bluuebunny/crossref_metadata_2025 huggingface dataset:
Haematopoietic Stem Cell Transplantation for Hepatitis-Associated Aplastic Anaemia: Clinical Characteristics, Outcomes and Risk Factors
Authors: Jia Li , Liu Yilin , Jiang Erlie , Feng Sizhou
Published: December 2023

Introduction:

Hepatitis-associated aplastic anaemia (HAAA) is a severe and life-threatening condition that requires effective treatment approaches. Hematopoietic stem cell transplantation (HSCT) has emerged as a promising therapeutic option for HAAA patients. In this retrospective study, we aimed to investigate the clinical characteristics, outcomes, and prognostic factors associated with HSCT in HAAA patients at a prominent Chinese blood disease hospital between 2008 and 2022.

Methods:

A comprehensive review of 35 HAAA patients who underwent HSCT was conducted, and clinical data were analyzed to determine the treatment's efficacy and safety. The patients were primarily categorized into those with severe AA (28.6%) and very severe AA (65.7%), with a predominant male representation (68.6%) and a median onset age of 23 years (range, 9-44). Both haploidentical and matched sibling donors were utilized in comparable proportions.

Results:

The study revealed a promising 5-year overall survival (OS) rate of 74.0% after HSCT, underscoring the effectiveness of this treatment approach in HAAA patients. However, certain factors significantly impacted survival rates and treatment outcomes. Prolonged intervals from diagnosis to HSCT (>=75 days), acute graft-versus-host disease (GVHD), and post-HSCT liver events, such as hepatic GVHD and a three-fold increase in aminotransferase or bilirubin, were found to worsen 5-year OS. Interestingly, higher doses of mononuclear cell count, CD34+, CD3+, CD4+, and CD8+ cells in allografts were associated with poorer OS or graft failure-free survival (GFFS), suggesting that pursuit of higher cell counts in certain cases may be counterproductive.

Furthermore, a close examination of the multivariate models revealed that recipients who received sex-matched grafts experienced better OS, while those with younger male donors had a lower incidence of grade II-IV acute GVHD. Moreover, a higher HLA matching degree (HLA >=7/10) between the recipient and donor emerged as an independent prognostic factor associated with better OS and GFFS.

Conclusion:

This study highlights the safety and efficacy of HSCT as a treatment modality for HAAA patients. Early transplantation, careful donor selection, and efforts to improve post-transplant liver events were identified as crucial factors to optimize treatment outcomes. While the use of certain cells in higher doses may have a negative impact on survival, the importance of achieving a high HLA matching degree cannot be overstated. Moving forward, these findings can serve as valuable insights in guiding the management and treatment decisions for HAAA patients undergoing HSCT. Further research and continuous improvement in transplantation protocols will undoubtedly advance the success rates and long-term prognosis for individuals afflicted with this challenging condition.
Thioredoxin-Interacting Protein (TXNIP) Binding to GATA1 May Affect GATA1 Transcriptional Activity in Megakaryocytes
Authors: Noh Jiyoon
Published: December 2023

GATA1 is a crucial transcription factor for both megakaryopoiesis and erythropoiesis. In our previous work, we established a murine megakaryocyte-erythroid progenitor (MEP) cell line, named G1ME2 cells, which can be maintained with doxycycline (dox)-induced anti- Gata1 shRNA. These cells can differentiate into erythroid cells or megakaryocytes upon removal of dox; however, their megakaryocyte differentiation potential showed alterations after prolonged cell culture (90-100 days). Notably, CD41 expression in G1ME2 cells declined during long-term culture, prompting us to investigate the disparities between CD41+ and CD41- G1ME2 cells using total RNA sequencing. Interestingly, the western blotting analysis revealed a significant enrichment of thioredoxin-interacting protein (TXNIP) in the late G1ME2 cells compared to the early cells. TXNIP is known for its role in regulating cellular metabolism, and recently, we observed age-associated thrombocytopenia in TXNIP knockout mice, suggesting its involvement in hematopoiesis. Additionally, previous studies have identified its role in erythropoiesis through diverse mechanisms. In our current study, we made a novel discovery that TXNIP directly binds to GATA1 during megakaryopoiesis. Furthermore, deletion of GATA1 C-terminal enhanced binding affinity, implying that this interaction might occur when GATA1 is not localized to DNA. Consistently, TXNIP exhibited the ability to suppress GATA1 promoter activity, leading to changes in GATA1 localization in the presence or absence of TXNIP. In conclusion, our findings demonstrate the involvement of TXNIP in megakaryopoiesis, partially through the regulation of GATA1 transcriptional activity.
CXCR4 Enriched T Regulatory Cells Preferentially Home to Bone Marrow and Decrease Inflammation
Authors: Huang Meixian , Lyu Mi-Ae , Zeng Ke , Sadeghi Tara , Masarova Lucia , Flowers Christopher R. , Parmar Simrit
Published: December 2023

Background and aim

Adoptive therapy with T regulatory cells (Tregs) is emerging as an important therapeutic strategy for several autoimmune and inflammatory diseases. Optimal homing and payload delivery of these anti-inflammatory cells remain a cornerstone of optimizing their therapeutic efficacy. CXCR4 cell surface expression has been shown to be critical for homing of hematopoietic cells to the bone marrow (BM). Also, CXCR4/CXCL12 axis plays a major role in the pathogenesis of BM inflammatory disorders including myelofibrosis. We hypothesize that enrichment of CXCR4 on the Tregs cell surface will enable their trafficking to BM and ensure a homogenous delivery.

