is a complex process leading to the production of 2.5 million red blood cells per second. Following commitment of hematopoietic stem cells to erythroid lineage, this process can be divided into two distinct stages: erythroid progenitor differentiation and terminal erythropoiesis. We recently resolved the heterogeneity of erythroid progenitors in human bone marrow into four different subpopulations termed EP1 to EP4. Here, we characterized the growth factor(s) responsiveness of the progenitor populations in terms of proliferation and differentiation. The four progenitor populations exhibited differential dose-dependent responses to SCF and EPO, while their growth and differentiation was IL-3 independent. Using mass spectrometry-based proteomics, we quantified the expression levels of ~5,500 proteins from EP1 to EP4 (Figure 1). Our functional analyses highlighted dynamic changes in cell cycle in these populations with an acceleration of the cell cycle during progenitor differentiation. The cell cycle is controlled at multiple levels; one of them being the G1 to S transition. Using EdU and DNA content to resolve different phases of the cell cycle, we noted a decrease in the percentage of cells in the G1 phase and a concomitant increase in S phase as EPs differentiated from EP1 to EP4. While the proportion of cells in G2/M progressively increased from EP1 to EP3, a significant decrease was noted as EP3 transitioned into EP4. Finally, by measuring the mean fluorescence intensity (MFI) of the S-phase, we found that the S phase speed increased from EP1 to EP4. Mechanistically, E2F4 has recently been identified in fast-cycling cells and in murine erythropoiesis, and we noted a specific 3-4-fold increase in E2F4 protein copy numbers from EP1 to EP4 (Figure 2), along with its binding to cell cycle-related genes at the EP3 stage, suggesting a potential role for this E2F member in the regulation of their proliferation.
Finally, we assessed the relative contributions of oxidative phosphorylation and glycolysis required to provide ATP to these fast-cycling cells. Both oxidative phosphorylation and glycolysis provide ATP needed for erythroid progenitors to proliferate and differentiate towards terminal stages.
Taken together, our data provide comprehensive insights into normal human erythroid progenitor biology and have implications for understanding ineffective erythropoiesis due to erythroid progenitor defects.
Use of Single-Arm Trials in FDA Approvals of Treatments in Relapsed or Refractory B-Cell Lymphoma
Authors: Zou Denise , Zhang Eileen , Wu Sherry , Rose Virgil 
Published: December 2023

Background: Single-arm trials are commonly used to evaluate therapies targeting life-threatening or rare diseases (e.g., late-stage cancers that are relapsed or refractory to prior interventions). A number of oncology products supported by single-arm trials have been approved by the United States Food and Drug Administration (FDA). The draft guidance issued by the FDA on 24 March 2023 recommended key considerations for trial design and analysis when a single-arm trial is considered appropriate to support accelerated approval. The objective of this study was to assess how often anticancer drugs approved by the FDA were based on single-arm trials in a typical relapsed or refractory indication and to evaluate the quality of trial design and data leading to approvals.
Methods: We reviewed past approvals between 2006 and 2023 listed on the FDA website, focusing on the indication of relapsed or refractory B-cell lymphoma. Of 11 approvals identified, eight were supported by single-arm trials and three were based on randomized controlled trials. Approvals based on single-arm trials were further investigated, and each trial was analyzed for the following criteria as recommended by the draft FDA guidance: 1) if the endpoint was based on response rate assessed using appropriate criteria; 2) if the sample size was sufficient to provide robust estimates of efficacy and safety profile; and 3) if the magnitude and duration of response were adequate.
Results: Among the eight approvals based on single-arm trials, one was approved in 2017, one in 2018, and two each in 2020, 2021, and 2023. Three were chimeric antigen receptor T-cell (CAR-T) therapies and were granted regular approval; the other five were non-CAR-T therapies which were granted accelerated approval. The primary endpoints adopted by all the trials were overall response rates (defined as complete and partial responders) and duration of response determined by an independent review committee. The magnitude of overall response rates reported by most trials ranged from 48% to 72% (with more than 50% of them with a complete response); one trial reported an overall response rate of 29% with complete response rate of 13%. A minimal median follow-up of six months after response was achieved in all trials. The estimated median duration of response varied from 10.3 to 21.7 months, with three trials reporting medians not reached. Based on the trial description, the analysis population was pre-specified for the evaluation of efficacy, ranging from 68 to 192 participants.
Conclusion: Submissions based on single-arm trials for oncology therapies in the relapsed and refractory setting tended to be well accepted by FDA for accelerated and regular approval when the key criteria of trial design and data quality were adequately met.
From in silico to in Vitro to In Vivo: Increasing Megakaryocytes to Treat Thrombocytopenia
Authors: Macias Alejandra , Cortes Mauricio , An Na , Li Na , Fulton Timothy , Simpson Allison , Miller Samuel , Sun Sunny , Fu Doris , Garcia Juan , Medicherla Satya , Steelman Scott , Thorarensen Atli , Goddeeris Matthew , Krishnamoorthy Sriram 
Published: December 2023

