bluuebunny/crossref_metadata_2025 huggingface dataset:
Cerebrospinal Fluid IL-6 Plus IL-10 As Effective Diagnostic Biomarkers for Secondary Central Nervous System Lymphoma
Authors: Gu Jiayan , Jin Jie
Published: December 2023

Background : Secondary central nervous system lymphoma (SCNSL) characterized by the involvement of the CNS at initial diagnosis of systemic lymphoma or in the setting of relapse. Accurate diagnosis and therapeutic response of SCNSL are particular challenges, which need to be improved. We aimed to evaluate the diagnostic and the post-therapeutic prognostic value of cerebrospinal fluid (CSF) cytokine in patients with SCNSL.

Methods: In this retrospective study involving 234 NHL patients and 53 other CNS patients, the NHL patients was divided into two sub-groups based on with and without CNS involvement: SCNSL (n=57) and non-SCNSL (n=177). The CSF cytokine profiles (interleukin (IL)-2, -4, -6, and -10, tumor necrosis factor-α, interferon-γ, and IL-17A) of these patients were measured by cytometric bead assay and evaluated the ability of CSF cytokine levels as diagnostic, therapeutic and prognostic biomarkers for SCNSL.

Results: CSF IL-6 and IL-10 levels were significantly higher in SCNSL patients than non-SCNSL and other CNS patients. CSF levels of IL-6 and IL-10 were significantly higher in patients with an stage at Ⅲ and Ⅳ, CSF WBC counts>5/µL, CSF protein ≥0.45g/L.Using a CSF IL-6 cut-off value of 10.13 pg/ml yielded a diagnostic sensitivity and specificity was 62.34% and 87.57%, respectively (AUC, 0.7798; 95% CI, 0.7083 to 0.8514); For a CSF IL-10 cutoff value of 7.82, the sensitivity was 76.23%, and the specificity was 98.31% (AUC, 0.8164; 95% CI, 0.7466 to 0.8862). Combined CSF IL-6 and IL-10 significantly improved the diagnostic effect of SCNSL (AUC, 0.8959; 95% CI, 0.8371 to 0.9547). The levels of IL-6 and IL-10 in cerebrospinal fluid of SCNSL patients who responded to intrathecal chemotherapy were significantly decreased during follow-up, while the levels of these two cytokines showed fluctuating changes in patients with progressive SCNSL. Moreover, poor Progression free survival (PFS) for patients with SCNSL was associated with the increased CSF IL-10 level at diagnosis with CNS involvement (P=0.035), but not with the increased CSF IL-6 level.

Conclusion: Our study confirmed CSF level of IL-6 and IL-10 were potentially effective diagnostic biomarkers for SCNSL, and poor progression free survival (PFS) for patients with SCNSL was associated with the increased CSF IL-10 level.
Efficacy of Allogeneic Hematopoietic Stem Cell Transplantation in Young and High-Risk Multiple Myeloma Patients
Authors: Pan Pan , Jiang Erlie , Pang Aiming
Published: December 2023

Objective: With the new therapeutic agents developed over last decades, the median survival of patients with multiple myeloma has approached 10 years. However, 15-20% of patients who are classified as High-risk multiple myeloma (HRMM) patients still have a median survival of less than 3 years, which leaves room for improvements. Simultaneously, transplantation related mortality (TRM) has been decreasing since progress of supportive care during these years. Thus, this study aims to evaluate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in young high-risk multiple myeloma (HRMM) patients and to conduct statistical analysis of various prognostic factors in the new era.

Method: This retrospective study analyzed clinical data from 14 young multiple myeloma patients with high-risk cytogenetic abnormalities or high-risk disease biological factors who underwent allo-HSCT at the Hematopoietic Stem Cell Transplantation Center of the Institute of Hematology & Blood Diseases Hospital between November 2016 and November 2022.

Results: There were 7 males and 7 females involved in this study, with a median age of 39.5 (31- 50) years at the time of transplantation. The median number of treatment lines before transplantation was 2 (1-6). Before transplantation, 42.9% (6/14) of the patients did not achieve complete remission (CR), with 35.7% (5/14) of the patients performing MRD (+). After transplantation, all patients could be evaluated for treatment response, in which the overall response rate was 100% (14/14). All 14 patients successfully underwent transplantation, with median engraftment times for neutrophils and platelets of 11 (10-14) days and 13 (9-103) days, respectively. Acute graft-versus-host disease (GVHD) of grades II-IV occurred in 5 patients (35.7%), and 2 patients (14.3%) developed moderate to severe chronic GVHD. The median follow-up time after transplantation was 18.93 (4.1-72.53) months, with the 2-year TRM of 7.1% (95%CI 0-19.7%) and the expected 2-year overall survival (OS) rate of 92.9% (95% CI 80.3-100%). Meanwhile, the expected 1-year and 2-year progression-free survival rates were 85.7% (95% CI 69.2-100%) and 45.7% (95% CI 20.9-99.9%), respectively, and the presence of chronic GVHD might influence the outcome positively.

Conclusion: Allogeneic hematopoietic stem cell transplantation is an effective treatment to improve the prognosis of young and high-risk multiple myeloma patients.
Prevalence of MGUS in Rural Indian Population: Results of Simple (IMAGe-002A) Study
Authors: Raphael Celine , Yanamandra Uday , Bobdey Saurabh , Sharma Vaibhav , Bhatti V K , Muthukrishnan J , Malhotra Pankaj , Nair Velu
Published: December 2023

Introduction: The incidence of monoclonal gammopathies is rising across the globe. Although the prevalence of monoclonal gammopathy of undetermined significance (MGUS) has been evaluated in two hospital-based studies in India, no data exists on its prevalence in community-based settings amongst the rural Indian population.

