Hematology Research Research Articles

Narrative Review: Advancing Dysbiosis Treatment in Onco-Hematology with Microbiome-Based Therapeutic Approach

This review explores the complex relationship between gut dysbiosis and hematological malignancies, focusing on graft-versus-host disease (GvHD) in allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients. We discuss how alterations in microbial diversity and composition can influence disease development, progression, and treatment outcomes in blood cancers. The mechanisms by which the gut microbiota impacts these conditions are examined, including modulation of immune responses, production of metabolites, and effects on intestinal barrier function. Recent advances in microbiome-based therapies for treating and preventing GvHD are highlighted, with emphasis on full ecosystem standardized donor-derived products. Overall, this review underscores the growing importance of microbiome research in hematology–oncology and its potential to complement existing treatments and improve outcomes for thousands of patients worldwide.

EXPERIENCE IN HEALING FARMS FROM LEUKEMIA OF CATTLE

This article presents the experience of rehabilitating two farms with initial infection with the bovine leukemia virus of 17.72% and 33.48%. A scheme for improving the health of a farm using the method of serological and hematological research, with the isolation of sero-positive animals and the subsequent elimination of animals with hematological leukemia, is described. This also shows the prevalence and incidence of oncornavirus infection in an uncontrolled epizootic process.

Optimization of pre-enrichment strategies for mouse hematopoietic stem cell isolation and metabolomic analysis

Blood cell production arises from the activity of hematopoietic stem cells (HSCs), defined by their self-renewal capacity and ability to give rise to all mature blood cell types. The mouse remains one of the most studied species in hematological research, and markers to define and isolate mouse HSCs are well-established. Given the very low frequency of HSCs in the bone marrow, stem cell pre-enrichment by red blood cell lysis and magnetic cell separation is often performed as part of the isolation process to reduce sorting times. Several pre-enrichment strategies are available, differing in their speed, degree of enrichment, final cell yield, and cost. In the current study, we performed a side-by-side comparison and provide a decision tree to help researchers select a pre-enrichment strategy for mouse HSC isolation depending on their downstream application. We then compared different pre-enrichment techniques in combination with metabolomics analysis of HSCs, where speed, yield and temperature during pre-enrichment are crucial factors, and found that the choice of pre-enrichment strategy significantly impacts the number of metabolites detected and levels of individual metabolites in HSCs.

Erythroferrone in focus: emerging perspectives in iron metabolism and hematopathologies.

Beyond its core role in iron metabolism, erythroferrone (ERFE) has emerged as a key player with far-reaching implications in various hematologic disorders. Its regulatory effect on hepcidin underlines its significance in conditions characterized by disrupted iron homeostasis. In β-thalassemia and myelodysplastic syndromes, its dysregulation intricately contributes to the clinical challenges of anemia and iron overload which highlights its potential as a therapeutic target. In anemia of chronic disease and iron deficiency anemia, ERFE presents a unique profile. In chronic kidney disease (CKD), the intricate interplay between ERFE, erythropoietin, and hepcidin undergoes dysregulation, contributing to the complex iron imbalance characteristic of this condition. Recent research suggests that ERFE plays a multifaceted role in restoring iron balance in CKD, beyond simply suppressing hepcidin production. The potential to modulate ERFE activity offers a novel approach to treating a spectrum of disorders associated with iron dysregulation. As our understanding of ERFE continues to evolve, it is poised to become a key focus in the development of targeted treatments, making it an exciting and dynamic area of ongoing research. Modulating ERFE activity presents a groundbreaking approach to treat iron dysregulation in conditions like iron deficiency anemia, thalassemia, and hemochromatosis. As new research unveils its intricate roles, ERFE has rapidly emerged as a key target for developing targeted therapies like ERFE agonists and antagonists. With promising studies underway, this dynamic field holds immense potential to improve patient outcomes, reduce complications, and offer personalized treatment options in hematology research. This comprehensive overview of ERFE's role across various conditions underscores its pivotal function in iron metabolism and associated pathologies.

