Red Cell Transfusion Research Articles

Outcomes of Neutropenic Patients with Severe Infections Treated with Granulocyte Transfusions: The USC Experience

Introduction Despite major progress in antimicrobial and supportive measures, prolonged disease-associated or treatment-related neutropenia is a leading cause of mortality and morbidity in patients after stem cell transplantation or with hematologic malignancies or bone marrow failure. Intuitively, a granulocyte transfusion should be as effective as a platelet or red cell transfusion. However, efforts to prove the effectiveness of granulocyte transfusions have been disappointing. Due to the lack of adequately powered randomized controlled trials, it is difficult to prove or refute the effectiveness of granulocyte transfusions in neutropenic patients. Therefore, they are likely underutilized or used inappropriately as a last resort. To better understand the utility of this treatment modality, we reviewed the outcomes of patients with hematologic conditions who received granulocyte transfusions at our institutions during the treatment of neutropenic sepsis. Methods We identified 20 patients with severe neutropenia who received a granulocyte transfusion at Norris Comprehensive Cancer Center or Keck Hospital of USC in Los Angeles, California between 2014-2022. 19 of these patients received a total of 166 transfusions for treatment of suspected or confirmed sepsis. One patient received a prophylactic dose of granulocytes and was excluded from this study. Granulocyte donors were mobilized with 480 mcg G-CSF and a minimum yield of 4 x 10 10 granulocytes was obtained. The primary outcome was attainment of “clinical improvement” defined as either safe discharge from the hospital or resolution of symptoms. Descriptive statistics were presented as counts and percentages for categorical variables and as means with standard deviations for continuous variables. Categorical variables were accessed by Chi-square test or Fischer's exact test when appropriate. For continuous variables, Kruskal-Wallis test was used. P-values of ±0.05 were regarded as significant and all tests were two-tailed and analyzed with SAS (9.4). Results In our cohort, 47.4% were male, ages ranged from 20-70, and 57.9% identified as Hispanic. The most common diagnosis was acute myeloid leukemia (AML) (n=13), followed by aplastic anemia (n=3), myelodysplastic syndrome (MDS) (n=2), and acute lymphoblastic leukemia (ALL) (n=1). The most common infections were fungal or presumed fungal (n=9) and mixed fungal and bacterial (n=7). Prior to the start of granulocyte transfusions, 63.2% of patients were neutropenic (ANC <500) for at least 30 days and 84.2% had an absolute neutrophil count (ANC) <100. In surviving patients, the average number of days from the first transfusion to clinical improvement was 17.5 days. 47.4% of patients had a reported reaction to the granulocytes, with the most common being fever and tachycardia. Of the 19 patients treated, seven (37%) died from infection. Of those who died, two (29%) died from E. coli bacteremia and five (71%) from mixed bacterial and fungal sepsis. All nine patients with acute fungal infections without concomitant bacterial infections survived. Conclusion In this cohort, granulocyte transfusions appear to have had a positive effect with 63% of critically ill patients attaining clinical improvement from infection. All patients who had a purely fungal infection achieved clinical improvement and all patients who died had bacterial or mixed infection. Time to transfusion from initial infection, number of transfusions, and demographic variables did not appear to impact overall survival. Granulocyte transfusion could be an effective therapeutic modality in the treatment of infection in patients with prolonged severe neutropenia, especially those with fungal infections. Adequately powered randomized controlled trials are the next step in determining the effectiveness of this treatment.

Pituitary and Pancreatic Iron Is Present at an Early Age in Transfused Patients with Alpha Thalassemia Major

Introduction Alpha thalassemia major is generally fatal in utero or shortly after birth without aggressive red cell transfusion therapy both pre- and postnatally. The Society for Maternal-Fetal Medicine has released evidence-based practice guidelines for the diagnosis and management of fetal anemia, largely focused on red cell alloimmunization, with intrauterine transfusion typically starting at 18 weeks gestational age.((SMFM) et al., 2015) Early initiation of intrauterine transfusion allows quick correction of anemia and a prolonged reduction of hemoglobin Bart's concentration leading up to birth.(Chmait, Baskin, Carson, Randolph, & Hamilton, 2015) Long-term transfusion regimens postnatally invariably lead to iron overload, which can lead to organ dysfunction.(Taher & Saliba, 2017) Minimal data is currently available regarding the pattern of iron overload in young patients with alpha thalassemia major given the rarity of the disease itself and variations in iron overload monitoring. Methods We reviewed patients with alpha thalassemia major treated at our center who had iron overload evaluations prior to 10 years of age. Diagnoses were confirmed by genetic testing. Iron deposition measurements were collected using multiple gradient echo pulse sequences on 1.5 Tesla magnets. Results Six subjects were included in the study. All but one subject received several intrauterine transfusions and were delivered at term. The last subject did not receive any intrauterine transfusions and was born at 24 weeks gestational age. Subjects were routinely transfused postnatally to keep fast migrating hemoglobin below 10%. Initial MRI was done prior to 4 years of age and as early as 1.5 years. All subjects had moderate to severe iron overload based on liver iron concentration (LIC) by R2* MRI, which decreased significantly after starting chelation in the four subjects who had subsequent MRI assessments (Figure 1A). Pancreatic iron was also elevated in all subjects, and none had cardiac siderosis. All but one subject had pituitary R2 by Z-score for age and sex greater than 2 in the coronal and sagittal planes, indicating pituitary siderosis, but none had pituitary volume loss (Figure 2B). Discussion This study shows significant extrahepatic iron deposition (pancreas, pituitary) at a very young age indicating exposure to toxic reactive ferrous iron (Fe 2+). The pancreas is known to load temporally before the heart.(Noetzli, Papudesi, Coates, & Wood, 2009) None of the subjects in this study had cardiac iron by MRI, probably reflecting slower cardiac loading and lowering of Fe 2+ by chelation. It is very rare to have cardiac loading from transfusion in patients less than 2 years old.(Berdoukas et al., 2013) It is reassuring that none of the patients had pituitary volume loss, which has been associated with pituitary dysfunction,(Noetzli et al., 2012) although we know very little about pituitary function and iron in very young children. Our observation that some subjects had loading at 1.5 years old suggests there is significant circulating Fe 2+ in this population and the pituitary may be at risk. Chelation is rarely started prior to 2 years of age due to concerns about theoretical effects of chelation on neurodevelopment.(Coates, 2019) Despite significant levels of iron overload seen in initial assessments, hepatic iron rapidly improved with chelation despite aggressive red cell transfusion in this cohort. There are many unknowns here, including effects of prenatal iron exposure from transfusion, the extent to which iron seen by MRI signifies organ damage, and response of extrahepatic iron to chelation. However, the effects of iron deficiency on brain development are well documented(Idjradinata & Pollitt, 1993; Lozoff et al., 2013) and the possible effects of chelators on the newborn/infant brain could be serious. Long-term pituitary iron data are still needed to support earlier implementation of iron chelation in this population.

