Pregnancy in Sickle Cell Disease (SCD), a severe hereditary genetic condition, highly prevalent in Sub-Saharan African countries, is associated with increased risk of complications and severe outcomes in pregnancy, like intrauterine growth restriction, low birth weight, premature birth, miscarriage, stillbirth, pre-eclampsia, and maternal mortality. Several factors have been identified as associated with the heterogeneity of SCD phenotypes, namely the hemoglobin subunit beta (HBB) haplotype and -3.7 kb α-thalassemia deletion. OBJECTIVE:This study aimed to identify pregnancy complications and severe outcomes, and their association with genetic variability in women with SCD. METHODS:In a cohort of 162 pregnant women followed at Maternidade Lucrécia Paim, Luanda, Angola, we collected clinical, hematological, biochemical, and genetic data (Sickle Cell Disease genotype, HBB haplotype, and -3.7 kb α-thalassemia). FINDINGS:The Central African Republic (CAR) haplotype was the most prevalent, being 87% of women homozygous. For the -3.7 kb α-gene deletion 11.7% of women were homozygous, and 36.4% were heterozygous. In this cohort, CAR/CAR women had over 9 times higher odds of having a premature birth, and homozygous women for the -3.7 kb α-thalassemia had over four times higher odds of having a livebirth than the other genotypes. Over 50% of babies were born with low birth weight, and 52,7% were considered premature. Severe maternal complications were registered in 68% of current pregnancies. CONCLUSION:These findings highlight the high burden of adverse outcomes in SCD pregnancy and the need for individualized and closer healthcare, especially in low and middle-income countries.
Background Sepsis mortality stems from infection-triggered immune dysregulation, causing multiple organ dysfunction. B cells, key to adaptive immunity, protect against infection and regulate inflammation through antibodies, cytokines, and cell signaling. Their dysfunction is linked to sepsis outcomes, but their dynamic differentiation and underlying mechanisms remain poorly understood. Methods Integrated single-cell B-cell trajectory analysis from four GEO datasets identified key fate regulators. Sepsis diagnostic hub genes were screened via XGBoost and LASSO, with subsequent immune and pathway analysis. Results Compared to healthy controls, sepsis patients showed a shift in B cell composition, with fewer naïve B cells and more early activated B cells, along with disrupted differentiation paths. Six hub genes (DDX17, SLC2A3, RPS4X, PCBP2, S100A9, PFDN5) demonstrated strong diagnostic value for sepsis. Notably, RPS4X was highly expressed in healthy individuals and correlated with higher B cell levels. These genes were primarily associated with DNA repair, oxidative phosphorylation, and IL6-JAK-STAT3 signaling pathways. Conclusions The six B cell-associated hub genes—DDX17, SLC2A3, RPS4X, PCBP2, S100A9, and PFDN5—hold promise as potential biomarkers for sepsis diagnosis and targeted therapy.
BACKGROUND:High oxygen-affinity hemoglobin (HOAH) variants are rare causes of familial secondary erythrocytosis. Hb Coimbra [HBB: c.300T>A p.(Asp100Glu)] is a rare variant that can mimic β-thalassemia traits on HPLC, leading to diagnostic delays. CASE PRESENTATION:We describe a three-generation Italian family affected by Hb Coimbra. The 5-year-old female proband presented with asymptomatic erythrocytosis (Hb 15.9 g/dL) and a spurious hemoglobin A2 (HbA2) peak of 39.6% on cation-exchange HPLC. Family screening revealed that her father was managed with erythroexchange (EEX), while her paternal grandfather suffered two ischemic strokes in the absence of traditional cardiovascular risk factors. Diagnosis was confirmed via DNA sequencing. CONCLUSIONS:This case highlights the importance of recognizing pathognomonic HPLC artifacts to avoid misdiagnosis. It also underscores the need for long-term vascular risk stratification in pediatric carriers, particularly when a family history of thrombotic events is present.