COVID-19 mRNA vaccination responses in individuals with sickle cell disease: an ASH RC Sickle Cell Research Network Study

AbstractChildren and adults with sickle cell disease (SCD) have increases in morbidity and mortality with COVID-19 infections. The American Society of Hematology Research Collaborative Sickle Cell Disease Research Network performed a prospective COVID-19 vaccine study to assess antibody responses and analyze whether messenger RNA (mRNA) vaccination precipitated any adverse effects unique to individuals with SCD. Forty-one participants received 2 doses of the Pfizer-BioNTech vaccine and provided baseline blood samples before vaccination and 2 months after the initial vaccination for analysis of immunoglobulin G (IgG) reactivity against the receptor binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 spike protein. Six-month IgG reactivity against the viral RBD was also available in 37 patients. Postvaccination reactogenicity was common and similar to the general population. There were no fevers that required inpatient admission. Vaso-occlusive pain within 2 to 3 days of first or second vaccination was reported by 5 participants (12%) including 4 (10%) who sought medical care. Twenty-seven participants (66%) were seropositive at baseline, and all 14 initially seronegative participants (34%) converted to seropositive after vaccination. Overall, mRNA vaccination had a good risk-benefit profile in individuals with SCD. This mRNA vaccine study also marks the first evaluation of vaccine safety and antibody response in very young children with SCD. This trial was registered at www.ClinicalTrials.gov as #NCT05139992.

Amino-modified polystyrene nanoplastics induced multiple response of Artemia hemocytes

Polystyrene polymers cause severe toxicity to aquatic animals. However, the process and mechanisms of innate immunity of invertebrates living at the bottom of the food chain to these pollutants remain unclear. In this study, the blood system responses of zooplankton Artemia were assessed through in vivo and in vitro exposure to amino-modified polystyrene nanoplastics (PS-NH2 NPs). The results indicated that the LC50 values of PS-NH2 NPs were 1.09 μg·mL−1 over 48 h and 0.42 μg·mL−1 over 7 d. Based on the five hemocyte subpopulations identified in Artemia, in vitro exposure assays revealed that phagocytosis was performed by plasmocytes and granulocytes with phagocytic rate of 22.64 %. TEM analysis further showed that PS-NH2 NPs caused cytoplasm vacuolization, swollen mitochondria, and lipid processing disorder. Gene expression pattern results demonstrated that Spatzle, Tollip, Hsp70, Hsp90, Casp8, API5and Pxn were significantly upregulated upon acute and chronic exposure (p < 0.05), while chronic exposure could induce significantly upregulation of ProPO (p < 0.05). Moreover, PS-NH2 NPs exposure remarkably varied the hemolymph microbiota and hemogram, particularly by increasing the proportion of adipohemocytes and phagocytes (p < 0.05). Our findings suggest that PS-NH2 NPs induce different responses in Artemia hemocyte, as primarily reflected by phagocytic processes, expression of immune and apoptosis relating genes, cell fates, hemogram and hemolymph microbiota variations. These findings support the possibility of using Artemia hemocytes as bioindicator to estimate nanoplastics pollution, thus contributing to hematological toxicity research in response to nanoplastics.

Diagnostic studies for enterotoxaemia in rabbits

The relevance of this study is highlighted by the occurrence of enterotoxaemia in rabbits, particularly caused by Clostridium perfringens, in the context of industrial rabbit farming. The research aimed to determine the causes of morbidity and increased mortality in young rabbits during the growing period. The study involved a stepwise analysis of feed samples, clinical examination of rabbits reared in an industrial rabbit farm, and laboratory examination of the obtained biological material. Additionally, to establish a diagnosis and provide the farm with appropriate recommendations regarding the maintenance and prevention of rabbit diseases, the health status of the rabbits, the causes of digestive disorders with symptoms of diarrhoea and increased mortality under production conditions were analysed. Clinical, haematological, pathological, microbiological, and statistical research methods were used. In diseased rabbits, an increase in rectal body temperature, symptoms of diarrhoea, and in some animals, seizures were established. Several animals succumbed to the disease. During laboratory blood tests of the rabbits, moderate anaemia was detected, with a decrease in the number of erythrocytes and a reduction in haemoglobin content. There was also a disturbance in the qualitative composition of erythrocytes, including the presence of poikilocytosis and altered erythrocyte forms: acanthocytes, echinocytes, schistocytes, dacryocytes, keratocytes, and drepanocytes. A decrease in the immune status of the rabbits was characterised by a reduction in the total number of leukocytes and lymphocytes in the blood and a low neutrophil-to-lymphocyte ratio. Biochemical analysis of rabbit serum revealed a decrease in glucose levels below the physiological range and an increase in the activity of the enzymes alanine aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase. Results of post-mortem examinations of the deceased rabbits indicated the presence of gas-filled small intestines, with a fluid content and no macroscopic signs of inflammation. The feed provided to the rabbits on the farm was found to contain sulphite-reducing clostridia, specifically Clostridium perfringens. Identifying the causes of morbidity and mortality in rabbits will enable the selection of appropriate methods for preventing deaths within specific farming conditions

Pharmacological composition based on bacteriocinnisin in experiments in vitro and in vivo.

