bluuebunny/crossref_metadata_2025 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.
Idiopathic Multicentric Castleman Disease Results in Endothelium Activation and Coagulation Dysregulation Via Tissue Factor and PAI-1
Authors: Lossos Chen , Brown Jenna , Chaturvedi Shruti , Naik Rakhi P P. , Francischetti Ivo M.B.
Published: December 2023

Idiopathic Multicentric Castleman disease (iMCD) is an inflammatory disease which may be associated with thrombosis. Diagnosis of this condition may be delayed with increase in morbidity and mortality. We investigated the cause of hypercoagulability in one patient with iMCD, who presented with shortness of breath, anasarca, abdominal distension and severe pulmonary hypertension with clinicopathologic findings consistent with iMCD with some features of TAFRO (thrombocytopenia, anasarca, fever, reticulin myelofibrosis, organomegaly) variant. Lab results detected high levels of inflammatory markers IL-6, CRP and ferritin. During the patient's hospital course, our patient developed evidence of consumptive coagulopathy, based on prolonged PT, aPTT, thrombocytopenia, high D-dimers and reduced fibrinogen levels, with associated microvascular thrombosis, limb gangrene, and biopsy-proven vascular fibrin formation. We examined several markers of coagulation activation in this patient to characterize the pattern of hypercoagulability in mCD. Cytokine and chemokine arrays and ELISA for selected coagulation factors were employed. We found an elevation in FVIIa/AT suggestive of in vivo upregulation of Tissue Factor (TF) as well as an increase in microparticles expressing functional TF (inhibited by Ixolaris). Our patient also demonstrated a marked increase in C5a and C3b. With respect to anticoagulants associated with the endothelium, both thrombomodulin and sEPCR were augmented in our patient, indicating endothelial cell injury and decreased protein C activation (APC). High levels of PAI-1, the main fibrinolysis inhibitor was evident. In addition, an increase in the chemoattractants for monocytes MCP-1, MCP-4 was noted. Similarly, we noted an increase in angiopoietin-2 and other markers associated with endothelial cell activation including adhesion molecules E-selectin, sICAM-1, sICAM-3 and sVCAM-1. Molecules affecting angiogenesis such as endoglin, IL-8, VEGF-A were also increased. Most markers were at normal levels when the patient was in remission. Our findings suggest that multiple pathways influence hypercoagulability in iMCD, including direct activation of coagulation through TF and complement, increase in thromboinflammation via monocytes, and endotheliopathy. Mechanistically, it is likely that the cytokine storm in iMCD, characterized by endothelial activation, components of immunothrombosis and complement activation result in TF-mediated coagulation dysregulation, shifting hemostasis towards a prothrombotic and antifibrinolytic state. Components of immunothrombosis, including inhibitors of TF, emerge as a therapeutic target in iMCD.
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.
A Multiomic, Single-Cell Measurable Residual Disease (scMRD) Assay for Phasing DNA Mutations and Surface Immunophenotypes
Authors: Murphy Charlie , Thompson Kate , Nousheen Lubna , Rao Divya , Druley Todd E.
Published: December 2023