Voxelotor, a hemoglobin S polymerization inhibitor, has been shown to reduce anemia in sickle cell anemia (SCA) but its effects on vaso-occlusive crisis frequency are limited. Its recent market withdrawal due to concerns over treatment-related excess mortality highlights the need for a better understanding of its mechanisms of action, especially with similar compounds under development.We investigated, in vitro, the effects of voxelotor on red blood cells (RBCs) from SCA patients, in hypoxia and normoxia. Parameters included: 1) RBC senescence markers including phosphatidylserine (PS) and CD47 exposure, calcium and reactive oxygen species (ROS) content measured with flow cytometry; 2) RBC aggregation assessed by light transmission; 3) RBC deformability using ektacytometry under shear stress, osmotic, and oxygen gradients.Voxelotor decreased RBC PS exposure in both hypoxia and normoxia but increased ROS levels. Voxelotor increased RBC aggregation and RBC deformability, decreased Ohyper (indicating decreased cell hydration state). The extent of RBC deformability increase with voxelotor was negatively correlated with fetal hemoglobin levels.While voxelotor demonstrated beneficial effects on RBC deformability and sickling propensity, its impact on RBC aggregation properties and ROS generation requires further investigation to better understand its different mechanisms of action.
Blood viscosity is recognized as a key determinant in the pathophysiology of vaso-occlusive crises (VOC) in sickle cell anemia (SCA). However, its longitudinal relationship with changes in VOC frequency over time remains poorly understood. The present study analyzed the associations between the changes in blood rheological parameters and the changes (delta) in the rate of VOC over time in 63 SCA patients. VOC rates (calculated over the 3 years preceding each blood sampling) and biological parameters were measured at two separate occasions, spaced by at least 3 yrs. (mean difference between the two measurements: 5.7 ± 2.0 years). A positive correlation was observed between the delta-VOC rate and the percentages of change in white blood cells, lactate dehydrogenase and blood viscosity. A multivariate linear regression was performed to test the independent association between the delta-VOC rate and the percentages of change in these biological parameters. The model was statistically significant (R2 = 0.47, p < 0.01), and blood viscosity was the only parameter independently associated with the percentage of change in VOC rate (β coefficient = 0.025, 95% CI: 0.009-0.040, p < 0.01). Our study demonstrated that steady-state blood viscosity is a valuable biomarker in SCA, given its associations with temporal variations of VOC frequency.
Sickle cell disease (SCD) is a genetic disorder strongly shaped by social, racial, and territorial inequalities. We identified, mapped, and characterized the population living with SCD in a state in Northeast Brazil and examined how fragmented institutional records affect the epidemiological visibility of the disease. We carried out a cross-sectional, multi-source study using institutional datasets on people with SCD. Records from these sources were consolidated and complemented by active tracing to update information. Adults were interviewed to describe their sociodemographic conditions, clinical profile, access to care, and experiences within the health system. Official counts substantially underestimated the affected population, reflecting unlinked information systems and limited record updating. The number of identified individuals increased from 266 to 471, corresponding to an estimated prevalence of 1.14 cases per 10,000 inhabitants. Interviewed adults (n = 119) showed high social vulnerability, with a predominance of Black participants, low educational attainment, low levels of income, and reliance on social benefits. A considerable proportion of participants were diagnosed only in adulthood. Specialized care was concentrated in the state capital and one additional urban center, creating major geographic barriers for residents of interior municipalities. Pain was frequent, severe, and functionally limiting, and many participants reported pain disbelief and stigmatizing labels during care. The gap between official records and identified cases indicates a significant weakness in health surveillance. Social vulnerability, geographically concentrated services, and routine delegitimization of pain suggest that historical inequalities, including racism, remain embedded even within a universal public health system such as Brazil's.
