bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of Innovative Laboratory Tests to Predict a Thrombotic Phenotype in a Family with Dysfibrinogenemia and a Novel FGG Mutation
Authors: Monard Amaury , Moenen Floor , Castoldi Elisabetta , De Simone Ilaria , Van Der Meijden Paola , Wichapong Kanin , van Doorn William , Spada Stefano , Hellenbrand Dave , Stork Alexander , Ten Cate Hugo , Beckers Erik AM , Henskens Yvonne 
Published: December 2023

Background:
Hypodysfibrinogenemia is a rare hereditary fibrinogen disorder characterized by quantitative and qualitative fibrinogen defects. These fibrinogen defects can cause thrombotic and hemorrhagic phenotypes. Unfortunately, predicting the phenotype in a specific patient is often not possible with routine coagulation tests.
Aims:
To characterize the phenotype and the genetic profile of a family with hypodysfibrinogenemia and to investigate the ability of innovative tests to predict bleeding and/or thrombotic phenotypes in asymptomatic family members.
Methods:
The proband, a 60-year-old woman with both bleeding and thrombotic complications who is currently on DOAC treatment, and two daughters were referred to our Hemophilia Treatment Center (HTC) for phenotypical and genotypical analysis of a congenital fibrinogen disorder (CFD). Extensive laboratory testing was done, as well as DNA-sequence analysis and molecular modelling. Thrombin generation and microfluidic testing were also performed to investigate their ability in phenotype prediction.
Results:
Fibrinogen activity and antigen levels led to the diagnosis of dysfibrinogenemia in the proband and hypodysfibrinogenemia in both daughters. In all three cases, the same heterozygous missense mutation in the FGG gene was uncovered. This likely pathogenic mutation leads to the p.(Tyr375Cys) amino acid change. Molecular modeling predicted possible conformational changes or covalent dimerization of the fibrinogen molecule. Thrombin generation was elevated in one daughter. Microfluidic testing showed enhanced fibrin formation in both daughters, regardless of the coagulation trigger.
Conclusion:
We described a family with hypodysfibrinogenemia in whom a novel heterozygous missense mutation in the FGG gene was found, possibly leading to conformational changes or covalent dimerization of the fibrinogen molecule. Furthermore we showed that microfluidic testing and thrombin generation can indicate a thrombotic phenotype in these patients, not detected with routine coagulation tests.
Clinical Analysis of Metagenomic Sequencing of Plasma Microbial Cell-Free DNA for Infected Patients in SAA
Authors: Jing Liping , Fan Huihui , Yang Wenrui , Xiong Youzhen , Zhang Li , Zhao Xin , Zhang Fengkui 
Published: December 2023

Backgroun d: As a high-risk group of clinical infection, patients with severe aplastic anemia (SAA) have high mortality due to agranulocytosis or immune function impairment. The current conventional detection methods such as blood culturetissue chulture etc. often cannot identify pathogenic bacteria. We evaluate the diagnostic performance of metagenomic next-generation sequencing (mNGS), and explore the application value of mNGS in infected patients in SAA, in order to provide help for precision therapy.
Methods: From Ocb 2020 to Dec 2022 in our center, we enrolled 70 SAA patients with infection and investigated the usefulness of mNGS of plamsa or tissue DNA for identification of infectious pathogens. The consistency of the results of mNGS and the results of routine etiological detection (blood culture(BC), swab/tissue culture) with clinical infection diagnosis and the influence of the results of mNGS on clinical infection treatment were compared.
Results: 70 SAA patients of mNGS were detected, including plasma-mNGS 66 cases and tissue-mNGS 4 cases. Among all patients, 48 cases(68.6%) were positive. In comparison to blood culture, the positive and negative agreement of mNGS were 100% (13/13) and 38.6% (22/57), respectively. In 48 mNGS positive cases, 36 patients (75%) underwent antimicrobials adjustment, resulting in positive impact on 30 patients. Fungi were detected in 29 cases, the detection rate was 41.4% (29/70) by mNGS, otherwise 2.8% (2/70) by BC. Of the 70 patients with SAA, 55 infections eventually impproved (78.6%).
Conclusion: The positive rate of mNGS detection and the consistent rate with clinical diagnosis are higher than that of conventional etiological detection, which provides a faster and more sensitive detection method for infected patients in SAA.