Antibiotic resistance is a global health problem related to the transmission of bacteria and genes between humans and animals. The development of new drugs with antimicrobial activity research is an urgent task of modern science. The article presents data of in vitro and in vivo experiments on new pharmaceutical composition based on nisin. The antimicrobial activity was studied on the mastitis pathogens. To identify microorganisms the Matrix-Assisted Lazer Desorption/Ionization Time-of-Flight (MALDI-TOF) (mass spectrometry) method was performed using. To determine sensitivity, the serial dilution method and the diffusion method were used. On laboratory animals, biochemical, hematological, and histological research methods were used. Female nonlinear white laboratory rats were used, which were divided into one control group and three experimental ones. "Duration" factor was statistically significant for the following indicators: hemoglobin, hematocrit, leukocytes, lymphocytes, erythrocyte sedimentation rate, and eosinophils. The "Dose" factor did not show significance for any indicator, which means that the effect was similar regardless of the dose chosen. When analyzing the biochemical indicators, significant differences were found in the "Duration" and "Dose" factors, in the direction of a decrease in the indicators of total protein, globulins, urea, and an increase in the concentration of alkaline phosphatase. When conducting histological studies in the first experimental group, it was established that there were no changes in the structural and functional units of the organs. In animals of the second experimental group, the presence of reversible pathological processes of a compensatory nature was noted. More profound changes in the structure of the studied organs were recorded in the third experimental group. An in vitro study on cell cultures showed that the pharmacological composition has high antimicrobial activity against isolates from the mammary gland secretion of cows with mastitis. An in vivo study on laboratory animals showed that the developed composition belongs to the IV class of substances "low-hazard substances". Histological examination made it possible to select the safest dose of the pharmacological composition of no more than 500 mg/kg.

Early histological transformation of follicular lymphoma to diffuse large B-cell lymphoma indicating adverse survival: A population-based analysis and validation.

The histological transformation (HT) of follicular lymphoma (FL) is a crucial biological event. The study aimed to evaluate the incidence, clinicial characteristics, prognosis and impact of HT time on survival of FL transforming to diffuse large B-cell lymphoma in population-based large-scale cohorts. A retrospective cohort study of FL with HT was performed in the Surveillance, Epidemiology, and End Results database. The Hematological Malignancy Research Network FL cohort and Aristotle study FL cohort were used to assess the external validity. Among 44,127 FL cases from the Surveillance, Epidemiology, and End Results database, 1311 cases were pathology-proven recorded to transform to diffuse large B-cell lymphoma. The cumulative rates of HT at 5, 10, and 15 years after FL diagnosis were estimated to be 1.19%, 2.93%, and 5.01%, respectively. Significantly worse overall survival and cancer-specific survival were exhibited in patients with HT than those without HT. Early HT (transformation of FL within 48 months after FL diagnosis [TOD48]) was an independent predictor for adverse overall survival of HT patients, regardless of treatment modalities before transformation. The adverse prognostic effect of TOD48 was validated in the Hematological Malignancy Research Network cohort and Aristotle study cohort. Older age (>75 years) and B symptoms within FL at diagnosis were the independent risk factors of TOD48. Furthermore, a novel prognostic model combining TOD48 with Follicular Lymphoma International Prognostic Index (TOD48-FLIPI) was constructed and validated for risk stratification. TOD48 was a risk indicator of HT, and the novel prognostic model "TOD48-FLIPI" for HT patients was proposed.

Pharmacological effects of the use of enzyme and probiotic feed additive for dry cows

