bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeted Lipid Nanoparticle Delivery of an RNA Gene Writer In Vivo Enables Generation of CAR-T Cells in a Humanized Mouse Model
Authors: Rodriguez Jason , De Iaco Alberto , Rottman James B. , Liu Donghui , Venugopal Kartika , Qiao Xin , Cao Yu , Levy Rebecca , Gupta Mohit , Palchaudhuri Rahul , Querbes Bill , Cotta-Ramusino Cecilia , Pujar Hari , Peshwa Madhusudan V , Holmes Michael C 
Published: December 2024

The FDA approval of autologous ex vivo Chimeric Antigen Receptor (CAR) T-cell therapies have added a powerful tool to treat relapsed and refractory leukemias and lymphomas. However, significant challenges limit access to these therapies including long vein-to-vein times for drug product delivery, viral vector supply chain limitations, cumbersome manufacturing processes and high costs. Allogeneic CAR-T therapies have not achieved the same level of therapeutic efficacy as conventional autologous T cell therapies highlighting the critical unmet need for same-day in vivo treatment options.
RNA Gene Writers leverage target-primed reverse transcription (TPRT) biochemistry that evolved from non-LTR retrotransposons to modify the genome of cells using RNA templates. Moreover, RNA Gene Writers can be engineered to catalyze a variety of editing reactions, such as writing or introducing gene-length DNA sequences, as well as rewriting or making single nucleotide substitutions or small insertions or deletions to the genome. These edits can be achieved by delivering all-RNA compositions to primary cells in vitro or in vivo, eliminating the need for viral vectors and DNA template-based genome editing.
Initial testing of T cell specific LNPs containing a Gene Writer with a GFP or CAR template in vitro led to an observed &gt;70% GFP+ and ~40% CAR+ T cells, respectively, in primary human T cells t. Moreover, these CAR+ T cells can clear antigen specific tumors in vivo in human xenograft mice mediating robust cytotoxic response and CAR-T cell expansion.
For in vivo applications, targeting and editing of resting T cells is a critical milestone to overcome. Optimization of our T cell specific LNPs and delivery to freshly isolated human primary T cells in vitro, without pre-activation, resulted in an average of 21% CAR expression. CAR+ cells generated in this manner are potent in their ability to drive cytotoxicity, cytokine production and CAR-T cell expansion in response to multiple tumor types.
In proof-of-concept studies, we achieved transient delivery of GFP mRNA to approximately 80% and 40% of T cells in humanized mouse and non-human primate (NHP) models, with minimal delivery to B cells. We next validated our in vitro CAR-T results in various in vivo models. Studies in humanized NSG mice achieved an average of 6% CAR writing in implanted human T cells when using T cell targeted LNPs. CAR writing in other relevant tissues, such as B cells and liver, was not observed. The ability to generate in vivo CAR-T cells using Gene Writers and our proprietary LNP delivery platform provides a unique opportunity to potentially eliminate the challenges associated with conventional ex vivo CAR-T cell therapies and viral based in vivo CAR-T gene therapies.
Frontiers in Myeloma: Barriers to Clinical Trial Participation in Multiple Myeloma
Authors: Kaiser Martin F , Ul-Hasan Saba , Lewis Jack , Mirto Federica , O'Neill Alicia , Gamertsfelder Elise , Biru Yelak 
Published: December 2024

Background
Clinical trials provide an opportunity for patients to access new treatments and receive expert medical care, which potentially improves the quality and length of their lives. By participating in trials, patients help further medical research, thereby improving treatments for others. However, patient participation in trials remains low due to multifaceted barriers related to trial design, patient factors, and attitudes of healthcare professionals.
Frontiers in Myeloma (FIM) is a multistakeholder initiative led by an international group of myeloma patient advocacy thought leaders who work to elucidate and describe the most pressing equality and equity issues that impact patients with myeloma. This study identified current barriers to clinical trial access in multiple myeloma (MM) across different jurisdictions and populations and provided practical recommendations for stakeholders to help facilitate access and participation.
Methods
Clinical trial registries (WHO, ClinicalTrials.gov, and EUCTR) were searched for clinical trials in MM that were registered between 2011 and 2021. Data were analyzed to determine the countries in which MM clinical trials recruited patients and how many trials took place. A systematic literature review was conducted in line with PRISMA recommendations to identify barriers and facilitators to enrollment in MM clinical trials. Three electronic databases were searched since their inception: MEDLINE, Embase, and APA PsycInfo.