【 Key words 】 Metagenomic Sequencing ; Plasma Microbial Cell-free DNA; Aplastic anemia; Infection; pathogen diagnosis
Familial Thrombocytopenia-Associated Germline ETV6 P214L Mutation Results in XPO1-Mediated Nuclear Export
Authors: McCONVILLE Michael , Thomas Toby , Valadez Catherine , Beckner Ryan , Chook Yuh Min , Chung Stephen , Liszczak Glen 
Published: December 2023

In recent years, a number of heterozygous germline mutations in ETV6 have been associated with familial thrombocytopenia and predisposition to hematologic malignancies. While most of these missense mutations occur in the DNA-binding-domain of ETV6, the recurrent P214L mutation is unique in that it occurs in the central disordered region of the protein. The ETV6 P214L mutation has previously been shown to disrupt the nuclear localization of ETV6 and the ability of ETV6 to act as a transcriptional repressor. Unexpectedly, we have found that the P214L missense mutation creates a recognition motif for the nuclear export protein XPO1. We have demonstrated in cellular and protein reconstitution assays that XPO1 directly engages the P214L but not the wild-type ETV6 construct. Consistent with this mechanism, we are able to fully rescue the nuclear localization of ETV6 P214L in mammalian cells with small molecule XPO1 inhibitors. We have also found that once localized to the nucleus, the P214L mutant construct exhibits functional consequences indistinguishable from wild-type ETV6 in Ba/F3 transformation assays.
When looking at the evolutionary conservation of the ETV6 amino acid sequence involved in the XPO1 recognition motif, we observed that the synonymous P216L mutation in the mouse ETV6 ortholog does not create a nuclear export signal. In order to study XPO1-mediated export of ETV6 P214L and its impact on hematopoiesis, we have generated a mouse model in which two point mutations (P216L and S211I) have been introduced into a single endogenous ETV6 allele, which precisely matches the sequence of this region in human P214L carriers. Using CRISPR-Cas9, we generated 3 transgenic mouse lines: Etv6 P216L, Etv6 S211I, and Etv6 S211IP216L. Immunofluorescence of fixed bone marrow shows that ETV6 is only mislocalized in megakaryocytes from ETV6 S211I P216L mice. Complete blood counts revealed a significant decrease in platelet counts in ETV6 S211I P216L heterozygotes (485 x 10 3/µL) compared to ETV6 S211I heterozygotes (1140 x 10 3/µL, p&lt;0.0001), ETV6 P216L heterozygotes (1065 x 10 3/µL, p&lt;0.0006), and wild-type littermates (1254 x 10 3/µL, p&lt;0.0001). Flow cytometry phenotyping of bone marrow from the three transgenic lines and wild-type littermates shows a significant decrease in the absolute numbers of hematopoietic stem cells (HSCs) and megakaryocyte precursors (MkPs) in ETV6 S211I P216L mice.
In summary, we have determined that XPO1-mediated nuclear export is the mechanistic basis for ETV6 P214L mislocalization. We developed a mouse model that recapitulates this export signal in the endogenous ETV6 allele and found that this model demonstrates a thrombocytopenic phenotype as seen in human patients. Reductions in HSCs and MkPs suggest both a decreased fitness of HSCs, as well as dysregulated megakaryopoiesis in this model.
Underrepresentation of Racial and Ethnic Minorities in Registrational Chimeric Antigen Receptor (CAR) T-Cell Therapy Trials in Hematologic Malignancies: An FDA Analysis
Authors: Mahat Upendra , Seam Pamela , Bouchkouj Najat , Sharma Poornima , Peredo-Pinto Helka , Elmacken Mona , Kaushal Megha , Wang Shu , Xu Zhiheng , Jia Yuxia , Richardson Nicholas , Kanapuru Bindu , Kasamon Yvette , Gormley Nicole 
Published: December 2023

Introduction: Equitable representation of diverse racial and ethnic groups in oncology clinical trials continues to be a major challenge. Numerous initiatives designed to enhance diversity in oncology clinical trials are ongoing, including efforts at the U.S. Food and Drug Administration (FDA) such as the Oncology Center of Excellence's Project Equity. Given the multiple recent FDA approvals of CAR T-cell therapies for relapsed or refractory hematologic malignancies, we evaluated the enrollment rates of racial and ethnic groups in the CAR T-cell registrational trials submitted to the FDA.
