bluuebunny/crossref_metadata_2025 huggingface dataset:
High Disease-Free Survival in Patients with High-Risk MDS Treated with Orca-T
Authors: Gandhi Arpita , Hoeg Rasmus T. , Patel Sagar S. , Waller Edmund K. , Pantin Jeremy M. , Oliai Caspian H. , McClellan J. Scott , Pavlova Anna , Agodoa Irene , Fernhoff Nathaniel B. , Meyer Everett H.
Published: December 2023

BACKGROUND

Allogeneic hematopoietic cell transplant (alloHCT) may be a curative treatment option for patients with myelodysplastic syndrome (MDS). The use of investigational therapies that potentially reduce transplant toxicity could improve outcomes in this patient population with difficult to treat disease. Per CIBMTR, survival after alloHCT for MDS using matched donors is approximately 65% at 1 year and relapse-free survival at 1 year was ~69% in MDS patients in the RICMAC trial who received myeloablative conditioning (MAC).

Orca-T is a high-precision cell therapy biologic that includes stem and immune cells, derived from allogeneic donors, that leverages highly purified, polyclonal donor regulatory T cells to control alloreactive immune responses. In this phase 1b study, Orca-T was evaluated in MDS patients who were eligible for transplant per 2017 International Expert Panel recommendations.

METHODS

As of 03/31/23, 16 patients with intermediate to very high risk (HR) MDS with ≤10% blasts in the bone marrow at the time of transplant had received Orca-T in the phase 1b study. IPSS-M prognostic scores were Low (6%), Moderate Low (6%), Moderate High (19%), High (44%), and Very High (25%). Baseline HCT-CI scores were 0 (n=6), 1 (n=3), 2 (n=2), 3 (n=3), and 4 (n=2). The median f/u was 19.1 months (range 6.3 - 32.2). Patients were aged 43 - 65 (median 59) and 56% female. Donors were 8/8 HLA-matched related (n=7) or unrelated (n=9). Patients received a busulfan-based myeloablative chemotherapy regimen prior to Orca-T, followed by single-agent GvHD prophylaxis with tacrolimus.

RESULTS

Orca-T was successfully manufactured in a single, centralized GMP manufacturing facility for all patients enrolled, distributed, and infused throughout the U.S. Orca-T was delivered within < 72 hours vein-to-vein (i.e. time between end of donor apheresis to start of recipient's Orca-T infusion).

Disease-free survival (RFS) at 12 months was 94% (Figure). At 1 year, no patients had experienced grade ≥ 3 acute GvHD and moderate to severe chronic GvHD had occurred in 2 patients. Modified GRFS (survival free of Grade 3-4 aGvHD, moderate-to-severe cGvHD and relapse) at 12 months was 75%. Non-relapse mortality (NRM) and overall survival (OS) were 0% and 94%, respectively, at 12 months.

Additionally, one MDS patient with very poor prognosis, characterized by mutated TP53 and complex karyotype, remains in CR without active signs of acute or cGvHD at 500 days post-alloHCT.

CONCLUSIONS

Orca-T demonstrated promising results in this HR-MDS patient population, including high RFS, low incidence of GvHD, and very high OS at 1 year. A randomized controlled phase 3 multi-center trial comparing Orca-T to SOC in patients with MDS, AML, and ALL is currently enrolling across the US (NCT05316701).
Early Initiation of Ketamine Infusion for Pain Management in Pediatric BMT Patients with Mucositis
Authors: Utley Kaylyn , Azadegan Ali , Rosenbaum Adam Rene , McGinn Brenna , Deis Amanda , Hamer Katie , Barthelmess Erin , David Alexandria , Southworth Debra , Trawick Cyndi , Tan Gee Mei , Fabrizio Vanessa A.
Published: December 2023

Background: As many as 90% of children undergoing hematopoietic stem cell transplant (HSCT) will experience painful mucositis. The mainstay of treatment for these children has been opioid analgesia which can be associated with sedation and respiratory depression. Additionally, these children may experience heightened opioid tolerance and physiologic dependence contributing to lengthy admissions. Ketamine is an anesthetic and analgesic which has been used successfully in conjunction with opioids in the treatment of painful mucositis. This study aimed to reduce total opioid use per day by 10% and overall pain scores by 2 in pediatric patients with mucositis undergoing HSCT through initiation of ketamine at the same time as an opioid patient-controlled analgesia (PCA).

Methods: Beginning April 2022, HSCT patients (both autologous and allogeneic) at Children's Hospital Colorado were started on ketamine infusion in conjunction with an opioid PCA for management of painful mucositis. Mucositis severity and duration, opioid consumption, pain and functional scores, time of initiation of ketamine therapy, ketamine side effects, and duration of PCA and ketamine therapy were recorded for a retrospective (April 2021- March 2022, n=6) and a prospective cohort (April 2022- April 2023, n=17). A 2-sample-T-test was used to calculate the difference in mean cumulative opioid use per day and the mean 24-hour average pain scores between the retrospective and prospective groups.

Results: 71% of children in the prospective group underwent ketamine initiation alongside an opioid PCA. Patients in this group had a 46% reduction in mean cumulative opioid use per day (CI 1.265-2.318, p=0.000). Reduction in 24-hour pain scores between the retrospective and prospective groups was 1.18, failing to meet our goal of 2.0. However, there was still a statistically significant difference in mean scores (CI 0.499-1.860, p=0.001). Additionally, this intervention was tolerated well and only 1 patient (16%) was intolerant of ketamine side effects in the prospective cohort.