Methods/results/conclusion

Magnetic enrichment of CD25 + cells was performed on umbilical cord blood on day 0, followed by dual enrichment on CXCR4 using CRANE TM (cord blood Tregs, activated and enriched) process on day 3 or 4 (Fig 1A). The enrichment process led to increased number of Tregs expressing CXCR4 on their cell surface (Fig 1B; Treg CXCR4) as well as a CXCR4 hi Treg cells when compared to Treg control (Fig 1C, p<0.0001; n=16). In a Transwell TM system, a higher number of Treg CXCR4 migrated to the bottom well in chemotactic response to SDF1α at 15 minutes as compared to the Treg control cells. Such migration was inhibited by CXCR4 antagonist, AMD3100 (Fig 1D). The migration superiority of Treg CXCR4 was maintained at 30 and 60 min (Fig 1E). In order to understand in vivo transit kinetics, Treg cells were injected into NSG mice and organs were harvested at 12 and 24 hours (Fig 1F). Treg CXCR4 showed preferential homing to BM (Fig 1G), where a higher expression of CXCR4 and CXCL12 than control, was observed (Fig 1H). Other markers including CD62L (homing and suppression), CD39 & CD73 (conversion of ATP to immunosuppressive adenosine), and CXCR5 (Follicular Treg) were also increased in Treg CXCR4 recipients. A decrease in biomarkers including TGFα and TNFβ (inflammation), IL-13 (fibrosis) (Fig 1I) with an increase in GM-CSF (increase Treg potency) (Fig 1J), was observed in Treg CXCR4 recipients with a corresponding decrease in TGFβ family (hematopoietic suppressor) (Fig 1K). We conclude that Treg CXCR4 shows superior homing to BM and is currently being studied in a clinical setting to treat myelofibrosis with suboptimal response to ruxolitinib (NCT05423691).
Oncogenic RAS Signaling Promotes an Immunosuppressive Niche in Acute Lymphoblastic Leukemia
Authors: Ferrao Blanco Mauricio Nicolás , Schweighart Elizabeth , Kester Lennart , Hehir-Kwa Jayne , Belderbos Mirjam E. , den Boer Monique L. , Heidenreich Olaf , Vormoor Josef
Published: December 2023

Emerging evidence suggests that genomic aberrations in cancer cells may impact the composition of the tumor microenvironment leading to a tumor-specific supporting niche. Previous genomic profiling studies have identified somatic alterations in RAS pathway genes that drive treatment resistance in B-cell acute lymphoblastic leukemia (B-ALL): yet it is unknown whether such genetic events may specifically hijack the leukemic niche.

Here, we analyzed bulk RNA-sequencing data of bone marrow mononuclear cells from 185 pediatric B-ALL patients entering our center and performed mutation calling of RAS pathway genes, namely KRAS, NRAS, BRAF and PTPN11. Transcriptomic analysis revealed attenuated immune responses and decreased T cell activation in samples harboring RAS mutations (n= 62) compared with wild type cases (n= 123). To further explore this finding at single-cell resolution, we sorted bone marrow mononuclear cells from 8 primary B-ALL pediatric patient samples (4 RAS mutant and 4 wild type), enriching on the hematopoietic and non-hematopoietic compartments, followed by single-cell RNA-sequencing using the 10x Genomics platform. Twelve cell populations were identified by the expression of bona fide markers, mapping different immune and stromal cells in the ALL landscape. Specifically, the RAS-mutant microenvironment was defined by dysfunctional CD8+ T-cells, characterized by high expression of TIM-3, TIGIT and LAG-3, as well as regulatory T cells, displaying expression of CTLA-4, ICOS and FOXP3, suggesting a RAS-mediated immunosuppressive mechanism.

In light of these changes, we examined flow cytometry data of bone marrow mononuclear cells collected at diagnosis from our cohort of 185 patient samples. Whereas the overall proportion of lymphoid subsets (T, B and NK cells) was not significantly changed among RAS mutant and wild type groups, we detected a decrease in myeloid subsets (neutrophils, eosinophils, and monocytes) in RAS mutant ALL, which was more prominent in monocytes. Cross comparison of the transcriptional state of monocytes in our single-cell data unravel that monocytes in the RAS mutant niche displayed a non-classical phenotype, characterized with high expression of the immune checkpoint TIM-3, which is a major driver of T cell exhaustion.

Taken together, the bone marrow microenvironment of ALL patients harbouring oncogenic RAS lymphoblasts is defined by dysfunctional immune cell populations. Further studies on the cellular dynamics in the niche will provide the basis for the development of therapeutic opportunities to target the immune micro-environment in RAS mutant ALL.
In Vivo CRISPR Screening Platform for Studying Myeloid Inflammation and Clonal Dynamics
Authors: Kim Peter Geon , Hergott Christopher B. , Loke Justin CT , Shin Wesley , McConkey Marie E , Ebert Benjamin L.
Published: December 2023