Appropriateness of Immunosuppression and Blood Product Utilization in Acquired Hemophilia A: A Multicentre Provincial Practice Audit

Introduction: Acquired hemophilia A (AHA) is a life-threatening bleeding disorder associated with significant morbidity and mortality. Despite publication of international consensus guidelines, quality of care and guideline adherence have not been examined. While some jurisdictions have reference centres for AHA management, in Canadian provinces outside of Quebec, AHA is managed in both hemophilia treatment centres (HTCs) and community hospitals. We performed a multicentre practice audit in a Canadian province to assess use of immunosuppressive therapy (IST), blood product utilization, and evaluate treatment burden and outcomes. Methods: We included adults diagnosed with AHA (January 2000-December 2021) in Alberta, Canada, and examined the following quality of care indicators: delayed IST initiation from diagnosis, corticosteroid use alone in high-risk AHA (FVIII <0.01 IU/ml or FVIII inhibitor >20 BU/ml), cyclophosphamide use in low-risk patients (FVIII ≥0.01 IU/ml and inhibitor ≤20 BU/ml), prolonged cyclophosphamide use (>6 weeks), and inappropriate use of plasma, intravenous immunoglobulin (IVIG) and hemostatic products. Outcome measures included response, length of stay (LOS) and 30-day readmission. We evaluated treatment burden including frailty syndromes, increased level of care and hospitalization for treatment-related adverse events. Results: Of the 38 patients diagnosed with AHA, 25 (66%) were female, the median age was 74 years (IQR 61-81), and median Charlson comorbidity index was 5 (IQR 3-7). Cardiovascular comorbidities were common, including venous thromboembolism (5; 13%), atrial fibrillation (5; 13%), and myocardial infarction (4; 11%). Fifteen (39%) had ≥1 frailty syndromes, including functional dependence (6; 16%), dementia (6; 16%), falls/fractures (5; 13%), and depression/anxiety (3; 8%). Five (13%) patients required higher levels of care (long term care placement) following discharge. Twenty-one (55%) patients had high-risk disease and 17 (45%) had low-risk. In those with low-risk disease, 11 (29%) had inhibitor titres 5-20 BU/ml and 6 (16%) had inhibitors <5 BU/ml. Despite diagnostic delays ≥7 days from bleeding in 15 (39%), most received prompt initiation of IST (median 0 days [IQR 0-3] from diagnosis). First-line IST included: prednisone (38; 100%), cyclophosphamide (23; 61%), and rituximab (13; 34%). IVIG was prescribed in 5 (13%), out of keeping with guideline recommendations. We observed very high rates of first-line cyclophosphamide use in low risk AHA (11/17; 65%), including 4/6 with inhibitor titres <5 BU/ml (Table 1). First-line cyclophosphamide was also used in 3 reproductive-aged women. Prolonged courses of cyclophosphamide >6 weeks and >6 months were used in 21/23 (91%) and 7/23 (30%) patients, respectively. Two high-risk patients did not receive cyclophosphamide or rituximab-containing therapy. Of the 58 episodes of bleeding events, 41 (71%) were ISTH major bleeds. Fifteen (26%) bleeding events did not receive hemostatic therapy, whereas 27 (47%), 19 (33%), and 3 (5%) were treated with recombinant factor VIIa, activated prothrombin complex concentrates, and porcine FVIII, respectively. Dosing of bypassing agents was in keeping with guidelines. DDAVP and human FVIII were inappropriately used in 3/58 (5%) and 1 (2%) bleeding episodes, respectively. Most patients (24; 63%) received red cell transfusions with a median of 2 (IQR 0-7) units each. Five (13%) patients received a total of 18 units of plasma. Clinical responses included: complete remission (CR) in 30 (79%), partial remission in 3 (8%), and unevaluable due to early deaths in 5 (13%). All 5 patients who relapsed (13%) achieved second CR. The median LOS was 38 days (IQR 19-57) across 95 hospitalizations. The 30-day readmission rate was 24%. Reasons for admission included bleeding (50/95; 53%), infections (14/95; 15%), and other treatment side-effects (13/95; 14%). Conclusion: In this provincial cohort of AHA patients with high comorbidity and frailty, we identified gaps in quality of care including overuse of cyclophosphamide in patients with low-titre inhibitors, prolonged duration of cyclophosphamide, and inappropriate use of IVIG and plasma. Aggressive cytotoxic therapy predisposes patients to unnecessary risks of infections and malignancies. Our findings highlight the need for centralization of care in specialized centres and education initiatives.

Optimizing Preoperative Iron Deficiency Anemia in Patients Undergoing Cardiac Surgery

Background: Preoperative anemia is common in patients undergoing cardiac surgery, with iron deficiency being the most common cause of anemia in these patients. Patients who are anemic in the preoperative setting have worse perioperative outcomes than patients with normal hemoglobin levels including higher rates of red cell transfusions, acute kidney injury, increased length of stay, and increased mortality. At the University of Virginia (UVA) Medical Center, there was no institutional process in place to identify and treat iron deficiency anemia in patients planned to undergo cardiac surgery. Objective: The UVA Perioperative Blood Management Committee is a newly-formed multidisciplinary group consisting of members from the departments of Hematology, Anesthesiology, Thoracic and Cardiovascular Surgery, Perfusion, and Blood Bank who work together to improve perioperative patient outcomes and blood product utilization. We set out to implement a testing algorithm with the purpose of improving the proportion of anemic patients undergoing cardiac surgery who have iron studies performed in the outpatient setting, thereby improving the percentage of patients receiving intravenous (IV) iron preoperatively. Methods: We adapted two published perioperative treatment algorithms (Lin, Hematology Am Soc Hematol Educ Program 2019; Munting and Klein, Anaesthesia 2019) which recommend iron therapy in patients with hemoglobin < 13 g/dL, ferritin < 100 µg/L, and/or transferrin saturation < 20% for our cardiac surgery preoperative clinic. In November 2022, we implemented a reflex test within Epic electronic medical record that automatically sent ferritin, iron, and transferrin for patients with hemoglobin < 13 g/dL to evaluate for iron deficiency. Patients meeting parameters for iron deficiency (ferritin < 30 ng/mL and/or transferrin saturation < 20%) were then scheduled for iron infusions. We compared the percentage of patients undergoing testing for iron deficiency, as well as the percentage of anemic patients receiving iron infusion preoperatively, to those from a six month period prior to our start date. Results: Between May 31, 2022 and October 31, 2022, 19% of cardiac surgery patients at UVA with hemoglobin values < 13 g/dL had iron studies drawn in the three months prior to surgery. After implementing outpatient reflex iron testing, that number increased to 42% between November 2022 and May 2023 (p < 0.001). In the six months prior to implementation, four iron deficient cardiac surgery patients received preoperative IV iron an average of one day prior to surgery. Between November 1, 2022 and May 29, 2023, fifteen patients received IV iron an average of 5.7 days prior to surgery. The patients who received IV iron had lower rates of blood transfusions than anemic patients who did not receive IV iron (53% vs. 65%) and fewer units of blood transfused per patient who received blood (3.1 vs. 5.7). Conclusions: Results from the first seven months of this project suggest that using a standardized testing methodology in the outpatient setting can increase the uptake of iron testing and thereby treatment of iron deficiency anemia in the preoperative setting. Patients who received IV iron had lower rates of blood transfusions and received fewer units of blood on average than anemic patients who did not receive IV iron. We next plan to implement an iron testing strategy in cardiac surgery inpatients to further improve rates of iron testing and iron infusions in this population. We have created an IRB-approved database to study more patient-centered outcomes such as acute kidney injury, stroke, length of stay, readmissions, and mortality to determine if we can detect a measurable impact from IV iron administration in our patient population.