Conclusions: Initiation of ketamine alongside an opioid PCA for the treatment of mucositis pain is safe and significantly reduces cumulative opioid exposure and 24-hour pain scores in children undergoing HSCT.
Comparison of Maintenance Therapy Regimens of Patients Treated for Multiple Myeloma
Authors: Wani Kashmira , Gutta Radhika , Mittal Aayush , Jamil Maria , Springer Kylie , Kuriakose Philip
Published: December 2023

Background: Despite significant progress in treatment modalities and improved survival and response rates in patients diagnosed with multiple myeloma (MM), it remains an incurable disease with a poor outcome, especially in high-risk groups. Though not all patients are eligible, autologous stem-cell transplantation (ASCT) remains an integral part of the treatment of patients with both newly diagnosed and relapsed MM. Regardless of whether patients receive a transplant, they do receive maintenance therapy, and recent evidence has demonstrated that maintenance therapies offer an advantage in progression free and overall survival. While Revlimid is the standard of care, data regarding the specifics of maintenance therapy in high-risk patients is limited and the overall impact of various regimens on survival needs to be further investigated. In our retrospective chart review we reviewed all adult patients with advanced MM within Henry Ford Cancer Institute in the last 10 years to determine the impact of maintenance therapy on progression free survival (PFS) and overall survival (OS).

Methods: We conducted a retrospective chart review of adult patients with MM who underwent ASCT between January 1, 2012 to December 31, 2022. Those who received maintenance chemotherapy after transplant were included in the study. Patients who were lost to follow-up or who had largely incomplete records were excluded. Data points including age, ethnicity, cytogenetic analysis, risk category, maintenance regimen after transplant, last chemotherapy regimen, last follow-up, and date of death (if applicable) were recorded. Maintenance chemotherapy regimens were recorded as Revlimid versus other. Patients were split into 2 categories based on risk - high risk and standard risk. Kaplan-Meier plots were used to illustrate overall survival and time to relapse between groups for various variables. Statistical significance was set at p<0.05

Results: 158 patients were included in the study of which 44 were considered high-risk based on cytogenetics, 106 were standard-risk and 8 were missing. Most of the patients (n=137, 87.3%) received Revlimid, while 20 (12.7%) received maintenance therapy other than Revlimid, and for 1 patient, the type of maintenance therapy was unknown. Within the high-risk group, no statistical significance in OS or PFS was found between patients that received Revlimid versus those that did not. Furthermore, there was no statistical significance in OS and PFS within high risk versus standard cytogenetic risk groups.

Conclusions: We did not see a difference in outcome based on risk and believe all patients would derive equal benefit from maintenance therapy We also did not see a difference in outcome between high and standard risk patients, and for the high-risk subgroup, there was a separation in curves suggesting that maintenance therapy has benefit compared to no maintenance.
IPSS-M Predicts Survival Outcomes Significantly Better Than IPSS-R in Patients Undergoing Allogeneic Hematopoietic Stem Cell Transplantation for Myelodysplastic Neoplasms
Authors: Yang Tingting , Jiang Bingqian , Luo Yi , Zhao Yanmin , Ouyang Guifang , Yu Jian , Lan Jianping , Lu Ying , Lai Xiaoyu , Ye Baodong , Chen Yi , Liu Lizhen , Xu Yang , Guo Qunyi , Shi Pengfei , Xiao Haowen , Hu Huixian , Fu Huarui , Ye Yishan , Wang Xinyu , Sun Jie , Zheng Weiyan , He Jingsong , Zhao Yi , Wu Wenjun , Cai Zhen , Wei Guoqing , Huang He , Shi Jimin
Published: December 2023

Background

The newly published molecular International Prognostic Scoring System (IPSS-M) represents a powerful risk stratification tool for treatment decision-making in myelodysplastic neoplasms (MDS); however, its utility in the context of allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains to be fully explored.

Methods

We retrospectively analyzed a large multicenter cohort of 347 MDS patients who underwent allo-HSCT between January 2017 and October 2022. The prediction accuracy of IPSS-M at diagnosis for 3-year survival outcomes was assessed and compared to that of the conventional revised International Prognostic Scoring System (IPSS-R).

Results

Among the 347 patients, median age at transplant was 48.3 years and 41.2% of patients were female. According to IPSS-M, patients were clustered as very low risk (1.2%), low risk (11.2%), moderately low risk (11.5%), moderately high risk (18.2%), high risk (33.4%), and very high risk (24.5%), resulting in a restratification of 49.3% of the entire cohort when compared with IPSS-R. Of these reclassified patients, 52.6% patients were upstaged and 47.4% were downstaged. Median follow-up time among the survivors was 28.6 (range, 4.7 to 76.6) months. With the IPSS-M model, overall survival (OS) and leukemia-free survival (LFS) discrimination was refined relative to the IPSS-R as evidenced by a 7.0 percentage- and 5.7 percentage-point increase in the concordance index (C-index), which was further supported by the lower Akaike Information Criterion and higher C-indexes in multivariate analyses. Among patients undergoing haploidentical HSCT, IPSS-M model also demonstrated significantly improved prognostic performance for LFS versus IPSS-R (C-index, 0.707 vs 0.604) which was validated by multivariate analyses. When restricting our analyses to younger patients (<49 years) and patients carrying detectable mutations, IPSS-M retained greater prognostic value with respect to OS and LFS; while it failed to stratify individual probability of OS and LFS in their counterparts.