Previous experimental approaches using CRISPR to study factors governing hematopoiesis in vivo have been hampered by heterogeneity in cellular expansion during hematopoietic reconstitution. To control for such ‘cellular jackpotting’, we developed an in vivo hematopoietic stem and progenitor (HSPC)-based CRISPR screening system with unique molecular identifiers (UMIs) and applied these techniques to study myeloid inflammation and clonal expansion of Tet2-mutated HSPCs. Transplantation of Cas9-expressing Lin -Sca1 +c-Kit + (LSK) cells transduced with high-diversity UMI-labeled guide-RNAs (gRNAs) enabled sufficient gRNA representation at a genome-wide level after lentiviral integration of one gRNA per cell and controlled for 10 log-fold differences in cellular expansion during hematopoietic reconstitution. Simultaneous comparison of barcoded CRISPR screening with non-barcoded screening revealed that barcoding decreases false-positive biological processes related to cell division while improving sensitivity for detecting biological processes related to transcriptional events and cellular differentiation. To study inflammatory mediators, we applied this CRISPR screening tool towards studying the recruitment of CD101 +Ly6G +CD11b + mature neutrophils during an inflammatory insult, which revealed the majority of genes involved in human congenital neutropenia (Figure A). To demonstrate the reproducibility of this screening system, we performed a targeted transcription factor (TF)-based CRISPR screen, which revealed concordance between the genome-wide screen and the targeted screen. Next, we also applied this screening system to studying dynamics of Tet2-mutated clonal hematopoiesis. To study master regulators of clonal expansion in Tet2 KO, we performed a UMI-based CRISPR screen using chromatic and transcription factors, which revealed Tet3 and Bcor, genes previously identified to cooperate with Tet2 KO to promote myeloproliferation (Figure B). Validation of additional novel genetic hits is currently ongoing. We hypothesize that this screening platform will be useful for dissecting many aspects of hematopoiesis including inflammatory mediators and clonal dynamics of HSPCs.
Static and Dynamic Assessment to Identify Ultra-High Risk Multiple Myeloma：Analysis for Patients with Overall Survival No More Than Three Years
Authors: Xu Yan , An Gang , Deng Shuhui , Sui Weiwei , Liu Wei , Huang Wenyang , Wang Tingyu , Yi Shuhua , Zou Dehui , Qiu Lugui
Published: December 2023

Background: The median overall survival (OS) in multiple myeloma (MM) is rapidly approaching 10 years. However, the prognosis of ultra-high risk MM (UHR-MM) remains very poor, the median overall survival (OS) is no more than 3 years. There is no consensus to define this population till now. Several prognostic factors with significant impact on survival have been discovered over time, with cytogenetic factors as the most potent. Nevertheless, there is little study to investigate the features of these UHR-MM, especially in Asian UHR-MM patients.

Patients and Methods: To understand the characteristic of UHR-MM and help identify these patients as early as possible, we retrospectively analyzed the data of patients whose OS is <3 years from a single Chinese center with static and dynamic assessment. Baseline characteristic and response dynamic were collected.

Results: From 2013.1 to 2019.12, 1270 patients with newly diagnosed MM (NDMM) were enrolled in a single Chinese center. In which, 265 (20.79%) patients deceased in 36 months. 271 survival patients with no more than 3 years follow-up were excluded. Compared with 734 patients survived more than 36 months, median OS were 18.48±10.05 months and 92.70±3.99 months, median progression survival (PFS) were 12.14±4.00 months and 53.66±2.43 months, respectively. These patients are older (median age: 56 vs. 61 years, p<0.0001), with higher percentage of genetic ultra-high risk (including del(17p)≥60%; or ≥2 high-risk cytogenetic abnormalities including TP53 mutation, del(17p)/P53 deletion, t(4;14), t(14;16), t(14;20), 1q21 gain/ amplification), more patients with ISS-III, R-ISS-III, R2-ISS-IV, more patients with primary refractory (including <MR after 2 cycles or <PR after 4 cycles standard first-line therapy) or early relapse (including lost best response within 6 months from the standard first-line therapy), and more patients with primary PCL history. With multivariate analysis, genetic ultra-high risk, primary refractory, early relapse and primary PCL history are independent factors for poorer PFS and OS.

Conclusion: With static and dynamic assessment, we can identify a group of UHR-MM with survival no more than 3 years as early as possible. For these patients, we need more potent therapy model to improve their survival.
Soluble CD163; A Possible New Biomarker in Multiple Myeloma
Authors: Gkioka Annita Ioanna , Gkiokas Alexandros , Alexandropoulos Alexandros , Papadatou Mavra , Koudouna Aspasia , Vasiliki Bartzi , Tryfou Thomai-Marina , Kafasi Nikolitsa , Kyrtsonis Marie-Christine
Published: December 2023

INTRODUCTION

Tumor associated macrophages (TAM) constitute a major part of the bone marrow microenvironment and play a pivotal role in growth of various hematologic malignancies, such as Multiple Myeloma (MM). Cytokines secreted by TAM, such as hemoglobin scavenger receptor CD163, can be detected in their soluble form (sCD163) and reflect the TAM burden. It was recently published that bone marrow TAMs protect neoplastic plasmacytes from chemotherapy-based apoptosis. In our study, we aimed to investigate the clinical utility of sCD163 in patients with Multiple Myeloma.

PATIENTS AND METHODS

One hundred and nineteen patients were included in the study of whom 80 had symptomatic MM, 30 appeared to have SMM and 8 had MGUS. Clinical and laboratory characteristics were reviewed, after patients' informed consent form. Their median age was 68 yrs (range, 30-89), 53% were men and 47%women; 39%, 27%, and 35% of MM patients were staged ISS1, ISS2 and ISS3 respectively. Serum sCD163 was tested in frozen sera collected at patients' diagnosis and in 30 healthy individuals (HI) . Measurements were performed by ELISA (Duo-Set R&D Quantiquine) according to the manufacturer's instructions. Median value of variables was used as the cut off point. Median time to treatment (TTT) was 0 months (range,0-248) in MM patients and median overall survival (OS) was 67 months (range, 0-248). Statistical analysis was performed with the SPSS v.26 software.

RESULTS

Median sCD163 was 26618 pg/ml (11831- 35548) in HI, 24366 pg/ml (13166 - 27429) in MGUS, 27665 (18266-39088) pg/ml in SMM, and 27870 (20720-37478) pg/ml in MM patients.Significantly lower levels of CD163 were observed between MGUS and HI (p=0.001) and HI and MM patients (p=0.015). Shorter time to treatment was observed in all MM patients including SMM with sCD163 levels above median (p = 0.034). Patients who became symptomatic after 6 months of diagnosis were studied separately, with levels above the median indicating a shorter time to therapy (p=0.003). Also, a trend of shorter overall survival was observed in all patients (MM, SMM, MGUS) with levels of CD163 above median (p=0.07).