Primary research was conducted through stakeholder interviews to identify barriers and initiate discussion on the types of recommendations and feasibility of their implementation. Interview questions were based on preliminary findings from the literature review; questions assessed participants' involvement in MM clinical trials and their views on the barriers and facilitators to patient enrollment. Key stakeholders involved in MM trials, including patients, patient advocates, clinicians, researchers, and clinical nurses, were identified and interviewed via Zoom.
Findings from the primary research were used to develop recommendations for stakeholders to address the identified barriers to clinical trial access and participation for patients with MM.
Results
Clinical trial analysis for regional data identified 3141 MM trials. Most trials were conducted in the US (61%), followed by Western Europe (24%), Southern Europe (14%), and East Asia (13%). Geographic regions in which the fewest trials were conducted were South Asia (&lt;1%) and Africa (&lt;1%).
Various barriers were identified in the systematic literature review, including clinical, patient, structural, and physical barriers. Stringent eligibility criteria and lack of geographic access were the most frequent barriers reported.
Funding: This study was sponsored byGSK. Editorial support was provided by to Nucleus Global, an Inizio Company and funded by GSK.
Blockade of Angiotensin II Signaling Ameliorates Cardiac Fibrosis and Improves Diastolic Dysfunction in Mice with Sickle Cell Anemia
Authors: Little Keara , Zhu Zhenqi , Gbotosho Oluwabukola , Malik Punam 
Published: December 2024

Introduction: Cardiopulmonary complications are the most common cause of death in adults with sickle cell disease (SCD). We previously reported a unique cardiomyopathy with restrictive physiology due to myocardial fibrosis, resulting in diastolic dysfunction and predisposition to arrhythmias, both in mice (Bakeer et al, PNAS 2016) and humans (Niss et al, Blood 2017) with SCD. However, the underlying mechanism resulting in myocardial fibrosis is unknown. We have also reported high Angiotensin-II (Ang-II) levels in mice and humans with SCD, in the absence of hypertension (Roy et al, AJH 2018). Herein, we hypothesized that increased Ang-II signaling promotes sickle cardiomyopathy and can be reversed by blocking this signaling in SCD mice.
Methods: We blocked Ang-II receptor-I (ATR1) signaling in 5-8-week-old Townes SCD mice with losartan mixed in drinking water, or vehicle control for 3 months, 6 months, or 9 months in different experiments. Cardiac parameters were measured using echocardiograms and electrocardiograms. IL-18 and Galectin-3 levels were measured as plasma biomarkers of cardiomyopathy by ELISA, and qRT-PCR was performed on myocardial mRNA for candidate genes. At sacrifice, cardiac histopathology was performed using H&E staining, as well as staining for fibrosis using Mason-trichrome and Sirius red.
Results: Echocardiograms showed that there was improvement in diastolic function and restrictive physiology with losartan treatment. There was a significant reduction in left atrium size (P=0.04), and left ventricular hypertrophy (P=0.0003), and an increase in isovolumetric relaxation time (P=0.0006) in losartan-treated SCD mice compared to vehicle-control SCD mice. Electrocardiograms showed significantly shortened QRS duration (P=0.001) with losartan treatment. There was a significant reduction in CTGF (connective tissue growth factor) mRNA expression (P=0.03), and plasma levels of Galectin-3 (P=0.07) and IL-18 (P=0.0005). Histopathology confirmed reduced intramyocardial fibrosis in losartan-treated mice, which was quantified using Aperio ImageScope64 software (P=0.01). All of these occurred despite more anemia in losartan-treated vs. control SCD mice, as ATR1 signaling enhances erythropoiesis. We also treated losartan and control groups of mice with biweekly blood transfusions and compared them to the corresponding non-transfused mice for 3 months. With this, we observed a similar reduction in IL-18 and Galectin-3, although the duration of treatment was not sufficient to show differences in myocardial fibrosis. Losartan treatment resulted in reduced oxidative stress, which may have contributed to the improved cardiomyopathy.
Conclusion: Ang-II signaling plays an important role in the development of cardiac fibrosis and its downstream complications in SCD patients, which can be reversed by blockade of ATR1.