Small Molecule Inhibitor Targeting Nemo-like Kinase Improves Erythropoiesis in Human and Mouse Models of Diamond Blackfan Anemia

Diamond Blackfan Anemia (DBA) is a rare inherited bone marrow failure syndrome characterized by anemia, congenital anomalies, and an increased risk of developing cancer. Approximately 80% of patients with DBA have a mutation in one of 20 different genes that encode ribosomal proteins, resulting in abnormal protein translation. The current standard of care for patients with DBA includes steroids, chronic red cell transfusions, or stem cell transplantation, but these are all associated with significant side effects including infections, iron overload, and the risk of graft versus host disease. Thus, the development of more effective and less-toxic therapies is needed to treat the anemia seen in patients with DBA. Our previous work demonstrated that Nemo-Like Kinase (NLK) is overly active in red cell precursors in models of DBA as well as in patient samples (Wilkes, et al., 2020). We hypothesize that NLK plays an important role in the pathogenesis of DBA, and is a potential target for DBA therapy. Human cord blood CD34+ hematopoietic stem and progenitor cells (HSPCs) were transduced with lentivirus co-expressing GFP and shRNA against RPS19/RPL11 or luciferase (control), and were differentiated in erythroid media in the presence of small molecule NLK inhibitors. After screening small molecule compounds that inhibit NLK as an off-target from previously approved or clinically advanced drugs, we found that a known MELK (maternal embryonic leucine zipper kinase) inhibitor, OTS167, improved erythropoiesis (CD71+ cell expansion) by 31% and 27% at a concentration of 50nM in RPS19- and RPL11-knockdown HSPCs, respectively. We also investigated the effect of OTS167 on the erythropoiesis in Rpl11-haploinsufficient mouse model of DBA ( Rpl11 wt/LoxpCre-ERT2 +, Morgado-Palacin, et al., 2015 and Rpl11 wt/LoxpMx-Cre + Liu,et al.,2023). Lineage-negative (Lin-) hematopoietic progenitors isolated from the mouse bone marrow were cultured in erythroid differentiation medium with OTS167. Our data showed that the erythroid progenitors (CD71+ cells) expanded from the cultured Lin- cells were increased by 20% (n=2, p=0.065) at an OTS167 concentration of 30nM. We confirmed that NLK activity was increased in erythroid progenitors from the Rpl11-haploinsufficient mice for in vitro phosphorylation of the substrate compared with tamoxifen-treated wild type mice. Moreover, we performed colony formation assay with OTS167 in the mouse bone marrow cells. OTS167 increased BFU-E colony numbers in a dose-dependent manner with the highest effect at a concentration of 50nM (n=3, p=0.003). OTS167 did not increased CFU-E colony number, suggesting that OTS167 affected the BFU-E stage and/or earlier stages of erythroid development. Also, OTS167 did not affected CFU-GM colony formation as myeloid lineage. The goal in treating DBA patients with NLK inhibitors is to sufficiently raise the hemoglobin to minimize the need for chronic red cell transfusions or treatment with steroids. Our in vitro data suggest that pharmacologic inhibition of NLK with OTS167 may achieve this goal. We have initiated studies to test the efficacy of OTS167 in vivo using Rpl11-haploinsufficient mouse models. Given that DBA is associated with a number of mutations in the ribosome, using one drug that targets NLK as a common therapeutic target could be a more practical treatment option for patients in contrast to gene therapy or gene editing followed by autologous stem cell rescue. Even if the drug targeting NLK does not completely rescue the anemia in ribosome-insufficient cell models of DBA, the improvement could be enough to help patients especially in combination with other drugs that are currently being used or studied .

A Preliminary Report of Clinical Study on Immunosuppressive Therapy Combined with Avatrombopag As the First-Line Treatment for Severe Aplastic Anaemia in Elderly Patients

Objective The objective is to evaluate the early hematologic response (HR) rate and safety of immunosuppressive therapy (IST) combined with avatrombopag as the first-line treatment for severe/very sever aplastic anaemia(V/SAA) in elderly patients. Methods Prospective，multi-center, single-arm, clinical research. IST treatment regimen: pig anti-human lymphocyte globulin (ALG-p, Wuhan Institute of Biological Products Co., Ltd. China) 20 mg/kg/d for five days and cyclosporin A (CsA) 3 mg/kg/d with adjusted concentration of 150-250 ng/mL, and oral avatrombopag 40 mg/d from the first day of IST initiation up to six months. The patients' treatment responses were evaluated and the major adverse events were collected. Results Ten patients have been enrolled in the study at our centre from June 2022 to October 2022.Among 10 patients, six had SAA and four had VSAA; male: female = 1:1, median age of 68 (60-82) years, and median time from diagnosis to treatment of 44 (20-480) days. The patients' treatment responses were evaluated after initiation of IST, at 3 months: 7 HR [1 complete response (CR), 3 good partial response (GPR), 3 partial response (PR), and 3 no response (NR)]; and at 6 months: 8 HR (1 CR, 5 GPR, 2 PR) and 2 deaths due to infection. The median time of red-cell transfusion independence was 60 (3-168) days in HR patients and that of platelets was 60 (16-153) days. No relapse and clonal evolution occurred within six months. Major adverse events within three months were infection in five cases, electrolyte disturbance in five cases, and cardiac insufficiency in three cases. Liver function abnormalities occurred in two cases, both of which showed elevated alanine aminotransferase up to 174 μ/L and 208.7 μ/L on day 8 and 20 of IST treatment, respectively. Two of the five infected patients had sepsis caused by Enterobacter cloacae. Except for infection, the remaining adverse events were of grade I-II. All the adverse events were effectively controlled. Table 1 summarises the characteristics and clinical status of patients before and after the treatment. Conclusion This preliminary study of IST combined with avatrombopag for the treatment of elderly patients with V/SAA showed the potential of the combination to improve haematologic response rate and quality in elderly patients with AA, and it is well tolerated as well. Keywords: immunosuppressive therapy, severe aplastic anaemia, avatrombopag, elderly

Efficacy of Luspatercept in Low-Risk Myelodysplastic Syndrome: A Systematic Review and Meta-Analysis

BACKGROUND Patients with anemia and lower-risk myelodysplastic syndromes (MDS) in whom erythropoiesis-stimulating agent therapy is not effective, inevitably become RBC-transfusion dependent, unless other lines of therapy are offered. Luspatercept, a recombinant fusion protein that binds transforming growth factor β superfamily ligands to reduce SMAD2 and SMAD3 signaling, has been FDA approved for use in MDS with ring sideroblasts (RS) and/or detection of an SF3B1 mutation. The efficacy of luspatercept in lower risk (LR) MDS (regardless of RS) patients has been evaluated in clinical and real-world (RWD) studies, suggesting conflicting data. OBJECTIVES This study aimed to conduct a systematic review and meta-analysis of clinical trials and RWD evidence to evaluate the efficacy of luspatercept among lower risk MDS patients. METHODS Prospective randomized controlled trials (RCTs) and phase II trials, retrospective cohort studies and case series of patients with MDS treated with single agent luspatercept were retrieved from PubMed, EMBASE and conference proceedings. References were screened until June 30 th 2023. Two reviewers appraised the quality of the studies and extracted data. Primary outcome was the rate of transfusion independence (for at least 8 weeks). Secondary outcomes included erythroid hematologic improvement (HI-e) rate and safety. Subgroup analysis was conducted for MDS-RS and SF3B1 mutation. Studies were critically appraised using Joanna-Briggs Institute checklists. All studies were included in the meta-analysis. RESULTS We screened 244 titles and eventually included 9 studies, conducted between 2013-2022, and including 839 MDS patients who were treated with luspatercept. Of them: 2 RCTs, one single arm phase 2, and 6 retrospective “real-world” (RWD) studies. Six were published in peer reviewed journals and three RWD studies published as abstracts. Median age of patients ranged between 68-75 years and 707/839 patients had MDS-RS. Three studies were of moderate-high quality (N= 389) and 6 were deemed moderate quality (N=450). Rates of transfusion independence for at least 8 weeks were highly variable ranged between 8-87% among all patients and between 42-87% among MDS-RS subgroup. The median duration of longest response ranged between 23.9-42 weeks. Rates of erythroid hematologic improvement (HI-e), ranged between 25-100%. Pooled estimate of achieving independence from red-cell transfusion for >= 8 weeks was 39.5% (95% CI 0.24, 0.55; I²=96.6%, p<0.001) by meta-analysis. Pooled estimate of HI-e was 54.4% (95% CI 0.44,0.65; I²=87.8%, p<0.001). The most common cause for drug discontinuation was lack or loss of response and disease progression. CONCLUSION The response rate to luspatercept is highly heterogenous among different studies. Overall, the response rate and duration in the RWD setting did not compare favorably with clinical trials. Of note, the vast majority of patients had MDS-RS and the role of luspatercept in the management of patients with MDS without RS is still unclear, even when data from several studies are pooled together. This study has many limitations arising from heterogeneity of patient characteristics and outcomes ascertainment. This highlights the need for standardized post-marketing studies of new drugs and the heterogeneity in RWD patient population.