CONCLUSION

Our findings reveal that serum sCD163 levels differed in patients with MGUS, SMM, MM, and HI, and could predict TTT in MM, indicating that cells of the monocyte-macrophage lineage may play a role in disease pathophysiology and progression. sCD163 could eventually prove to be a significant prognostic factor in MM.
Long Noncoding RNA NEAT1 Regulates Acute Graft-Versus-Host Disease By Promoting Macrophage M1 Polarization through JNK Pathway
Authors: Zhang Ran , Huang Zhenli , Wang Li , Cao Weijie , Small Donald , Zhu Ruiqi , Shi Wei , Jiang Zhongxing , Xia Linghui
Published: December 2023

Introduction

Acute graft-versus-host disease (aGVHD) is a complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). The function of macrophages in aGVHD is a hot point in current research. Long noncoding RNA nuclear enriched abundant transcript 1 (lncRNA NEAT1) is closely related to immune-related diseases and affects the function of macrophages. However, the role of LncRNA NEAT1 in aGVHD is unclear in current study.

Methods

Peripheral blood mononuclear cells (PBMCs were collected from the patients with or without aGVHD after allo-HSCT. The expression of lncRNA NEAT1 in PBMCs was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The diagnostic value of lncRNA NEAT1 in aGVHD was analyzed by receiver operating characteristic curve (ROC) and area under correlation curve (AUC). We transfected RAW264.7 cells and bone marrow-derived macrophages (BMDM) with NEAT1 lentiviral vector or NEAT1 small interfering RNA to change the expression level of NEAT1. The function of NEAT1 and its effect on JNK pathway were analyzed by flow cytometry, qRT-PCR, western blotting and ELISA. Finally, aGVHD mouse model was constructed to evaluate the function of JNK inhibitors in aGVHD.

Results

Compared with non-aGVHD patients, LncRNA NEAT1 was significantly up-regulated in PBMCs of aGVHD patients. ROC and AUC analysis confirmed that the expression level of lncRNA NEAT1 was correlated with the occurrence of aGVHD, which may have certain diagnostic value in aGVHD. We found the expressions of NEAT1 in BMDM and RAW264.7 were remarkably increased after being stimulated with lipopolysaccharide (100ng/ml). The overexpression of lncRNA NEAT1 in RAW264.7 could significantly up-regulate the expression level of iNOS, and induce the differentiation into M1 type. Knockdown of lncRNA NEAT1 in BMDM could significantly down-regulate the gene expressions (iNOS and NLRP3), reduce the protein levels (iNOS, NLRP3 and p-JNK), decrease the proportion of M1 macrophages and inflammatory cytokines secretion (TNF-α and IL-1β), and inhibit the JNK pathway. We found that JNK inhibitor could perform the same function as knocking down lncRNA NEAT1 in BMDM. In addition, JNK inhibitor could improve the survival and pathology, decrease the inflammatory cytokines secretion (TNF-α and IL-1β), and reduce the infiltration of M1 macrophages into the target organs in aGVHD mouse model.

Conclusions

Our results showed that lncRNA NEAT1 may regulate the M1 polarization of macrophages and the secretion of inflammatory cytokines by regulating the JNK pathway. JNK inhibitor may reduce the inflammation in aGVHD mouse models by blocking the function of NEAT1 in macrophages. This study provides a promising therapeutic target for aGVHD.
Pharmacokinetic-Targeting and Dose-Adjustment of I.V. Busulfan for Myeloablative Conditioning in Allogeneic Hematopoietic Cell Transplantation
Authors: Khanolkar Rutvij A. , Kangarloo Shahbal B , Li Na , Khan Faisal , Storek Jan
Published: December 2023

Introduction: Pharmacokinetically (PK)-targeting intravenous busulfan improves outcomes following allogeneic hematopoietic cell transplantation (HCT). However, the optimal busulfan area under the curve (AUC) likely differs based on the specific conditioning regimen, and the target busulfan exposure is unknown in combination with fludarabine and low-dose total body irradiation (TBI).

Methods: This study included 1019 adult HCT recipients that received myeloablative conditioning including fludarabine, busulfan, anti-thymocyte globulin, and low-dose (4cGy) TBI. Busulfan was administered as a total dose of ~3.2mg/kg given equally from days -5 to -2 pre-transplant. Total AUC was estimated using measurements of serial serum samples. Multivariate Cox and Fine-Gray regression were used for comparison of AUC subgroups. The primary outcomes of interest were relapse-free survival (RFS) and overall survival (OS).

Results: Median AUC was 62.3 mg∙hr/L (range: 39.4-128.0 mg∙hr/L). Total AUC exposure of 49.3-57.5 mg∙hr/L was associated with greater RFS (67% vs. 47%, HR=1.82, P=0.014) and OS (71% vs. 46%, HR=1.99, P=0.008) compared to patients with higher AUCs of 57.5-73.9 mg∙hr/L. Although very low (<49.3 mg∙hr/L) or very high (>73.9 mg∙hr/L) AUCs trended towards worse RFS and OS compared to 49.3-57.5 mg∙hr/L, this analysis was limited by the small number of patients with extreme AUCs and did not reach statistical significance. Except potentially for patients with a high/very high HCT disease risk index, 49.3-57.5 mg∙hr/L appeared to be the optimal AUC regardless of patient sex, age, or primary disease.