An Inactivating Mutation in Rat IL-34 Reveals Redundant and Nonredundant Functions of CSF1-IL-34 Signalling in Homeostasis and Inflammation
Authors: Huang Stephen , Patkar Omkar , Schulze Sarah , Carter-Cusack Dylan , Ranpura Ginell , Maxwell Emma , Keshvari Sahar , Millard Susan , Cowin Gary , Kurniawan Nyoman , Pettit Allison , Irvine Katharine M , Hume David A 
Published: December 2024

Colony stimulating factor 1 receptor (CSF1R) is required for the differentiation and maintenance of many mononuclear phagocyte lineage cells and is a definitive marker of this lineage in humans and mice. CSF1R signaling is activated by two cognate ligands CSF1 and IL34. In mice, the expression of CSF1 and IL34 is spatially regulated in homeostasis and increased in inflammatory conditions. Whereas CSF1 expression is ubiquitous, IL34 expression is predominantly observed in the epidermis and central nervous system. Accordingly, IL34-deficient mice lack Langerhan's cells and grey matter microglia. CSF1-deficient mice lack many tissue macrophage populations, but microglia and Langerhan's cells are present. In other species, including humans and rats, IL34 is more broadly expressed, raising the possibility that IL34 plays distinct roles, or that there is more redundancy between CSF1R ligands, in species other than mice.
We generated and characterised a novel IL34 knockout rat. IL34 KO rats are phenotypically indistinguishable from wildtype littermates from birth up to 2 years of age. Similar to mice, grey matter microglia are significantly reduced in IL34-deficient animals in a dose dependent manner (+/+ &gt; +/- &gt; -/-). The loss of grey matter microglia was not associated with any observable histological pathology or behavioural abnormalities. IL34 KO rats also have a reduction in the number of Langerhan's cells, but do not completely lack them, unlike IL34 KO mice. Blood and bone marrow monocytes and all other tissue macrophage populations examined were indistinguishable between IL34 KO rats and littermate controls. To investigate the role of IL34 in tissue injury we utilised an adenine diet-induced chronic kidney disease (CKD) model, since Il34 is relatively highly expressed in the rat kidney. Both Il34 and Csf1 were induced in chronically injured kidneys. IL34 KO rats had reduced macrophage numbers in injured kidneys, but the KO did not affect pathology at peak injury or recovery after cessation of the adenine diet. We conclude that IL34 is largely redundant for the maintenance of monocyte-macrophage lineage cells and normal physiological homeostasis at steady state in rats.
Towards Remote Monitoring of Sickle Cell Disease for Heart Failure: A Single Lead ECG-AI Model
Authors: Karabayir Ibrahim , Davis Robert L. , Chinthala Lokesh , Rai Parul , McGuire Dan , Hankins Jane S , Akbilgic Oguz 
Published: December 2024

Background
Cardiopulmonary dysfunction, including heart failure (HF), is a leading cause of death in patients with sickle cell disease (SCD). Early detection of HF in SCD patients is critical for timely intervention and improved outcomes. Conventional diagnostic modalities such as echocardiograms and cardiac MRI are often difficult to obtain in resource-limited environments or remote settings. There is a need for low cost and widely accessible tools to assist with early recognition of HF among SCD patient.
Hypothesis
We hypothesize that artificial intelligence (AI) models using single lead ECG as an input can accurately detect SCD patients with HF.
Methods/Approach
An ECG-AI model based on convolutional neural networks was previously developed to classify HF patients, utilizing a large ECG repository from Wake Forest Baptist Health (WFBH). This model was trained on 1,078,198 digital Lead I ECG of standard 12 lead 10 seconds clinical ECG records from 165,243 patients, with a demographic distribution of 73% White, 19% Black, and 52% female individuals, and a mean age (SD) of 58 (15) years. An AUC of 0.84 was achieved in identifying HF patients from controls in a hold-out validation - unseen subset of WFBH. In the current study, the ECG-AI model was externally validated on data from SCD patients at the University of Tennessee Health Science Center (UTHSC). Additionally, a logistic regression (LR) model was constructed for the UTHSC cohort, incorporating basic demographic variables along with the outcomes of the ECG-AI model.
Results/Data
The external validation cohort from UTHSC comprised data from 2,107 SCD patients, including 188 with HF and 1,919 without HF. The cohort was predominantly Black (98%), with 72% female individuals and a mean age of 39 (SD 14) years. Despite notable demographic differences-such as a higher proportion of Black patients and a younger average age compared to the derivation cohort-our single lead ECG-AI model yielded an AUC of 0.80 (0.77-0.82) in detecting HF within the UTHSC SCD population. When the ECG-AI outcome, representing an ECG-based risk score between 0 and 1, was combined with age and sex in a logistic regression (LR) model, a moderate improvement in AUC was observed (DeLong Test, p&lt;0.01), reaching 0.82 (0.79-0.84) with a sensitivity of 0.73 (0.70-0.76) and a specificity of 0.73 (0.71-0.76).
