bluuebunny/crossref_metadata_2025 huggingface dataset:
Lactate Metabolism-Related Genes to Predict the Clinical Outcome and Molecular Characteristics of Multiple Myeloma
Authors: Shu Lingling , Zhu Ziang , Li Yang , Huang Hanying , Chen Xiaoqin , Xia Zhongjun , Liang Yang
Published: December 2024

Background Metabolic reprogramming is a hallmark of cancer progression and holds great significance for the tumor microenvironment (TME). Lactate, as an abundant metabolite, has been found to play a critical role in the development of cancer and in the immunosuppression of TME. However, the potential role of lactate metabolism-related genes in multiple myeloma (MM) remains unclear.

Methods RNA sequencing data and clinical information of multiple myeloma (MM) were obtained from The Cancer Genome Atlas (TCGA) database. Lactate metabolism-related genes (LMRGs) were sourced from Molecular Signature Database v7.4, and then compared with the candidate genes from TCGA to obtain the final gene set. Prognostic genes were screened using univariate analysis and Least Absolute Shrinkage and Selection Operator (LASSO) Cox regression. A lactate metabolism-related risk profile was constructed using multivariate Cox regression analysis. The signature was validated by time-dependent ROC curve analysis and Kaplan-Meier analysis. Gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway functional analysis were performed to compare high-risk groups with low-risk groups. CCK8 assays were utilized to assess the proliferation ability of MM cells. Transwell assay was employed to detect their migration ability. Intracellular lactate content as well as glucose content was measured to evaluate lactate metabolism levels. Loss or gain-of-function studies were carried out using shRNA knockdown or lentiviral transduction strategies in order to investigate the precise role of LMRGs in MM.

Results We developed a risk signature based on 18 LMRGs. Kaplan-Meier curves confirmed that the high-risk group exhibited a poorer prognosis compared to the low-risk group. Subsequently, a nomogram was constructed to predict the probability of MM survival. Additionally, we conducted GO enrichment analysis and KEGG pathway functional analysis between the high and low-risk groups, which revealed significant associations with immune regulation and energy metabolism. Furthermore, we demonstrated that one of the lactate metabolism-related genes, JAMIP2, could promote myeloma cell proliferation, migration, and lactate metabolism via activating PI3K-Akt signaling pathway.

Conclusion In conclusion, this study identified and constructed LMRGs in MM, which showed high diagnostic accuracy in predicting OS in MM patients and linked to immunity and drug sensitivity. JAMIP2, which was identified as a novel molecule mediating lactate metabolism, promoted MM cell proliferation, migration and lactate metabolism through PI3K-Akt signaling pathway. Future studies are expected to validate the utility of the constructed LMRGs in MM patients.
CD163+ Macrophages Orchestrate Tumor Immunity By Spatially Fortifying Follicular Lymphoma Against CD8+ T Cells Attack during Early Progression
Authors: Lin Zhijuan , Yu Xingxing , Hong Yan , Liu Long , Li Zhifeng , Zha Jie , Xu Bing
Published: December 2024

B ack ground: The tumor immune microenvironment (TIME) plays a pivotal role in the development and progression of follicular lymphoma (FL). However, the evolution of TIME during progression of disease within 24 months (POD24) remains poorly understood.

Methods: We employed imaging mass cytometry with a panel of 36 meta-tagged antibodies to examine the immune architecture of lymph nodes from 13 FL patients, both at diagnosis and during POD24.

Results: Our analysis revealed a decrease in immune infiltration from the peri-follicular regions towards the follicular centers in FL. Notably, there was an increase in CD163+ macrophages within the follicles, accompanied by an upregulation of PD-1 ligands during POD24. Spatial analysis showed that FL cells tend to evade immune attacks not merely through spatial separation from CD8+ T cells but by proactively recruiting immunosuppressive cells prior to encountering these T cells. Furthermore, during POD24, more CD8+ T cells in proximity to FL cells were found interacting with macrophages rather than regulatory T cells (Tregs). There was also an observed increase in the cooperation between macrophages and Tregs, which collectively worked to subdue cytotoxic T cell activity, thereby creating a more immunosuppressed environment.