Methods: We conducted a pooled, patient-level analysis of all registrational trials of CAR T-cell therapies submitted to the FDA to support approval for hematologic malignancies between 2017 and 2021. Datasets from approved licensing applications were pooled, and demographics and efficacy outcomes were analyzed descriptively.
Results: A total of 13 registrational trials involving 6 autologous CAR T-cell products were analyzed, comprising 1,319 patients with hematologic malignancies. The median age at enrollment was 60 years (range: 3 to 86 years). Of the 1,319 patients, 81% (1068) were White, 5% (66) were Black, 5% (65) were Asian, 4% (48) were reported as Other, and 8% (110) were Hispanic or Latino. Race and ethnicity information was not reported for 5% (72) and 10% (127) of patients, respectively. The majority of the patients enrolled in the registrational trials were from the United States (80%), followed by Europe (11%) and Australia (2%). The hematologic malignancies included large B cell lymphoma (59%), multiple myeloma (15%), follicular lymphoma (13%), B cell acute lymphoblastic leukemia (9%), and mantle cell lymphoma (5%). Response rates by race, ethnicity, and histology are shown in Table 1 and Table 2.
Conclusions: Despite the majority of patients being enrolled in the U.S., racial/ethnic subgroups other than White and non-Hispanic/Latino were underrepresented in CAR T-cell registrational trials. This highlights the need to understand and overcome the continued barriers to CAR T-cell clinical trial enrollment across racial and ethnic groups. Although clinically meaningful efficacy was observed across these racial/ethnic groups and diseases, more data are needed to evaluate if efficacy outcomes vary across these subgroups. Work to describe safety and tolerability, particularly cytokine release syndrome (CRS) and neurotoxicity, in these racial and ethnic subgroups is ongoing. More effort is needed to better enhance diversity in CAR T-cell trials, especially given the expanding use of CAR T-cell therapies in the treatment of patients with hematologic malignancies.
Tern-701 (HS-10382) Is a Potent Inhibitor of BCR::ABL1 and Is Synergistic with Active Site Tyrosine Kinase Inhibitors
Authors: M Parsons Benjamin , Jasper Jeffrey R. , Jones Christopher 
Published: December 2023

CONCLUSIONS
Although tyrosine kinase inhibitors (TKIs) targeting the ATP-binding site of the BCR::ABL1 oncoprotein are effective therapeutics for chronic myeloid leukemia (CML), patients often develop drug resistance due to ATP-site mutations that inhibit drug binding. TERN-701 is a potent and highly selective inhibitor of BCR::ABL1 that is designed to bind the allosteric myristoyl site on the kinase, circumventing resistance to active site mutations, with potential for synergistic combination with active site TKIs.
Competition binding screens were used to assess the selectivity of TERN-701 on more than 450 targets spanning the human kinome. The potency of TERN-701 against the proliferation of wild type and mutant CML cell lines, as well as a panel of 102 cancer cell lines, was assessed using CellTiter-Glo®. Synergy between TERN-701 and active site TKIs was assessed in vitro with interactions quantified using Bliss, combination, and curve-shift analyses. Prospective mutagenesis experiments were conducted by treating BaF/3 cells engineered to overexpress the BCR::ABL1 kinase with various concentrations of both TERN-701 and the TKI imatinib, followed by observation of clonal outgrowth of treated cells over time.
We have previously shown that TERN-701 is a potent inhibitor of native CML cell lines such as K562. Here, we show that TERN-701 inhibited cell proliferation in various clinically relevant mutant CML cell lines (including T315I) with IC 50s ranging from 5 to 26 nM and 70 to 1200 nM against active site and myristoyl site mutations, respectively. TERN-701 was highly selective for BCR::ABL1, with no appreciable activity (&gt;50%) against &gt;450 purified kinase targets. Against a panel of cancer cell lines, TERN-701 was more selective than the comparator allosteric BCR::ABL1 inhibitor asciminib while maintaining similar potency. In vitro combination studies previously revealed that TERN-701 works synergistically with multiple TKIs in the non-mutant K562 cell line. Here, expanded studies using KCL22-s and BCR::ABL1-T315I mutant cell lines identified strong synergistic interactions between TERN-701 and active site TKIs, including ponatinib and dasatinib. Furthermore, prospective mutagenesis studies showed that the combination of TERN-701 and imatinib resulted in greater inhibition of the outgrowth of resistant clones relative to either compound alone.