Conclusion
The ECG-AI model has demonstrated moderately high accuracy in detecting HF in SCD patients using single Lead I of clinical 12-lead ECGs. Knowing that this is the lead mimicked by smartwatches and other wearables with ECG functionality, this study shows feasibility of remote monitoring of SCD patients for HF risk. Future studies will focus on calibration and testing of our ECG-AI models specifically for SCD patients and smartwatch ECGs.
Iron Deficiency in Pregnancy Is Ubiquitous
Authors: Fein Steven , Alonso Dayne , Campos Gloria , Lewkowitz Adam K , Van Doren Layla N , Auerbach Michael , Peralta Gina , Peralta Nicole 
Published: December 2024

Background: Iron deficiency anemia (IDA) is highly prevalent in pregnancy. It has been associated with increased maternal morbidity, peripartum complications, and impaired cognitive development in the offspring. Yet little attention has been given to IDA's antecedent, iron deficiency (ID).
The American College of Obstetricians and Gynecologists (ACOG) recommends screening anemic pregnant patients for iron deficiency (ID) but recommends against screening for ID among non-anemic pregnant women. Nevertheless, symptoms such as fatigue that are typically attributed to pregnancy physiology may in fact be attributed to ID.
In this study, we sought to quantify the prevalence of ID and fatigue symptoms in pregnant women at 23-29 weeks' gestation and to examine whether ID was associated with increased risk of fatigue symptoms compared to those without ID.
Methods: We conducted a cross-sectional study of low-risk pregnant women receiving care at two large community obstetric practices. 75% identified as Hispanic/Latino, and 6% identified as African-American. Pregnant women were approached at their laboratory appointment for gestational diabetes screening (23-29 weeks' gestation). Consenting patients had complete blood count, ferritin, and iron saturation measured, and each completed the Functional Assessment of Chronic Illness Therapy (FACIT)-Fatigue scale to evaluate for ID-related fatigue. IDA was defined as ferritin &lt;30 ng/mL or iron saturation &lt;20%, and hemoglobin &lt;11 g/dL. Iron deficiency without anemia (IDWA) was defined as ferritin &lt;30 ng/mL with hemoglobin &gt;11 g/dL.
Results: Among 122 approached patients, 110 (90%) consented. Ferritin &lt;30 ng/mL was found in 98/110 (89%) pregnant women and ferritin &lt;15 ng/mL was found in 76/110 (69%) pregnant women. IDA and IDWA were present in 24/110 (22%) and 76/110 (69%). The rate of screening positive for fatigue on the FACIT-Fatigue scale was similarly high among patients with IDA (10/24; 42%) and IDWA (42/76; 55%), compared to 20% (2/10) in patients without iron deficiency. Only nine participants (8%) had neither anemia nor ID, and only one patient with anemia did not have ID.
Conclusion: In this cross-sectional study of low-risk pregnant women at 23-29 weeks' gestation receiving prenatal care from community-based obstetricians, we found that iron deficiency (ID) is ubiquitous in pregnancy, with nearly 90% of pregnant women exhibiting low ferritin levels. This would frequently be missed if anemia were prerequisite for iron testing, as recommended by ACOG guidelines. We also found that pregnancy-associated fatigue is common in women with ID, including many of those who have IDWA. These findings highlight the need for routine iron deficiency screening in pregnancy to ensure timely intervention and optimize maternal and fetal health outcomes.
High HLA Evolutionary Divergence in Donor HLA-B Locus Impacts Transplant Outcomes after Haploidentical Hematopoietic Cell Transplantation with Post-Transplant Cyclophosphamide
Authors: Solh Melhem M. , Solomon Scott R. , Roark Christina , Aubrey Michael T , Morris Lawrence E , Holland H Kent , Bachier-Rodriguez Lizamarie , Freed Brian , Bashey Asad 
Published: December 2024

The human leukocyte antigen (HLA) divergence allele advantage hypothesis states that an HLA genotype with two alleles that are more divergent results in more diverse presentation of immunopeptidomes. Higher divergence in the recipient after transplantation from a matched donor, will allow more tumor associated antigen s (TAA) and potentially lower risk of relapse. On the contrary, low HLA evolutionary divergence (HED) in HLA class I and HLA-DR were found to be strong predictors for worse overall survival after matched related donor transplantation. In the haploidentical transplantation (HIDT) setting with post-transplant cyclophosphamide (PTCY) , the level divergence in the donor can have an impact on the ability of donor cells to recognize recipient TAAs and potentially affect transplant outcomes. Additionally, Antigen presenting cells from donor have an impact on graft versus tumor effect post allogeneic transplantation (Jiang Ming Li, 2004). To study the effect of HED in donor HLA-B and its impact on transplant outcomes, we studied 322 consecutive recipient/donor pairs who received their transplant at our center with a median follow up of 69.2 months (range 30-208). HED was calculated as pairwise differences using Grantham distance metric by perl script (https:// sourceforge.net/projects/granthamdist/). We hypothesized that a higher HED score in donor HLA-B will improve survival post HIDT.
