bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adiponectin Promote TPO Response for Treating ITP By Relating Rab6A-myosin9 Trafficking Vehicle Carrying c-Mpl to Cell Surface
Authors: Zhao Xin , Ma Huixian , Li Guosheng , Sun Tao , Ji Chunyan 
Published: December 2023

Impaired megakaryocyte maturation is one of the pathogenic mechanisms of primary immune thrombocytopenia (ITP). Exploring the regulatory mechanisms of megakaryocyte maturation is important for the clinical treatment of ITP. The thrombopoietin (TPO) receptor c-Mpl on the membrane surface of megakaryocytes plays a critical role in the cellular reception of TPO signals and megakaryocyte maturation. But how the distribution of c-Mpl on the membrane surface of megakaryocytes influences the pathogenesis of ITP and the specific regulatory mechanisms are still unclear.
In this project, we found that the distribution of c-Mpl on the membrane surface of megakaryocytes of ITP patients was significantly reduced, confirming that the abnormal distribution of c-Mpl is associated with the pathogenesis of ITP. Further, we applied immunoprecipitation and protein profiling to find the intracellular c-Mpl transport-related protein molecules. Motor protein myosin 9 and G protein-associated protein Rab6A were screened out. Next, we down-regulated the expression of myosin 9 and Rab6A, proved that aberrant expression of both leads to reduced transportation of the c-Mpl to membranes and impaired megakaryocyte maturation, identifying the regulatory complex for c-Mpl transport to membranes was composed of myosin 9 and Rab6A.
Moreover, we confirmed that the formation of the above complex is dependent on GTP activation of Rab6A by transient overexpression of wild-type Rab6A (wt), dominant-negative (T27N) and GTPase-deficient (Q72L) Rab6A, clarified the working mechanism of how the Rab6A-myosin9 trafficking vehicle regulate c-Mpl transport to membranes. At the same time, we demonstrated that the abnormally low expression of Adiponectin in the plasma of ITP patients affects the transportation of the c-Mpl to membranes by inhibiting the interaction between Rab6A and myosin 9, thereby reducing the responsiveness of megakaryocytes to TPO and inhibiting the activation of its corresponding intracellular signaling pathways.
Finally, using a mouse model of ITP disease, we demonstrated that Adiponectin could mediate the transportation of the c-Mpl to membranes by regulating the Rab6A-myosin9 complex and can improve the efficacy of TPO in the treatment of ITP. In conclusion, the present study identified a novel trafficking complex mediating the transportation of the c-Mpl to membranes, clarified the abnormal mechanism of this complex in ITP, and confirms that Adiponectin can regulate this complex to improve the efficacy of TPO, which provides a new idea for the clinical treatment of ITP.
Post-Marketing Observational Study of the Safety and Efficacy of Fibrinogen Concentrate in Congenital Fibrinogen Deficiency in Patients of All Ages
Authors: Peyvandi Flora , Pezeshki Gita , Solomon Cristina 
Published: December 2023

Background and Significance: Congenital fibrinogen deficiency (CFD) is a rare coagulation disorder characterized by a lack or low levels of functional fibrinogen leading to inadequate clot formation. Patients with CFD may experience severe or frequent bleeding episodes (BEs), either spontaneously or following trauma or surgery. Human fibrinogen concentrate (HFC; Fibryga® [Octapharma]) can restore hemostasis, and its efficacy and safety has been demonstrated in interventional clinical studies of adult and pediatric patients. The primary objective of this planned observational study is to collect real-world evidence to further assess the safety profile of this HFC, in patients of all ages with congenital fibrinogen deficiency, by characterizing the risks of thrombosis and anaphylaxis following its administration for treatment of bleeding (Figure 1). The observational data collected in this study will provide a more comprehensive safety profile of this HFC in patients of all ages following its marketing authorization.