Conclusions

IPSS-M was confirmed to increase prognostic discrimination at the individual level and is applicable to transplant-specific settings, which also had an advantage for subjects carrying mutations and younger patients.
Transient CDK9 Inhibition with AZD4573 Effectively Induces Apoptosis in Burkitt Lymphoma As a Monotherapy in Pre-Clinical Models
Authors: Potter Danielle S , Ott India , Saeh Jamal , Fawell Stephen , Drew Lisa , Roderick-Richardson Justine
Published: December 2023

Burkitt Lymphoma (BL) accounts for almost two thirds of all B-cell non-Hodgkin lymphoma cases in children and adolescents. It may affect the jaw, bones of the face, bowel, kidney, ovaries and in some cases spread to the central nervous system. Nearly all types of BL are characterized by c-MYC gene translocation of chromosome 8 with immunoglobulin genes at chromosome 2, 14 or 22 resulting in high levels of the transcription factor. Recently, MCL-1 was shown to be overexpressed in BL and is a critical survival factor in pre-clinical BL models using genetic and chemical approaches.

CDK9, a serine/threonine kinase, regulates transcription elongation through phosphorylation of RNA polymerase II at serine 2 (pSer2-RNAPII). AZD4573 is a highly potent and selective CDK9 inhibitor that results in downregulation of short-lived transcripts and labile proteins such as c-MYC and key survival proteins including MCL-1 and BFL-1, resulting in apoptosis in several hematological malignancies (Cidado et al. 2020 & Boiko et al. 2021)

We selected several preclinical BL models to assess the rapid apoptogenic potential of AZD4573 in vitro and in vivo. To measure apoptosis, we evaluated cleaved caspase-3 (CC3) induction following acute treatment (6h) using Caspase-Glo 3/7. Using this assay, we found that 3 BL models were sensitive to CDK9 inhibition (EC50 < 100nM; max. CC3 > 50%) and 2 were resistant (EC50 > 100nM; max. CC3 < 50%). Treatment with AZD4573 in these sensitive BL cell line models decreased pSer2-RNAPII, and downregulation of MCL-1 and c-MYC within 6 hours. In AZD4573 resistant lines we observed relatively higher levels of anti-apoptotic proteins BCL2 and BCL-xL, compared to the sensitive cell lines. These increased levels potentially lead to the decreased sensitivity to AZD4573, which does not reduce levels of BCL-2 and BCL-xL due to their longer half-life. Using CRISPR genetic knockouts we demonstrate that BL cell lines are dependent on c-MYC and MCL-1 and that loss of these protein decreases caspase induction after AZD4573 treatment, highlighting the importance of these proteins in driving the efficacy of AZD4573. In vivo AZD4573 dosed on a once weekly schedule resulted in tumor growth inhibition in aggressive BL cell line xenografts Namalwa and Ramos (40-60%) and decreased pSer2-RNAPII, MCL-1 and c-MYC and increased CC3 8 hours after treatment. AZD4573 is currently in a phase 2 study (NCT05140382) to assess the efficacy, safety, and PK of AZD4573 in patients with relapsed or refractory Peripheral T-cell lymphomas (pTCL). Our findings demonstrate that targeting CDK9 with AZD4573 can effectively induce apoptosis in pre-clinical BL models and could be an effective therapy for BL patients.
CHD4 Promotes Tumor Formation, Survival, and Resistance to Genotoxic Therapeutics in TP53 Mutant T-Cell Acute Lymphoblastic Leukemia Cells through the NuRD Chromatin Remodeling Complex
Authors: Shang Shengzhe , Aijaz Javeria , Ginder Gordon
Published: December 2023

T-Cell acute leukemia(T-ALL) though generally responsive to current chemotherapy remains incurable in over 50% of adult patients. Importantly, the survival for patients with TP53 mutant T-ALL disease is less than 10%. We have previously shown the role of the Methyl Binding Domain protein 2-Nucleosome Remodeling and Deacetylase (MBD2-NURD) chromatin remodeling complex in triple negative breast cancer cell growth and the role of the NuRD component, Chromatin-helicase-binding protein 4(CHD4), in tumor colony formation and genotoxic resistance in acute myeloid leukemia.

In the present study, we have examined the role of specific NuRD complexes and CHD4 in T-cell acute leukemia cells with mutant TP53. The prototypical NuRD complex possesses both a histone deacetylase (HDCC) sub-complex and an ATPase chromatin remodeling sub-complex containing CHD4 linked to MBD2 or MBD3 of the HDCC. CHD4 is linked to the MBD2/3 anchors of the NuRD complex, via GATAD2A or GATAD2B. Depletion of CHD4 was found to increase the rate of apoptosis in several TP53 mutant T-ALL cell lines including Jurkat, MOLT4, RPMI and CCRF by up to 5 to 10-fold. Likewise, depletion of both MBD2 and MBD3 also increased apoptosis and sensitization to the standard genotoxic chemotherapeutic agent, cytosine arabinoside (Ara-C) but had only minimal effects in normal human CD34+ hematopoietic progenitor cells. Depletion of GATAD2B and GATAD2A by shRNA also resulted in a significantly increased survival of 60 days compared to 20 days in Scramble shRNA controls, in a xenogeneic mouse model of T-ALL. Depletion of CHD4 resulted in a profound (10 to 100-fold) decrease in colony formation of the TP53 mutant T-ALL cell lines in methylcellulose. RNA seq analyses identified the apoptosis, ferroptosis and mitophagy pathways as potential mechanisms for the striking effect of NURD disruption on colony formation.