The scientific research conducted worldwide on the use of probiotics in animal husbandry demonstrates only a positive effect on the functioning of the macroorganism, but questions persist about the various pharmacological effects of multicomponent microbial preparations in combination with enzymes important in the digestion of certain types of productive animals, which makes it important for veterinary medicine and farmers to obtain environmentally friendly and high-quality products. The purpose of this study was to investigate the effect of a multicomponent enzyme and probiotic supplement on the duration of calving stages, prevention of postpartum complications in cows and some haematological parameters. Clinical, haematological, and statistical research methods were employed in this study. The research was conducted in a private dairy farm with 195 cows. The housing conditions and diet of the animals in the control group (n=6) and the experimental group (n = 17) were analogous, and the enzyme and probiotic supplement “ProActivo” (containing enzymes, Bacillus subtilis strain AX20, B. licheniformis strain EA22, and Enterococcus faecium) was added to the diet of the experimental cows at the rate of 10-14 g/animal, once a day, for 21-30 days before calving. A positive effect on the body of pregnant cows was found – elimination of diarrhoea symptoms in 4 days in 100% of diseased animals, and no changes in the control group. The study proved that the prenatal course of probiotic therapy of cows provided 95% prevention of afterbirth retention, a significant (P ≤ 0.001) reduction in the duration of the stage of afterbirth separation in cows of the experimental group (253.9±3.6 min) compared to the control group (326.8±8.9 min). A significant (P≤0.01) increase in the count of lymphocytes by 3.25±0.2 G/l, monocytes by 0.43±0.03 G/l, neutrophils (P≤0.001) by 1.6±0.28 G/l in cows treated with an enzyme-protein supplement was recorded, which is associated with the optimisation of the functioning of the primary immune system and the pharmacosynergistic effect of the multicomponent supplement. The findings obtained are valuable for veterinarians and dairy herd keepers, as the addition of an enzyme and probiotic supplement to the diet of dry cows will facilitate the calving stages and prevent afterbirth retention

Hematological manifestations of paraneoplastic syndromes in dogs

Hematological paraneoplastic syndromes are common but poorly studied complications in veterinary oncology, manifested by a variety of hematological changes associated with the system effects of malignant neoplasms on the animal's body. The article presents the results of studying hematological paraneoplastic syndromes, the research involves 131 dogs with spontaneous malignant neoplasms. We used a comprehensive methodological approach, including clinical, instrumental, morphological and hematological research methods. During the morphological study we found out that carcinomas were diagnosed in 33.6% of cases in dogs, of which breast carcinoma was registered in 24.4%, in other cases lung, liver, kidney and abdominal lesions were noted. We also discovered: sarcomas in 22.1% of cases, including hemangiosarcoma of the spleen in 7.6%; and gastrointestinal lymphoma in 16.8% of cases and lymph nodes; in 16.8% of mastocytomas. in 4.6% of melanomas of the skin and mucous membranes; squamous cell carcinoma in 6.1% of cases. As a result of hematological studies, it was revealed that anemia syndrome (18.4%), lymphopenia (16.8%) and thrombocytopenia (15.4%) are the most frequent changes in clinical blood analysis in dogs with oncological diseases. Neutrophilic leukocytosis, leukopenia and thrombocytosis had an identical incidence, which was 12.2% of cases. Monocytosis and lymphocytosis were diagnosed less frequently, in 4.6% of cases. Leukogram changes were observed in 75.4% of cases: in 66.2%, an increase in the percentage of neutrophils was found, with a shift to the left in 38.5% of cases, a decrease in the percentage of lymphocytes in 18.5% of cases, monocytes in 6.2% of cases, eosinophils in 4.6% of cases. The frequency of hematological changes in the general clinical blood analysis of dogs depended on the histogenesis of the tumor. Hematological paraneoplastic syndromes can be used as an early marker of the oncological process, and they can also prevent timely prescription of treatment. Anemia and thrombocytopenia are absolute contraindications for chemotherapy and radiation therapy.

Raman Spectroscopy as a Research and Diagnostic Tool in Clinical Hematology and Hematooncology.

Raman spectroscopy is a molecular spectroscopic technique able to provide detailed information about the chemical structure, phase, crystallinity, and molecular interactions of virtually any analyzed sample. Although its medical applications have been studied for several decades, only recent advances in microscopy, lasers, detectors, and better understanding of the principles of the Raman effect have successfully expanded its applicability to clinical settings. The promise of a rapid, label-free diagnostic method able to evaluate the metabolic status of a cell in vivo makes Raman spectroscopy particularly attractive for hematology and oncology. Here, we review widely studied hematological applications of Raman spectroscopy such as leukocyte activation status, evaluation of treatment response, and differentiation between cancer and non-malignant cells, as well as its use in still unexplored areas in hematology. We also discuss limitations and challenges faced by Raman spectroscopy-based diagnostics as well as recent advances and modifications of the method aimed to increase its applicability to clinical hematooncology.