Study Design and Methods: FORMA-07 (NCT03793426) is a prospective, non-interventional, multicenter, observational, post-marketing study. Patients of all ages with a documented diagnosis of CFD (i.e., congenital afibrinogenemia or hypofibrinogenemia) requiring on demand HFC for BE treatment are eligible for inclusion (Table 1). Exclusion criteria include other coagulation disorders, history of anti-fibrinogen inhibitors, and participation in concurrent clinical studies. This study will examine a highly purified, plasma derived, double virus inactivated/eliminated, lyophilized HFC, which will be used according to individual country approval and labelling, and individually dosed as per the locally approved package insert. Independent ethical committee approval will be sought prior to study start and freely given, written informed consent obtained from all participants, or legal guardians in the case of pediatric patients. This study will be conducted in accordance with the ethical principles laid down in the Declaration of Helsinki. The primary endpoint is the incidence of thromboembolic adverse drug reactions (ADRs) . Serious ADRs, ADRs of special interest (including thromboembolic events and anaphylaxis), and thromboembolic monitoring using the Wells Score will be documented. Hemostatic efficacy will be assessed as a secondary endpoint by the study investigator using a robust 4-point objective scale from 2 to 24 hours following treatment.
FORMA-07 is ongoing and actively enrolling patients of all ages with CFD. The study aims to enroll ~25 patients over 7 years, collecting data from 105 BEs (≥10 major BEs in 10 patients). Results of HFC safety and efficacy will be continually monitored throughout the study. Completion is expected Q4 2027.
The Plasma Proteomic Landscape of Thrombotic Events across Multi-Center Cohorts
Authors: Kreft Iris charlotte , Smit Eva , Camilleri Eleonora , Burggraaf Louise , Van Rein Nienke , Van Vlijmen Bart , Hulshof Anne-Marije , Van Bussel Bas , van Rosmalen Frank , van den Berg Tom , Henskens Yvonne , Ten Cate Hugo , Coutinho Jonathan , Kruip Marieke , Eikenboom Jeroen , Hoogendijk Arie , Cannegieter Suzanne , Van Den Biggelaar Maartje 
Published: December 2023

Background: Diagnosis and risk prediction of thrombosis is challenging due to nonspecific symptoms, diverse risk factors, symptom overlap with other conditions and the variability in thrombosis presentations. Little is known on the impact of thrombosis on the plasma proteome across the spectrum of its clinical representations.
Objective: The aim of this study was to explore the plasma proteomic landscape across multi-center thrombosis cohorts. The focus was on identifying protein signatures representative of thrombosis in general or among patients at risk of thrombosis, those with a history of thrombosis, those at acute phase and those with COVID-19 associated thrombosis.
Method: Using state-of-the-art mass spectrometry-based proteomics, we analyzed 420 plasma proteome samples both cross-sectional and longitudinal from 211 patients, including those at risk of thrombosis, in the acute thrombotic phase (cerebral venous sinus thrombosis patients) and with COVID-19-associated thrombosis (Figure 1). Linear models and unsupervised co-expression clustering were used to explore common and distinct protein signatures among the cohorts.
Results: Across the analyzed samples 475 proteins were quantified, with 86% showing a CV below 30%, and we found a strong correlation between protein levels quantified by MS and ELISA. Plasma proteomic alterations exhibited variation across cohorts, with the most pronounced changes observed in COVID-19 patients. While co-expression analysis unveiled distinct protein clusters associated with SARS-CoV-2 signaling pathways (CRP, SAA1) and Vitamin K-dependent (VKA) coagulation proteins (F9, F2, F10, PROS, PROC, PROZ, C4BPA), we did not identify a specific protein cluster related to thrombosis (Figure 2). Notably, VKA-therapy was associated with decreased abundance of VKA-dependent coagulation proteins, with F10 and PROZ showing significant alterations between patients who experienced a thrombosis compared to healthy controls. We observed an inverse correlation between the International Normalized Ratio and abundance of VKA-dependent coagulation proteins, with an increased correlation of F2 and F9 in patients who experienced a thrombotic event. Additionally, our analysis also highlighted potential variations in multi-center cohorts during sample handling, emphasizing the importance of standardized protocols in ensuring the reliability and comparability of results.
Conclusion: Using a comprehensive MS-based proteomics approach, we identified clear plasma proteomic signatures associated with VKA-treatment and COVID-19 , but no distinct thrombosis signature across multi-center thrombosis cohorts.