Conclusion: Within the evaluated AUC range, 49.3-57.5 mg∙h/L appeared to be associated with the most favourable survival. Pharmacokinetic-targeting to this range may improve outcomes.
PRT2527, a Novel Highly Selective Cyclin-Dependent Kinase 9 (CDK9) Inhibitor, Has Potent Antitumor Activity in Combination with BTK and BCL2 Inhibition in Various Lymphoid Malignancies
Authors: Fultang Norman , Schwab Ashley M , McAneny-Droz Sophia , Heiser Diane , Scherle Peggy A , Bhagwat Neha
Published: December 2023

CDK9 is a master regulator of transcription that modulates transcription elongation via phosphorylation of RNA polymerase II. Short-term inhibition of CDK9 depletes short-lived transcripts and labile proteins such as MCL1, BFL1 and MYC to promote cancer cell death.

We previously described PRT2527, a novel, potent, highly selective CDK9 inhibitor with anti-leukemic activity in various preclinical models. PRT2527 is currently under evaluation in a Phase I clinical trial in patients with relapsed/refractory hematologic malignancies (NCT05159518).

Here we demonstrate PRT2527 potently inhibits cancer growth and induces cell death in various models of Diffuse Large B Cell Lymphoma (DLBCL), Mantle Cell Lymphoma (MCL) and Chronic Lymphocytic Leukemia (CLL), through depletion of MCL1, BFL1 and MYC. We also show that combination of PRT2527 with BTK inhibitors (BTKi) or BCL2 inhibitors (BCL2i) leads to more robust and durable responses in multiple pre-clinical models. In these studies, the combination of PRT2527 with BTKi or BCL2i was well tolerated.

Mechanistically, we show that in cell lines, BTK inhibition upregulates BMF, an endogenous inhibitor of BCL2 and BCL-xL, driving cells towards MCL1 and BFL1 dependency. Concurrent depletion of MCL1 and BFL1 with PRT2527 led to complete inhibition of Bcl-2- family mediated survival signaling, potently inducing cell death. In a similar fashion, Venetoclax-driven BCL2 inhibition also potentiated the anti-tumor activity of PRT2527. Consistent with our findings in vitro, we confirmed potent combinatorial inhibition of tumor growth by PRT2527, administered once weekly, and BTKi or BCL2i, administered daily, in CDX and PDX in vivo models of DLBCL and MCL. We observed reduced BFL1 and MCL1 as well as increased BMF in tumors of mice treated with BTKi and PRT2527 suggesting complete Bcl-2 family inhibition. We also demonstrated enhanced induction of apoptosis by PRT2527 in primary CLL patient samples simultaneously treated with BTKi or BCL2i.

Additionally, we demonstrated that PRT2527 potently inhibits growth in Ibrutinib-resistant MCL cell lines. Further work characterizing PRT2527 as monotherapy and in combination with BTKi and BCL2i in in vivo models of BTKi-resistant MCL and CLL is currently ongoing.

Altogether these data provide a rationale for evaluating PRT2527 in combination with BTK and BCL2 inhibitors for the treatment of patients with relapsed/refractory hematologic malignancies.
Improved Risk Prediction in DLBCL By Combining Clinical and PET Features with Interim PET Assessment
Authors: Hanoun Christine , Heymans Martijn , Wiegers Sanne , Bes Annelies , Duehrsen Ulrich , Huettmann Andreas , Kurch Lars , Barrington Sally F , Mikhaeel George , Lugtenburg Pieternella , Ceriani Luca , Zucca Emanuele , Gyorke Tamas , Czibor Sandor , Zwezerijnen Gerben , Boellaard Ronald , Zijlstra Josée M. , Eertink Corinne
Published: December 2023

Background: Accurate detection of patients at high risk of treatment failure following frontline immunochemotherapy in diffuse large B-cell lymphoma (DLBCL) is of paramount importance as these patients might benefit from early treatment escalation. Recently, we introduced the IMPI prognostic model based on metabolic tumor volume (MTV), age and stage that outperformed the international prognostic index (IPI). However, radiomic features such as Dmax bulk and SUV peak as well as an early treatment response at interim PET (i-PET) as a measure of chemosensitivity using ΔSUVmax may have additional predictive value. We tested different models for risk prediction aiming at a dynamic risk tool in the era of evolving radiomic features in functional imaging.

Methods: All patients within the PETRA database with newly diagnosed DLBCL, who were treated with R-CHOP and had available clinical data, baseline PET and i-PET scans were included.

The optimal transformation of Dmax bulk, SUV peak and ΔSUV max was determined by choosing the best fitting Cox regression model with 3-year PFS as outcome, with highest R2 and lowest Akaike Information Criterion (AIC), while the cross-validated c-index was obtained as a measure for discrimination.

Subsequently, risk models were developed using clinical, baseline PET and i-PET data. The best risk model was compared to the IMPI model and our subsequent ClinicalPET model, also incorporating radiomic features (MTV, IPI, age, SUV peak and D maxbulk) by determination of risk re-classification rates and by generating kaplan-meier (KM)-curves based on 60-30-10 PFS risk groups.

Results: 1014 patients were included in the analyses. Adding i-PET reponse (ΔSUV max) to the IMPI model markedly improved outcome prediction (AIC 3177.44, c-index 0.72) and was superior to IMPI model alone (AIC 3247.09, c-index 0.68). By adding D maxbulk outcome prediction was further improved (AIC 3143.23, c-index 0.74), while SUV peak did not show significant impact on outcome (p=0.07). Compared to the IMPI and the ClinicalPET model, the new model combining baseline features (MTV, age and D maxbulk) with i-PET reponse (ΔSUV max) led to a sharper segregation of KM-curves with an improved rate of correct progression risk classification (22%; 95% confidence interval 12.1-31.1%).