Data were collected from our database where it was prospectively entered. Patient diagnosis included AML (n=122), ALL (n=68), MDS (n=41), NHL (n=50), other (n=41). Baseline recipient characteristics included a median age of 50 years (19,80), 53% white, HCT-CI≥3 in 50%, Disease risk (intermediate 53%, high/very high 33%), PBSC graft 80% and myeloablative conditioning 49%. Donor characteristics were median age 37 years (13,73), CMV positive 61%, male 61% and female donor to male recipient 21%. Pairwise divergence of donor HLA -B showed the median HED score of 7.93 (0-14.33). On univariate analysis using median cutoffs, a high HED (&gt;7.93) in donor HLA-B was associated with better OS ( HR 0.71, p=0.05), DFS ( HR 0.71, P=0.05), non-relapse mortality NRM (HR 0.51, p=0.03) and similar relapse ( HR 0.84, p=0.4) compared to lower HED scores. A multivariable analysis adjusting for factors such as disease risk, sex, comorbidity score and race, a high HED in donor HLA-B was significant for lower risk of non-relapse mortality (HR 0.509, p=0.04) compared to lower HED.
In conclusion, our data show that among HIDT recipients, a high HED in donor HLA-B has significant impact on transplant outcomes and survival by improving NRM. The benefit of higher HED in HLA-B among donors can be used to select the best donor to lower transplant mortality
Single-Cell Analysis Reveals an Exhaustion of TEM and CTL CD8+ T Cells in Hemophagocytic Lymphohistiocytosis with EBV Infection
Authors: Luo Xiaodan , Lai Wenpu , Liu Jianbo , Shi Boyun , Tang Jianli , Yuan Danyun , Chen Ao , Huang Jiayu , Tan Huo , Zheng Runhui 
Published: December 2024

Background
Hemophagocytic lymphohistiocytosis (HLH) is a relatively rare and lethal syndrome characterized by uncontrolled hyperinflammatory reaction, often associated with Epstein-Barr virus (EBV) infection, which significantly complicates treatment approaches, posing challenges in studying T cell function across different stages of disease. T cell exhaustion is well-documented in chronic viral infections, yet its specific role remains unidentified in HLH with persistent EBV infection and inflammation.
Methods
We analyzed peripheral blood single-cell RNA sequencing (scRNA-seq) data from 15 individuals, including 3 healthy controls and 6 HLH patients with EBV infection, categorized into acute and remission phases. In total, 125, 737 cells including 30 cellular subtypes were analyzed. EBV-DNA level in different lymphocyte subsets was determined by flow cytometry and quantitative polymerase chain reaction (qPCR).
Results
We found that EBV infected various cellular subtypes in hematopoietic system including a minor population of hematopoietic stem cells (HSCs). Compared to individuals with EBV-infected B cells or CD4+ T cells, more cells expressing EBV-related genes were found in those with EBV-infected natural killer (NK)/NKT cells. Interestingly, there was a rapid increase in cells expressing EBV-related genes in individuals experiencing HLH relapse compared to those at initial diagnosis, particularly among those who progressed to lymphoblastic leukemia, where copy number variation (CNV) in lymphocyte cells often occurs. Specifically, CD8+ T effector memory (TEM) cells and cytotoxic T cells (CTL) in patients exhibited terminal exhaustion state characterized by increased expression of PDCD1, TIGIT, LAG3, HAVCR2, and TOX in HLH with EBV infection as compared to controls. Although improvement of exhaustion state was observed during disease remission in some cases, levels of exhaustion markers remained significantly higher than in healthy individuals. Furthermore, lymphocytopenia and a relative increase in myeloid cells persisted even after disease remission. Notably, myeloid cell subtypes, particularly monocytes and dendritic cells, exhibited suppressed antigen presentation and reduced cytokine secretion.
Conclusion
Through comprehensive analysis of PBMCs in HLH, we found that EBV may induce CD8+ TEM and CTL exhaustion and suppress the function of antigen presentation cell, contributing to HLH pathogenesis. Persistent or recurrent EBV elevation emerges as a pivotal factor in HLH relapse and progression. These insights offer crucial data for understanding and potentially targeting the mechanisms underlying this rare disease.