Objectives: To determine the prevalence of MGUS in a rural Indian population in a community-based setting.

Methods: A cross-sectional study was conducted in an agrarian village in India covering a perimeter of 24.76Km and 4588.3 hectares with an 89.2% population coverage for adults more than 45y spending 12096 man-hours as part of SIMPLe ( Screening Intervention for Myeloma and Prevention of Lifestyl e diseases) study under the aegis of Indian Myeloma Academic Group e (IMAGe) registered vide CTRI/2023/03/051220. For the complete study population, demographic details, medical history, and blood samples were collected after a thorough medical examination. Hemogram, biochemistry (Liver and renal function tests), and serum protein electrophoresis (SPEP) were universally performed in all individuals. Serum immunofixation electrophoresis (SIFE) was performed for those with abnormal SPEP graphs (those with M spike and those with abnormal bulges in beta-2 or gamma region suspicious of MGUS). Patients with monoclonal protein on SPEP or SIFE were further assessed for any features of smoldering/ multiple myeloma. The data was analyzed using JMP ver. 16.0.0.

Results: The prevalence of MGUS in the study population was 1.75%. The mean age of individuals with MGUS was 63.4±5.2y with male preponderance (60%). The mean Hb was 12.39±1.8g/dL with none of the patients having any renal dysfunction. None of the patients fit into the criteria for smoldering or multiple myeloma using IMWG criteria. Among those with MGUS, the distribution of Heavy and light chains was IgG - 83.33%, IgA - 16.67% with none having IgM/ IgD paraprotein, Kappa - 50%, and Lambda - 50%. Two individuals only had light chain MGUS.

Conclusion: Our study provides a comprehensive description of the prevalence of MGUS in a rural community setting in India. The background prevalence is much less compared to the Western (both USA ~3% and European~5%) population.
Acute Myeloid Leukemia in Elderly, Unfit Patients: Analysis of Turkish AML Prospective Registry Database, on Behalf of Acute Leukemia Working Group of Turkish Society of Hematology
Authors: Urlu Selin Merih , Cengiz Seval Guldane , Erdogan Yücel Elçin , Mehtap Ozgur , Yenihayat Emel Merve , Polat Merve Gokcen , Malkan Umit Yavuz , Ozbalci Demircan , Yigit Kaya Sureyya , Durusoy Salih Sertac , Akdeniz Aydan , Kirkizlar Onur , Dogan Ali , Pinar Ibrahim Ethem , Kacmaz Murat , Afacan Ozturk Hacer Berna , Atas Unal , Deveci Burak , Korkmaz Akat Gulten , Guven Zeynep Tugba , Sevindik Omur Gokmen , Demirkan Fatih , Alacacioglu Inci , Karakus Volkan
Published: December 2023

Objective: Low dose Ara-C and hypomethylating agents (HMAs) - decitabine (DEC) and azacitidine (AZA) - have made it possible to treat more elderly patients with acute myeloid leukemia (AML). Both HMAs demonstrated efficacy in monotherapy and in combination with targeted therapies. We analyzed the outcomes of elderly and unfit patients with newly diagnosed AML in our population.

Methods: Analyses of our database revealed 222 eligible AML patients from diagnosed between January 2013 to December 2020. All patients were received low intensity treatments including AZA +/- venetoclax, DEC and low dose Ara-C. The effects of cytogenetic risk group, response rate, treatment type on overall survival (OS) and PFS (progression-free survival) were evaluated. Survival analyses were performed with the Kaplan-Meier method and log-rank test. A p-value of <0.05 was considered statistically significant.

Results: Median age was 73 years (51-89). Eastern Cooperative Oncology Group (ECOG) performance score was <2 and ≥2 for 75 patients (36.8%) and 147 patients (63.2%), respectively. Of the patients 8.2% had favorable, 76.9% intermediate and 15.0% adverse risk according to European Leukemia Network (ELN) risk classification. Extramedullary involvement was present in 4.5%. Low dose treatment regimens included AZA in 119 patients (53.6%), DEC in 38 (17.6%), low dose ARA-C in 26 (11.7%) and AZA + venetoclax in 39 (17.1%). Complete remission was achieved in 61 patients (27.5%), whereas 39 patients (17.6%) had partial remission (PR) and 64 (28.8%) had stable/progressive disease (SD/PD). Overall response rate was 48.1%. CR rate was higher with AZA + venetoclax (44.7%, p=0,005) and other regimens had similar outcomes. Median OS was 10.3 months. No significant difference difference in PFS was observed between the regimens. However, the lowest PFS was observed with low-dose Ara-C (p=0.33). ELN risk groups revealed no significant effect on PFS and OS (p=0.77 and p=0.82, respectively). Infections were the most common cause of death (42.7%).

Conclusions: The prognosis of AML in the elderly remains poor. HMA monotherapy is effective among elderly and unfit patients. However, combination therapies may be more effective and safe. The incorporation of targeted therapies may provide additional benefit in this patient group. However, the accessibility of several novel targeted drugs is still very limited in many countries.
The Important Role of Egfr Change in Patients with Multiple Myeloma
Authors: Liu Xiang , Tang Wenjiao , Niu Ting
Published: December 2023

Background. Renal impairment is a common complication in multiple myeloma (MM) patients and related to a higher mortality. However, whether the change of estimated glomerular filtration rate (eGFR) is associated with prognosis is not clear currently.