The small population of cancerous cells that remain following treatment, known as measurable residual disease (MRD), is the major cause of relapse in acute myeloid leukemia (AML). Usually, these refractory cells have gained additional resistance mutations or changed their surface immunophenotypes in ways that preclude detection and phasing by current gold standard flow cytometry or bulk next-generation sequencing assays. For this reason, a multiomic single-cell MRD (scMRD) assay could offer a more comprehensive indicator of relapse and the potential for faster response. Here, we present a new scMRD assay with a 0.01% limit of detection that provides single-cell clonal architecture and immunophenotyping to not only identify residual leukemia cells, but also identify putative DNA or protein targets for salvage therapy. The assay enables rare-cell detection on a standard Mission Bio Tapestri run by adding (i) an upfront bead-based protocol to enrich for blast cells, (ii) a DNA and protein panel specifically designed for AML MRD diagnosis and treatment, and (iii) a new, automated analysis pipeline to evaluate single-cell multiomics output. By utilizing Mission Bio's single-molecule DNA sensitivity for single cells, this pipeline can identify and correlate co-occurring de novo variants, thereby reducing false positive rates over bulk assays that do not correlate variants. It furthermore can create phylogenetic trees of the detected MRD cells and present their surface protein signature and arm-level copy number. In addition, the multiplexing of up to three patient samples combined in one run via germline identification further reduces per sample costs and increases throughput. To demonstrate these features on 0.01% MRD, samples were constructed by titrating diseased cells into healthy bone marrow cells before processing them with the scMRD assay. We detected 0.01% spike-in (CD34+) and 0.1% spike-in (CD117+) in 6 of 6 samples, with an average enrichment of 41x and 13x, respectively. Further testing detected CD34+ 0.1% spike-ins in 10 of 10 samples (32x average enrichment). We applied the scMRD assay to banked bone marrow aspirate samples from 3 AML patients. The scMRD assay resolved the clonal architecture identifying multiple leukemic clones with co-occurring mutations. The assay readily distinguished pre-leukemic from leukemic clones thereby increasing the specificity of MRD results. The integration of genotype and immunophenotypic further enhanced MRD detection by identifying genotype-specific protein expression patterns. By combining high sensitivity with multiomics, this assay offers a potential scalable solution for comprehensive MRD detection that guides therapeutic decision-making.
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 >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.
Post-Transplant Maintenance in AML and MDS Patients Undergoing Non-Myeloablative Peripheral Blood Stem Cell Transplant with Post-Transplant Cyclophosphamide
Authors: Lei Lan , Bashey Asad , Solomon Scott R. , Morris Lawrence , Holland H Kent , Solh Melhem M. , Jackson Katelin C , Zhang Xu , Bachier Rodriguez Lizamarie
Published: December 2023

Disease relapse remains a major cause of mortality for patients with AML and MDS undergoing allogeneic hematopoietic stem cell transplant (HSCT). Several trials have evaluated the use of hypomethylating agents (HMA) like azacitadine or decitabine as post-transplant maintenance with its use not currently recommended. Here we review our experience with AML and MDS patients undergoing non-myeloablative (NMA) peripheral blood stem cell transplant using post-transplant cyclophosphamide (PTCy) as GVHD prophylaxis and in whom post-transplant maintenance was used.

We retrospectively reviewed AML and MDS patients who had a NMA HSCT from January 2015 until December 2022. All patients received fludarabine, cyclophosphamide and TBI200 or TBI300 as conditioning regimen with tacrolimus, mycophenolate mofetil and PTCy as GVHD prophylaxis (Table 1). Sixty patients received TBI200 and 28 patients received TBI300. Median age at transplant was 65.5 years in the TBI200 group vs 69 years in the TBI300 group. Haploidentical donors were used in 59 (98%) of patients in the TBI200 group and 18 (64%) of patients in the TBI300 group. Minimal residual disease (MRD) was present 35 (58%) of patients receiving TBI200 and 12 (43%) of patients receiving TBI300. Post-transplant therapy was given in 46 (77%) of patients in the TBI200 group and 23 (82%) in the TBI300 group. The majority of patients in both groups received azacitadine. The median cycles of HMAs was 4 (range 1-12).

We found no difference in OS, PFS, relapse, or NRM comparing Flu/Cy/TBI200 and Flu/Cy/TBI300 (not shown). We evaluated the use of post-transplant maintenance on survival endpoints and found no difference in endpoints. In addition, post-transplant maintenance had no effect on OS, PFS, relapse, NRM or grade II-IV aGVHD regardless of MRD status at time of transplant. While, this is a small retrospective series, it warrants further investigation into the use of post-transplant maintenance in the setting of NMA HSCT using PTCy. While the use of HMA's did not increase NRM, their efficacy and perhaps combination with other drugs needs to be more closely studied in NMA PTCy transplants to decrease relapse.
Blockade of PD-L1/CD80 Interactions Augments Function of Activated Donor T Cells Results in Augmenting Gvhd Induced By Donor Naïve CD8 + t Cells and GVL Effect Mediated By Donor Memory CD8 + T Cells
Authors: Song Qingxiao , Zhang Yuankun , Cassady Kaniel , Li Qinjian , Martin Paul J. , Zeng Defu
Published: December 2023