TERN-701 is a potent and highly selective allosteric inhibitor of BCR::ABL1 that can act synergistically with active site TKIs in vitro, even against the T315I gatekeeper mutation that confers resistance to all approved active site TKIs except ponatinib, which has known safety liabilities. These data support the continued development of TERN-701 for the treatment of CML using both monotherapy and combination approaches.
A Comprehensive Echocardiographic Portrait of Adult Sickle Cell Disease Patients in the Modern Era
Authors: Chan Kok Hoe , Liao Zean , Idowu Modupe 
Published: December 2023

Background
Cardiovascular complications, especially pulmonary hypertension, are a known cause of mortality in patients with sickle cell disease (SCD). With advances in medicine and the introduction of various disease-modifying medications, the incidence and prevalence of cardiovascular complications are poorly documented.
Aim
To determine rates of cardiovascular complications in patients with SCD in the modern era of disease-modifying medications in a single center.
Method
We performed a retrospective observational single-center study of patients with SCD: all genotyped and ≥16 years old who had a recent echocardiogram performed during the study period of 2019-2023. Demographics, echocardiogram results, and laboratory findings were collected and analyzed. Descriptive statistics are presented in percentages, medians, and interquartile ranges. An unpaired Student's t-test and Fisher's exact test were used to determine, where appropriate, the level of significant differences among continuous and categorical variables, respectively, with p value &lt;0.05 considered statistically significant. Data was analyzed using GraphPad Prism version 9.0.2.
Results
As of June 2023, 80 patients (38 male and 42 female, median age 30 years) had undergone a routine transthoracic echocardiogram. Sixty-five patients had the HbSS genotype, 8 HbSC, and 7 HbSB null. Sixty-one patients had received at least one treatment, and 19 (24%) were on concurrent therapy. Hydroxyurea was the common SCD-modifying therapy that the patients received (51 patients), followed by L-glutamine and voxelotor (11 each) and crizanlizumab (9 patients).
In total, 56 (70%) patients had at least one cardiac abnormality. The most common cardiac abnormality was left atrial dilatation (41%), followed by left ventricular hypertrophy (33%), right atrial dilation (24%), and left ventricular dilatation (23%). Pulmonary hypertension was present in 30 (38%) patients with tricuspid regurgitation velocity (TRV) ≥2.5 msec (Table 1).
The cohort of patients with pulmonary hypertension tended to be older (median age 31 years, p=0.0357), have a higher creatinine level (median 0.8, p=0.0414), and have coexisting echocardiographic abnormalities such as left atrial dilatation (70% vs. 24%, p&lt;0.0001) or right atrial dilatation (40% vs. 14%, p=0.0136) than SCD patients without pulmonary hypertension (Table 2).
Conclusion
We present a comprehensive echocardiographic assessment of patients with SCD. Despite advances in the treatment of SCD, there remains a high prevalence of cardiac involvement in SCD patients. We found out that 70% of the patients we reviewed had at least one cardiac abnormality on echocardiography. Left atrial dilatation and left ventricular hypertrophy were the most common cardiac abnormalities noted. Pulmonary hypertension was observed in 37.5% of patients with co-existing right and left atrial enlargement. Larger studies are needed to corroborate these findings.
The Molecular Prognostic Scoring System for Normal Karyotype Myelodysplastic Syndromes
Authors: Wang Wei , Zhang Yudi , Yang Wenli , Han Yueyuan , Ge Zheng , Tong Hongyan 
Published: December 2023

Purpose: Myelodysplastic syndromes (MDS), is characterized by clinical heterogeneity and varying prognoses. Reliable prognostic models are essential to stratify MDS patients and optimize clinical management. Although incorporating molecular information into MDS prognostic models improves their predictive ability, limitations of next-generation sequencing panels and potential heterogeneity among MDS patients with different karyotypes hinder their clinical translation.
Methods: In total, 237 normal karyotype (NK) MDS patients from the First Affiliated Hospital of Zhejiang University School of Medicine were used as the derivation set to investigate the molecular frequency and prognostic spectrum. A parsimonious molecular-clinical model was developed with overall survival (OS) as the endpoint and compared with other models using the concordance index as the primary evaluation index. An external independent validation set from the International Working Group for Prognosis in MDS cohort(691 NK MDS patients) was used to evaluate the model generalization ability.