Patients and Methods. A total of 1552 myeloma patients in West China Hospital from July 1, 2008 to September 1, 2022 were retrospectively included in this study, patients with dialysis or complicated with other malignancy were excluded. All patients had at least 3 measurements of creatinine and eGFR between 6 months and 2 years after baseline. The 1-year change in eGFR (slope) was calculated by linear regression, patients were divided into 3 groups by the eGFR slope (sustained group: eGFR slope was -5 ~ 5 ml/min per 1.73 m2 per year over 2 years; increased group: eGFR Slope > 5 ml/min per 1.73 m2 per year over 2 years; decreased group: eGFR Slope < -5 ml/min per 1.73 m2 per year over 2 years ), the Kaplan-Meier survival analysis, multiple Cox regression model and Restricted cubic spline (RCS) analysis were applied to explore the association of eGFR slope with the best efficacy and progression of myeloma patients.

Results. After a median follow-up of 21 months, 977(62.95%) patients had reached VGPR or CR (the best efficacy), 24.1% patients experienced progression, the survival status is currently being followed up. The eGFR decline more than 5 ml/min per 1.73 m2 per year over 2 years (decreased group) was showed to have a higher risk of disease progression (P=0.01, Figure 1a) and to be an independent risk factor for disease progression, both in patients with (eGFR<60 ml/min per 1.73 m2) or without (eGFR>60 ml/min per 1.73 m2) renal failure (Table 1). The RCS analysis showed that the decline of eGFR within 2 years was positively related to the risk of disease progression, while the eGFR rise with a certain range could reduce the risk of progression (0-6.4 ml/min per 1.73 m2 per year) (Figure 1b). Inversely, the eGFR increased group showed a higher possibility of acquiring very good partial remission (VGPR) or complete remission (CR) (Figure 1c, P<0.001), and was a beneficial factor of reaching VGPR or CR for MM patients (HR 1.3, 95%CI 1.05-1.62, P=0.015, Table 1). The RCS analysis showed that eGFR slope was linearly correlated with patients' VGPR or CR rate (Figure 1d). Notably, in patients without renal insufficiency at baseline (eGFR>60 ml/min per 1.73 m2), eGFR improvement did not increase the rate of VGPR or CR (Table 1).

Conclusions. eGFR decreasing >5 ml/min per 1.73 m2 per year over 2 years could increase the risk of disease progression in MM patients with or without renal impairment, while eGFR increasing at a certain range could reduce progression risk. For myeloma patients with renal impairment, eGFR increasing may improve the rate of VGRP or CR in MM patients.
Retinoic Acid and Ascorbate Remodel the Epigenome of Leukemia Cells to Improve Therapeutic Efficacy By Enhancing TET Activity
Authors: Leesang Tiffany , Yap Yoon Sing , Brabson John P , Klimas Abigail , Ahearn Emily L , Barbieri Daniela A , Lam Minh Quang , Lyon Peter D , Figueroa Maria Ken E. , Beckedorff Felipe , Cimmino Luisa
Published: December 2023

Ten-eleven translocation (TET) enzymes are commonly mutated in leukemia, leading to aberrant DNA methylation and impaired hydroxymethylation. All-trans retinoic acid (ATRA) is a vitamin A derivative and cofactor of retinoic acid receptors (RARs) which, in combination with arsenic trioxide, can curatively treat acute promyelocytic leukemia. ATRA has also been shown to upregulate expression of Tet2 and Tet3 in murine embryonic stem cells, and retinol and ascorbic acid (vitamin C) have been shown to enhance pluripotent stem cell reprogramming through increasing hydroxymethylcytosine and promoting DNA demethylation. Our previous work has demonstrated ascorbate can slow disease progression in vivo and, combined with the PARP inhibitor Olaparib, promote cell cycle dysregulation and induce cellular differentiation in acute myeloid leukemia (AML) models both in vitro and in vivo.

Here, we show ATRA can induce TET2 and/or TET3 expression in AML cells, and, combined with ascorbate, increase genome-wide oxidized methylcytosine in a TET-dependent manner. Combination treatment with ATRA and ascorbate also induces transcriptional reprogramming, leading to upregulation of DNA repair genes and pathways leading to increased expression of myeloid differentiation markers. Additionally, we observe chromatin remodeling associated with an enrichment for ETS motifs including genes and regions regulated by the transcription factor PU.1, an essential modulator of hematopoietic cell fate. We also show enrichment of retinoic acid receptor A (RARA) at the TET2 locus and at several RAR elements in response to ATRA in AML cells. Using very low dose ascorbate and ATRA, we show that the combination treatment can enhance the efficacy of Olaparib and Venetoclax to alter the cell cycle, promote differentiation, and induce apoptosis of AML cells. These data demonstrate the potential for ATRA and ascorbate to increase TET activity and drive myeloid differentiation and death of AML cells that can be exploited as an adjuvant therapy for the treatment leukemia.
Clinically Significant CMV Infection in Allogeneic Stem-Cell Transplant Recipients: A Single Center Experience
Authors: Gulied Amaal , Alasmar Aya , Bakr Mohammad , Shawayli Hawra , Ghasoub Rola , Fadul Afraa Mustafa Sulieman , Gameil Amna , Mudawi Deena SIDEEG , Hamad Anas Ahmad , Gaziev Javid
Published: December 2023

Introduction:

Cytomegalovirus (CMV) infection is one of the major causes of concern in allogeneic stem cell transplantation (allo-HSCT). Here we report the clinically significant CMV infections among allo-HSCT recipients at the National Center for Cancer Care and Research (NCCCR) in Qatar.

Methods:

A retrospective review of the electronic health records of all malignant hematology patients who underwent allo-HSCT at NCCCR was performed from September 2017 to December 2022.