Assessing Iron Deficiency Anemia in Obese Adolescents and Identifying Contributing Factors

Objective: Clinical outcomes can be significant for obesity and iron deficiency, two public health issues for which adolescents are especially vulnerable. The aim of this study was to assess the iron parameters and blood vitamin B12 levels in obese adolescents and identify the contributing variables to the development of anemia. Material and Method: The present study involved a retrospective evaluation of 260 children (130 obese-130 control) who were admitted to the Ankara Pediatrics Haematology Oncology Training and Research Hospital, Pediatric Outpatient Clinics, between March 2013 and May 2015. Body mass index (BMI) above the 95th percentile for age and gender was required for inclusion in the study group, which included children aged 12 to 18 years without acute or chronic illnesses. Data from patient files were used to collect information on physical examination findings, sociodemographic characteristics, daily dietary status, and level of physical activity. All patients had evaluations for CRP, iron parameters, vitamin B12, and complete blood count. Results: The study revealed that the obese group had significantly higher serum ferritin levels (p=0.002) and lower serum iron and vitamin B12 levels (p=0.036 and 0.047, respectively) as compared to the control group. In the obese population, elevated BMI has been demonstrated to be correlated with elevated CRP levels. Conclusion: In obese children, low serum iron and high ferritin levels can be regarded as typical reference values. Furthermore, it was discovered that low vitamin B12 levels were not associated with nutrition in obese children, although they were at risk for developing low levels

The Role of New Morphological Parameters Provided by the BC 6800 Plus Analyzer in the Early Diagnosis of Sepsis

Background: Late diagnosis of sepsis is associated with adverse consequences and high mortality rate. The aim of this study was to evaluate the diagnostic value of hematologic research parameters, that reflect the cell morphology of blood cells, available on the BC 6800 plus automated analyzer (Mindray) for the early detection of sepsis. Materials and Methods: A complete blood count (CBC) was performed by Mindray BC 6800 Plus Analyzer in 327 patients (223 with a confirmed diagnosis of sepsis following sepsis-3 criteria, 104 without sepsis), admitted at the Intensive Care Unit of the Novara’s Hospital (Italy) and in 56 patients with localized infection. Results: In univariate logistic regression, age, Hb, RDW, MO#, NMR, NeuX, NeuY, NeuZ, LymX, MonX, MonY, MonZ were associated with sepsis (p &lt; 0.005). In multivariate analysis, only RDW, NeuX, NeuY, NeuZ, MonX and MonZ were found to be independent predictors of sepsis (p &lt; 0.005). Morphological research parameters are confirmed to be predictors of sepsis even when analyzing the group with localized infection. Conclusions: In addition to already established biomarkers and basic CBC parameters, new morphological cell parameters can be a valuable aid in the early diagnosis of sepsis at no additional cost.

Hemato Keeps You Updated on the Research in Hematology

Hemato (ISSN 2673-6357) is an open access, peer-reviewed journal that publishes original articles and reviews highlighting important advances in the fundamental areas of Hematology [...]

Interest and barriers to research in obstetric haematology – findings from a national survey in the United Kingdom

Introduction In 2021, the steering committee members of British Society of Haematology Obstetric Haematology Special Interest Group noted difficulties in opening research studies. This led to the development of a survey to further evaluate this issue. Method An electronic survey was distributed to all members of the British Society of Haematology Obstetric Haematology Special Interest Group and to relevant specialty leads of the National Institute for Health and Care Research Clinical Research Network for further dissemination within these networks. Results Responses were received from 65 participants (73% consultant grade); mainly haematologists (52%) or obstetricians (39%). Less than a third of participants reported dedicated time for research in their job plan, with only five participants reporting no challenges in opening research studies in obstetric haematology. Discussion The survey confirmed significant interest in obstetric haematology research, with barriers to participation. We propose further actions to facilitate increased research.

BinaryClust2: An Integrated Computational Pipeline for Systematic Mining of Mass Cytometry Data to Assist Deep Immune Profiling in Haematological Research