Conclusions: FL cells exhibit more aggressive evasion tactics and exist within a more immunosuppressive microenvironment during POD24. These findings suggest that immunotherapeutic strategies targeting PD-1, macrophages, and CTLA-4 may hold promise for improving outcomes in FL patients experiencing early progression.
Pro-Inflammatory Monocytes As Key Effector Cells in Peripheral Circulatory Inflammatory Storms Driven By IFN-γ and IL-6 Pathways in Imcd
Authors: Yin Xuejiao , Liu Yi , Tong Hongyan , You Liangshun
Published: December 2024

Objective: Dysfunctional cytokine-driven inflammatory storms in patients with idiopathic multicentric Castleman disease (iMCD) are a recurring issue affecting both symptoms and mortality, yet a thorough understanding of the involved immune cells remains incomplete. A comprehensive and detailed peripheral immune landscape is crucial for effectively treating iMCD.

Design: We employed single-cell RNA sequencing of 150936 cells from a cohort of 19 individuals, including 15 iMCD patients with mild or severe disease, as well as 4 healthy controls. For validation, we also enrolled a second cohort of 108 newly diagnosed iMCD patients with available IFN-γ data from 2015 to 2024.

Results: The activation and mobilization pervasive across all immune cells subtype during flares contribute to widespread systemic inflammatory storm in iMCD. Peripheral memory B C2 cells were the main source of plasma B and activated B cells in iMCD. Additionally, this cell subpopulation is where IL-6 mRNA is predominantly found in peripheral blood mononuclear cells (PBMCs). PADI/CCL/IL7R monocytes may be the primary effector cells in peripheral circulatory inflammatory storms. These cells exhibit the highest inflammatory scores, show the greatest activation of IL-6 signaling pathways, and overproduce VEGF-A, thereby being classified as pro-inflammatory monocytes. Furthermore, pro-inflammatory monocytes stimulated the overproduction of IL6 through promoting memory B C2 cell proliferation and transitions via BAFF signaling pathway, thus forming a vicious cycle that constantly stimulates the generation and expansion of inflammatory storms. In the cell-cell communication analysis, we identified IFN-II as a crucial signaling pathway, uniquely enriched in iMCD, with the highest relative strength index in both count and weight signaling patterns. Ligand-receptor analysis showed IFNG was mainly expressed in KLCRC2 NK, GZMK NK, NKT, and CD8TEX cells, whereas IFNGR1 and IFNGR2 were mainly expressed in pro-inflammatory monocytes. Besides, the level of IFNG expression was observed to correlate with iMCD severity. In the second iMCD cohort used for validation, IFN-γ levels were positively correlated with disease severity but negatively correlated with treatment response and progression-free survival (PFS).

Conclusion: This study provided an important resource for in-depth understanding the pathogenesis of iMCD and identify additional therapeutic strategies.
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 >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 (<1%) and Africa (<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.
Control of Macrophage Fusion By Sensing the Physical Properties of Fibrin(ogen) Matrices
Authors: Podolnikova Nataly P. , Balabiyev Arnat , Hur Woosuk S. , Flick Matthew J. , Ugarova Tatiana P.
Published: December 2024