Perioperative transfusion of red cell products in patients operated at general surgical units of National Hospital of Sri Lanka

Indications for red cell transfusion among general surgical patients have changed over time. Despite a scarcity in evidence, many international guidelines advocate a restrictive transfusion protocol.Objectives: To describe the red cell transfusion practices including transfusion thresholds in general surgical wards of a tertiary hospital in Sri Lanka.Methods: A descriptive cross-sectional study was done in all general surgical units of the National Hospital of Sri Lanka, Colombo. All patients transfused with red cell products for a 3-month period from 1/5/2017-1/8/2017 were included in the study. Indications for transfusion, haemoglobin at transfusion and the number of units transfused was the primary data recorded.Results: A total of 196 patients were included in the study. Most transfusions 79.08% (n=155) were for elective reasons. Most elective transfusions were to facilitate chronic extremity wound healing (52.2%, n=81). Most emergency transfusions were for intraoperative bleeding (31.7%, n=13). The majority (73.5%, n=114) of elective transfusions occurred at a Hb of 7-10 g/dL. The mean number of RBC units transfused in elective circumstances for a Hb of 7-10 g/dL was 1.59. A pre-transfusion blood picture was not available in over 85% (n=132) of elective transfusions. Only a single febrile non-haemolytic transfusion reaction was reported.Conclusions: The study revealed substantial divergence of red cell transfusion thresholds in perioperative practice when compared with international guidelines, nonadherence to rational practice such as obtaining a blood picture prior to elective transfusion and therefore the need to formulate local guidelines. It also highlighted the transfusion burden of diabetic foot disease.

Evaluation of GBT021601 As a Therapeutic Agent to Restore Bone Marrow Health and Effective Erythropoiesis in a Sickle Mouse Model

Introduction: Hypoxia-mediated damage resulting from vaso-occlusion in sickle cell disease (SCD) leads to abnormal angiogenesis, such as occurs with moyamoya disease and proliferative retinopathy. Abnormal angiogenesis in mouse models of SCD has been observed to impact the bone marrow niche, resulting in a structurally abnormal vascular network. A dysfunctional bone marrow environment results in ineffective erythropoiesis and can damage hematopoietic stem cells (HSCs). Healthy HSCs are essential to the maintenance of hemoglobin (Hb) levels regardless of the rapid turnover of red cells and because of their potential as material for gene therapy. Recently, red cell transfusions were shown to reverse pathogenic angiogenesis in a sickle cell mouse model. However, chronic red cell transfusions are difficult to sustain in individuals with SCD due to complications including iron overload, risk of alloimmunization, and the time and cost involved. GBT021601 is a potent sickle Hb polymerization inhibitor that improves the rheology of red blood cells (RBCs) and has been shown to increase total Hb by as much as 3 g/dL. Here, we explore the effectiveness of GBT021601 for restoring bone marrow health and decreasing proangiogenic factors while maintaining the stemness of marrow cells to preserve erythrocyte maturation. Understanding the impact of GBT021601 on aberrant angiogenesis and the function of HSCs will indicate if it has a role in improving the bone marrow niche and HSC fitness in an era of expanded cell-based therapies for SCD. Methods: Nine-week-old, humanized Townes HbSS SCD mice were split into 3 cohorts (n=8 per group: 4 male, 4 female) and for 12 weeks were fed either standard chow (control) or chow containing 0.1% or 0.2% of GBT021601. After 12 weeks, the mice were phlebotomized and euthanized. Spleens were collected and weighed. Whole bone marrow was collected via centrifugation from both tibias and one femur; the other femur was saved for subsequent 3D confocal microscopy. Bone marrow was analyzed using flow cytometry to measure erythroid cell maturation (Annexin V, TER119, and CD44) and HSC markers (Lin, Sca-1, c-kit). Immunoblot analysis was performed on peripheral plasma to measure hypoxia-induced signaling and markers of angiogenesis (VCAM-1, VEGF-A, ANG-1, and -2). Results: Hb increased by 3 g/dL on average with administration of GBT021601, and spleen weights were significantly reduced ( P=0.0023 and P=0.0015 for control vs 0.1% and 0.2%, respectively). Flow cytometry of the extracted bone marrow revealed a significant increase in mature RBCs and a decrease in erythroid progenitors at all stages ( P<0.001 for RBCs and basophils; P=0.04, P=0.002, and P=0.001 for pro-, poly-, and ortho-chromatic erythroblasts, respectively) (Figure 1). HSCs, defined as Lin-Sca-1 +c-kit +, were not significantly different between groups (control vs 0.1%, control vs 0.2%, and for 0.1% vs 0.2%). Immunoblot analysis revealed a reduction in VCAM-1 and ANG-1 markers with administration of GBT021601 relative to control. However, no difference between ANG-2 and VEGF-A markers was observed between treated and control mice (Figure 2). Conclusions: Treatment with GBT021601 was effective at reducing extramedullary hematopoiesis as determined by the reduction in spleen size compared with control mice. VCAM-1 and ANG-1 are increased in HbSS compared with HbAA mice; treatment with GBT021601 reduced factors that contribute to pathogenic angiogenesis. Treatment with GBT021601 did not change ANG-2 and VEGF-A expression; notably, VEGF-A levels are similar between HbSS and HbAA mice. Our findings support GBT021601 as a sustainable treatment to reduce ineffective erythropoiesis and abnormal angiogenesis-which can otherwise damage the bone marrow niche-thereby suggesting a potential benefit for individuals with SCD participating in future cell-based therapies with curative intent and improved outcomes. More work is planned to examine the ability of GBT021601 to normalize the bone marrow environment and reduce the risk of deleterious effects on allogenic or autologous edited HSCs.

Fucosyltransferase VII (FUT7) Activity in Patients with Sickle Cell Disease and Healthy Controls