Introduction: The key role of immune system in modulating treatment efficacy and clinical outcomes in the settings of haematological diseases has been well recognized through the application of flow cytometry technologies over years. Mass cytometry (MC) is a next generation cytometry platform to simultaneously profile over 40 protein markers on single-cell resolution, which facilitates clinical research on immune monitoring and biomarker discovery. Nonetheless, the high-dimensional data generated present a daunting challenge for clinical investigators. Therefore, we describe BinaryClust2 (https://github.com/JingAnyaSun/BinaryClust2), an R-based computational framework integrated with the state-of-the-art algorithms and novel cell lineage characterisation method to assist comprehensive exploration of liquid MC data. Method: Here, we present the implementation of BinaryClust2 to three example MC datasets: In-house dataset from peripheral blood mononuclear cells (PBMCs) of 9 baseline myeloproliferative neoplasm (MPN) patients (~4 million cells), previously published data from 11 MPN patients receiving influenza vaccine (~0.2million cells), and published dataset of 59 covid patients and 23 healthy donors (~2million cells). BinaryClust2 has a streamlined analytical workflow, which comprises the following steps (Figure 1A): Step 1: Quality control, batch effect evaluation and correction Data quality control is a separate step before downstream analysis which includes diagnostic plots and batch effect examination. Algorithms CytofRUV and CytoNorm are available in the pipeline to remove unwanted variations caused by batch effects. Step 2: Semi-supervised identification of main cell types BinaryClust2 adopts a knowledge-based semi-supervised approach to predict main cell types. Users are required to provide a simple marker expression matrix of pre-defined cell types along with fcs files and metadata to construct a SingleCellExperiment (SCE) object, then the embedded algorithm can automatically classify cell populations without manual annotation. Step 3: In-depth interrogation and differential testing Specific population can be further extracted from whole cells and subject to in-depth exploration using unsupervised algorithms for subpopulation discovery. Dimensionality reduction tools UMAP and TSNE, unsupervised clustering methods Phenograph and flowSOM, and various data visualization plotting functions are offered in BinaryClust2. For statistical analysis, multiple study group comparison (n&amp;gt;2) of cell abundance and functional marker expression is supported via Kruskal Wallis test with multiple testing correction and post hoc analysis. Results: The performance of the semi-supervised classification function was tested independently in the MPN and influenza PBMC datasets, 7 main cell lineages were identified with accuracy comparable to manual gating by human experts (Figure 1B): average F-measure reached 0.93 and 0.98 respectively. Moreover, taking manual gating as ground truth reference, BinaryClust2 outperformed the unsupervised approach flowSOM concerning accuracy (F-measure: 0.93 vs. 0.70) and speed (140s vs 339s) in MPN dataset, while remaining equivalent to the well-performing semi-supervised approach LDA in accuracy (F-measure: 0.93 vs. 0.93) but faster in runtime (140s vs 595s), as shown in Table 1. Application to covid-19 dataset by Chevrier et al. achieved reproducible results and additional discoveries. 13 main cell types were characterised, abundance of B cells, Basophils, cDCs, DN T cells, Monocytes, Neutrophils, NK cells, pDCs, CD8 T cells obtained statistical significance (all P&amp;lt;0.05) among study conditions (healthy, mild covid, severe covid). We also grouped markers reflecting functional status of immune cells and found Granzyme B expression was significantly increased in the majority of main immune cells of covid patients. Phenograph was further applied in neutrophils and monocytes and returned 14 and 16 subsets respectively. Conclusion: Overall, BinaryClust2 incorporates expert's prior biological knowledge in a semi-supervised fashion to accurately deconvolute well-defined main cell lineages, while also preserving the potential of unsupervised approaches to discover novel cell subsets and providing a user-friendly toolset to remove the analytical barrier for high-dimensional immune profiling.

Clinical Factors but Not Somatic Mutations Predict for Survival in Patients with Myelofibrosis Undergoing Allogeneic Hematopoietic Cell Transplant: Analysis of the North American Myelofibrosis Transplant Outcome (NAMTO) Study