Circadian rhythms regulate immune cell activity, but their impact on hematopoietic stem cell infusion remains unclear. This retrospective study evaluated 177 severe aplastic anemia (SAA) patients undergoing unrelated donor hematopoietic stem cell transplantation (URD-HSCT) (2012-2023) to assess impacts of circadian infusion timing. No significant differences emerged between early and late infusion groups in cumulative incidence (CUI) of neutrophil/platelet engraftment, acute/chronic graft-versus-host disease (aGVHD/cGVHD), or CMV/EBV viremia. Notably, CUI of grade III-IV aGVHD was higher in the early group (5.7%, 95% CI: 1.8%-13.4%) versus 0% in the late group (95% CI: 0%-3.4%, P = 0.03). Five-year overall survival (OS: 87% vs. 93%, P = 0.1871), failure-free survival (FFS: 85% vs. 92%, P = 0.1389), and graft-versus-host disease/relapse-free survival (GRFS: 76% vs. 82%, P = 0.2781) showed no statistical disparities. Subgroup analysis revealed older patients (>30 years) receiving late peripheral blood HSCT had superior OS (78% vs. 97%, P = 0.0154) and FFS (78% vs. 94%, P = 0.0390). These data suggest that the timing of infusion may affect the risk of GVHD and the survival of older patients, for whom delayed infusion may confer a survival benefit.
Sickle Cell Disease (SCD), a genetic blood disorder, claims the lives of thousands of young children in Africa every year. The World Health Organization estimates that 75% of the diagnosed cases of the disease are within sub-Saharan Africa (SSA), with the mortality rate of children under five years of age soaring up to 90% and an estimated 80% of the affected children remain undiagnosed. Early diagnosis and early preventive treatment have been effective in easing the disease's life-threatening manifestations. However, this requires the availability of simple, accurate and affordable diagnostic tools suitable for resource-limited environments where access to proper diagnostics and clinical care remain a challenge and where reliable data on the true prevalence rates of SCD is still needed. Here, we present the results of a large-scale clinical study in Nigeria utilizing a portable platform that pairs a novel hemoglobin variant microchip electrophoresis testing technology and wireless networks to overcome this issue. This platform has great potential to expedite diagnosis, clinic referrals and patient tracking and follow-up. The platform also produces reports for all cases of SCD, which can be used to develop a comprehensive medical informatics architecture that facilitates local and national responses.
BACKGROUND:Heavy menstrual bleeding (HMB) a common manifestation of bleeding disorders in adolescents contributes to anemia, transfusions, and impaired quality-of-life (QOL). Evidence supporting practical management strategies in resource-limited settings remains limited. METHODS:This observational study in adolescents with confirmed bleeding disorders with HMB at a tertiary hemophilia treatment centre evaluated menstrual blood loss and QOL after a standard regimen of progesterone, tranexamic acid and iron; blood products or clotting factor concentrates were reserved for refractory bleeding. RESULTS:Twenty adolescents were included (2020-2025); von Willebrand disease (n = 11), chronic immune thrombocytopenia (n = 4), congenital aplastic anemia (n = 3), afibrinogenemia (n = 1), and factor VII deficiency (n = 1). Median PBAC score at presentation was 300 (range 190-500). Eighteen patients (90%) had anemia and prior transfusion exposure. After therapy, median PBAC declined to 100 (range 60-150). Hemoglobin improved steadily and no patient required further red cell transfusion during a median follow-up of four years. Hemostatic products were required only in two patients while on this regimen. Quality-of-life scores improved across physical, emotional, and social domains. CONCLUSIONS:A low-cost regimen combining progesterone, antifibrinolytic therapy, and iron effectively controls HMB, corrects anemia, reduces transfusion and factor utilization, and improves QOL in adolescents with bleeding disorders in resource-constrained settings.