Background:Sickle cell disease (SCD) is a severe monogenic disorder with a majority of the affected individuals living in Africa. Vaso-occlusion is a significant cause of morbidity and mortality in SCD, and selectin mediated intercellular adhesion is recognized as an important target for therapy. As such Anti-P-selectin agents are in clinical use to reduce the frequency of vaso-occlusive crises (VOCs) in SCD patients. Sialyl lewis x is a crucial ligand common to all members of the selectin family and the final step in the biosynthesis of sialyl lewis X antigen (fucosylation), is catalysed by Fucosyltransferase VII (FUT7). This enzyme is most expressed in leukocytes and endothelial cells. We hypothesized that FUT7 enzyme activity is abnormal in SCD patients and differs by VOC severity. We sought to measure plasma levels of FUT7 in SCD patients and healthy controls, and investigate their association with clinical and laboratory features, such as the frequency of VOCs. Methods:This wasa single centre cross-sectional analytical study at the University of Nigeria Teaching Hospital, Enugu. 38 steady state adult SCD patients and 40 HbAA age- and sex-matched healthy control subjects were enrolled. SCD patients were stratified by clinical severity into: Group A- 19 subjects with mild disease (0-1 crises and no disease complication such as stroke, chronic leg ulcers, nephropathy, or avascular necrosis in the previous year), and Group B- 19 patients with moderate to severe disease (2 or more crises in the previous year, with or without complications). We excluded patients with red cell transfusion within 4 months, on hydroxyurea, or other agents that may affect vaso-occlusive crisis frequency, and co-existing chronic disorders unrelated to SCD, such as asthma and diabetes mellitus. We performed a CBC and plasma FUT7 enzyme concentration measurements (ELISA kit, MyBiosource Inc, USA). Statistical analysis was performed with SPSS version 22.0 for Windows. A 2-tailed P value of <0.05 was used. Results:Our subjects were predominantly female (SCD 52.5%, HbAA 55%), with median age 28 years (range 19-46 and 19-45 for SCD and HbAA, respectively). Median frequency of VOCs in patients was 1 (range 0-5). Avascular necrosis (32.5%) was the most common complication, while stroke, acute chest syndrome, and eye disease were the least common. As expected, SCD patients had lower haemoglobin levels than HbAA (7.9±1.3g/dL vs. 12.6±1.4g/dL, P<0.001). Significantly higher WBC, ANC, and platelet counts were observed in SCD patients compared with HbAA controls (P<0.001). No significant correlations were found between FUT7 and haematocrit (r =0.06), WBC count (r = -0.08), neutrophil count (r = -0.12), and platelet count (r =0.1), with P values of 0.7, 0.6, 0.5, and 0.6, respectively. We found no difference between the median and IQR values of plasma FUT7 of SCD (2.1; 1.8-2.5) and controls (2.2; 1.8-2.8), P = 0.7. FUT7 activity was higher in SCD subjects in Group A (2.2; 1.9-2.9) than in Group B (2.0; 1.2- 2.4), with a trend towards significance P = 0.1. Discussion:In this study, FUT7 activity did not differ between SCD patients and controls. We also did not find correlations between FUT7 and blood cell counts, suggesting that plasma FUT7 activity is not increased at baseline in SCD and that the degrees of anemia or leukocytosis are not associated with changes in FUT7. Nonetheless, a mild decrease in FUT7 levels was found in more severe SCD patients and should be confirmed in a larger study. Our study did not evaluate patients in crises or who received hydroxyurea and/or other SCD-modifying therapies. Since FUT7 catalyses selectin expression, variation in its activity may be more relevant in subgroups of SCD patients on anti-selectin therapies. Future studies should be carried out in such patients to explore FUT7 activity as a marker of acute vaso-occlusion and as a modifier of response to therapy.

Autologous STEM Cell Collection after Daratumumab, Bortezomib, Thalidomide and Dexamethasone Versus Bortezomib, Thalidomide and Dexamethasone in NEWLY Diagnosed Multiple Myeloma: A Real-Life Monocentric Italian Experience

Introduction In newly diagnosed multiple myeloma (NDMM) patients (pts), induction followed by high-dose chemotherapy and autologous stem cell transplantation (ASCT) remains the standard of care. The addition of an anti-CD38 antibody, daratumumab, to an established frontline protocol based on bortezomib, thalidomide and dexamethasone (D-VTd), improves efficacy but could hamper peripheral blood stem cell collection (PBCS). Methods The aim of our study is to evaluate the possible impact of the addition of daratumumab to induction therapy prior to ASCT on PBCS and hospitalization parameters a real-world population with NDMM. We hereby, retrospectively analyzed 28 NDMM pts who received 6 cycles of D-VTd as induction therapy before ASCT and compared them with a consecutive series of 32 VTD-treated NDMM pts who were matched for cytogenetic abnormalities and age, managed at the Hematology Department of Sapienza University in Rome. Results Patient baseline characteristics were equally distributed among both groups, as summarized in Table 1. After induction therapy, a combination of cyclophosphamide (CTX) with granulocyte colony stimulating factor (G-CSF) at standard dose of 10 mg/sqm/day was used as the first mobilizing regimen attempt in both groups. The median time between the last day of induction therapy and the first day of mobilization therapy was 30 days for D-VTd groups and 22 days for VTD groups (p=0.041). Overall, 94% of pts treated with VTd met the desired collection goal after first mobilization therapy, compared to 68% of pts treated with D-VTd (p=0.01). In the VTd arm, 1 pts required a second mobilization attempt, and 1 pts required a further one. In the D-VTd arm, 5 pts required a second mobilization attempt, and 4 pts also required a further one. Overall, 3 pts in the D-VTd arm did not meet the collection goal. In pts with impaired mobilization, plerixafor was used in 4/32 pts in the VTd arm (13%), compared to 29% of pts treated with D-VTd (p=0.195). In the D-VTd arm, 12/28 pts (43%) had received a higher dose of CTX as mobilization therapy (3 g/m 2) compared to the standard dose of 2.4 g/m 2 in 16/28 pts (57%). Nevertheless, no difference was observed in the rate of PBSC collection at the first attempt (p=0.7). No baseline clinical or disease characteristics were observed to have negative impact on the PBSC at first mobilization attempt, including treatment response at the end of induction therapy. Nevertheless, a significantly lower pre-mobilization level of platelet was detected in the group of D-VTd treated pts (173.000/mm 3 vs 210.000/mm 3, p=0.012) that did not meet the desired collection goal after first mobilization therapy. The median PBSC collection was 9.1x10 6 CD34+ cells/Kg in the whole cohort, with no significant difference in the two groups. A lower pre-mobilization level of neutrophils (p=0.028) and platelets (p=0.004) was observed in the D-VTd arm compared to the VTd arm, possibly reflecting prolonged daratumumab-related hematological toxicity. No significant difference was observed in the leucocyte (WBC) azimuth value in the two arms. However, the two groups reached this value on two different days. In the VTd arm the median day was 8, and in the D-VTd arm it was 11 (p=0.034). The use of G-CSF for more days did not result in an increase in CD34+ and the median azimuth value of CD34 was significantly lower in the D-VTd group than in the VTD group (p<0.001). The median collection volume from apheresis devices was significantly higher in the D-VTd group (372 ml) compared to VTd arm (301 ml) (p=0.05), possibly related to a higher WBC azimuth in D-VTd arm, resulting in a prolonged apheresis procedure. Pts receiving D-VTd had slightly delayed hematopoietic neutrophil (11 vs 14 days, p=0.030) and platelet (15 vs 14 days, p=0.018) engraftment, but no requiring a higher number of red cell (p=0.6) and platelet (p=0.3) transfusions and a longer hospitalization (20 vs 17 days, p=0.2) after the first ASCT, compared to pts treated with VTd. No difference in the two groups regarding rate of febrile neutropenia (65% vs 45%, p=0.2) and use of antibiotics (53% vs 57%, p=0.8) were observed. Conclusions In conclusion, frontline induction daratumumab-based therapy is likely to affect PBSC collection after first mobilization attempt, even if not clinically relevant, and potentially mitigated with higher use of plerixafor. Furthermore, the inclusion of daratumumab in pretransplant therapy regimens did not impair ASCT.

Trial in Progress: Venetoclax in Combination with ASTX727, an All-Oral Therapy for Chronic Myelomonocytic Leukemia and Other MDS/MPN with Excess Blasts (VICTORY MDS/MPN): A Randomized, Phase 2 Trial