In summary, CHD4, through the NuRD complex, promotes tumor cell formation, survival and therapeutic resistance of TP53 mutant T-ALL cells. Current efforts are aimed at validating these findings in primary T-ALL cells and structure-function studies to advance development of tool compounds for disrupting the NuRD complex, with the long-term goal of developing improved small molecule therapeutics for treatment resistant acute leukemia.
Apoptotic Pathway Inhibition in Cold-Stored Platelet Concentrates Facilitates Retained Ability to Form Thrombi
Authors: Kirschall Johanna , Marini Irene , Pelzl Lisann , Witzemann Andreas , Wolska Nina , Bakchoul Tamam
Published: December 2023

Introduction: Platelet concentrates (PCs) are used to treat or prevent bleeding in patients with impaired platelet function or after injury. However, the standard storage of PCs at room-temperature (RT) increases the risk of bacterial contamination and is associated with reduced platelet functionality due to storage lesions. Our group reported that cold-stored PCs have better functionality compared to RT-stored PCs but reduced in vivo circulation time due to cold-induced apoptosis. In order to investigate cold-stored PCs hemostatic functionality with and without apoptosis inhibition, we established an ex vivo model simulating physiological conditions of blood flow to evaluate the contribution of cold-stored PCs in thrombus formation.

Methods: Apheresis-derived PCs were stored for 1, 4, 7 and 10 days under agitation at 4°C or RT either with or without the apoptosis inhibitor G04 (RhoA GTPase inhibitor). Next, PCs were stained with CD41 antibody, recalcified and incubated with TRAP6 (thrombin receptor-activating peptide 6) to initiate thrombus formation. Platelet-depleted whole blood samples from healthy donors were spiked-in with PCs and administered to the microfluidic channels of the ex vivo system (Bioflux), under physiological shear pressure. After five minutes of perfusion the resulting thrombi were imaged and analyzed. Ability to form thrombi was further assessed by thromboelastography and impedance aggregometry.

Results: Platelets from 24h storage PCs showed stable thrombus formation upon activation with TRAP6. Unstimulated platelets did not form any thrombi during the entire period of perfusion. To evaluate the effect of cold storage on platelet functions we analyzed the extent of thrombus formation of PCs stored at RT and 4°C upon CD41 staining. Our results indicate that cold-stored platelets form larger clots under flow conditions and maintain higher functionality compared to PCs stored at RT. Moreover, incubation of cold-stored PCs with the apoptosis inhibitor G04 improved ex vivo thrombus formation compared to PCs stored without the inhibitor, without inducing any adverse effects (Figure 1).

Conclusion: The results indicate that our ex vivo assay, which simulates PC transfusion in thrombocytopenic patients, is suitable to test the hemostatic functions of PCs under physiological flow conditions. We demonstrated that PCs stored at 4°C show better thrombus formation ability upon stimulation by agonists compared to RT-stored PCs and that the ability to form clots under shear stress is still preserved after incubation with the apoptosis inhibitor.

To conclude, cold-storage of PCs in combination with apoptosis inhibition is a promising stragedy to reduce the risk of bacterial contamination and prolong in vivo survival time while preserving platelet's hemostatic functionality.
A SAGA Complex-Specific Dependency in Multiple Myeloma Orchestrates Oncogenic Transcriptional Programs
Authors: Chen Ying-Jiun C , Badiger Bhaskara Govinal , Lu Yue , Dent Sharon
Published: December 2023

Multiple myeloma (MM) is a malignancy of antibody-producing plasma cells that reside within the bone marrow. Epigenetic factors contribute to the initiation, progression, relapse and clonal heterogeneity in MM, but our knowledge of epigenetic regulators involved in MM development is far from complete. Results from the Cancer Dependency Map (DepMap) Project revealed that several components of the SAGA complex are particularly important for MM cell growth and survival. The SAGA complex serves as a coactivator in transcription and also catalyzes histone acetylation and deubiquitination via its lysine acetyltransferase (KAT) and deubiquitinase (DUB) modules, respectively. Our analysis showed that each SAGA complex component is overexpressed in at least 50% of MM patients. To investigate SAGA-specific functions, we centered on ADA2B, a subunit in the KAT module that specifically functions in the SAGA complex.

We observed reduced cell viability upon ADA2B knockdown and CRISPR/Cas9-mediated knockout in MM cell lines. To identify the underlying transcriptional pathways that ADA2B regulate, we performed transcriptome sequencing using MM cell lines with and without ADA2B depletion. Our results showed several oncogenic pathways, including MYC, E2F and MM-specific MAF oncogenic programs, are tightly regulated by ADA2B. We also discovered that the protein but not transcript levels of MAF and the KATs in SAGA, GCN5 and PCAF, as well as the global level of histone H3 lysine 9 acetylation (H3K9ac) are all prominently decreased upon ADA2B depletion. To investigate whether the global reduction in H3K9ac would alter chromatin accessibility, we performed assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) and found decreased accessibility at many cancer marker and cancer-promoting genes.

To decipher the protein domains that are essential for ADA2B function in MM, we generated and expressed mutated versions of ADA2B in MM cell lines. Excitingly, mutations of core hydrophobic amino acid residues in the SANT domain disrupt interactions with GCN5 and PCAF and negatively affect protein stability of ADA2B. Hence, drug inhibitors that target the SANT domain could achieve suppression of ADA2B and MM cell growth.

Our findings uncover novel epigenetic and transcriptional mechanisms controlling MM development, and suggest potential for new epigenetic inhibitors targeting SAGA-specific ADA2B as future treatment modalities for MM.
The Efficacy and Safely of Anticoagulation for the Management of Gonadal Vein Thrombosis
Authors: Brown Cameron , Ng Sarah , Zarka Farah , Delluc Aurélien , Carrier Marc
Published: December 2023

Background

Gonadal vein thrombosis (GVT) is an uncommon condition that has been associated with different risk factors (e.g., post-partum period, cancer, recent pelvic surgery, etc.). The optimal management of GVT remains unclear. Anticoagulation therapy may help to mitigate the risk of thrombosis extension and recurrent venous thromboembolism (VTE) but may also be associated with bleeding complications.We sought to assess the efficacy and safety of anticoagulation therapy in patients with GVT.