A recombinant VWF D’D3 albumin fusion protein (rD'D3-FP) has been developed to extend the half-life of co-administered coagulation factor VIII (FVIII) for the treatment of haemophilia A. We examined the impact of albumin fusion of rD'D3 on the Stabilin-2 (STAB2) clearance receptor pathway and rD'D3-FP recycling via engagement of the neonatal Fc receptor FcRn as two potential mechanisms for rD'D3-FP half-life extension. Binding and intracellular trafficking of fluorescently labelled rD'D3-FP was examined using human 293-F cells overexpressing STAB2 or in combination with FcRn by flow cytometry and confocal microscopy. Pharmacokinetics of rD'D3-FP in FVIII knock out (KO), FcRn KO and hFcRn Tg32 transgenic mice was also assessed. We show that the interaction of rD'D3-FP and his-tagged rD'D3 with STAB2 is optimal at neutral pH but dissociates under acidic conditions. Whilst albumin fusion appears to hinder interaction of rD'D3-FP with STAB2, STAB2-mediated internalisation and lysosomal degradation of rD'D3-FP could be observed in an in vitro system. Rescue of rD'D3-FP via diversion into Rab11+ recycling endosomes was achieved with the co-expression of STAB2 and FcRn. A significant improvement in rD'D3-FP pharmacokinetics was observed compared to his-tagged rD'D3, however, only a modest improvement in rD'D3-FP half-life extension was observed in hFcRn Tg32 mice compared to FcRn KO and wild-type mice. Whilst FcRn may play a minor role in rD'D3-FP half-life extension in mice, albumin fusion may potentially reduce other rD'D3-FP clearance mechanisms. We thus hypothesise several independent mechanisms that may contribute to rD'D3-FP half-life extension in higher order species.
We quantified hepatic fat fraction (FF) by magnetic resonance imaging (MRI) in non-transfusion-dependent thalassemia (NTDT) patients, and we evaluated its associations with demographic, clinical, and biochemical parameters, tissue iron overload, and alterations of glucose metabolism. 169 NTDT patients (45.75 ± 13.41 years, 56.8% females) enrolled in the Extension-Myocardial Iron Overload in Thalassemia Network underwent standardized MRI assessments of hepatic, pancreatic, and cardiac iron and of hepatic FF. We also considered 301 transfusion-dependent thalassemia (TDT) patients and 25 healthy controls. To ensure reliable FF quantification, patients with severe hepatic iron overload (R2* > 333 Hz) were excluded. Glucose metabolism was assessed using an oral glucose tolerance test (OGTT). Both regularly transfused (RT) and never/sporadically transfused (NT) NTDT patients had hepatic FF values comparable to those of healthy controls, but significantly lower than those observed in TDT patients. RT-NTDT patients exhibited lower hepatic FF and hepatic iron levels than NT-NTDT patients. Independent of transfusional status, hepatic FF was unrelated to age, sex, splenectomy, ferritin, hepatitis C infection, or pancreatic and cardiac iron. In NT-NTDT, hepatic FF correlated positively with liver iron concentration. In RT-NTDT, hepatic FF and pancreatic iron showed similar diagnostic performance in identifying abnormal OGTT results. NT-NTDT patients exhibited higher hepatic FF than RT-NTDT patients, linked to hepatic iron levels. Hepatic fat accumulation and pancreatic iron overload may jointly contribute to altered glucose metabolism in RT-NTDT.
Sickle cell disease (SCD) is a monogenic blood disorder in which vaso-occlusive crises (VOC) are the main cause of hospitalization. Along with vascular adhesion, inflammation plays a central role in VOC. This study compared inflammatory and hematological markers in SCD patients during VOC and steady state, examined their association with outcomes such as hospital stay, acute chest syndrome (ACS), and mortality, and evaluated the impact of hydroxyurea. A total of 109 VOC patients admitted to the Department of General Medicine, VIMSAR, and 40 steady-state patients attending the outpatient clinic were included. Hematological (HbF, HbS, hemoglobin, WBC, neutrophils, platelets), biochemical (bilirubin, AST, ALT, urea, creatinine), and inflammatory markers (IL-6,CRP, ferritin, LDH, ESR) were measured. Data were analyzed using GraphPad Prism; comparisons were made by unpaired t-test and correlations by Pearson analysis. VOC patients showed significantly lower hemoglobin and higher WBC and neutrophil counts compared to controls. Inflammatory markers were markedly elevated during VOC: IL-6 (41.95 vs2.1 pg/mL), CRP (61.34 vs3.2 mg/L), ferritin (847.6 vs116.7 ng/mL), and LDH (1151.7 vs719.5 U/L). Platelet count and ESR did not differ significantly. Hydroxyurea use was associated with lower inflammatory marker levels. Inflammation strongly influences VOC severity and outcomes, and hydroxyurea mitigates this effect. Routine monitoring of inflammatory markers may support predicting course and management in SCD.