Introduction: Myelofibrosis (MF) is a clonal hematologic malignancy characterized by constitutive JAK-STAT activation and aberrant cytokine signaling. Allogeneic hematopoietic stem cell transplantation (allo-HCT) remains the only potentially curative therapy in MF. The contribution of disease-, patient- and donor-related factors to allo-HCT outcomes remains to be fully elucidated, as well as the presence of somatic mutations in genes such as ASXL1, EZH2, SRSF2, U2AF1 and IDH1/2 which are associated with worse prognosis in MF patients (pts). In this large multi- center retrospective analysis, we aimed to study the impact of clinical and molecular mutations on transplant outcomes. Methods: This analysis including 498 pts from 11 centers in North America and Canada. Univariate proportional-hazard Cox regression analysis stratified by center was used to assess the association of patient, transplant, and disease characteristics on overall survival (OS). Baseline patient demographic data and disease characteristics were selected as covariates in the multivariate analysis. Results: Pts transplanted between 2002-2021 were included in this analysis. Baseline and transplant characteristics are summarized in table 1. The 1- and 2-year OS was 77% (95% CI; 73-81) and 70% (95% CI; 66-74) and non-relapse mortality (NRM) 19% (95% CI: 17-21) and 23% (95% CI: 19-27), respectively. The cumulative incidence (CMI) of relapse at 1- and 2-years was 12% (95% CI; 10-14) and 16% (95% CI; 12-20). The CMI of acute graft versus host disease (aGVHD) grade II-IV was 31% (95% CI; 27-35) at 3 months and 42% (95% CI; 38-46) at 1-year post-transplant and grade was III-IV 13% (95% CI; 11-15) and 18% (95% CI; 14-22), respectively. The CMI of neutrophil engraftment at 1 and 3 months was 90% (95% CI: 87-93) and 97% (95% CI: 95-95) and for platelets 55% (95% CI: 50-59) and 80% (95% CI: 76-83). Univariate analysis included pts (age and gender), transplant (donor type and conditioning regimen intensity) and disease characteristics (DIPSS, mutations, cytogenetics (CG), blood counts, and spleen size). Hemoglobin &amp;gt; 10 g/dL (HR .5, 95% CI .34 -.73 p=3 x 10 -4) and platelets &amp;gt;50× 10 9/L (HR .59, 95% CI .42-.81, p=.0013) prior to transplant were associated with improved OS. High risk DIPSS was associated with decreased OS (p=0.0002). Mutations in ASXL1, EZH2, IDH1/2, U2AF1 and SRSF2 were not associated with decreased OS in this analysis (Figure 1). However, mutations in ZRSR2 were associated with inferior OS (HR 2.07, 95% CI 1.08-3.95, p=.0252). Lastly, JAK2 which had the most frequent mutations was not associated with OS (HR .0.93, 95% CI .65-1.31). There was no association between the CG risk groups and OS. Splenomegaly, measured radiographically, had no impact on OS but was strongly associated with engraftment (better engraftment defined as HR&amp;gt;1). For platelets, spleen size of 15.6-18.9 cm HR of 0.69 (95% CI .47-1, p=.0489) and size &amp;gt;18.9-22.5 cm HR of 0.48 ( 95% CI .33-.7, p=1x10^-4). For neutrophils, spleen size &amp;gt;19.9-22.5 cm had a HR of 0.68, (95% CI .48-.96, p=.0268) and size&amp;gt;22.5 cm had a HR of 0.65 (95% CI .46-.91, p=.0114). Mismatched unrelated (MMURD) and haplo identical donors had no impact on survival but were associated with delayed platelet engraftment; MMURD (HR .67, 95% CI .47-.96, p=.0298) and haploidentical donors (HR .56, 95% CI .39-.8, p=.0017). In a multivariate analysis adjusted for age, gender, highest DIPSS, donor type and conditioning regimen, platelets &amp;gt; 50× 10 9/L (HR .61, 95% CI .44-.87, p=.0053) hemoglobin &amp;gt;10 g/dL (HR .6, 95% CI .4-.9, p=.0244) and lower DIPSS at the time of transplant were associated with improved OS. Conclusions: In this large multicenter analysis only clinical variables; hemoglobin and platelets, were found to be associated with OS after allo-HCT in MF patients. Somatic mutations associated with poor prognosis were not associated with OS suggesting that transplant can overcome their negative impact. Splenomegaly was associated with delayed engraftment but not OS. Interestingly, using alternative donors (MMURD and haplo identical) didn't have a negative effect on OS, possibly reflecting the use of post-transplant cyclophosphamide for GVHD prophylaxis in recent years. These results highlight the importance of biologic factors in the prognostication of outcomes in MF pts undergoing allo-HCT independent of somatic mutations. Supported by the American Society of Hematology (ASH) and MPN Research Foundation