Methods

A systematic search of MEDLINE, EMBASE and PubMed for randomized controlled trials and observational studies was performed . Studies with patients presenting with GVT defined as ovarian vein thrombosis or testicular vein thrombosis and reporting at least one of the desired outcomes were included in the analysis.The primary efficacy outcome was recurrent VTE, and the primary safety outcome was major bleeding episode. Secondary outcomes included clinically relevant non-major bleeding and overall mortality. Incidence rates of the outcomes were pooled using the random effects model and expressed as event per 100 patient-years with its associated 95% confidence intervals (CI) using R software.

Results

The search strategy identified 7084 article records. A total of 13 observational studies (1039 patients) with GVT met the inclusion criteria and were included in the analysis. Only one study reported the incidence of testicular vein thrombosis. Hence, 96% of the total patient population were females with ovarian vein thrombosis. Overall, the incidence rate per 100 patient-years for recurrent VTE was 3.60 (95% CI: 2.12-6.13; I-square 75%; Figure 1). A large majority of studies did not report if recurrent events occurred during or following discontinuation of anticoagulation. Similarly, most studies did not report on major or clinically relevant non-major bleeding events, or overall mortality preventing the possibility of conducting pooled analyses for these outcomes.

Conclusion

GVT seems to be associated with a relatively low risk of recurrent VTE. However, it remains unclear if anticoagulant therapy is associated with a favorable risk/benefit ratio. Future studies (e.g., registries, population-based studies) are needed to investigate the efficacy of anticoagulant and whether it outweighs the risk of bleeding in this patient population.
Risk of Developing Post Thrombotic Syndrome after Deep Vein Thrombosis with Different Anticoagulant Regimens, a Systematic Review and Pooled Analysis
Authors: Brown Cameron , Tokessy Lauren , Delluc Aurélien , Carrier Marc
Published: December 2023

Background

Post-thrombotic syndrome (PTS) is a chronic condition that arises in up to 20-50% of patients with deep vein thrombosis (DVT) of the lower extremity. Symptoms range from skin color changes and limb heaviness to edema, chronic pain and ulcers. PTS has a significant impact on patients' quality of life. Hence, it is important to optimize DVT management to minimize the incidence and severity of PTS. It is unclear if different anticoagulant therapies (e.g. vitamin K antagonists (VKA), direct oral anticoagulants (DOACs) or low molecular weight heparin (LMWH)) are associated with different risks of PTS.

Objective

We sought to assess the incidence rates of PTS development after a proximal DVT of the lower extremity managed with different anticoagulation regimens.

Methods

A systematic search of MEDLINE, EMBASE and PubMed as well as conference proceedings (inception to June 2023) was performed. Studies were screened and selected if they met the criteria for patients (age ≥18) receiving anticoagulation therapy for a minimum of 3 months after the diagnosis of a proximal DVT of the lower extremity. The primary outcome was development of PTS defined by the Villalta or PRV score of ≥ 5, or as defined by the individual studies. Incidence rates (overall and per type of anticoagulant regimens) were pooled using the random effects model and expressed as event per 100 patient-years with its associated 95% confidence intervals (CI) using R software.

Results

Out of the 2837 identified studies, 77 were reviewed in full text and 21 (n= 4342 patients) were included in the analysis (7 randomized controlled trials, 14 observational studies). There were 17 studies (2834 patients), 12 studies (1212 patients) and 2 studies (296 patients) reporting the outcomes for VKA, DOAC and LMWH, respectively. The crude incidence rates for PTS development are depicted in Figure 1. After adjusting for duration of anticoagulation, the overall incidence of PTS was 16.3 per 100 patient-years (95% CI: 11.1-24.12). The incidence of PTS was 15.1 per 100-patient-years (95% CI: 8.7-26.1), 18.2 per 100 patient-years (95% CI: 9.4-35.1), 24.6 per 100 patient-years (95% CI: 9.2-65.5) for VKA, DOAC and LMWH, respectively. The most used DOAC was rivaroxaban (incidence rate of 12.3 per 100 patient-years (95% CI: 1.6-96.4)).

Conclusions

PTS is a common complication among patients with proximal DVT of the lower extremity. Incidence of PTS development may differ based on the anticoagulant regimen. DOACs seems to be associated with lower incidence PTS development compared to VKA and LMWH. Future clinical trials are needed to confirm these findings.
NFκB Dysregulation in the Bone Marrow Niche Drives Epigenomic Reprogramming of Hematopoietic Stem Cells and Myeloid Bias
Authors: Chia Jennifer J , Singh Apeksha , Lin Yu-Sheng Eason , Tran Tiffany , King Jennifer , Rao Dinesh S. , Hoffmann Alexander
Published: December 2023

Introduction

Aging and chronic inflammation are known to skew hematopoietic output by favoring myelopoiesis and disfavoring lymphopoiesis. In addition, the bone marrow niche plays critical roles in supporting steady-state hematopoiesis, and alterations in the microenvironment may similarly influence disordered hematopoiesis. The NFkB system is a central, pleiotropic regulator of inflammatory signaling in many cell types. Thus, we reasoned that experimental perturbations of NFκB signaling may be used to study how inflammatory dysregulation affects hematopoiesis.