Macrophage fusion is a salient and unwanted characteristic of implanted biomaterials. We have recently shown that macrophage fusion requires fibrinogen, as fibrinogen-deficient mice do not support this process. We also found that this is not fibrinogen itself but rather its clotting product, the fibrin polymer deposited on the surface of biomaterials, which drives macrophage fusion. This is evidenced by the data showing that macrophage fusion on biomaterials implanted into FibAEK mice expressing a mutated form of fibrinogen incapable of thrombin-mediated polymerization was strongly reduced. This phenotype was similar to fibrinogen-deficient mice, even though FibAEK is structurally similar to wild-type fibrinogen and normally adsorbs on the surface. It is well established that the physical properties of substrates control cellular functions through integrin-mediated cell adhesion and signaling. Fibrinogen adsorption on various surfaces at its physiological concentrations is known to reduce integrin-mediated adhesion and signaling in leukocytes and platelets by forming the nanoscale extensible multilayer matrix. To test the possibility that the fibrinogen multilayer cannot support macrophage fusion due to its inability to mediate firm adhesion and, consequently, transduce a competent mechanotransduction response in fusing macrophages, we examined the matrices prepared from wild-type fibrinogen and FibAEK. Adhesion, spreading, and podosome formation in macrophages were strongly reduced on both fibrinogens adsorbed on the biomaterial surface. We also used immunostaining to examine the localization of YAP, a sensor, and a molecular reporter of the ECM rigidity. On rigid substrates, YAP undergoes dephosphorylation and relocalization from the cytoplasm to the nucleus, and this regulation requires the tension of the actomyosin cytoskeleton developed in spread cells. We found that YAP localized exclusively in the cytoplasm of poorly spread macrophages attached to the fibrinogen multilayer matrix. In contrast, YAP was redistributed to the perinuclear region in macrophages that firmly adhered and spread on the fibrinogen monolayer, a model rigid substrate prepared by adsorption of low-density fibrinogen. IL-4-induced macrophage fusion exhibited a similar pattern with a lack of fusion in cells seeded on the fibrinogen multilayer and robust fusion in cells adherent to the monolayer. Furthermore, YAP remained in the cytoplasm in the non-fused mononucleated cells attached to the multilayer matrix while fully localized to the nucleus in multinuclear macrophages undergoing fusion on the monolayer. These data indicate that fibrinogen adsorption as a nonadhesive multilayer matrix on the surface of biomaterials discourages macrophage fusion, which has implications for controlling the foreign body reaction by modulating the mechanical properties of biomaterials.
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 (+/+ > +/- > -/-). 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<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 <30 ng/mL or iron saturation <20%, and hemoglobin <11 g/dL. Iron deficiency without anemia (IDWA) was defined as ferritin <30 ng/mL with hemoglobin >11 g/dL.

Results: Among 122 approached patients, 110 (90%) consented. Ferritin <30 ng/mL was found in 98/110 (89%) pregnant women and ferritin <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.
Heterozygous ddx41 Mutation Drive Age-Dependent Hematopoietic Stem and Progenitor Cell Dysfunction
Authors: Feliz Norberto Maria , Wang Yanhua , Wang Qing , Nizhnik Anastasia , Ulloa Bianca A , De Oliveira Sofia , Bowman Teresa V.
Published: December 2024

Myelodysplastic syndromes (MDS) are a group of blood cancers characterized by hematopoietic stem and progenitor cell (HSPC) expansion, ineffective hematopoiesis, myeloid cell dysplasia, cytopenia, and inflammation. Driving events for MDS are largely unknown, but clinically relevant to improve monitoring for at-risk patients and generate preventative therapies. Heterozygous, loss-of-function mutations in the ATPase DEAD-box Helicase 41 (DDX41) gene are common in inherited late-onset adult MDS, providing a compelling model to study the mechanisms preceding MDS. We established ddx41 heterozygous loss-of-function mutant zebrafish as an in vivo model that develops age-associated MDS features to study progressive changes in the hematopoietic system leading to disease. By 12-14 months, ddx41 heterozygous animals displayed erythroid dysplasia, HSPC expansion, and increased NFkB-active myeloid cells compared to wildtype siblings. Based on prior work, we hypothesized that the innate immune sensing cyclic GMP-AMP synthase (cGAS) - stimulator of interferon genes (STING) pathway that can respond to many damage-associated molecular patterns could be driving HSPC expansion. Indeed, HSPC frequency in 14-month-old ddx41 heterozygous; cgas homozygous mutants was significantly reduced compared to ddx41 heterozygous; cgas heterozygous mutants. To identify early arising deregulation that contributes to later MDS features in ddx41 heterozygous animals, we performed scRNA-seq on total marrow cells from 3-, 6-, and 14-month-old ddx41 wildtype and heterozygous animals. We identified gene signatures indicative of elevated cellular stress in HSPC in ddx41 heterozygous animals as early as 3 months. Our findings suggest that Ddx41 insufficiency promotes cellular distress that triggers age-associated cGAS-mediated HSPC expansion. These results have the potential to uncover new ways to prevent MDS progression and will also provide valuable insight into fundamental mechanisms controlling HSPC homeostasis.
Successful Generation of iPSC-Derived CD4+ and CD8+ CAR T Cells with Αβ-like T Cell Function, Including Antigen-Dependent Expansion and IL-2 Production
Authors: Heinze Dar , Sottoriva Killian , Dettloff Kristen , Chin Melissa , Gallagher Justin , Galuppo Alexa , Fread Kristen , Fang Justin , Hasselkus Ryan , Thomas Sarah , Ma Eric , Helou Ynes , Bauer David , Kurtz Jonathan , Whitney Dustin , Cowan Chad , Welstead G. Grant
Published: December 2024