Results:

Fifty-one malignant hematology patients have been transplanted during the data collection period. Patient characteristics are summarized in Table 1. The median age was 34 years (range 14-53 years). CMV-serostatus was positive in fifty recipients (98%) and all donors. Thirty-two patients (62.7%) have received matched-related grafts. The most frequent indication for HSCT was acute myeloid leukemia, followed by acute lymphoblastic leukemia. Forty-six patients (92%) underwent myeloablative conditioning therapy. Graft-versus-host disease (GvHD) prophylaxis used in the non-haploidentical group was cyclosporine and short-course methotrexate while haploidentical recipients received cyclosporine, mycophenolate mofetil, and post-transplant cyclophosphamide. Letermovir prophylaxis was accessible for use for high-risk patients from February 2022. CMV-PCR was monitored twice per week for all patients until day +100. The clinically significant CMV infection rate was 51%. The cumulative incidence of CMV infection in haploidentical and non-haploidentical groups was 70% and 30%, respectively (figure 1). The median CMV-PCR viral load was 1783.5 IU/mL (range 133- 9600 IU/mL), with the earliest CMV reactivation on day -1 of transplantation. The clinically significant CMV infection incidence was significantly higher in patients who did not receive letermovir prophylaxis. Preemptive antiviral therapy was initiated to prevent CMV disease in all indicated recipients. Foscarnet was the preferred first-line pre-emptive treatment option for patients during cytopenia. No CMV disease or CMV-related mortality occurred in our population.

Conclusion:

This is the first report from Qatar to report CMV infection post-allogeneic stem cell transplantation. Future in-depth analysis of our population and treatment responses is needed.
Impact of Genetic Variations on Thrombotic Risk in Saudis with Sickle Cell Disease
Authors: Algahtani Farjah , Alshabeeb Mohammad , Alzahrani Mohsen , Alwadaani Deemah , Alajmi Hala , Abohelaika Salah , Alsaeed Hussain , Rashid Mamoon , Alsomaie Barrak , Alzayed Abdullah , Daly Ann
Published: December 2023

Background

Sickle cell disease (SCD) is a Mendelian disease characterised by multigenic phenotypes. Previous reports indicated a higher rate of venous thromboembolism (VTE) in SCD patients. A number of candidate polymorphisms in certain genes (e.g. FVL, PRT, and MTHFR) were previously reported as risk factors for VTE in different clinical conditions. This study aimed to genotype these genes and other loci predicted to underlay VTE in SCD patients.

Methodology

A genome-wide association study involving Saudi SCD adult patients with a history of VTE (n=65) and control patients without VTE history (n=285) was performed. Genotyping used the 10x Affymetrix Axiom array which includes 683,030 markers.

Results

No associations were identified between the targeted single nucleotide polymorphisms (SNPs) [FVL (rs6025), PRT (rs1799963), and MTHFR (rs1801133)] and VTE episodes in SCD patients. In contrast, GWAS analysis showed two genome-wide significant signals (P<5×10 -8) on chromosome (Chr) 11. These SNPs, rs35390334 and rs331532, are in strong linkage equilibrium (LD, r 2=0.97). A total of 35 other markers had an association threshold of P<5×10 -6. Four of these variants are located in olfactory receptor genes [OR51B5 (rs147062602), OR51V1 (rs7933549), OR51A1P (rs80034548), and OR52A5 (rs2472530)].

Conclusion

No association between FVL, PRT, and MTHFR genes and VTE episodes was seen in Saudis with SCD. In contrast, two markers (rs35390334 and rs331532) showed significant genome-wide associations and less significant associations (P<5×10 -6) were identified for 35 other variants (24 of them are located on Chr 11). Further replication of these findings is needed.
Loss of 12-Lipoxygenase Improves the Post-Transfusion Function of Stored Platelets
Authors: Larsen Hannah , Byrne Daire , Ozpolat Hasan Tahsin , Chauhan Aastha , Bailey Shawn , Rhoads Nicole , Reed Franklin , Stolla Massiel Chavez , Adili Raymond , Holinstat Michael , Fu Xiaoyun , Stolla Moritz
Published: December 2023

Objective Platelets for transfusion are stored for 5-7 days. During storage, platelets undergo numerous detrimental functional changes. Previous studies have shown that HETE levels in the storage bag negatively correlate with platelet performance in vivo suggesting that the dysregulation of bioactive lipid mediators may contribute to the storage lesion. In the current study, we sought to understand how genetic deletion and pharmacologic inhibition of 12-lipoxygenase (12-LOX) affects platelets during storage and after transfusion.

Approach and Results Platelets from12-LOX+/+ (wild-type [WT]) and 12-LOX-/- mice were stored for 24 and 48 hours and profiled using LC-MS/MS-MRM. Polyunsaturated fatty acids increased significantly in stored platelets from 12-LOX-/- mice, while oxylipin concentrations were significantly higher in WT platelets. Baseline αIIbβ 3 integrin activation was significantly lower before and after 24 hours of storage in 12-LOX-/- platelets than in WT platelets. Surprisingly, after transfusion to thrombocytopenic mice, we observed significantly more baseline αIIbβ3 integrin activation in 12-LOX-/- platelets than in WT platelets. Stored platelets from 12-LOX-/- mice occluded vessels significantly faster than stored WT platelets in thrombocytopenic recipients challenged with a FeCl 3-induced carotid artery injury. In hemostasis models, significantly more stored 12-LOX-/- than WT platelets accumulated at the site of venous injury leading to reduced blood loss. Inhibition of cyclooxygenase-1 (COX-1) with acetylsalicylic acid abrogated both increased integrin activation and thromboxane generation in stored 12-LOX-/- platelets, highlighting a critical role of this pathway for improved post-transfusion function.