Returning Secondary Genomic Findings to a Sickle Cell Disease Cohort

Introduction: As gene therapy for sickle cell disease (SCD) continues to advance, there is growing potential for genomic screening to be utilized in determining eligibility for this therapy. With this comes the ethical obligation to look for secondary findings (SF) in clinical exome and whole genome sequencing. The American College of Medical Genetics (ACMG) has issued recommendations for returning SF in clinical care and annually updates a list of pathogenic variants. This abstract reports on our experience with returning SF to a SCD study population in research. Methods: The INSIGHTS Study (NCT02156102) is a cross-sectional perspective research study examining phenotypic variation in adults living with SCD. All study participants consented to whole genome sequencing (WGS) and agreed to have secondary genomic findings returned. Participants completed the study between 2014-2020. To date, 264 samples have undergone WGS, performed at the National Institutes of Health Intramural Sequencing Center. Results: In accordance with the guidelines issued by ACMG, 12 participants (4.5%) were found to have genetic variants on the current list (ACMG SF v3.2) that were eligible for return (Table 1). Additionally, one participant was found to have a sex chromosome aneuploidy, and after a full ethics board review, it was determined this result should also be returned based on its clinical actionability and the risk/benefit analysis. Of these 12 participants, three were deceased. One participant was already aware of the secondary finding. We contacted the remaining eight participants and as of July 2023 five participants provided a new sample for CLIA confirmation. Results were returned in consultation with the Secondary Findings Consult Service at the National Human Genome Research Institute, which provided a genetic counselor. Participants met with a clinical member of the study team and the genetic counselor either by telephone or in person. The consultation consisted of sharing the result and associated risk of disease, obtaining pertinent medical history and detailed family history, and providing recommendations and referrals. Discussion: With the expansion of genomics into hematological and sickle cell disease research, it is paramount that the implications of its integration continue to be examined. The responsibility of the research team to facilitate and/or provide ancillary care to their study participants to best support them in receiving SF results should be clearly defined. In addition, each patient will have unique needs that should be considered and may require specialized assistance. Questions on how the SF affects the patient's primary diagnosis will need to be addressed, which will require a multidisciplinary approach to care. These consultations will require a high level of patient coordination and clinical support. It is important that these patient encounters 1) involve a known and trusted medical professional; 2) provide accurate and understandable information on the genomic finding, usually by a genetic counselor; and 3) refer the patient to an appropriate clinical provider who is an expert in that disease.

Spplaash Study: A Randomized Study to Evaluate the Incidence of Postdural Puncture Headache with the Use of Atraumatic Needles in Hematology

Introduction - Postdural-puncture headache (PDPH) is the most common complication after lumbar puncture, probably due to leakage of cerebrospinal fluid (CSF) from the dural defect into the epidural space. Most cases of PDPH occur within 8 days of a lumbar puncture and may be associated with other symptoms, such as photophobia, nausea or neurological complications. In this context, PDPH is a debilitating condition that significantly impacts quality of life. While several risk factors have been proposed to participate in PDPH etiology, no preventive or therapeutic measures have robustly proven their efficacity . Among PDPH-related risk factors, needle tip shape has been shown to influence the incidence of PDPH, especially in the context of diagnostic LP. The purpose of this study was to assess the incidence of PDPH using conventional (cutting-type) or atraumatic (pencil-point) spinal needles in hematological patients. Methods -This was a randomized, double-blind study of 68 adult patients scheduled for a diagnostic and/or therapeutic lumbar puncture (LP) (SPPLAASH study, NCT03126045). Subjects were randomly assigned to receive one of two needles : 22-gauge cutting (n=36) and 26-gauge atraumatic (n=32). LP and CSF collection were performed in accordance with standardized procedures. The main outcome was the occurrence of severe headache (≥4 out of 10 on the numerical rating scale (NRS)) as measured by interviews for 8 days after LP, repeated until headache resolution. Secondary outcomes included maximum intensity and duration of PDPH as well as procedural failure. Results - The median age at the time of LP was 60 years (Q1-Q3, 46-70) and 40% of patients were women. Demographic and procedural characteristics are summarized in Table 1. No significant difference was statistically observed between the two treatment groups, except for LP indication. The overall rate of severe PDPH was 19.7% (95% CI - 10.9-31.3). Interestingly, the incidence of severe PDPH was statistically reduced from 31.3% (95% CI - 16.9-49.3) in the conventional needle group to 6.5% (95% CI - 0.8-21.4) in the atraumatic group (p=0.011). High body mass index (OR 0.72, 95% - CI 0.57-0.91) and male sex (OR 0.29, 95% CI 0.08-1) were factors associated with a lower risk of PDPH. Means of maximum intensity of PDHP at 8 days were estimated to be 2.3 (sd 3.1) in the conventional needle group and 0.7 (sd 1.31) in the atraumatic group, with a statistically difference when considering median intensities (p=0.032) and number of days with NRS ≥4 (p=0.030). We used Sankey flow diagram to map the trajectories of PDPH intensities during the 8-day follow-up (Figure 1). First attempt LP success, failure rate, mean number of attempts, and the frequency of analgesic prescritions did not statistically differ between the two needle groups. Conclusion - Lumbar puncture is an essential and common procedure in hematologic clinical care and research. Our data indicate that atraumatic needles were associated with a decrease in the incidence of severe PDPH as well as in the duration of PDPH event. Moreover, the use of atraumatic needle was safe and feasible. Given that PDPH has a major impact on quality-of-life already altered by hematological treatments, and that headache might be caused by a differential diagnosis (e.g. cerebral sinus venous thrombosis due to Asparaginase in acute lymphoblastic leukemia), the use of atraumatic needles should be encouraged in this population, which often benefits from multiple LP.