Background and Significance: The rare nature and diagnostic difficulty associated with myelodysplastic/myeloproliferative neoplasms (MDS/MPN) has limited the inclusion of patients (pts) with these diseases, other than chronic myelomonocytic leukemia (CMML), in relevant clinical trials. This has resulted in the complete lack of approved agents for the treatment of pts with non-CMML MDS/MPN. Moreover, ASTX727 (oral decitabine-cedazuridine) was approved for CMML based on a study of only 16 pts with CMML. A larger study of CMML and non-CMML MDS/MPN is needed. MDS/MPNs appear to be BCL2-dependent (Bogenberger et al., 2015; Jilg et al., 2016), but data for venetoclax combination therapy are derived from retrospective analyses of relapsed/refractory disease (Cortes et al. 2019; Hammond et al., 2020) or single-arm studies not dedicated to MDS/MPN (NCT04655755, Venugopal et al. 2021). Venetoclax exposures are not affected by coadministration of ASTX727 and predict responses similar to historical data (Mannis et al. 2021). Based on these promising early data and the critical need, we proposed a randomized study of ASTX727 alone or in combination with venetoclax for pts with MDS/MPN and >5% blasts. Study Design and Methods: This is a National Cancer Institute-sponsored, multi-center, international, randomized phase 2 study (NCT05600894, NCI 10538) designed to evaluate the efficacy of ASTX727 alone or in combination with venetoclax in pts with newly diagnosed MDS/MPN with >5% marrow blasts. Key eligibility criteria include allowing hydroxyurea cytoreduction to a WBC <10,000/mm 3 for proliferative disease and excluding pts with need for emergent disease-directed therapy as well as prior therapy for myeloid malignancy other than hydroxyurea or thrombopoietic/erythropoietin stimulating agents. Pts will be randomized 1:1 to receive either ASTX727 alone (Days 1-5) or in combination with venetoclax, initially administered with a 3-day dosing ramp-up and given for 14 days per 28-day cycle (Figure 1). Pts are stratified by disease subtype, specifically CMML and non-CMML MDS/MPN. The primary endpoint is the rate of complete remission (adapted from the MDS/MPN-specific response criteria developed by Savona et al. 2015). Secondary endpoints include the overall response rate, overall survival, progression-free survival, allogeneic hematopoietic stem cell transplantation rate, clonality at time of hematologic remission, number of red cell and platelet transfusions required and toxicity of ASTX727 versus ASTX727 + venetoclax. Pts randomized to the ASTX727 monotherapy arm and without response after 4 cycles of therapy, or with progressive disease after an initial response, cross over to the ASTX727 + venetoclax arm. A futility analysis is planned for each cohort (first 18 and 13 pts in each treatment arm for the CMML and non-CMML cohorts, respectively). We assume that 85% and 90% of pts in the CMML and non-CMML MDS/MPN cohorts, respectively, randomized to the ASTX727 monotherapy arm will not achieve the primary endpoint and will cross over to the ASTX727 + venetoclax combination arm. The estimated accrual time of a total of 70 pts with CMML and 50 pts with non-CMML MDS/MPN will require about 2.9 and 2.1 years, respectively. Planned integrated studies include the use of an apoptosis multiplex immunoassay to evaluate how the addition of a BCL-2 inhibitor will influence the ability of ASTX727 to induce apoptosis and if particular thresholds of apoptosis are predictive of clinical response. Planned exploratory studies include the use of: 1) RNA-seq to evaluate if differences in baseline transcriptional programs appear to be predictive of response to ASTX727 and ASTX727+venetoclax, 2) next generation (Oncomine Comprehensive Assay (OCAv3)/whole exome sequencing to uncover biomarkers predictive of response/outcome, validate risk modeling to stratify these pts and detect/monitor measurable residual disease (MRD) on serial marrow samples that can predict depth of response with translation in improvement in event-based endpoints, and 3) circulating tumor DNA as a non-invasive means (i.e., without need for a marrow evaluation) of prospective MRD detection for prognostic use. This ongoing, randomized study may inform frontline therapeutic options, provide prospective data for venetoclax combination after HMA failure, and generate striking correlative science for pts with “high risk” MDS/MPN.

Survival and Causes of Death in Patients with Alpha and Beta-Thalassemia

Background: Thalassemia is the most common hereditary anemia worldwide. The prevalence is high in the malaria-endemic areas, including Southeast Asia, and Thailand. Severe thalassemia can result in shortened survival due to disease-related complications. In the last three decades, with regular red cell transfusions and iron chelators, the survivals of patients with thalassemia, especially for TDT, have significantly improved. The most distinctive change was the decrease in the iron-overloaded cardiomyopathy. Both alpha and beta-thalassemia are prevalent in Thailand. We aimed to study the survival, and the causes of death in the patients with thalassemia across the disease severity from NTDT to TDT, and to explore the factors influencing the survival. Methods: A retrospective review of patients with thalassemia who were followed up at Chiang Mai University Hospital was conducted. The survival status was determined from medical records and the Thai National database (Official statistics registration systems). The survival and clinical factors associated with survival were analyzed. Results: Our study cohort comprised 789 patients with thalassemia, who were registered in the Departments of Internal Medicine and Pediatrics. The median age (IQR) at the end of follow-up was 18∙8 (11∙7) years. There were 301 patients (38∙1%) with Hb H disease, 279 patients with Hb E/beta-thalassemia (35∙4%), and 209 patients (26∙5%) with beta-thalassemia major, respectively. Half of the patients (395, 50∙1%) had transfusion-dependent thalassemia (TDT). Sixty-five (8∙2%) patients had died. The mean age at death was 17∙0 ± 0∙9 years. Multivariable analysis showed that TDT (adjusted HR 3.68, 95%CI 1.39-9.72, p 0∙008) and mean serum ferritin level ≥3,000 ng/mL (adjusted HR 4∙18, 95%CI 2.20-7.92, p<0∙001) were independently associated with poorer survival. The two major cause of death were infection-related causes (36∙9%), cardiac complications (27∙7%), thalassemia/anemia (13∙8%), trauma (6∙2%), and other causes (15∙4%). Conclusions: TDT and mean serum ferritin level ≥3,000 ng/mL were the main factors associated with poorer survival in thalassemia patients. Infections and cardiac complications were the two major causes of death.

Use of Conventional Transfusion Thresholds Contributes to Ongoing Iron Overload Issues in Patients with Homozygous (SS) Sickle Cell Disease

Introduction: Homozygous sickle cell disease is characterized by chronic anemia which contributes to a clinical course characterized by recurrent pain crisis episodes. These episodes are usually managed with the use of packed red blood cell products. Randomized data is currently available that supports lower transfusion thresholds in non-sickle cell related conditions including but not limited to patients undergoing orthopedic procedures, cardio-vascular interventions and those needing intensive care. Due to lack of similar randomized trials in patients with sickle cell disease, recommendations from other trials are applied to patients with sickle cell diseases as well. However, this has the potential of worsening iron overload states that patients with sickle cell disease usually suffer from. Methods: Patients with homozygous SS sickle cell disease were evaluated with regarding to transfusion support given during hospitalization for vaso-occlusive pain crisis episodes. The hemoglobin threshold that was used for deciding whether or not to give packed red cell transfusion was studied in patients who were previously getting periodic red blood cell exchange procedures for chronic sickle cell related pain. Serum ferritin levels during the period of exchange transfusion support was compared with levels during simple transfusions. 8 patients were selected from the cohort of patients with homozygous sickle cell disease who had been receiving elective packed red blood cell exchanges. Patients with ferritin levels persistently remaining above 1000 ng/ml were then evaluated with regard to transfusion thresholds used for simple transfusion support during hospitalization for pain crisis episodes. Results: Six patients had persistently high ferritin levels. Thresholds for simple transfusion in this group were studied. A total of 30 units of packed red blood cells were found to be transfused over a 6-month period. During this period only one patient required an exchange transfusion session. 2 out of 30 units were given for a Hgb level above 7 g/dl. The majority, representing 24 out of the total 30 pack red blood cell units were given above a Hgb threshold of 6.0 g/dl. Only 4 units were given using a Hgb threshold less than 6 g/dl. Mean ferritin level before and after the period of simple transfusion support was 6097 ng/ml and 6076 ng/ml respectively. Mean Hgb level in while on exchange transfusion was 7.4 g/dl and it was 6.9 g/dl in those receiving a simple transfusion protocol. Conclusions: Transfusion threshold used in patients with homozygous SS sickle cell disease appears to be similar to patients without sickle cell disease. Current practice based on studies including TRICC and TRISS is to keep the Hgb level above 7 g/dl. This has been shown to be safe and in fact even improves patient outcomes. However, in patients with sickle cell disease this may not be appropriate since their baseline Hgb levels are lower to begin with. Also, transfusions don't appear to improve Hgb levels considerably. These patients also tend to have iron overload at baseline which likely will not improve unless all transfusions are limited. Therefore, applying a lower transfusion threshold would be recommended to avoid acute transfusion related issues and long-term iron overload problems in patients with SS disease.