Consistent with our mouse studies, human platelets stored with the 12-LOX inhibitor, VLX-1005, showed increased integrin activation compared to vehicle-treated platelets upon transfusion in NOD/SCID mice.

Conclusion Deleting 12-LOX improves the post-transfusion function of stored murine and human platelets by increasing thromboxane generation through a COX-1-dependent mechanism. Future studies should determine the feasibility and safety of 12-LOX-inhibited platelets transfused to humans.
Bleeding and Thrombosis in Patients with Cancer and Acute Venous Thromboembolism Requiring Urgent Procedures
Authors: Owusu Tony , Derkach Andriy , Burge Miranda , Nemirovsky David , Zwicker Jeffrey I.
Published: December 2023

BACKGROUND

There are limited data of outcomes in patients who require anticoagulation interruption for a surgical procedure shortly after an acute VTE, especially in patients with cancer who are at higher risk for both hemorrhage and recurrent thrombosis. Our goal was to assess the incidence of recurrent thrombosis or major hemorrhage at 60-days following an urgent surgical procedure in patients with acute VTE (within one month prior to surgery).

METHODS

We performed a retrospective cohort study of patients with cancer at Memorial Sloan Kettering Cancer Center (from January 1 st 2021 to December 31 st 2022). Patients were eligible for inclusion if they were diagnosed with a new acute deep vein thrombosis or pulmonary embolism and underwent surgical procedure within 30-days following the acute VTE diagnosis. Recurrent VTE was defined as symptomatic new deep-vein thrombosis or pulmonary embolism, incidental new deep-vein thrombosis or pulmonary embolism involving segmental or more proximal pulmonary arteries, or fatal pulmonary embolism or unexplained death for which pulmonary embolism could not be ruled out as the cause. Major hemorrhage was characterized according to ISTH definition. Competing risk framework was used to estimate cumulative incidences of major hemorrhage, VTE and death at 30 and 60-days.

RESULTS

A total of 90 patients were included in this analysis. The median age was 73 years and 62% were female (Table 1). Surgical procedures were classified by their risk of bleeding with 64 patients undergoing lower bleeding risk procedures and 26 high-risk procedures (Table 1). There were 10 recurrent or new VTE within 60-days of surgery (60-day cumulative incidence of 8.9%, 95% CI 4.1-16%). The recurrent sites were deep venous (N=7) and new PE (N=3). There were 12 major hemorrhagic events within 60 days (cumulative incidence of 12%, 95% CI 6.5-20%). The most common sites of hemorrhage were the lower gastrointestinal tract and genitourinary system. The competing risk framework is shown graphically in Figure 1. Among patients who received IVC filters the incidence of recurrent VTE was 1.1% and major hemorrhage was 2.2%. The 60-day cumulative incidence of death following surgical procedure was 19% (95% 12-28%). There were no fatal VTE and 3 fatal hemorrhagic events.

CONCLUSION

Among patients with cancer undergoing surgical procedures within 30-days of an acute VTE, the rates of recurrent VTE and major hemorrhage are substantively higher than reported in non-surgical acute VTE cohorts. Placement of temporary IVC filters may improve short-term bleeding and thrombotic outcomes.
Safety and Efficacy of New Drugs Combinations in Newly Diagnosed Multiple Myeloma with Renal Failure at Onset: A Retrospective Multicentric Study
Authors: Frolli Antonio , Cerchione Claudio , Staiano Riccardo , Guglielmelli Tommasina , Nappi Davide , Mattei Agnese , Marini Silvia , Cilloni Daniela , Parvis Guido Eugenio
Published: December 2023

Renal injury at onset of newly diagnosed multiple myeloma (NDMM) is associated with poor outcome. Patients with severe renal failure due to MM are excluded from clinical trials with new drugs. The aim of this retrospective study was to describe the clinical characteristics, the treatment choices, the efficacy and the incidence of treatment-related toxicities of NDMM patients with renal insufficiency. We enrolled all NDMM patients with renal failure due to the disease treated at our Centers from 2012 to present. Fifty-one patients with a median age of 75 years were included; 76% had an International Staging System (ISS) of 3; 76% had anemia, 74% bone lesions and 30% had hypercalcemia; 31% had micromolecular myeloma. Thirty seven percent of patients had severe renal failure requiring dialysis treatment. Thirty-five percent of patients received treatment according to a VMP (Bortezomib, Melphalan and Prednisone) scheme, 20% received a triplet including Bortezomib and an immunomodulatory drugs (IMIDs) and 33% received a regimen including Daratumumab. The remaining 12% received a douplet (lenalidomide-dexamethasone or melphalan-prednisone). Thirty one percent of patients underwent an autograft procedure. Overall response rate (ORR) was 94% with a complete response (CR) rate of 17%. In terms of recovery of renal function, 50% achieved at least minimal renal response according to IMWG criteria, with 40% obtain a complete recovery of renal function. Patients who received more innovative treatment (regimens containing Daratumumab or Bortezomib in combination with IMIDs) achieved a response rate superior than very good partial response (≥ VGPR) of 74% with hematologic, neurologic, and infectious toxicities grade ≥ 3 of 22%, 7%, and 26%, respectively. Patients who received therapy with VMP or douplet had response rates ≥ VGPR of 25% with hematologic, neurologic, and infectious toxicity grade ≥ 3 rates of 29%, 4%, and 21%, respectively. Complete renal response rates in these two groups of patients were 45% and 20%, respectively. No death due to toxicity was registered. Patients with NDMM with renal failure at onset had a high burden of disease with unfavorable prognostic features. Although the small numbers of the cohort and the retrospective nature of this study it can be concluded that these patients can received new drugs combinations treatments with excellent response rates and without increased toxicity.
First Real-World Experience Administering Etranacogene Dezaparvovec Gene Therapy for People with Hemophilia B
Authors: Ryan Matthew L. , Miller Andrea , O'Hearn Patrick , Desai Vidhi Parikh
Published: December 2023