Conclusions: Adding i-PET reponse to baseline clinical and PET parameters optimizes risk classification in DLBCL enabling individualized risk assessment in early phase of frontline treatment and outperforms our previous IMPI model and ClinicalPET models.
Prospective MRI and Neurocognitive Assessment of “Chemobrain” in Acute Myeloid Leukaemia: The Mineral Study
Authors: Scott Ashleigh , Lo Ada , Thompson Kate , Xia Ying , Bodimeade Harriet , Fazlollahi Amir , Curley Cameron , Lane Steven W. , Morris Kirk Lachlan , McNamara Caroline Jane , Pillai Elango S. , Weber Nicholas , Salvado Olivier , McMahon Katie L , Kennedy Glen A
Published: December 2023

CONCLUSIONS

“Chemobrain” is commonly reported by patients treated for Acute Myeloid Leukaemia (AML), potentially attributable to chemotherapy or iron overload. We aimed to determine whether “chemobrain” could be objectively detected in AML patients.

We performed a prospective, observational, longitudinal cohort study in adults commencing chemotherapy for AML, powered to capture a potential 25% incidence of “chemobrain.” Participants underwent neurocognitive assessment (NCA) and brain magnetic resonance imaging (MRI), including iron quantitative susceptibility mapping (QSM), as a biomarker of neuro-inflammation, at three timepoints: pre-treatment (T1), end of treatment (T2), and 6 months later (T3). Longitudinal changes of minimum 1.64 SD were considered significant. The primary outcome was change in NCA score between T1-T2, with secondary correlation analyses between NCA, MRI, QSM and clinical data. This study was funded by a RBWH Foundation 2018 Diamond Care Grant.

Between January 2019 and June 2021, 20 patients were enrolled; of these, 15 and 10 remained alive/enrolled for evaluation at T2 and T3 respectively. Overall NCA scores for the cohort between T1-T2 were similar; however, four (27%) participants performed significantly worse (-1.64SD) in Language or Verbal Memory, with three returning to baseline at T3. Objective NCA score did not correlate with subjective symptoms, and quality of life and physical fatigue improved T1-T2. Cohort brain volumes were similar T1-T2; however, correlation analysis identified interesting potential associations between worse Language performance and volume loss in grey matter (GM; p=0.021) and Wernicke's area (p=0.02), and suggested increased iron susceptibility in frontal lobe (p=0.041) and possibly cortical GM (p=0.055).

Despite patient-reported symptoms, objective neurological toxicity appears uncommon in AML patients, and those with measurable impairment appear to subsequently return to baseline.
Transient Anti-Interferon Auto Antibodies in the Airway Are Associated with Recovery in Mild COVID-19
Authors: Babcock Benjamin R , Kosters Astrid , Roan Nadia R , Lee Sulggi , Ghosn Eliver E.B.
Published: December 2023

Pre-existing blood anti-interferon autoantibodies have been associated with susceptibility to severe COVID-19. However, nothing is known about the autoantibody response in the airway mucosa of mild to severe COVID-19 patients. We profiled longitudinal samples collected from the airway and matching blood of more than 100 donors over >18 months, spanning from mild COVID-19 to hospitalized patients in the ICU. We found that transiently induced anti-type I interferon autoantibodies in the nasal mucosa are a common feature and are associated with recovery from mild and moderate COVID-19.

To quantify the breadth of antibody response in blood and the airway site of infection, we developed flowBEAT, a novel flow cytometry-based assay capable of simultaneously measuring all 8 human antibody isotypes against 22 SARS-CoV-2 structural and non-structural antigens, including autoantigens. We identified the induction of nasal anti-type I interferon autoantibodies within the first two weeks of infection, waning with disease recovery. Notably, there was an inverse association between nasal and blood anti-interferon responses, while nasal anti-interferon was linked to recovery in cases of mild/moderate disease. In contrast, near-ubiquitous anti-interferon in the endotracheal aspirates of critical cases correlated with heightened airway hyper-inflammation. In both blood and mucosa, anti-interferon positively correlated with a broad anti-viral antibody response, including non-structural proteins. While nasal anti-interferon was universally transient, we identified a group of donors sustaining long-term blood anti-interferon autoantibodies who may be at risk for a more severe subsequent viral infection.

Within our longitudinal cohort, we assessed 500 infected-recovered and vaccinated samples and showed that nasal anti-SARS-CoV-2 antibodies are sustained over 18 months by repeated vaccination and boosting. We identified isotype signatures associated with disease severity and an autoantibody signature predictive of vaccine non-response. Thus, our findings emphasize the importance of continued vaccination/boosting to sustain nasal immunity and help prevent ongoing community spread. More importantly, our studies reveal predictive blood markers of protective nasal immunity and highlight a protective role for nasal anti-interferon autoantibodies in the immunopathology of COVID-19.
Stag2a or Stag2b Knockout Zebrafish Are Viable, Yet Loss of Stag2a Is Associated with Abnormal Expression of Early and Late Myeloid and Lymphoid Markers
Authors: Anderson Rebecca A , Oyarbide Usua , Corey Seth J
Published: December 2023

Introduction:Mutations in Stromal antigen 2 ( STAG2) are frequently found in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). In mice, loss of Stag2 is embryonic lethal. Conditional knockout of Stag2 in adult mice results in MDS-like hematopoietic anomalies. Zebrafish harbor two paralogues of STAG2 ( stag2a and stag2b). To date, a complete stag2-null zebrafish model organism has not been developed. Recently reported, a stag2b-/- zebrafish demonstrated downregulation in primitive erythropoiesis. However, any hematopoiesis beyond 2 days post fertilization (2 dpf) was not detailed. Furthermore, a stag2a mutant zebrafish has not been reported.