Methods

We generated a mouse model of inflammatory immune dysregulation, the IκB - mouse ( Nfkbia +/- Nfkbib -/-Nfkbie -/-), and then compared the peripheral blood and bone marrow inflammatory proteins and cell compositions betweenyoung WT, young IκB -,and aged WT mice. Using bone marrow transplantation, we next assayed the effect of an inflamed IκB - recipient niche on the hematopoietic output of WT donor bone marrow. Finally, we performed single cell RNA sequencing (scRNA-seq) and assay for transposase-accessible chromatin sequencing (ATAC-seq) on sorted hematopoietic stem and progenitor cells (HSPC) to characterize the epigenomic and transcriptional changes occurring in WT HSPC from an immune dysregulated environment.

Results

We observed an increase in inflammatory cytokines in both young IκB - and aged WT mice compared to young WT controls, as well as myeloid bias in the peripheral blood leukocytes and bone marrow HPSC. Transplantation studies showed that inflammatory dysregulation in the radio-resistant microenvironment resulted in myeloid-biased hematopoiesis. However, this myeloid bias was lost upon secondary transplantation of bone marrow into a WT recipient. Performed on HSPC from an inflamed microenvironment, our scRNA-seq studies showed myeloid bias at the transcriptional level, and ATAC-seq studies further revealed epigenomic changes, with an increased transcription factor (TF) binding motif accessibility for myeloid-priming TFs in long term HSC.

Conclusions

Our studies establish the IκB - mouse as a new model of chronic inflammatory dysregulation that can be used to study the effects of an inflamed bone marrow microenvironment on hematopoiesis. Furthermore, our findings indicate that dysregulated NFκB signaling in the bone marrow microenvironment leads to myeloid bias in HSPC, and is associated with epigenomic reprogramming at the stem cell level. However, despite this epigenomic reprogramming, we observed that myeloid bias could be reversed upon bone marrow re-transplant into a non-dysregulated niche. Together, these data suggest that further understanding niche-derived inflammatory dysregulation may be a useful in targeting disordered hematopoiesis in chronic inflammatory states such as aging.
Preventing Kidney Failure in Hemolytic Anemias with a Novel Heme and Hemoglobin Scavenger
Authors: Lucas Daniela , Cabrales Pedro , Munoz Carlos J , Gopal Srila , Muller Cynthia R
Published: December 2023

Pathophysiological conditions characterized by chronic hemolysis, such as genetic and acquired hemolytic diseases, lead to increased levels of circulatory cell-free hemoglobin (fHb), free heme, and iron. The toxicity of free acellular hemoglobin has been a subject of intense study to understand its impact on the human body. fHb's small size allows it to extravasate into various organs, causing damage. The kidney is particularly vulnerable to Hb toxicity as it can filtrate through the kidneys, affecting glomerular filtration rate and leading to kidney failure.

To address this, our group has developed a novel technique using the dorsal window chamber model in Golden Syrian hamsters. This method allows us to non-invasively measure glomerular filtration rate (GFR) using FITC-Sinistrin. The advantage of this approach is that it can assess renal function in awake animals, eliminating the need for anesthesia and minimizing its impact on the assessment.

We also tested the function of ApoHb-Hp, a scavenger for Hb and heme, developed by our group, to see how it protects kidney function from mild and severe hemolysis. Our data involved two sets of experiments. In the first set, we injected groups with 10% of their blood volume, either with Hb alone or along with ApoHb-Hp. In the second set, we performed a 20% volumetric exchange of Hb or ApoHb-Hp with Hb. The results indicated that the group infused with Hb showed a sharp 60% decrease in half-life after 2 hours, but it returned to baseline after 24 hours. On the other hand, the group infused with ApoHb-Hp only showed a 30% decrease, suggesting that Hb is responsible for a more pronounced acute decrease, rather than just the volume change. Furthermore, when comparing the exchange data, both the Hb and ApoHb-Hp groups exhibited a 55% decrease in half-life after 2 hours compared to baseline, attributed to the change in volume. However, at the 24-hour mark, the ApoHb-Hp group returned to baseline, while the Hb group's half-life continued to decrease, reaching an 86% decrease compared to baseline.

In conclusion, our preliminary data indicates that continuous exposure to Hb has a more prolonged effect on the kidney's filtration rate, potentially causing prolonged kidney impairment and irreversible damage. Further research is needed to understand the full implications of these findings and to explore the protective role of ApoHb-Hp in mitigating Hb-induced kidney damage.
Epigenetic and Transcriptional Consequences of PIM Kinase Activity and Inhibition in Diffuse Large B-Cell Lymphoma (DLBCL)
Authors: Debek Sonia , Komar Dorota , Garbicz Filip , Zolyniak Aleksandra , Pawlak Michal , Jablonska Ewa , Szydlowski Maciej , Skrzypczak Magdalena , Radomska Sylwia , Bajor Małgorzata , Serin Nazli , Chapuy Bjoern , Juszczyński Przemysław
Published: December 2023

The PIM kinase family consists of 3 proto-oncogenic proteins: PIM1, PIM2 and PIM3, expressed in numerous malignancies, including DLBCL. PIM kinases regulate crucial processes, such as proliferation, apoptosis, metabolism, or migration, therefore their inhibition is of great interest as a potential therapeutic strategy. Although recent studies have shed new light on the biological role of PIMs in lymphoid cancers, the details and mechanisms of PIM's oncogenic effects and the consequences of their inhibition in DLBCL remain insufficiently understood. Earlier studies have demonstrated a potential epigenetic role of PIM through histone H3S10 phosphorylation, however, described only at a single locus. The broad genomic role of PIM in modulating transcription in lymphoma cells remains unclear.