This study aims to assess the effect of endurance training on cardiorespiratory adaptations and health-related quality of life (HRQoL) of patients with sickle cell disease (SCD). Three databases (Pubmed, Embase and Web of Science) were searched from inception to September 2025. Randomized controlled trials (RCTs), Interventional Non-RCTs and case series studies assessing endurance training in SCD were included. Study selection, data extraction, and risk-of-bias assessments (using RoB 2) were performed by two independent reviewers, with discrepancies resolved by a third. Certainty of evidence was evaluated using GRADE. This review included seven studies with 176 patients with SCD engaged in different interventions of endurance training with follow-up lasting six to twelve weeks. Exercise training has favorable impact on functional capacity domain (MD 6.94; 95%CI 1.97 to 11.92) with trivial to no-effect on mental health and general health status domains of HRQoL. Exercise training seems to enhance functional capacity as demonstrated by increases in cardiorespiratory function and blood lactate levels at the first ventilatory threshold and peak oxygen consumption in RCTs and non-RCTs. Two case-series studies identified increases in the oxygen consumption at first ventilatory threshold in 15 patients (MD 1.75 ml/kg/min; 95%CI 0.73 to 2,77) after exercise training; and two RCTs identified improvements in the six-minutes walking test (MD 6.61%; 95%CI 1.42 to 11.79). Endurance exercise training improves cardiorespiratory function and HRQoL related to physical function in patients with SCD. These findings support the role of endurance exercise as complementary therapy to attenuate health risks and cardiovascular deterioration in SCD.
Sickle Cell Disease (SCD) is an autosomal recessive disorder with a substantial global burden. Despite its particularly high incidence in sub-Saharan Africa, early diagnosis remains limited in many countries. The objective of this study was to implement a newborn screening programme for SCD in one of the largest maternity hospitals in Angola and to support the subsequent pediatric follow-up of the affected children. Between June 2023 and December 2024, all children born in or attending the main hospital for vaccination were screened after parental or guardian consent. Blood was collected by heel-prick onto filter paper, and haemoglobin electrophoresis was performed by isoelectric focusing. Samples identified as HbSS were confirmed by PCR-RFLP, and atypical electrophoretic patterns were further analysed by DNA sequencing. In a total of 13,256 samples analysed the prevalence of HbSS was 1.38% (n = 183), and 20.31% (n = 2692) were HbAS. Other variants identified (n = 44) included HbE, HbC, and α-globin gene alterations. Of the infants diagnosed with SCD, 106 (58%) families were successfully contacted, but only 76 (42%) children initiated regular medical follow-up and prophylactic treatment (penicillin, multivitamins, and vaccinations. 30 families (28%) declined treatment and just one declined follow-up. These findings confirm the high prevalence of SCD in Angola and demonstrate the capacity of newborn screening programmes in reducing early morbidity and mortality. However, the substantial proportion of families refusing follow-up highlights the need for strengthened community health education to improve understanding of SCD.
PURPOSE:To assess the prevalence and severity of transfusion-associated iron overload (IO) in childhood cancer survivors (CCS). PATIENTS AND METHODS:Serum iron, total iron binding capacity, percent iron saturation and ferritin were measured in 75 survivors of childhood cancer. In addition, blood bank records were reviewed to determine the total volume of packed red blood cells (pRBCs) administered during cancer therapy. Patients who received ≥120 mL/kg of pRBCs or had a ferritin ≥1000μg/L underwent hepatic R2 and cardiac T2* MRI for iron quantification, echocardiogram, assessment of liver and endocrine function, and genetic analysis for hereditary hemochromatosis. RESULTS:Forty-nine patients qualified for second level studies. Of these, 35 completed the MRI scans. Fifteen patients had a liver iron concentration (LIC) >3 mg iron/g (moderate hepatic iron overload), including eight patients who had an LIC greater than 7 mg iron/g (severe hepatic iron overload), with a mean LIC of 4.3 mg iron/g (0-15.6 mg iron/g). LIC correlated with total volume of pRBCs and ferritin. No patient had cardiac iron loading by MRI. Eleven patients were heterozygous and one was homozygous for mutations associated with hereditary hemochromatosis. There was no correlation between iron overload and hereditary hemochromatosis gene status. CONCLUSION:There is a high prevalence of transfusion-associated iron overload among survivors of childhood cancer. This is concerning given the overlap between organ toxicities associated with cancer treatment and those known to be associated with iron overload. The tight correlation between LIC and ferritin suggests ferritin may be a reliable indicator of iron load in this patient population.