ABO-Incompatible Platelet Transfusion Is Associated with Ischemic Lesions on Brain MRI in Patients with Intracerebral Hemorrhage
Authors: Carvalho Poyraz Fernanda , Ridha Mohamed , Simonetto Marialaura , Kumar Aditya , Hess Evan , Magid-Bernstein Jessica , Connolly E. Sander , Ghoshal Shivani , Agarwal Sachin , Park Soojin , Claassen Jan , Hod Eldad A. , Roh David 
Published: December 2023

Background: We have identified that major ABO-incompatible platelet transfusions are associated with poor intracerebral hemorrhage (ICH) outcomes. The underlying drivers for the relationship of these incompatible platelet units on outcome are unknown, yet do not appear to be related to impaired hemostasis. Conversely, ICH patients are known to encounter remote, “silent” ischemic lesions, thought to be from microthrombosis, during their hospitalization. Though these ischemic lesions are well known to associate with poor ICH outcomes, risk factors for its formation are unknown. We explored whether major ABO-incompatible platelet transfusions are associated with acute ischemic lesions on brain MRI during an ICH hospitalization.
Methods: Consecutive spontaneous ICH patients enrolled into a single-center, prospective cohort study between 2009 and 2018 were assessed. Patients who received a single platelet transfusion within 24 hours of ICH admission and had a brain MRI during the hospitalization were included in the analysis. Major ABO incompatible platelet transfusion (vs minor mismatch/identical) was defined as the exposure. Presence of remote ischemic lesions, defined as diffusion restriction &gt;10 mm away from the hematoma detected on brain MRI, was the outcome. Regression models were used to assess relationships between major ABO-incompatible platelet unit exposure and acute ischemic brain lesions, adjusting for time from ICH onset to MRI or ICH severity.
Results: We identified 40 ICH patients receiving an acute platelet transfusion and having a brain MRI performed during the ICH hospitalization. The mean age was 67.1 (SD 14.1), 37.5% were female, and 20% received a major ABO incompatible platelet unit. Patients receiving a major incompatible platelet unit were more likely to encounter ischemic lesions (62.5% vs 21.9%). We also identified an association of major ABO-incompatible platelets units with acute ischemic brain lesions in our regression models after adjusting for time from ICH symptom onset to MRI (OR 5.45, 95%CI 1.01-29.30, p=0.048) and ICH severity (OR 9.15, 95%CI 1.37-62.67, p=0.024). Additional sensitivity analysis adjusting for patient blood type did not alter this relationship (OR 17.1, 95%CI 1.34-217.9, p = 0.029).
Conclusions: Among ICH patients receiving acute platelet transfusions and MRI neuroimaging, major ABO incompatible units were associated with remote ischemic lesions on brain MRI. Further work is needed to address the generalizability of our data and to clarify relationships of platelet unit characteristics, cerebral microthrombosis, and clinical outcomes after ICH.
Deep Immunophenotyping Using High Complexity Flow Cytometry in Acute Myeloid Leukemia
Authors: Pearl Jessica , Bruni Elena , Hsu Haoting , Walling Brandon , Schiavini Paolo , Ritchie Michael , Heidemann Karin abarca , Mancini Maria 
Published: December 2023

Acute Myeloid Leukemia (AML) is a highly heterogenous disease with a diverse range of abnormal myeloid subpopulations resulting in challenges in clinical diagnosis and treatment of disease. While sequencing may provide precise information about mutations, the turnaround times make it impractical for routine tracking of disease progression. In contrast, flow cytometry utilizes the expression patterns of cellular markers to determine AML subpopulations with high accuracy. This, coupled with the limited turnaround time, makes flow cytometry an excellent tool for the identification and tracking of disease progression. AML lineage characterization and disease progression are found to be a predictor of both residual disease and the likelihood of relapse further emphasizing the need for effective and accurate characterization of AML populations.
Champions Oncology has developed a high-complexity 18-color flow cytometry panel to identify AML subpopulations at high resolution, named PhenoSeek AML ®. Clinical samples may be shipped to a central location for processing which requires sample populations to be retained during transit. Utilizing Champions' bank of primary AML samples, we have evaluated the stability of key AML subpopulations and found stability to be retained for up to 72 hours after collection. In addition to sample stability, assessing the lower limit of detection is critical for minimum residual disease and relapse screening. Flow cytometry provides an excellent tool for these tests due to its single cell nature, which allows for the interrogation of individual populations with as little as hundreds of cells. Indeed, the dilution of primary AML subpopulations was detected with as little as 10 3-10 4 cells.