We confirmed the effect of PIM inhibition on histone modifications using small molecule inhibitors as well as genetic silencing of PIM kinases in DLBCL cell lines. PIM kinase inhibition changed the global amounts of histone modifications (including histone H4 pan-acetylation), accompanied by lower phosphorylation of RNA polymerase II, suggesting an involvement of PIMs in the RNA polymerase II pause release/elongation phase of transcription.

Local histone acetylation changes in the enhancer (H3K27ac) and promoter (H3K9ac) regions in response to treatment with the pan-PIM inhibitor were also identified, coupled with gene expression profiling in DLBCL cell lines. PIM inhibition decreased the expression of genes controlled by super-enhancers (SE), i.e. wide regulatory regions responsible for the high level of expression of the most important genes for the cell, including oncogenes. The integrated results of epigenomic and transcriptomic analyses explain the changes in expression of genes associated with many pathways, including transcription, inflammatory response, apoptosis, epigenetic mechanisms or DNA damage. The induction of the latter was further experimentally confirmed by the examination of DNA breaks using γH2AX, the activating phosphorylation of CHK2 or comet assays following PIM inhibition.

The conducted studies document an epigenetic function of PIM kinases, and suggest that inhibition of PIM activity leads to alterations in the DLBCL epigenetic landscape and associated changes in transcription. They also indicate that PIM inhibitors can be a therapeutic option in DLBCL.

Study supported aby Foundation For Polish Science TEAM Grant # POIR.04.04.00-00-5C84/17 and Polish National Science Centre Grant # 2018/29/B/NZ5/01471.
Habit Efficacy Trial: A Multi-Site Randomized Controlled Trial of Community Health Worker Support to Increase Hydroxyurea Adherence of Youth with Sickle Cell Disease
Authors: Green Nancy S. , Manwani Deepa , Aygun Banu , Appiah-Kubi Abena , Smith-Whitley Kim , Jia Haomiao , Smaldone Arlene
Published: December 2023

Introduction: Despite disease-modifying effects of hydroxyurea (HU) on sickle cell disease (SCD), poor adherence among affected youth commonly impedes treatment impact. The HABIT multi-site randomized controlled efficacy trial aimed to increase HU adherence.

Methods: Youth with SCD ages 10-18 years with impaired adherence were identified through laboratory evidence, primarily flagging levels of treatment-induced fetal hemoglobin (HbF). Eligible youth were enrolled as dyads with their primary caregivers for one year. We tested a 6-month phase of semi-structured supportive, multi-dimensional dyad intervention led by community health workers (CHW) with 4-5 visits addressing information, social issues/referrals, barriers, habit formation using goal-setting and cues, augmented by daily tailored automated text message reminders, compared to standard care. This phase was followed by a 6-month sustainability phase (months 7-12) of observation and, for the intervention group, 1 CHW booster visit. Primary intervention outcomes were 1) HbF levels compared to enrollment; and 2) proportion of days covered (PDC) for HU versus the pre-trial year. The secondary outcome was sustainability of these changes up to month 12.

Results: Starting in 2020, the COVID-19 pandemic disrupted enrollment and clinic-based procedures. The trial enrolled 50 dyads, missing target enrollment. Compared to enrollment levels, both HbF level and PDC significantly improved within the intervention group at month 6 (p=. 03 and . 01, respectfully), with parallel increased MCV (p=.05). Increased HbF did not reach signifance compared to controls (p=.07). No significant within- or between-group differences were found at month 12.

Conclusions: Within-group results suggest that our CHW-based dyad intervention with text message reminders improved HU adherence at 6 months but there were no between-group differences. Effects did not endure through the subsequent 6-month sustainability phase. These findings suggest that improved HU adherence for youth with SCD though our community-based, multi-modal support for youth-caregiver dyads temporarily improved usage. Durability of impact on adherence may require integrating CHW-led support and text reminders into clinical care.
Acute Cardiovascular Complications of Disseminated Intravascular Coagulation in Acute Myeloid Leukemia
Authors: Araji Ghada , Wei Chapman , Niazi Muhammad , Rajavel Anisha , Sharma Rubal , Grovu Radu , Siddiqui Fasih Sami , Mustafa Ahmad
Published: December 2023

Background: Disseminated intravascular coagulation (DIC), an acute or chronic systemic process with multifactorial etiology, can present as a life-threatening emergency with the potential for causing thrombosis and hemorrhage. DIC may be seen with all leukemia subtypes, but it is an especially prominent feature of Acute Myeloid Leukemias (AML). DIC complicating AML can lead to a variety of complications and has been implicated in worse clinical outcome, however, its association with acute cardiovascular complications has not been reported before.

Methods: National Inpatient Sample (NIS) Database 2016 -2018 was used to procure individuals with AML, and baseline demographics and comorbidities were collected using ICD-10-DM codes. Patients with missing data or those receiving immunotherapy, chemotherapy, or radiation during hospitalization were excluded. Patients were stratified into those with and without DIC. Greedy propensity matching using R was performed to match the two cohorts in 1:1 ratio on age, gender, medications (aspirin/ antiplatelet/ anticoagulation) and fourteen other baseline cardiovascular comorbidities. Univariate analysis pre and post-match along with binary logistic regression analysis post-match were used to analyze outcomes. P-value of <0.05 was considered statistically significant.