Research in gene therapy for hemophilia B has been ongoing for decades. Etranacogene dezaparvovec-drlb (CSL Behring) is the first gene therapy for hemophilia B to be approved by the United States Food and Drug Administration (2022) and the European Medicines Agency (EMA) (2023). The gene therapy treatment journey is a multistep process and requires coordination between multiple stakeholders within a hemophilia treatment center (HTC). The first 2 patients to receive etranacogene dezaparvovec-drlb since approval were successfully infused at the Hemophilia Outreach Center in Green Bay, Wisconsin. Here we describe the process by which the Hemophilia Outreach Center prepared their center and patients for the administration of etranacogene dezaparvovec-drlb.

Early education, both for HTC stakeholders and patients, was critical in addressing key steps in the treatment journey, including patient eligibility, preparation, administration, and post administration monitoring and management. The education was iterative, involved multiple stakeholders (physician, pharmacist, nurse, social worker, and patient) and required an open channel of communication among the HTC stakeholders, the patients, and manufacturer when appropriate. Although no certification process was required, the manufacturer provided education to the HTC stakeholders in the format of ongoing trainings and a detailed handbook describing the gene therapy process as it relates to etranacogene dezaparvovec-drlb. This education provided a foundation for the HTC stakeholders to develop their own center-specific protocols, checklists, and worksheets. These protocols provided clear guidance on how to manage every step of the process in addition to providing a contingency plan for different scenarios (e.g., infusion reactions and transaminitis), ensuring the center's preparedness for administration.

By implementing early and iterative education and developing center-specific protocols, the Hemophilia Outreach Center was able to successfully administer etranacogene dezaparvovec-drlb. These elements are crucial to ensure smooth execution and optimal patient outcomes.
NR2F1 : A Novel Candidate for Gamma Globin Activation
Authors: Wu Ruiqiong , Yi Siqi , Li Yichao , Shrestha Dewan , Zheng Minzhang , Zhang Jingjing , Cheng Yong
Published: December 2023

The reactivation of human gamma globin holds tremendous promise as a potential treatment for hemoglobinopathies, including sickle cell disease and beta-thalassemia. Despite identifying various repressors of fetal globin genes in adult erythroid cells over the past decade, evidence concerning stage-specific factors that activate human fetal globin prior to the switch to adult beta-globin remains inadequately understood.

In our study, we constructed an erythroid-specific gene regulatory network (GCN) by integrating multiple RNA-seq datasets from erythroid cells at different stages of differentiation and development. Our analysis unveiled both known HbF activators, like HIC2, and novel candidates. Among the novel contenders, NR2F1, a member of the steroid/thyroid hormone receptor superfamily, emerged as a prominent candidate. To validate our predictions, we overexpressed NR2F1 in Hudep2 cells and observed more than a four-fold increase (28% vs. 6%) in the proportion of F cells and a three-fold rise in HbF levels as measured by HPLC. In addition, the overexpression of NR2F1 did not appear to impact beta-globin expression or cell differentiation. Intriguingly, we observed a significant reduction in the protein level of BCL11A, a master regulator of HbF, despite no major change in the total RNA level of BCL11A. This observation suggests that NR2F1 plays a role in the post-transcriptional modification of BCL11A. Additionally, NR2F1 overexpression led to an upregulation of Lin28B expression in Hudep2 cells. Lin28B is known to directly bind BCL11A mRNA and prevent its effective translation without disturbing BCL11A transcription. Consequently, the observed upregulation of Lin28B in NR2F1-overexpressing cells implies a potential NR2F1-Lin28B-BCL11A axis involved in controlling human hemoglobin switching.

In conclusion, our findings propose NR2F1 as a novel activator of gamma globin, representing an attractive target for gamma globin reactivation. Additional investigation is required to explore the molecular mechanisms through which NR2F1 regulates gamma globin expression and its role in the NR2F1-Lin28B-BCL11A axis.
PACSIN2 Orchestrates Actin Reorganization during Megakaryocyte Maturation
Authors: Boyd Emily , Gooding Gabrielle , Hoffmeister Karin M. , Falet Herve
Published: December 2023