Finally, utilizing Champions' cryopreserved never-passaged AML models characterized by both flow cytometry and sequencing, we investigated how the cryopreservation processes impacted the expression of markers associated with specific AML subpopulations. We observed similar expression and detection of AML populations following cryopreservation indicating that these models can be utilized as an excellent tool for further studies.
Collectively, our data indicates that the panel developed at Champions Oncology provides robust detection and deconvolution of AML subpopulations and can be an excellent tool for the tracking of AML disease progression.
Outcomes of Newly Diagnosed Hodgkin Lymphoma Patients in 4 Academic Centers in Paraguay. the Impact to Access to PET-CT Scanning in Survival
Authors: Von Glasenapp Seisha Alana , Enciso Arrua Maria Elvira , Quiroz Alfredo , Paats Aline Nicole , Morel Laura , Jiménez Leticia Rocio , Andino Lidiane , Salinas Victor Ladislao , Valcarcel Bryan , Malpica Castillo Luis Enrique 
Published: December 2023

Background: Hodgkin's lymphoma (HL) is a highly curable lymphoproliferative neoplasm. Assessment of HL at baseline (pre-therapy), intermediate (iPET) and end-of-therapy (EOT) by positron emission tomography/computed tomography (PET/CT) is considered a “gold standard” procedure and has become the main tool for clinical decision making in the management of HL. However, inequity in access to diagnostic medical technology in Paraguay has not allowed clinicians to manage these patients as recommended by international guidelines. We describe here the clinical characteristics, access to PET/CT study and outcomes of patients with HL treated in Paraguay.
Methods: We conducted a retrospective cohort study of patients aged ≥17 years with newly diagnosed HL in 4 academic centers in Paraguay between 2015 and 2022, with follow-up until July 2023. Medical records were manually reviewed and data were abstracted in a standardized form. A historical analysis was performed to evaluate the utilization of PET/CT in all stages of HL treatment (baseline, intermediate and EOT). Survival probabilities were estimated using the Kaplan-Meier method.
Results: A total of 145 patients were identified; 101 had sufficient data for analysis (Table 1). Patients were young, with a median age of 29 years (16-79) and a slight male vs. female predominance (56.4% vs. 43.6%). Nodular sclerosis (76.6%) and mixed cellularity (20.2%) were the most frequent HL subtypes. More than half of the patients (58%) had ECOG 0, 50% advanced stage and 55.9% B symptoms. The majority of patients (97%) received an ABVD regimen as induction therapy. Sixty-nine percent of patients did not undergo PET/CT at baseline, most of them (97%) due to lack of insurance coverage, 16.5% underwent iPET and 58.4% underwent PET/CT at EOT. With a median follow-up of 44 months, overall survival (OS) at 4 years was 77% and progression-free survival (PFS) was 46%. In a subgroup analysis to investigate the short 4-year PFS observed in our cohort, the PFS of patients who underwent PET/CT at EOT versus those who did not was 84 versus 36, respectively (Figure 1).
Conclusions: To our knowledge, this is the first study to characterize the clinical characteristics and outcomes of patients with HL in Paraguay. It is striking to see that lack of access to PET/CT had a significant impact on PFS in our patient population. HL is a highly curable disease affecting young people, therefore, minimizing late side effects in this patient population (e.g., secondary comorbidities and malignancies) remains a high priority. The authors' conclusion is to raise awareness of the importance of improving access to diagnostic tools such as PET/CT to provide optimal care for our patients, especially those who do not have access to PET/CT.
Assessment of Ultra-Deep Mass Spectrometry Based Proteomics Compared to Flow Cytometry and RNA-Based Methods for the Discovery and Validation of Therapeutic Targets in Immune Cells
Authors: Au Yu-Wah , Rigter Gemma , Yu Wingyan , Satijn David , Mehnert Martin , Lachaud Amaury , Vowinckel Jakob , Neijssen Joost , Bon Helene 
Published: December 2023

Immuno-oncology (IO) has substantially improved the survival of cancer patients over the past several years encouraging the discovery of novel IO targets which are typically proteins expressed on the surface of immune cells. Sensitive quantification of proteins in complex biological samples is routinely achieved by immunoassays that use antibodies specific to target proteins. Such approaches can be a limitation in IO drug discovery since the development of de novo antibodies is associated with long lead times, high costs, and high failure rates, often resulting in the substitution of the protein readout by a surrogate transcript readout using RNA sequencing methods.