Results: Out of a total of 37344 patients with AML, 996 had DIC. DIC patients were younger, predominantly males, and had lower prevalence of baseline cardiovascular comorbidities. On pre-match univariate analysis, DIC patients had statistically significant higher mortality (30.2% vs 7.8%), acute myocardial infarction (5.1% vs 1.8%), acute pulmonary edema (2.3% vs 0.7%), cardiac arrest (6.4% vs 0.9%), and acute DVT/PE (6.6% vs 2.7%). Multivariate logistic regression model after matching showed similar outcomes with higher mortality (OR:4.63), acute myocardial infarction (OR:2.64), acute pulmonary edema (OR:2.75), cardiac arrest (OR:3.77), acute DVT/PE (OR:2.98), and acute heart failure becoming statistically significant (OR:1.89) in DIC patients.

Conclusion: DIC was significantly associated with higher mortality and acute cardiovascular complications in patients with AML. The underlying mechanisms involve a complex interplay of procoagulant factors, anticoagulant deficiencies, cytokine release, and endothelial dysfunction. Additionally, leukemic blasts and their interactions with platelets and endothelial cells contribute to microthrombi formation and subsequent organ damage. These findings highlight the importance of close cardiovascular monitoring and prompt recognition of complications in AML patients with DIC. Further studies are needed to corroborate these findings and develop targeted interventions for prevention and management of these complications.
A Novel Approach: Nanopore Sequencing of Native Cell-Free DNA in Diffuse-Large B-Cell Lymphoma Patients
Authors: Erblich Thomas , Sauer Carolin , Muyas Francesc , Ficz Gabriella , Cortes-Ciriano Isidro , Gribben John G.
Published: December 2023

Introduction: Nanopore sequencing (NS), a third generation long-read sequencing technology, is increasingly being used to study cancer genomes. NS has many advantages over next-generation sequencing, including the capability to simultaneously extract base sequence as well as base modifications in the same run without any further library modification. Notably, its portability and ability to provide real-time data make it a potentially powerful tool for clinical applications. This study explores the use of NS to directly sequence native cell free DNA (cfDNA) from patients with diffuse-large B-cell lymphoma (DLBCL), the most common type of non-Hodgkin lymphoma.

Methods: For this exploratory study, 20 patients with either a new diagnosis or relapsed DLBCL were enrolled. We adapted NS methods for cfDNA and sequenced cfDNA before each chemotherapy cycle using both an Illumina platform (as a control) and the Nanopore platform. For differential methylation, we also sequenced plasma cfDNA from five healthy donors on the Nanopore.

Results: Our findings demonstrate that we can directly sequence cfDNA on the Nanopore platform, obtaining a whole genome coverage (average 1.2x) with deeper coverage in some regions and also observe ~60% global DNA methylation level in CG context. From this data we are now analyzing regions that are differentially methylated (DMRs) between normal and DLBCL, determining SNV/Indel profiles, and comparing copy number variants (CNVs). These results will be available and presented at the time of the meeting.

Conclusions: NS is capable of detecting cfDNA and measuring methylation levels, offering a promising and potentially clinically applicable new approach for the study of cfDNA.

Future directions: As we continue this research, our vision is to leverage the portability and real-time data capabilities of NS for clinical applications. By simultaneously combining multiple genomic features with machine learning, we aim to develop a method for measuring lymphoma cfDNA-based MRD in DLBCL patients to be used in clinical settings.
Title: MLLT1 Is Required to Maintain Normal B-Lineage Hematopoiesis and Mitochondrial Function
Authors: Prakash Janani , Achille Nicholas , Zhang Shubin , Hemenway Charles S. , Zeleznik-Le Nancy J.
Published: December 2023

MLLT1 (aka ENL) is an epigenetic reader protein that binds to crotonylated/acetylated lysine residues of histone 3 through its N-terminal YEATS domain. Multiple different positive and negative regulatory complexes compete for binding to the MLLT1 C-terminal domain. MLLT1 has been implicated in leukemic stem cell survival in MLL-rearranged cell lines but its role in normal hematopoiesis has not been investigated. Mllt1 is expressed in multiple bone marrow cell subsets, with high expression levels in multipotent and B-lineage progenitors. We generated conditional Mllt1 knockout ( Mllt1 del) mice and observed a B-lineage defect in bone marrow of Mllt1 deleted mice compared to control Mllt1 fl/fl mice. Mitochondrial activity has been linked to hematopoietic lineage fate decision choices. We performed seahorse mitochondrial stress test on lineage negative (stem and progenitor) bone marrow cells and found that lineage negative cells of Mllt1 del mice have lower mitochondrial activity (as a measure of oxygen consumption rate) than control Mllt1 fl/fl mice. In addition, we observed mitochondrial mass (as a quantitative measure of ratio of mtDNA gene CytB / nDNA gene ActB) to be decreased in lineage negative cells of Mllt1 del mice when compared to control Mllt1 fl/fl mice. Surface marker labeling was used to sort more defined subpopulations from bone marrow of Mllt1 del and Mllt1 fl/fl mice. We observed decreased mitochondrial mass in MPP4s and MEPs of Mllt1 del mice when compared to control Mllt1 fl/fl mice. Flow cytometry-based cell surface labeling to distinguish different bone marrow progenitors, plus mitochondria staining using MitoTracker Green (for mitochondrial mass) and MitoTracker Red CMXRos (for mitochondrial membrane potential) showed altered mitochondrial function of B-lineage progenitors in Mllt1 del mice when compared to control Mllt1 fl/fl mice. Thus, Mllt1 is essential for normal mitochondrial function, which may be necessary to maintain specific bone marrow hematopoietic subpopulations. Ongoing studies include gene expression profiling of sorted lymphoid progenitor populations and analysis of direct Mllt1 target genes that contribute to the altered phenotype in Mllt1 del mice.