Preventing graft-versus-host disease (GVHD) while preserving graft-versus-leukemia/lymphoma (GVL) activity remains an elusive goal for allogeneic hematopoietic cell transplantation (HCT). It was reported by others that while donor naïve CD8 + T cells mediate both GVHD and GVL activity, donor memory CD8 + T cells could mediate GVL activity without GVHD. We recently reported that in vivo specific blockade of PD-L1/CD80 interactions augment tumor immunity mediated by memory CD8 + T cells (Zhang et al: PNAS 2023). In the current studies, we evaluated the impact of blockade of PD-L1/CD80 on GVHD induced by donor naïve CD8 + T cells and GVL activity-mediated by donor memory CD8 + T cells. Sorted naïve CD8 + T or memory CD8 + T cells and TCD-BM cells from C57BL/6 donors were transplanted into lethal TBI-conditioned BALB/c recipients with or without bearing ALL cancer cells. 4 days after HCT, the recipients were injected I.P. with anti-PD-L1 mAb (43H12) that specifically blocks PD-L1/CD80 interactions without inferring PD-L1/PD-1 interactions or control IgG. We observed that naïve but not memory CD8 + T cells severely infiltrated GVHD target tissues, and the memory CD8 + T cells expanded mainly in the lymphoid tissues. In vivo blockade of PD-L1/CD80 interactions augmented GVHD induced by the naïve T cells and GVL activity mediated by the memory CD8 + T cells. In addition, blockade of PD-L1/CD80 interactions increased production of granzyme B and inflammatory cytokines (IFN-g and TNF-α) of activated donor CD8 + T cells. Since cytolytic activity of effector CD8 + T cell is reported to be associated with its metabolic fitness, we evaluated the effect of blockade PD-L1/CD80 interactions on metabolism of alloreactive donor CD8 + T cells. And blockade of the interactions enhanced the fitness of mitochondria, as indicated by electronic microscopy analysis. The blockade also increased ECAR, OCR, and ATP production of the activated CD8 + T cells, with increased influx of glucose and fatty acids; and the blockade augmented mitophagy of the activated CD8 + T cells, as indicated by reducing mitochondria mass and dysfunctional mitochondria in the CD8 + T cells. The results indicate that 1) PD-L1/CD80 interactions play an important role in regulating the mitochondria function of activated alloreactive CD8 + T cells; 2) Blockade of PD-L1/CD80 interactions can augment GVL activity mediated by donor memory T cells without augmenting GVHD. This is different from blockade of PD-L1/PD-1 interactions that resulted in lethal GVHD.

*Zhang and Song are co-first authors of this abstract
Monoclonal Protein Concentration and Risk of Multiple Myeloma: Is 0.5 g/DL the New 1.5?
Authors: Salehi Mahta , Wang Mei , Sanfilippo Kristen M. , Thomas Theodore , Schoen Martin William , Chang Su-Hsin
Published: December 2023

Introduction : Monoclonal Gammopathy of Undetermined Significance (MGUS) is an asymptomatic clonal plasma cell disorder with a risk of progression to multiple myeloma (MM). To provide an accurate prognosis, monoclonal protein (M-protein) concentration has been used to assess risk of progression to myeloma. Current risk models developed from stored serum samples in a predominately white population of patients indicated a low risk of progression (less than 1% per year) in patients with M-protein of <1.5 g/dL. Little is known about M-protein concentration and risk of progression in patients tested during routine clinical care in a more diverse population. We assessed the outcomes of MGUS five years after diagnosis, and determined the association between baseline characteristics and the rate of progression to MM.

Methods: We used data from national Veterans Health Administration system to identify patients diagnosed with MGUS from 1999-2021 and were followed from MGUS diagnosis until progression, death, or the date 7/11/2023 was reached. MGUS diagnosis and progression to MM was confirmed utilizing a natural language processing algorithm. We included patients of black and white race and those with IgG, IgA, or light chain MGUS. The exposure was M-protein concentration at MGUS diagnosis, stratified by ≤0.5, >0.5-≤1.0, >1.0-≤1.5, >1.5 g/dL. We plotted the cumulative incidence function curve and used a multivariable Fine-Gray sub distribution hazard model with progression at 5 years as the event of interest, and death as a competing event. Covariates included sex, race, MGUS type, age, body mass index (BMI), diabetes, renal disease, and other comorbidities summarized by Charlson Comorbidity Index at MGUS diagnosis.