Thrombocytopenia, defined by a platelet count below 150,000/µL, poses significant mortality risks, primarily due to bleeding. Blood platelets are produced by bone marrow megakaryocytes (MKs) in unique cellular processes that require polyploidization and extensive intracellular membrane rearrangements leading to the formation of the demarcation membrane system (DMS), the surface-connected membrane reservoir for future platelets. The membrane-deforming F-BAR protein PACSIN2 is one of the most abundant and conserved BAR proteins in platelets, and exomechip analysis and genome-wide association studies have associated PACSIN2 single nucleotide variants with platelet count and mean platelet volume in humans. Our previous work has shown that PACSIN2 localizes with the initiating DMS in MKs in a process regulated by the cytoskeletal and scaffolding protein filamin A (FlnA). Recently, we have found that Pacsin2 -/- mice display mild thrombocytopenia with slightly enlarged platelets and severe thrombus formation defects due to increased activation of platelet integrin β1. Here we investigated the role of PACSIN2 in MK maturation and platelet production. Cultured fetal liver cell-derived Pacsin2 -/- MKs formed proplatelets normally compared to controls. Pacsin2 -/- bone marrow sections displayed MK count comparable to control mice, and freshly isolated bone marrow Pacsin2 -/- MKs had normal ploidy, indicating normal MK maturation. The observed effects of PACSIN2 deficiency led to a defective formation of the DMS within MKs. In PACSIN2 deficient MKs, the DMS exhibited a less distinct structure, making it challenging to delineate clear platelet territories. Interestingly, the lack of PACSIN2 also impacted the spatial distribution of GPIbα and integrin β1 membrane receptors. These receptors, typically dispersed within the cytoplasm in normal MKs, were concentrated at the cell periphery in Pacsin2 -/-MKs. A similar relocation was observed for polymerized actin. This finding underscores the pivotal role of PACSIN2 in actin reorganization within MKs, which in turn influences the rearrangement of GPIbα and integrin β1. The data imply that receptor and actin cytoskeleton rearrangements are critical to DMS formation and subsequent platelet production in vivo.
Blood Collection Workflow for Interrogation of Translational Samples with Single Cell Technologies
Authors: Walter Dagmar , Abousoud Jawad , Drennon Tingsheng , Kunihiro Connor , Martin Kelly , Li Yina , Raman Nandhini , Rodriguez Veronica , Herschleb Jill Lynden
Published: December 2023

Blood is a valuable and easily accessible sample type for clinical researchers that provides a snapshot of the organ systems at the time of sample collection. Readouts obtained from blood help inform clinical treatments, investigate disease mechanisms, evaluate drug response, and monitor outcomes. However, the impact of existing blood collection workflows on data output from new and high resolution technologies, like single cell RNA sequencing (scRNA-seq) remains a challenge. Recent scRNA-seq studies have shown that changes in gene expression can occur in blood cells in as little as a few hours post collection1. While bulk measurements rely on sample fixation to preserve the sample during transport, this approach does not preserve intact cells and hence, is not compatible with single cell technologies. With the recent availability of fixation compatible scRNA-seq assays comes the need to preserve blood while retaining single cell information. In this study, we evaluated various PBMC isolation and blood preservation methods which preserve single cell information while easing sample logistical constraints.

Our initial experiments compared various PBMC isolation methods. scRNA seq analysis of these samples demonstrated that all isolation methods captured single cell gene expression with some variability in cell types along with user-variability. Changes in cell type proportions were observed in as little as 4 hrs post blood draw, emphasizing the need for an effective blood preservation method to retain single cells during transport and blood cell isolation.

Next, we compared different methods of blood preservation including cryopreservation and fixation. Healthy and diseased whole blood samples were collected, preserved, stored followed by isolation of blood samples at different time points. scRNA-seq analysis of the samples demonstrated that blood can be preserved, transported, and stored (weeks to months) before blood cell isolation. Blood cells isolated from these cells can then be stored long term for scRNA-seq analysis. This blood collection and preparation workflow also eases blood transportation logistical constraints, allowing for batch shipping of samples from distributed collection sites. Coupled with automatable cell isolation methods and multiplexing single cell sequencing solutions, these workflows can be adapted for large scale translational research studies.

References: Massoni-Badosa, R., Iacono, G., Moutinho, C. et al. Sampling time-dependent artifacts in single-cell genomics studies. Genome Biol 21, 112 (2020).

Keywords: single cell RNA-seq, blood preservation
Efficacy of Low Dose Vs Standard Dose of Thalidomide in Patients with Transfusion-Dependent Thalassemia (TDT): A Non-Inferiority Trial
Authors: Kakkar Shruti , Sharma Ratna , Mahajan Amita , Radhakrishnan Nita , Dewan Priyanka , Manglani Mamta , Pandey Amit , Goel Manish , Parakh Nupur , Singh Varinder , Arora J S , Chandra Jagdish
Published: December 2023

Background

Thalidomide has been demonstrated to induce global gene expression hereby increasing the proliferation of erythroid cells. Recent studies have revealed encouraging data regarding the efficacy and safety of thalidomide in both transfusion dependent (TDT) and non-transfusion dependent (NTDT) thalassemia. However, the optimal dose of thalidomide remains unclear. The study was designed to compare the efficacy and safety of thalidomide at low-dose (1 mg/kg/d) vs standard-dose (2 mg/kg/d).

Methods

Patients with TDT > 12 years of age with no recent (previous 6 months) intake of Hb enhancers were enrolled in a non-inferiority trial across 4 sites in India. Patients with hypersplenism, prior history of thromboembolism, HIV, active hepatitis B/C infection, or any other known systemic illness like neurological, renal, significant hepatopathy, and sexually-active females unwilling to use contraceptives/undergo medical termination of pregnancy in case of accidental pregnancy while on the drug were excluded. Patients were randomised to receive low-dose or standard-dose thalidomide for 6 months. Pre-transfusion Hb, transfusion frequency and volumes, and adverse effects were recorded at each visit. Response was graded based on reduction in transfusion requirement at 24 weeks: Good (> 50 %), moderate (25-50 %), or minimal (<25%).

Results:

A total of 188 patients with mean age of 18.1 ± 5.2 years and M:F ratio of 2.1:1 were enrolled. 85.6% (80 in low-dose and 81 in standard-dose) participants completed the study period. 14.4% discontinued thalidomide due to various reasons. The mean Hb and baseline transfusion requirement were 8.6 ± 0.98 g/dl and 35.91 ml/kg in the preceding 12 weeks. The overall response rate was 55.9%. There was no statistically significant difference in the response rates in 2 groups (63% in low-dose vs 48% in standard-dose) (p:0.135). Good, moderate, and minimal responses were seen in 19.3%, 36.6 %, and 44.1% of the participants respectively. Response rate was not affected by age, sex, transfusion requirement or serum ferritin at baseline. The commonest adverse events were somnolence (n=40), constipation (n=29), weight gain (n=25), reversible cytopenia's (n=22), and peripheral neuropathy (n=14). 3.7% participants experienced Grade 2/3 events. There were no grade 4 events.