Successful Implementation Trial to Improve Stroke Risk (TCD) Screening in Children with Sickle Cell Anemia: A Roadmap to Care Enhancement

Stroke remains a devastating complication of sickle cell anemia (SCA). The STOP (Stroke Prevention Trial in Sickle Cell Anemia) protocol adopted in 2014 by the National Heart Lung and Blood Institute (NHLBI) guidelines includes stroke-risk screening using transcranial Doppler ultrasound (TCD) and prevention with chronic red cell transfusion therapy (CRCT). Theserecommendations were reinforced by the American Society of Hematology guidelines.However, despite the evidence supporting TCD screening in stroke prevention, the DISPLACE study (Dissemination and Implementation of Stroke Prevention Looking at the Care Environment) showed that real-world implementation was insufficient. To evaluate and improve sickle stroke screening, we conducted the NHLBI-funded multi-center DISPLACE study to identify barriers to TCD implementation and test novel methods for improving outcomes. DISPLACE is a 4-part study: 1) Retrospective calculation of TCD screening rates; 2) Assessment of barriers and facilitators; 3) Implementation trial to improve TCD screening and 4) Comparison of implementation sites to controls. Importantly, DISPLACE is the largest prospective study of children with SCA. This study employed a multi-level, multi-faceted cluster-randomized study of interventions to improve implementation rates of TCD. Methods: DISPLACE is a 28-center US consortium representing a range of: regions (urban vs rural), size (patient number), and institution type (academic vs community). In Part 1, sites performed a rigorous retrospective review of TCD screening from 2012-2016 to collect baseline rates. Part 2 included a barriers and facilitators assessment to TCD screening to inform the trial interventions: 1) Re-branding TCD as Sickle Stroke Screen, a meaningful name for SCD stakeholders; 2) Using a Single Coordinator (SC) for each site's TCD scheduling, and follow-up; 3) A custom designed provider-focused application, ProviderMinder TM (PM) created to remind providers to track TCD appointments and follow-up on results. When a TCD was due, PM alerted the provider to enter scan results or reason(s) missed. Once the TCD results (normal/conditional/abnormal) were entered in PM, providers were prompted to enter the next TCD date and/or intervention plan (if not normal). If a result was not entered, PM alerted the provider q3 days until the TCD was recorded as completed or rescheduled. The implementation trial (parts 3 and 4 of DISPLACE) included 16 sites that had the lowest TCD rates in part 1. The implementation trial included children with SCA aged 2-7 years (the group at highest risk for an abnormal TCD). Cluster randomization was used to allocate sites to PM alone (8 sites) or PM + SC (8 sites). The primary outcome was comparison of TCD rates between PM and PM+SC sites. Secondary outcomes included comparisons between 1) Part 3 sites' TCD rates and Part 1 baseline rates 2) Part 3 sites and control sites (to assess the impact of the educational rebranding of TCD). Results: Part 1 included 28 centers with 5246 children with SCA. Of the 16 intervention sites, 3 sites could not implement the interventions due to administrative issues but remained in the study and were re-designated as “control sites” and a 4th control site was added (they were not informed prospectively to avoid bias). Thus, there were 13 evaluable sites for the primary endpoint including 776 patients; 484 in sites using PM and 292 in sites using PM+SC. Control sites included 185 patients. The trial demonstrated superiority of the PM arm compared to the PM + SC arm (P<.0001 year 1 and p=0.002 year 2) in both years of implementation and in comparison, to the control sites (P<.0001 year 1, p=.009 year 2) with an overall improvement in mean TCD screening from part 1 by 25% (table 1). Conclusions: This was a highly successful implementation study in SCA. This study emphasizes the importance of performing barrier and facilitator assessments when introducing new guideline-recommended screenings into clinical practice. These findings also highlight the importance of ongoing quality assurance in SCA to ensure prevention practices are maintained. Despite the benefit of TCD (sickle stroke screening) in children with SCA, implementation was poor prior to these interventions. Results from this study will guide ongoing optimization of TCD screening and demonstrate the importance of adopting these interventions in the real world.

Adults with Transfusion-Dependent Thalassemia Have Variable Clinical Profiles Depending on Where They Receive Care: Thalassemia Treatment Center Vs Community

Introduction: Advances in the treatment of transfusion-dependent thalassemia (TDT) have decreased complications and improved survival. Established TDT guidelines have improved outcomes through regular red cell transfusions to maintain a pre-transfusion hemoglobin > 9-9.5 g/dL and chelation to maintain a serum ferritin < 1500 ng/mL, liver iron content < 5 mg/g dry weight and a cardiac T2*> 20 ms. Increased lifespan of individuals with TDT now necessitates adult hematology providers with capacity to manage uncomplicated thalassemia. While it is recommended that thalassemia care be provided through a multidisciplinary thalassemia treatment center (TTC), there are a limited number of TTCs in the United States. As a result many adults with TDT receive care in the community from hematologists with variable knowledge and experience with thalassemia. In this study, we thus sought to evaluate the prior care of patients referred to an adult TTC in order to determine if there was a significant difference in the care provided by a pediatric TTC compared to community hematologists. Methods: We evaluated adult patients (> 18 years old) with a diagnosis of TDT referred to the Penn Comprehensive Adult Thalassemia Program. Patients were either referred from the Children's Hospital of Philadelphia TTC via a dedicated transition program, or from community hematologists (Comm). Data collection was performed via manual chart review. Data collected included the mean of three consecutive pre-transfusion hemoglobin levels prior to the initial visit, serum ferritin, and most recent liver and cardiac iron quantitation by MRI. Results: Of the 94 patients with thalassemia referred to the adult program between 2013 and 2023, 54 had a diagnosis of TDT. Twenty-seven patients were referred each from the TTC and the community. At the initial adult thalassemia center visit, the median age was 38.5 years (TTC) and 28 years (Comm). More patients in the TTC group maintained a pre-transfusion Hgb > 9 g/dL (78% vs 52%). Serum ferritin trended lower in the TTC group (1725 vs 2629 ng/ml, p-value = 0.056). While 67% of the TTC group had a serum ferritin < 1500 ug/mL compared to 41% in the Comm group, there was no significant difference in the liver iron content between both groups (TTC 10.2 vs Comm 12.2 mg/g dry weight). However, 60% of the TTC group compared to 48% in the Comm group had a LIC < 5. Those followed at the TTC possibly have more cardiac iron with 4 vs 1 patient in the TTC vs Comm group having a T2* < 20 ms, likely reflective of more complicated patients being referred to the TTC. Conclusion : Patients managed by a TTC were more likely to be adequately transfused with serum ferritins < 1500 ng/mL. Maintaining a pre-transfusion hemoglobin > 9-9.5 g/dL and a serum ferritin < 1500 ng/mL are known to reduce long-term complications of chronic anemia, ineffective erythropoiesis and iron overload in thalassemia. Thus in order to improve long-term outcomes in patients with TDT, we propose that all adult hematology providers, regardless of care setting should aim to gain knowledge on the management of individuals with thalassemia and establish collaborative care relationships with experts in thalassemia care.