Results: We included 15,520 patients with MGUS. The mean age at the diagnosis was 71.1. IgG subtype was more common (83.2%) than IgA (12.7%), and light chain (4.1%). 19.6% and 39.5% of patients had renal disease and diabetes, respectively. Black veterans represented 5600 (36.1%) of the cohort. At 5 years, 26.6% of veterans with an M-protein of >1.5 g/dL were diagnosed with myeloma, compared to 11.7% with 1.1-1.5 g/dL, 6.9% with 0.6-1.0 g/dL, and 3.3% with ≤0.5 g/dL. In multivariable models an M-protein >1.5 was associated with a greater risk of progression compared to ≤0.5 (multivariable-adjusted Hazard Ratio [aHR] 9.76, 95% Confidence Interval [CI] 8.37-11.40). Overall, increasing age was shown to have lower risk of transitioning to MM at 5 years with aHR of 0.983 (95% CI 0.978-0.988).

Conclusion: In this modern cohort, only an M-protein of ≤0.5 g/dL was associated with a less 1% per year risk of developing MM. Conversely, 26.6% of veterans with an M-protein level >1.5 g/dL developed MM at 5 years. Risk of MM in patients clinically evaluated for MGUS may be higher than previously thought and new models of MM progression should be developed.
Hepcidin and Erythroferrone Levels across Human Pregnancy, and the Effect of IV Iron Infusion - a Revamp Substudy
Authors: Fielding Katherine , Mwangi Martin , Moya Ernest , Ataide Ricardo , Mzembe Glory , Randall Louise , Braat Sabine , Harding Rebecca , Phiri Kamija S. , Pasricha Sant-Rayn
Published: December 2023

Hepcidin is known to be suppressed in human pregnancy; and while it's known that intravenous iron infusion increases hepcidin levels, it is unknown to what degree this occurs during pregnancy.

The REVAMP trial was an open label, individually randomised controlled trial, in 862 Malawian pregnant women in the second trimester, with moderate to severe anaemia (haemoglobin<10.0g/dL). Participants were randomised equally to either intravenous Ferric Carboxymaltose (FCM), or standard of care (Oral iron), and followed up to 28 days post partum.

To evaluate hepcidin and erythroferrone levels, a total of 250 participants were randomly selected. This sample was balanced with the main trial with regards to trial arm (FCM 50%, Oral iron 50%), baseline iron deficiency (42.2%), anaemia (96.4%), parity (nulliparous 55.6%), and HIV status (HIV positive 15.3%). Hepcidin and erythroferrone levels were measured by ELISA, on serum samples collected longitudinally across 3 pregnancy timepoints (baseline, 28-days post randomisation, 36-weeks gestation), delivery, and 28-days postpartum.

Serum hepcidin levels (reported as: median [IQR]) were not different between the trial arms at baseline (FCM: 11.91ng/ml [5.56-20.53], Oral iron: 12.79ng/ml [6.65-20.40]). At 28 days post randomisation, hepcidin was higher in those treated with FCM (31.92ng/ml [14.80-59.84]) than oral iron (5.85ng/ml [2.50-15.12]), p<0.001. Hepcidin remained significantly increased at 36 weeks gestation (FCM: 21.14ng/ml [11.84-41.62], Oral iron: 10.86ng/ml [8.34-18.03], p<0.001). At delivery levels were higher overall, remaining higher in those who received FCM treatment (80.56ng/ml [25.11-149.6]) than Oral iron (37.39ng/ml [16.35-91.85], p=0.001). At 28days postpartum, hepcidin remained higher in the FCM arm 39.29ng/ml [19.52-76.37] than the Oral arm 17.67ng/ml [11.05-38.21], p<0.001.

Erythroferrone levels were not statistically different between arms at any timepoint; values were highest at baseline (FCM median 0.92ng/ml [0.42-1.45], Oral iron 1.06ng/ml [0.52-2.04]).