Hematopoietic stem cells undergo a multi-step differentiation program through several precursor stages to give rise to megakaryocytes (Mk), which ultimately release platelets. Thrombocytopenia can manifest in various clinical settings and can adversely affect a patient's quality of life through easy bruising and risk of bleeding. An example is chemotherapy-induced thrombocytopenia (CIT) which occurs when myelosuppressive chemotherapy is given, especially with certain agents such as platinum analogs and cyclophosphamide. We used a single-cell RNA-sequencing approach to study the differentiation of hematopoietic stem cells to Mk progenitors (MkP) and delineated the trajectory that defines the transition of cells to the megakaryocytic lineage. Using our proprietary platform which pairs new approaches in computation and machine learning in drug discovery, we predicted small molecule interventions that could modulate the hematopoietic lineage direction to increase the differentiation towards MkP and subsequent increases in platelet counts. Using an in vitro 5-day CD34-based Mk differentiation assay we demonstrated that some of our predicted interventions increased generation of MkP from stem cells. Analysis of early MkP (CD34-CD235a-CD71+CD41b+CD42b-) and late MkP (CD34-CD235a-CD71-CD41b+CD42b+) by flow cytometry elucidated differential mechanisms of action such as induction of early or late MKP or both. We performed an 8-day study in healthy mice, which revealed that a selected set of compounds increased MkPs in vivo, consistent with our predicted cell behavior and in vitro findings.
To evaluate a possible application for this MkP and platelet increases we tested the efficacy of one of our hit compounds CP3 in a CIT mouse model using Carboplatin. In this model, we recorded a &gt;70 % reduction in platelet counts on day 9 post carboplatin administration, with platelet numbers recovering on day 13 and return to basal levels by day 21. To assess the efficacy of CP3, mice dosed first with this compound were challenged with carboplatin. The primary endpoint of this study was the ability of the compound to rescue mice from nadir of thrombocytopenia and accelerate recovery to baseline. Prior to carboplatin dosing, 7 days of CP3 dosing showed a dose dependent increase in platelet counts confirming our initial studies. However, CP3 dosing did not rescue platelet nadir or accelerate recovery to baseline in the CIT model. This study demonstrated the capacity of CP3 to increase platelets in a healthy BM environment but not under conditions of myelosuppression as seen in the CIT mouse model.
Taken together, these findings highlight the benefits of our platform to predict compounds that modulate lineage differentiation towards MkP, and a resultant increase in platelet numbers in an animal model.
Trial in Progress: An Open-Label Expansion Trial Evaluating the Safety, PK/PD, and Clinical Activity of Emavusertib (CA-4948) + Ibrutinib in R/R Primary CNS Lymphoma
Authors: Grommes Christian , Nowakowski Grzegorz S. , Rosenthal Allison C. , Lunning Matthew A , Ramchandren Radhakrishnan , Regales Lucia , Fowle Meaghan , Lane Maureen , Wang Catherine , Omuro Antonio , Leslie Lori A. , Soussain Carole , Dabrowska-Iwanicka Anna Paulina , Ferreri Andrés José María , Tun Han 
Published: December 2023

Background:
Primary central nervous system lymphoma (PCNSL) is a rare and aggressive form of NHL isolated to the central nervous system or vitreoretinal space. Despite high initial rates of responses to available therapies, most PCNLS relapse in two years, presenting a very poor prognosis. Combination with novel therapies may overcome resistance (Guidetti 2023a). Upregulation of the B-cell receptor (BCR/NF-κB) and toll-like receptor (TLR)/NF-κB) signaling axis have been identified as key pathways in the pathogenesis of PCNSL. MYD88 L265P, a widely expressed somatic mutation in more than 70% of PCNSL patients, activates NF-κB signaling through stimulation of BTK and IRAK4. Emavusertib (CA-4948) is a novel, potent oral inhibitor of interleukin 1 receptor-associated kinase 4 (IRAK4), with additional inhibitory activity against FMS-like tyrosine kinase 3 (FLT3) and CDC-like kinases (CLK1/2/4). Targeted inhibition of IRAK4 with emavusertib in PCNSL xenografts indicated sufficient blood brain barrier penetration of emavusertib, target inhibition, and antitumor activity (von Roemeling 2022). Further preclinical studies have demonstrated that emavusertib synergizes with ibrutinib and can re-sensitize and overcome resistance to BTK inhibitors (Guidetti 2023a and 2023b). Initial clinical data have reported responses in BTK-resistant PCNSL and good tolerance of the combination (Iqbal 2022; Grommes 2023).
Study Design:
This is an ongoing multi-center, open-label trial (NCT03328078) in adult patients with hematologic malignancies to evaluate the safety, PK, and anti-cancer activity of oral administration of emavusertib alone or in combination with ibrutinib. Part Bis an open trial expansion cohort with BTKi relapsed or refractory PCNSL. About 20 evaluable PCNSL patients will be enrolled globally. Patients will receive oral CA-4948 at 100 or 200 mg BID in combination with ibrutinib at 560 mg QD in 28-day cycles until disease progression or intolerable toxicity. The primary endpoint is expanded safety/tolerability; secondary endpoints include ORR, DOR, PFS, OS, and pharmacokinetics. Key inclusion criteria for this expansion cohort: pathologically confirmed diagnosis of PCNSL with evidence of disease progression, measurable disease, relapsed or refractory with no more than 3 lines of prior anti-PCNSL therapy (including BTKi), and absence of residual Grade ≥2 toxicity from prior therapy. Key exclusion criteria: intraocular PCNSL without brain lesion or CSF involvement or significant co-morbidity. Key clinical assessments include brain imaging, CSF- and ocular examinations.