Objective To investigate the relationship of procalcitonin (PCT), C-reactive protein (CRP), and interleukin-6 (IL-6) with postoperative delirium (POD) in intensive care unit (ICU) patients, and to identify influencing factors. Methods Based on POD occurrence, 120 postoperative ICU patients were split into delirium (n = 72) and non-delirium (n = 48) groups. Clinical data and post-operative 24-h serum PCT, CRP, and IL-6 levels were gathered. Pearson correlation assessed relationships between these inflammatory markers, delirium incidence, and DRS-R-98 scores. Multivariate logistic regression identified predictors, and ROC curves evaluated the predictive value of the inflammatory markers. Results Delirium patients were older with higher APACHE II scores, more frequent mechanical ventilation, and longer ICU stays (P < 0.05). Their serum PCT, CRP, and IL-6 levels were higher and positively correlating with POD incidence and severity (DRS-R-98 scores) (r > 0, P < 0.05). These markers were identified as independent POD predictors (OR > 1, P < 0.05), demonstrating good predictive value with AUCs of 0.936 (CRP), 0.894 (PCT), and 0.820 (IL-6). Conclusion Postoperative inflammation serves as an independent risk factor for POD in ICU patients. Serum PCT, CRP, and IL-6 levels facilitate early detection and risk assessment, supporting preventive strategies and mitigating POD-related complications.
Hematopoietic stem and progenitor cells (HSPCs) are valuable for therapies and research due to their self-renewal and differentiation abilities. The ex vivo culture of these cells necessitates conditions that maintain their unique properties and provide a niche-like environment. Human platelet lysate (HPL), a growth factor-rich substance used in stem cell studies, influences various biological pathways. This study investigates HPL's effects on HSPCs' proliferation and erythroid-megakaryocytic differentiation, addressing the need to improve HSPC culture conditions for clinical and research applications. HPL was prepared, and isolated peripheral blood-derived HSPCs were cultured for 7 days in control and 10 % HPL-supplemented groups. Cell counts, viability, and population doubling time (PDT) were measured. Erythroid and megakaryocytic differentiation was assessed using qPCR and flow cytometry. Colony-forming cell (CFC) assays confirmed multilineage differentiation capacity. HPL significantly increased HSPCs proliferation, resulting in a 1.96-fold increase by day 7 and a reduced PDT. CFC assays confirmed that the cells maintained their ability to differentiate into various blood cell types. Flow cytometry showed a decrease in the CD71 marker without affecting the overall red blood cell population. Gene analysis revealed increased β-globin production while other red blood cell-related genes remained stable. Markers and genes associated with megakaryocyte development showed no significant changes. HPL as a culture medium supplement markedly enhances the proliferation of peripheral blood-derived hematopoietic stem cells during cell culture without inducing unwanted bias into the erythroid and megakaryocytic lineages.