This is the first study examining the effects of modern intravenous iron infusions on hepcidin levels during pregnancy, and demonstrates that there is a statistically significant increase in hepcidin levels after ferric carboxymaltose, throughout pregnancy, persisting to four weeks post partum.
The linear ANRIL transcript P14AS regulates the NF-κB signaling to promote colon cancer progression
Authors: Ma Wanru , Hu Junhua
Published: December 2023

Background

The linear long non-coding RNA P14AS has previously been reported to be dysregulated in colon cancer, but the mechanistic role that P14AS plays in colon cancer progression has yet to be clarified. Accordingly, this study was developed to explore the regulatory functions of ANRIL linear transcript-P14AS in cancer.

Methods

The expression of P14AS, ANRIL, miR-23a-5p and their target genes were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Cell supernatants of IL6 and IL8 were measured by Enzyme linked immunosorbent (ELISA) assay. Dual-luciferase reporter assays, RNA immunoprecipitation, or pull-down assays were used to confirm the target association between miR-23a-5p and P14AS or UBE2D3. Cell proliferation and chemosensitivity of NF-κB inhibitor BAY 11-7085 were evaluated by cell counting kit 8 (CCK8).

Results

When P14AS was overexpressed in colon cancer cell lines, enhanced TNF-NF-κB signaling pathway activity was observed together with increases in IL6 and IL8 expression. The Pita, miRanda, and RNA hybrid databases revealed the ability of miR-23a-5p to interact with P14AS, while UBE2D3 was further identified as a miR-23a-5p target gene. The results of dual-luciferase reporter, RNA pull-down, and RNA immunoprecipitation experiments confirmed these direct interactions among P14AS/miR-23a-5p/UBE2D3. The degradation of IκBa mediated by UBE2D3 may contribute to enhanced NF-κB signaling in these cells. Moreover, the beneficial impact of P14AS on colon cancer cell growth was eliminated when cells were treated with miR-23a-5p inhibitors or UBE2D3 was silenced. As such, these findings strongly supported a role for the UBE2D3/IκBa/NF-κB signaling axis as a mediator of the ability of P14AS to promote colon cancer progression.

Conclusions

These data suggested a mechanism through which the linear ANRIL transcript P14AS can promote inflammation and colon cancer progression through the sequestration of miR-23a-5p and the modulation of NF-κB signaling activity, thus highlighting P14AS as a promising target for therapeutic intervention efforts.
Dancing the Nanopore limbo – Nanopore metagenomics from small DNA quantities for bacterial genome reconstruction
Authors: Simon Sophie A. , Schmidt Katharina , Griesdorn Lea , Soares André R. , Bornemann Till L. V. , Probst Alexander J.
Published: December 2023

Background

While genome-resolved metagenomics has revolutionized our understanding of microbial and genetic diversity in environmental samples, assemblies of short-reads often result in incomplete and/or highly fragmented metagenome-assembled genomes (MAGs), hampering in-depth genomics. Although Nanopore sequencing has increasingly been used in microbial metagenomics as long reads greatly improve the assembly quality of MAGs, the recommended DNA quantity usually exceeds the recoverable amount of DNA of environmental samples. Here, we evaluated lower-than-recommended DNA quantities for Nanopore library preparation by determining sequencing quality, community composition, assembly quality and recovery of MAGs.

Results

We generated 27 Nanopore metagenomes using the commercially available ZYMO mock community and varied the amount of input DNA from 1000 ng (the recommended minimum) down to 1 ng in eight steps. The quality of the generated reads remained stable across all input levels. The read mapping accuracy, which reflects how well the reads match a known reference genome, was consistently high across all libraries. The relative abundance of the species in the metagenomes was stable down to input levels of 50 ng. High-quality MAGs (> 95% completeness, ≤ 5% contamination) could be recovered from metagenomes down to 35 ng of input material. When combined with publicly available Illumina reads for the mock community, Nanopore reads from input quantities as low as 1 ng improved the quality of hybrid assemblies.