Results: In NK MDS, the gene mutation frequency spectrum is similar to the overall MDS population, the differences mainly lie in the prognostic spectrum. Genes such as TP53, previously associated with MDS prognoses have lost their prognostic value. Therefore, a seven-parameter molecular-clinical prognostic model called the Molecular Prognostic Scoring System for NK MDS (NK-PSS-M) was developed based on a specific molecular prognostic spectrum of NK MDS. The model includes age, hemoglobin, platelet count, blast percentage, CEBPA, RUNX1, and U2AF1. The NK-PSS-M model allows for personalized risk assessment based on patient characteristics, with higher scores indicating a greater risk and poorer prognosis. In addition, we developed a five-class classification system based on these scores, which differentiates patients across risk categories and predicts outcomes, such as OS and LFS(Figure 1 A and B). The performance of NK-PSS-M was comparable to that of the Molecular International Prognostic Scoring System(IPSS-M), and it significantly outperformed IPSS-R and other molecular prediction models based on it. The above results were confirmed in the external independent validation cohort(Figure 1C). Our transcriptomics analysis revealed that the low-risk group defined by NK-PSS-M demonstrated significant activation of the inflammatory pathway, and immune activation, which is consistent with the current understanding of the immunological landscape of MDS.
Conclusion: The NK-PSS-M model improved the risk stratification of non-molecular models and provided a reliable alternative to the IPSS-M. In addition, we demonstrated the molecular heterogeneity of MDS across different cytogenetics and emphasized the importance of developing cytogenetics-specific prognostic models.
Long-Term Hematopoietic Dysfunction in Patients with Single Large-Scale Mitochondrial DNA Deletion Syndromes
Authors: Greenberg-Kushnir Noa , Barg Assaf Arie , Schiby Ginette , Mardoukh Corine , Varda-Bloom Nira , Bar Diana , Anikster Yair , Bielorai Bella , Toren Amos , Jacoby Elad 
Published: December 2023

Pearson syndrome (PS) is the only single large-scale mitochondrial DNA deletion syndrome (SLSMDs) presenting substantial hematologic manifestations, usually a severe though transient cytopenia of childhood. Recently, progression to myeloid malignancies has been described among PS patients. Older PS patients, as well as patients with SLSMDs presenting with Kearns-Sayre syndrome (KSS) or with progressive external ophthalmoplegia (PEO), do not display hematologic abnormalities. Since the underlying mitochondrial DNA deletion is similar in all SLSMDs, we hypothesized that bone marrow (BM) dysfunction is a common and persistent manifestation in these patients.
We studied 16 patients with molecular-confirmed SLSMDs: 13 with PS, and three with KSS. The median age at diagnosis was 25 months (range 2-136 months). Seventy-five percent of patients presented with cytopenia in infancy, and 56% required blood products. Resolution of cytopenia occurred in 92% of this subgroup, at a median age of 24 months (range 12-39 months).
BM studies were performed in all patients, and 56% of patients had multiple BM evaluations. All BM biopsies examined showed signs of marrow dysfunction, including in patients without a history of cytopenia. Median BM cellularity was 50% (range 30-80%) at a median age of 5 years. BM samples showed a paucity of precursor myeloid cells, signs of dyserythropoiesis, and abnormal or missing megakaryocytes. We report low hematopoietic stem and progenitor cell functionality in patients with SLSMDs: The median CD34+ count was 0.68% (range 0.27-1.92%, expected range 1-2%). BM colony forming unit (CFU) capacity was reduced in SLSMD patients: the median CFU in patients was 280 per 10 6 cells compared to 1090 per 10 6 cells in healthy controls (p=0.004). Conventional cytogenetic analysis was done on 10 patients (62%), and FISH analysis was done on 15 patients (93%). Monosomy 7 was identified in seven patients (44%): in one patient the monosomy 7 resolved in a follow-up BM assay. Another patient with signs of myelodysplasia and 30% of the BM cells having monosomy 7 was referred to an allogeneic bone marrow transplantation.
To conclude, long-term BM dysfunction is evident in all patients with SLSMD and includes a risk of developing clonal evolution and monosomy 7. This data, combined with other recent publications, highlights the ongoing marrow failure and suggest that SLSMD syndromes should be included under inherited BM failure syndromes.