Methods and Results: Using CRISPR/Cas9 genome editing, we created a stag2a mutant anda stag2b mutant zebrafish. Both mutations result in a frame shift and premature protein truncations. Unlike the murine Stag2 -/- mutant, zebrafish stag2a -/- mutants and stag2b -/-mutants survive to adulthood, with no obvious morphological phenotypes. Detection of hemoglobin, using O-dianisidine staining, reveals anemia at 31 hours post fertilization (31 hpf) in stag2a +/- mutants (10/29), stag2a -/-mutants (10/15), stag2b +/-mutants (6/11), and stag2b -/-mutants (5/7); wildtype siblings appear unaffected. Intriguingly, at 4 dpf, stag2a maternal-zygotic mutants show increased expression levels of early and late myeloid and lymphoid markers. ( Figure 1) Work is ongoing to analyze blood cell lineages from zebrafish whole kidney marrow, the equivalent of mammalian bone marrow, in adult stag2a and adult stag2b mutants. In addition, we are characterizing the zebrafish stag2a;stag2b double mutants, which we will present.

Conclusions: In zebrafish, stag2a and stag2b contribute to proper primitive and definitive hematopoiesis. Our organismal model of Stag2 deletion in the zebrafish will yield new insight into how loss of STAG2 may lead to MDS/AML.
Real World Data Venetoclax Based Therapies in a Single Center in Latin America
Authors: Rocha Juliana Dall Agnol , Arcuri Leonardo Javier , Kojima Rodrigo Seiti , Hamerschlak Nelson , Santos Fabio P. S. , Ennes Amanda Inácio Dias , Datoguia Tarcila Santos , Nascimento Jade Zezzi , Marques Carolina Perrone , Chaves Felipe Galvão Batista , Junior Luiz Frederico Bezerra Honorato , Galvao Barbara Ferreira Cordeiro
Published: December 2023

Introduction: Venetoclax-based therapies have changed the outcomes of acute myeloid leukemia, in combination with hypomethylating agents, low-dose cytarabine or in older patients not eligible for intensive regimens. Combinations with venetoclax with target therapy and with standard chemotherapy are being tested to improve outcomes in specific subgroups. In Latin America, the lack of clinical trials increases prescriptions not yet approved in international guidelines (off-label use). In this paper, we present data from a private reference center in Latin America for the treatment of acute myeloid leukemia and high-risk myelodysplasia.

Methods: This is a unicentric, retrospective, and non-interventional cohort study performed at Brazilian Hospital Albert Einstein. Inclusion criteria were adult patients (aged ≥18 years) diagnosed with AML between May 2019 and January 2023, who were treated with a venetoclax-based regimen. Independent variables analyzed were age, gender, prognostic risk according to ELN 2022, indication of bone marrow transplantation, and molecular and cytogenetic risk at diagnosis.

Results: Sixty-three (N= 63) patients were included, with a median age of 63.1 years old. Forty-eight patients received a combination of venetoclax and a hypomethylating agent, two were associated with low doses of cytarabine, eight were associated with target therapy (inhibitor of FLT3), and five with intensive chemotherapy. The median follow-up was 303 days, and the median use of venetoclax in those who did not suspend treatment was 352 days. The main cause of suspension was hematological toxicity. In the univariate analysis, the following onco-hematological karyotype variables, age, type of regimen combined with venetoclax, and decision for transplant did not show statistical significance.

Conclusion: Real-world data show that venetoclax-based therapies are effective in patients with AML, and their combinations have been increasingly explored.
Albumin: Globulin Reversal As a Screening Tool for Monoclonal Gammopathy (Targeted Screening) in Hospitalized Setting
Authors: Raphael Celine , Yanamandra Uday , Kumar Abhinav , Kotwal Narendra , Malhotra Pankaj , Nair Velu
Published: December 2023

A

Introduction: There is an unmet need to discover novel targeted screening guidelines for Qmonoclonal gammopathy of undetermined clinical significance (MGUS) in the Indian scenario which can help in early detection and diagnosis. Liver function tests (LFTs) are routinely performed in hospital settings for a variety of indications, with many having albumin-globulin (A:G) ratio reversals. We have designed this study to gauge the prevalence of MGUS in individuals above the age of 50 years with A: G ratio reversal in hospitalized settings. The objectives of the study were to measure the incidence of A: G ratio reversal in total LFT samples run in the hospital for various indications and to determine whether targeted screening of unexplained A: G ratio reversal in hospitalized patients is a fruitful strategy.

Materials and Methods:

This is a single-center, prospective observational study carried out from 01 May 2022 to 30 May 2023 at tertiary care center in Western India. All the LFTs of the admitted patients over the study period to the medical wards were included in the study. Patients with A:G reversal were subjected to detailed clinical examination and investigations to rule out non-myelomatous causes for the reversal of the ratio. Samples with unexplained A:G ratio reversal and unresolving A:G reversal despite of three months of observation were subjected to Serum Protein Electrophoresis (SPEP). Data was analysed using JMP ver. 16.0.0.

Results:

A total of 5148 LFTs were screened during the study period, of which 300 (5.82%) had A:G reversal. The median age of the study population with A:G reversal was 59 years (IQR - 14.3) with a male preponderance (59%). Etiologically amongst patients with A:G reversal 54% had infection, 15% had inflammation, 13% suffer from Chronic Liver Disease and 9% have Chronic Kidney Disease. Only 9% of patients (n-27) had no evident secondary causes, and of these 11% (n-3) have an M-spike on SPEP. Of all patients with A:G reversal, 1.1% have evidence of MGUS, all being IgG Lambda variants and none having any features of smoldering or multiple myeloma based on IMWG guidelines.