Conclusion

Our results show that the recommended DNA amount for Nanopore library preparation can be substantially reduced without any adverse effects to genome recovery and still bolster hybrid assemblies when combined with short-read data. We posit that the results presented herein will enable studies to improve genome recovery from low-biomass environments, enhancing microbiome understanding.
Acute pancreatitis in intraductal papillary mucinous neoplasm: a single-center retrospective cohort study with systematic review and meta-analysis
Authors: Xu Ji-Hao , Ni Chu-Yan , Zhuang Yan-Yan , Li Li , Lin Ying , Xia Zhong-Sheng , Wu Wei-Rong , Chen Qi-Kui , Zhong Wa
Published: December 2023

Background

Intraductal papillary mucinous neoplasm (IPMN) is a cystic tumor of the pancreas arising from abnormal papillary proliferation of ductal epithelial cells, and is a precancerous lesion of pancreatic malignancy. This study aimed to evaluate associations between acute pancreatitis (AP) and histologic subtypes of IPMN.

Methods

In the clinical study, patients with IPMN confirmed by surgical resection specimens at our institute between 2009 and 2021 were eligible for inclusion. Associations and predictive accuracy of AP on the presence of HGD were determined by logistic regressions. In addition, a systematic review and meta-analysis was conducted through literatures upon search in PubMed, Embase, CENTRAL, China National Knowledge Infrastructure (CKNI), and Wanfang database, up to June, 2023. Pooled effects of the associations between AP and HGD and intestinal epithelial subtype subtype, shown as odds ratios (ORs) with 95% confidence intervals (CIs), were calculated using random effects model.

Results

The retrospective cohort study included 47 patients (32 males, 15 females) diagnosed with IPMN at our center between 2009 and 2021, including 11 cases with AP (median 62 years) and 36 cases (median 64.5 years) without. Accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of AP in predicting HGD were 78.7%, 57.1%, 82.5%, 36.4%, and 91.7%, respectively. Univariate logistic regression analysis showed that AP group had greater odds of presence of HGD (OR: 6.29,95% CI: 1.14–34.57) than non-AP group. Meta-analysis of five case-control studies in the literature included 930 patients and showed that AP-IPMN patients had higher odds for HGD (OR: 2.13, 95% CI 1.38–3.29) and intestinal epithelial subtype (OR: 5.38, 95% CI: 3.50–8.27) compared to non-AP IPMN.

Conclusions

AP is predictive of malignancy in patients with IPMN.
Mediation role of anxiety on social support and depression among diabetic patients in elderly caring social organizations in China during COVID-19 pandemic: a cross-sectional study
Authors: Zhao Lanlan , Xu Fuqin , Zheng Xin , Xu Ziwen , Osten Benjamin , Ji Kai , Ding Shuo , Liu Guoqing , Yang Shufan , Chen Ren
Published: December 2023

Background

Diabetes has become a prominent global public health problem, which is an important cause of death, disease burden, and medical and health economic burden.

Previous studies have reported that majority of persons diagnosed with diabetes later presented with psychological and mental health diseases. The study aimed to explore the mediation role of anxiety on social support and depression among diabetic patents in elderly caring social organizations (SOs).

Methods

A multi-stage stratified cluster random sampling method was used in this cross-sectional study, and a questionnaire consisting of demographic questionnaire, MSPSS, GAD-7, and CES-D-10 was utilized to gather data. SPSS 22.0 and MPLUS 7.4 were used for statistical analysis. Spearman correlation analysis was employed to investigate correlations of key variables. A generalized linear model was used to exam factors associated with depression. Finally, the mediation effect among study variables was investigated by structural equation modeling (SEM).

Results

The average scores of social support, anxiety, and depression were 58.41 ± 14.67, 2.95 ± 3.95, and 7.24 ± 5.53, respectively. The factors of gender, social support, and anxiety were identified as significantly influential factors related to depression among diabetic patients in elderly caring SOs. The effect of social support on depression was significantly mediated by anxiety (β = -0.467, 95%CI: -0.813 to -0.251). Furthermore, anxiety partially mediated the relationship between family support and depression (β = -0.112, 95%CI: -0.229 to -0.012), and anxiety functioned as a complete mediator in the effect of significant others' support and depression (β = -0.135, 95%CI: -0.282 to -0.024).

Conclusions

The indirect effect of social support on depression through anxiety among diabetic patients in elderly caring SOs was elucidated. Social support played a key role in maintaining and regulating their mental health, particularly from family and significant others. Social support provided by both family and significant others exerted an important influence on maintaining and regulating their mental health. In light of this pathway, the elderly caring SOs should enhance the magnitude of social support from these two sources, thereby diminishing the likelihood of experiencing anxiety and depression.