Conclusions:

The low-dose thalidomide was as efficacious as the standard dose in reducing transfusion requirements. Thalidomide is well tolerated and can be considered as an adjunct to optimal transfusion-chelation regimen under close monitoring in a resource constrained setting.
Daratumumab Combinations in US Veterans: Patient Characteristics and Outcomes
Authors: Prasad Sonika , Yin Ming , Lynch Julie A , Madison Cecelia J , Friedman Daphne R , Schoen Martin William
Published: December 2023

Background: Daratumumab is an anti-CD38 antibody with efficacy both in relapsed and newly diagnosed multiple myeloma (NDMM).Clinical trials evaluating daratumumab have demonstrated improved outcomes when combined with lenalidomide (DRd) or bortezomib (DVd) and with dexamethasone, as well as using all four drugs (D-VRd). We evaluated characteristics and outcomes of veterans treated with daratumumab combination in NDMM.

Methods: This retrospective, cross-sectional study analyzed electronic medical record data from the Veterans Affairs (VA) Corporate Data Warehouse (CDW). We identified veterans by evidence of daratumumab administration in prescription records in combination with a second agent within 30 days and who did not receive any anti-myeloma agents prior to 30 days. Age, race, Quan's Charlson Comorbidity Index (CCI), the time to next treatment (TTNT), and overall survival (OS) were assessed. Patient characteristics were compared using ANOVA or chi-square and overall survival was estimated using Kaplan-Meier. Analyses were conducted using SAS 9.x.

Results: There were 428 veterans who received daratumumab containing regimens. Mean age was 69.5 years and 36.2% self-identified as African American. There were 147 patients who received DRd, 190 DVd, and 91 D-VRd. Veterans who received DRd were significantly older than veterans receiving DVd (mean 73.0 vs. 69.1 years) and D-VRd(73.0 vs. 64.9 years, p<0.001). Use of daratumumab combinations has increased yearly since FDA approval (see table). There was no significant difference in use of regimens by race or comorbidity, measured by CCI. TTNT displayed distinct variations among the regimens, with DRd showing the most extended duration at a median of 316 days, followed by D-VRd at 231 days, and DVd at 40 days. These observed differences in TTNT may be attributed to changes in regimen, such as starting lenalidomide after 1-2 cycles of DVd. Median overall survival with DRd at 45 months (95% CI 38-not reached (NR)) and with DVd was 28 months (95% CI 20-39). Median survival in D-VRd was not reached.

Conclusion: Daratumumab combinations are increasingly used to treat multiple myeloma in the VHA. DRd is used more commonly in older veterans and D-VRd in younger veterans. DVd is used more frequently than other combinations and veterans may transition or other treatments.
A Novel, Probe-Based and Instrument-Free Method for Improved Ultrahigh-Throughput Single-Cell Gene Expression Analysis of Peripheral Blood Mononuclear Cells
Authors: Eastburn Dennis , White Kevin , Camiolo Salvatore , Montis Gioele , Martin Nicole , Seligmann Bruce
Published: December 2023

Single-cell gene expression assays are critical tools for identifying functional subtypes of immune cells, as well as changes within cells resulting from pathology or treatment. However, most current methods are expensive, low in sample throughput, rely on reverse transcription and lack the sensitivity to consistently measure low expressed genes from single cells - preventing measurements of many key immune biomarkers and molecular pathways. Another problem with existing single-cell sequencing methods is that many of the cell-associated reads either fail to map to transcripts or map to uninformative ribosomal and mitochondrial genes. Some methods also require the use of live cells, which means the single-cell assay has to be carried out at the specific time of treatment or when culture is terminated. The aforementioned limitations have contributed to most single-cell sequencing studies being restricted to small numbers of samples and have prevented their wider scale adoption in biopharma, clinical, translational and applied applications.

Here we report on the development and performance of a novel combinatorial split-pool-based workflow, TempO-LINC™, that adds cell-identifying molecular barcodes onto high-sensitivity gene expression probes within fixed cells. Cells can be fixed at the end of a treatment, or when obtained from a patient and stably stored for months until they are ready to be processed with other samples through the TempO-LINC assay. All probes within the same fixed cell receive an identical barcode, enabling the reconstruction of single-cell gene expression profiles across as few as several hundred cells and up to 100,000 cells per run. TempO-LINC simultaneously processes up to 96 fixed cell samples per run, has a simple workflow that is amenable to automation and as we will show, has a higher gene detection rate than existing single-cell platforms. We will also show that TempO-LINC has an observed doublet rate of less than 0.5% for 12,500 cells analyzed. Critically, TempO-LINC reduces sample sequencing costs and although we show the assay accurately profiles the whole transcriptome (23,000 transcripts), it can also be targeted to measure only genes/pathways of interest - dramatically reducing sequencing costs yet further. We applied TempO-LINC to assay PBMCs and a human hepatocyte model for toxicology and demonstrated the ability to correctly identify unique cell states and molecular pathways from these populations. We will also present time course and dose response data from compound treatment of cells. All of these features position TempO-LINC as a potential disruptive technology that hematology researchers, clinicians and drug developers can use for new large-scale applications/studies using single-cell sequencing across thousands of samples while obtaining improved data quality.