Red blood cells (RBC) from patients with sickle cell disease (SCD) are continuously exposed to high levels of oxidative stress, which impacts RBC deformability. In this study, we applied a novel technique (oxidantscan) to measure deformability of RBCs under shear stress while exposed to oxidative stress. Using RBCs collected from a cohort of patients with SCD and healthy volunteers, we measured different parameters including T-POD (Time to initiate Oxidant-induced change in Deformability) and EIMin (the minimum deformability reached during the test). T-POD was significantly shorter in patients with HbSS (n = 21, 729 ± 410 s) and HbSC (n = 19, 1031 ± 225 s) when compared to healthy controls (n = 20, 1739 ± 328 s, p < 0.05). Also EIMin was significantly lower in patients with HbSS (0.26 ± 0.10) and HbSC (0.30 ± 0.10) compared to healthy controls (0.54 ± 0.02). These results indicate that RBCs from patients with SCD are more susceptible to oxidative stress, which can be measured in a reproducible and quantifiable way. We also demonstrate that Oxidantscan outcome parameters correlated with sickling behavior and markers of hemolysis, indicating a possible link between these key pathophysiological features and the ability to withstand oxidative stress (all p < 0.05). Finally, we provide preliminary evidence for the potential of this technique to evaluate antioxidant therapy with l-glutamine as SCD RBCs showed a significant improvement in T-POD (11.5 %, p = 0.01) and EIMin (46.9 %, p = 0.03) upon ex vivo treatment with l-glutamine. The Oxidantscan is a promising technique for evaluating the response of SCD RBCs to oxidative stress, providing new insights in disease pathophysiology and potential novel treatment strategies.
Complete blood counts (CBCs) are commonly measured in clinical practice and are associated with different diseases and traits. Statistical adjustment for blood cell abundance or related traits in these metabolomic epidemiology studies is not a common practice. However, it is plausible that common and cheaply measured blood cells would capture some of the variation also captured by metabolomic measures. Here, we assessed the association between metabolites and blood cell traits with a multi-ethnic meta-analysis of named metabolites from untargeted panels in the JHS, MESA, ARIC, and the HCHS/SOL cohorts. Among 2859 metabolite and blood cell trait pairs measured in at least two cohorts, 734 (25.6 %) were significantly associated. We also tested metabolite-blood cell trait pairs in the UK Biobank with a targeted NMR-based metabolomics panel. A large percentage (92 %) of the tested pairs was significant, and adjustment for blood cells altered relationships between metabolites and age and sex. We evaluated metabolite effect size changes for metabolite association with different incident diseases adjusting for CBC traits using the UK Biobank. Most notably, the metabolomic effect sizes on COPD were downweighted after adjustment. Blood cell trait adjustment showed similar but not overlapping effect size changes when compared to adjusting for eGFR, which is a common covariate in metabolomics analyses to account for kidney function. Our results show that statistical adjustment for blood cell traits may reduce confounding and clarify additional predictive power above existing clinical biomarkers in metabolomic studies.
Platelet-targeted gene therapy for hemophilia entails modifying a patient's hematopoietic stem cells (HSCs) ex vivo to produce platelets containing coagulation factors, offering a potential cure by localized factor release at injury sites. However, the associated bone marrow transplantation carries significant risks, underscoring the critical need for low-toxicity conditioning regimens to enable clinical translation. This study evaluated G-CSF and 5-FU as conditioning regimens to facilitate the engraftment of HSCs expressing platelet-targeted FIX Padua. Hemophilia B (HB) mice were conditioned with G-CSF, 5-FU alone, or a combination of G-CSF, plerixafor, and 5-FU. Bone marrow mononuclear cells (BM-MNCs) from transgenic donors (2bF9-R338L) expressing platelet-stored FIX Padua were transplanted. G-CSF conditioning enabled long-term engraftment with 1 × 108 BM-MNCs; however, the efficacy declined significantly with lower cell doses. A single-dose of 5-FU (150 mg/kg) administered one day pre-transplant achieved optimal FIX expression and minimal toxicity. Critically, a combined regimen of G-CSF/plerixafor plus timed 5-FU (day -1) yielded stable, long-term platelet FIX expression and phenotypic rescue. The success of 5-FU underscores the importance of creating a transient niche vacancy for donor HSC engraftment. These results demonstrate that this mobilization-based, non-myeloablative conditioning strategy is a promising approach for platelet-targeted gene therapy for hemophilia.