Conclusion:

G reversal compounded with good clinical history and examination can be an effective tool for targeted screening of MGUS. A majority of A:G reversals in hospital settings are due to secondary causes, requiring deliberate effort for exclusion.
Transcriptome analysis reveals temporally regulated genetic networks during Drosophila border cell collective migration
Authors: Burghardt Emily , Rakijas Jessica , Tyagi Antariksh , Majumder Pralay , Olson Bradley J.S.C. , McDonald Jocelyn A.
Published: December 2023

Background

Collective cell migration underlies many essential processes, including sculpting organs during embryogenesis, wound healing in the adult, and metastasis of cancer cells. At mid-oogenesis, Drosophila border cells undergo collective migration. Border cells round up into a small group at the pre-migration stage, detach from the epithelium and undergo a dynamic and highly regulated migration at the mid-migration stage, and stop at the oocyte, their final destination, at the post-migration stage. While specific genes that promote cell signaling, polarization of the cluster, formation of protrusions, and cell-cell adhesion are known to regulate border cell migration, there may be additional genes that promote these distinct active phases of border cell migration. Therefore, we sought to identify genes whose expression patterns changed during border cell migration.

Results

We performed RNA-sequencing on border cells isolated at pre-, mid-, and post-migration stages. We report that 1,729 transcripts, in nine co-expression gene clusters, are temporally and differentially expressed across the three migration stages. Gene ontology analyses and constructed protein-protein interaction networks identified genes expected to function in collective migration, such as regulators of the cytoskeleton, adhesion, and tissue morphogenesis, but also uncovered a notable enrichment of genes involved in immune signaling, ribosome biogenesis, and stress responses. Finally, we validated the in vivo expression and function of a subset of identified genes in border cells.

Conclusions

Overall, our results identified differentially and temporally expressed genetic networks that may facilitate the efficient development and migration of border cells. The genes identified here represent a wealth of new candidates to investigate the molecular nature of dynamic collective cell migrations in developing tissues.
Development and validation of risk of CPS decline (RCD): a new prediction tool for worsening cognitive performance among home care clients in Canada
Authors: Guthrie Dawn M. , Williams Nicole , O’Rourke Hannah M. , Orange Joseph B. , Phillips Natalie , Pichora-Fuller M. Kathleen , Savundranayagam Marie Y. , Sutradhar Rinku
Published: December 2023

Background

To develop and validate a prediction tool, or nomogram, for the risk of a decline in cognitive performance based on the interRAI Cognitive Performance Scale (CPS).

Methods

Retrospective, population-based, cohort study using Canadian Resident Assessment Instrument for Home Care (RAI-HC) data, collected between 2010 and 2018. Eligible home care clients, aged 18+, with at least two assessments were selected randomly for model derivation (75%) and validation (25%). All clients had a CPS score of zero (intact) or one (borderline intact) on intake into the home care program, out of a possible score of six. All individuals had to remain as home care recipients for the six months observation window in order to be included in the analysis. The primary outcome was any degree of worsening (i.e., increase) on the CPS score within six months. Using the derivation cohort, we developed a multivariable logistic regression model to predict the risk of a deterioration in the CPS score. Model performance was assessed on the validation cohort using discrimination and calibration plots.

Results

We identified 39,292 eligible home care clients, with a median age of 79.0 years, 62.3% were female, 38.8% were married and 38.6% lived alone. On average, 30.3% experienced a worsening on the CPS score within the six-month window (i.e., a change from 0 or 1 to 2, 3, 4, 5, or 6). The final model had good discrimination (c-statistic of 0.65), with excellent calibration.

Conclusions

The model accurately predicted the risk of deterioration on the CPS score over six months among home care clients. This type of predictive model may provide useful information to support decisions for home care clinicians who use interRAI data internationally.
Comprehensive analysis of cancer of unknown primary and recommendation of a histological and immunohistochemical diagnostic strategy from China
Authors: Ren Min , Cai Xu , Jia Liqing , Bai Qianming , Zhu Xiaoli , Hu Xichun , Wang Qifeng , Luo Zhiguo , Zhou Xiaoyan
Published: December 2023

Background

Previous studies on cancer of unknown primary (CUP) mainly focus on treatment and prognosis in western populations and lacked clinical evaluation of different IHC markers, so this study aimed to evaluate characteristics of CUP and recommend a diagnostic strategy from a single center in China.

Methods and results

Data of 625 patients with CUP were retrospectively collected and reviewed. The patients ranged in age from 20 to 91 years, with a female-to-male ratio of 1.3:1. The predominant histological type was poor or undifferentiated adenocarcinomas (308; 49.3%). The results of Canhelp-Origin molecular testing for the identification of the tissue of origin in 262 of 369 patients (71.0%) were considered predictable (similarity score > 45), with the most common predicted primary tumor site being the breast (57, 21.8%). Unpredictable molecular results correlated with more aggressive clinical parameters and poor survival. Thee positivity rates of several targeted antibodies (GATA3, GCDFP15, TTF1, Napsin A, and PAX8), based on the clinically predicted site, were lower than those reported for the corresponding primary tumors. Nonetheless, TRPS1 and INSM1 were reliable markers of predicted breast carcinoma (75.0%) and neuroendocrine tumors (83.3%), respectively. P16 expression, as well as HPV and EBER testing contributed significantly to the diagnosis of squamous cell carcinomas. Survival analysis revealed that older ages (> 57), ≥ 3 metastatic sites, non-squamous cell carcinomas, bone/liver/lung metastases, unpredictable molecular results, and palliative treatment correlated with poor overall survival.

Conclusions

We recommend a CUP diagnostic strategy involving the use of targeted antibody panels as per histological findings that is potentially applicable in clinical practice. The markers TRPS1, INSM1, and P16 expression, as well as HPV and EBER testing are particularly valuable in this aspect. Molecular testing is also predictive of survival rates.