Protein quantification using mass spectrometry (MS) is agnostic to any capture agent and removes the barriers of availability or specificity of antibody-based assays. Furthermore, MS proteomics workflows can support large scale discovery studies with wide proteome coverage, and also represent an attractive antibody-free alternative in targeted quantitative studies for protein measurement.
The main purpose of this work was primarily to assess the sensitivity of DIA-based MS-proteomics platform for the deep proteome profiling of human primary immune cells by measuring the maximum protein coverage obtained across cell inputs ranging from 2 million down to single cells ( performance assessment). The secondary purpose of this work was to assess the accuracy of the quantification of targeted MS proteomics workflow in comparison with the gold standard QIFI® flow cytometry approach, and other methods using mRNA readouts in immortalized human hematological cell lines ( accuracy assessment).
We found that our MS-based proteomics workflows achieve high sensitivity in detection of immune cell markers down to few immune cells as input. Additionally, we report a strong correlation of the quantitative data derived from our MS-based proteomics workflows compared to the flow cytometry gold standard (r 2= 0.7354). Notably, we observed that this correlation was significantly higher than using transcriptomics approaches, emphasizing the benefit of substituting immunoassays for MS-based proteomics platforms in IO target discovery and validation.
Transcription Factor Redistributors Pharmacologically Actuate Non-Canonical Gene Networks to Drive AML Differentiation
Authors: Taylor Samuel , Stauber Jacob , Bohorquez Oliver , Schwenger Emily , Sundaravel Sriram , Farahat Abdelbasset , Bartholdy Boris , Boykin David , Poon Gregory , Steidl Ulrich 
Published: December 2023

Aberrant transcriptional networks are hallmarks of aging and cancer, yet our ability to target these aberrations is poor. PU.1 is one such transcription factor (TF) who's transcriptional networks are corrupted in disease, including in &gt;50% of Acute Myeloid Leukemia (AML) cases. In this study we investigate an unappreciated pharmacological approach to target the aberrant PU.1 network, by employing novel small molecules which competitively inhibit PU.1:DNA interactions.
We performed an extensive multiomics-driven, molecular characterization of AML cells following exposure to PU.1-DNA binding inhibitors (e.g. DB2115). Unexpectedly, we found that such compounds not only led to a reduction of many canonical PU.1 transcripts, but also (seemingly paradoxically) caused increases in alternative PU.1-driven transcripts. We discovered that this two-sided response was a result of robust cistromic repositioning of PU.1 chromatin binding rather than global inhibition. From kinetic CUT&Tag and ATAC sequencing studies, we identified that PU.1 redistribution involved rapid PU.1 loss (~1hr) followed by delayed PU.1 gain (~4-12hrs), causing subsequent opening of the chromatin, thus demonstrating endogenous PU.1's powerful pioneering ability. Through development of CLICK-on-CUT&Tag, we identified exclusive drug binding at displaced PU.1 binding sites, and identified unique, sequence-specific ETS motifs dictating PU.1 gain or loss. By employing these motifs in eGFP reporter assays and CRISPRd systems, we identified direct PU.1-mediated transcriptional control of known and novel functionally relevant genes, including MYC, POMP and STRAP. Finally, we establish that primary AML samples are sensitive to pharmacological PU.1 redistribution upon exposure to DB2115, and the rewiring of the PU.1 network in these cells drives differentiation down the myeloid lineage (Figure 1).
Overall, we uncover a novel biological phenomenon we describe as: Pharmacological transcription factor (TF) redistribution. “TF Redistributors”, such as DB2115 and next generation derivatives, are an unprecedented investigative and pharmacological tool which allow the study of complex and fast transcription factor dynamics without disrupting the structure or levels of the TF itself. Furthermore, we discover that pharmacological binding-site restriction and consecutive repositioning of the PU.1 network, cascades into an extensive rewiring of transcriptional circuits which ultimately drives myeloid differentiation of AML cells, potentially providing a novel therapeutic intervention for PU.1-corrupted AML.