Successful Use of Haploid Hematopoietic Stem Cell Transplantation for Severe Glanzmann's Thrombasthenia

Objective: Glanzmann's thrombasthenia (GT) is a rare autosomal recessive disorder, which characterized by the clinical manifestations from purpura to severe bleeding. It was caused by mutations of the ITGA2B or ITGB3 gene on chromosome 17, which lead to deficits or reduced expression of the platelet membrane glycoprotein GPIIb/IIIa. We report 5 pediatric patients with severe Glanzmann's thrombasthenia who underwent allogeneic hematopoietic stem cell transplant, to observe the outcomes of allo-HSCT in children with severe Glanzmann's thrombasthenia. Methods: We retrospectively summarized the clinical and genetic profiles of five patients with Glanzmann's thrombasthenia who underwent allo-HSCT from June 2021 to July 2023. All patients were reduced intensity conditioning (Bufulsan 3.2-6.4mg/kg, CTX 120mg/kg, FLU 180mg/m 2, ATG 10mg/kg). After conditioning chemotherapy, all the patients received stem cells collected via bone marrow harvest and/or peripheral blood apheresis (PBSC) with CD34 dose of 2.89-7.2×10 6/kg and received graft vs. host disease (GvHD) prophylaxis with cyclosporine (CSA) and methotrexate (MTX). Results: There were five children were involved in this observation, one patient suffered from recurrent epistaxis bleeding which was life-threatening; the second suffered from gastrointestinal bleeding and could only accept liquid diet to 10-years-old; other patients also suffered from epistaxis bleeding, gum bleeding or gastrointestinal bleeding. For all the patients, regular red cell transfusions were required, platelet transfusions and recombinant factor VIIa were also used in severe and life-threatening bleeding events. Two of the five developed antiplatelet antibodies before HSCT. All the patients were successfully treated with allo-HSCT. One case received non-relative matched donor of 10/10 HLA, one case with sibling matched donor, and 3 cases with haploid transplantation of HLA 5-6/10 matched. The mean time to neutrophil engraftment was 11 days (11-12 days), and the mean time to platelet engraftment was 16 days (15-18 days). One case had CMV reactivation, one case had HHV-6b infection, one case had grade II acute GVHD of the skin which was controlled rapidly. There was no evidence of chronic GVHD for all the patients. The platelet disorder was corrected to a normal level after allo-HSCT, with no significant complication related to the transplantation. Comparing with HLA matched donor, either from sibling or non-related, the patients receiving haploid transplantation with 5-6/10 HLA matched got similar outcome. Conclusions: We proposed haploid transplantation treated with the reduced intensity conditioning regimens produced similar outcomes as HLA matched did which could be recommended as an alternative donor for severe Glanzmann's thrombasthenia.

The impact of anemia and blood transfusion on mortality after open abdominal surgery in the elderly

BackgroundMajor abdominal surgery is associated with considerable mortality in the elderly. Anemia has been linked to increased mortality in other types of surgery, such as hip and cardiac surgery. This study aimed to assess the impact of preoperative anemia on mortality in the elderly undergoing major abdominal surgery, and how allogeneic red cell blood transfusion influences mortality in these patients.Materials and methodsWe conducted a single-center, register-based retrospective study on patients, who were aged beyond 60 years and underwent one of 81 open abdominal surgical procedures. Patients operated on during the period from January 1, 2000, to May 31, 2013, were consecutively identified in the Danish National Patient Registry. Plasma hemoglobin was measured within 30 days prior to surgery and the primary endpoint was 30-day postoperative mortality. Information about patient transfusions from the hospital blood bank was available from 1998 to 2010.ResultsA total of 3199 patients were included of whom 85% underwent emergency surgery. The total mortality after 30 days was 20%. The median preoperative hemoglobin value of survivors was 7.7 mmol/L vs 6.9 mmol/L in those who died. The difference in hemoglobin values, between those who survived or died, decreased from the pre- to the post-operative phase. The 30-day postoperative mortality was 28%, 20%, and 12% in patients with a preoperative hemoglobin level in the lower, median, and upper quartile respectively. Transfusion therapy was associated with higher postoperative mortality, except in patients with very low hemoglobin values.ConclusionPreoperative anemia has a clear association with surgically related mortality. The distribution of hemoglobin values in patients with a fatal outcome differs significantly from that of survivors. Red cell transfusion is associated with increased mortality, except in patients with very low hemoglobin values which supports recent guidelines suggesting a restrictive transfusion strategy.

Diagnosis, management and prevention of Pediatric Acute Hemolytic Anemia: Egyptian adapted evidence-based clinical practice guidelines

BackgroundAcute hemolytic anemia (AHA) is a common emergency in pediatric emergency departments, hence the need to adapt pre-existing high-quality practice guidelines for the diagnosis, management, and prevention of AHA to be available for national use.MethodsThe guideline panel used the adapted ADAPTE methodology. The panel prioritized the health questions and recommendations according to their importance for clinicians and patients. The procedure included searching for existing guidelines, quality appraisal, and adaptation of the recommendations to be used in low-resource countries.ResultsThe guideline provided approach to a child with AHA: laboratory diagnosis of glucose-6-phosphate dehydrogenase (G6PD) deficiency, autoimmune hemolytic anemia (AIHA), and hemolytic uremic syndrome (HUS); treatment of AHA including indications for red cell transfusion, medical treatment, plasma exchange, and indications of antibiotic in HUS; how to avoid further episodes of hemolysis; and when to refer to a hematologist. Implementation tools included a checklist for history and examination, lists of differential diagnoses, flow charts for the diagnosis of AHA, and a list of medications and food to be avoided in patients with G6PD deficiency.ConclusionThis adapted guideline will aid decision-making related to the diagnosis, management, and prevention of AHA.

Diversifying cardiac intensive care unit models: Successful example of an operating surgeon-led unit

ObjectiveThe intensivist-led cardiovascular intensive care unit model is the standard of care in cardiac surgery. This study examines whether a cardiovascular intensive care unit model that uses operating cardiac surgeons, cardiothoracic surgery residents, and advanced practice providers is associated with comparable outcomes. MethodsThis is a single-institution review of the first 400 cardiac surgery patients admitted to an operating surgeon-led cardiovascular intensive care unit from 2020 to 2022. Inclusion criteria are elective status and operations managed by both cardiovascular intensive care unit models (aortic operations, valve operations, coronary operations, septal myectomy). Patients from the surgeon-led cardiovascular intensive care unit were exact matched by operation type and 1:1 propensity score matched with controls from the traditional cardiovascular intensive care unit using a logistic regression model that included age, sex, preoperative mortality risk, incision type, and use of cardiopulmonary bypass and circulatory arrest. Primary outcome was total postoperative length of stay. Secondary outcomes included postoperative intensive care unit length of stay, 30-day mortality, 30-day Society of Thoracic Surgeons–defined morbidity (permanent stroke, renal failure, cardiac reoperation, prolonged intubation, deep sternal infection), packed red cell transfusions, and vasopressor use. Outcomes between the 2 groups were compared using chi-square, Fisher exact test, or 2-sample t test as appropriate. ResultsA total of 400 patients from the surgeon-led cardiovascular intensive care unit (mean age 61.2 ± 12.8 years, 131 female patients [33%], 346 patients [86.5%] with European System for Cardiac Operative Risk Evaluation II <2%) and their matched controls were included. The most common operations across both units were coronary artery bypass grafting (n = 318, 39.8%) and mitral valve repair or replacement (n = 238, 29.8%). Approximately half of the operations were performed via sternotomy (n = 462, 57.8%). There were 3 (0.2%) in-hospital deaths, and 47 patients (5.9%) had a 30-day complication. The total length of stay was significantly shorter for the surgeon-led cardiovascular intensive care unit patients (6.3 vs 7.0 days, P = .028), and intensive care unit length of stay trended in the same direction (2.5 vs 2.9 days, P = .16). Intensive care unit readmission rates, 30-day mortality, and 30-day morbidity were not significantly different between cardiovascular intensive care unit models. The surgeon-led cardiovascular intensive care unit was associated with fewer postoperative red blood cell transfusions in the cardiovascular intensive care unit (P = .002) and decreased vasopressor use (P = .001). ConclusionsIn its first 2 years, the surgeon-led cardiovascular intensive care unit demonstrated comparable outcomes to the traditional cardiovascular intensive care unit with significant improvements in total length of stay, postoperative transfusions in the cardiovascular intensive care unit, and vasopressor use. This early success exemplifies how an operating surgeon-led cardiovascular intensive care unit can provide similar outcomes to the standard-of-care model for patients undergoing elective cardiac surgery.