Off-the-shelf CAR-T cell therapies promise to build on the efficacy of their autologous counterparts while providing patients with a well-defined, scalable, and cost-effective drug product that can be administered on-demand. Their successful development requires complex engineering to prevent graft vs host disease (GvHD) and rejection by the patient's immune system in addition to providing the cells with anti-tumor capabilities, such as tumor targeting. Induced pluripotent stem cells (iPSCs) can self-renew and have the potential to make any cell in the adult body. For this reason, the iPSC platform offers the opportunity to precisely engineer cells with desired characteristics that can be expanded, characterized, and banked for future differentiation to any desired cell type. To date, a major challenge for the application of this approach for making CAR T cell therapies has been the lack of a robust, scalable method to generate CAR T cells that function like healthy donor ab T cells. We report the first demonstration of an iPSC-derived CAR T cell that has αβ-like T cell function. iPSCs were engineered with a BCMA or CD19 CAR and differentiated using a novel, feeder-free method designed to mimic normal human T cell development. By successfully generating a pure CD4+/CD8+ double positive T cell population through controlled delivery of Notch ligands and other important developmental cues, we enabled the subsequent induction of both CD4+ and CD8+ T cells that have αβ-like properties. At the end of this process, a mix of CD4+ and CD8+ iPSC CAR T cells displayed Ag-dependent cytotoxicity, proliferation, and cytokine production, including IL-2 that was comparable to primary CAR T cell levels. Importantly, these attributes resulted in robust and prolonged control of tumor cells in vitro without the need for exogenous cytokines as well as tumor control in standard in vivo tumor models. These data provide support for the use of this platform for the development of allogeneic CAR T cell medicines.
Donor T-Derived Memory Progenitors Differentiated into CD103+/- CXCR6+CD4+ Tissue-Resident T Cell Subsets Driven Cutaneous Chronic Graft-Versus-Host Disease
Authors: Kong Xiaohui , Wang Bixin , Wu Xiwei , Fu Weijia , Li Qinjian , Zheng Moqian , Pillai Raju , Nakamura Ryotaro , Martin Paul J. , Zeng Defu
Published: December 2024

Cutaneous chronic graft versus host disease (cGVHD) is the major clinical manifestation of cGVHD patients. Tissue-resident memory T (Trm) cells and TCF1+ T progenitors locally in GVHD target tissues play important roles in maintaining GVHD. However, the mechanisms regulating TCF1+ progenitor differentiation into Trm in the skin remain largely unknown. In the current studies, with cGVHD murine models (C57BL/6 donors and BABL/c recipients) and humanized models of human HLA-A2-DR4- PBMC to MHC-/-HLA-A2+DR4+ NSG mice, we observed with flow cytometry analysis that most (>90%) of the injected donor CD4+ T cells in the skin tissues were CD69+ Trm, and the TCF1+ progenitors were less than 4%, which was markedly lower than in the liver and lung (>20%). The majority of Trm cells in the cGVHD skin were CD103+CXCR6+Foxp3- conventional CD4+ T cells that produce large amount of pro-inflammatory IFN-γ and GM-CSF, while CD103+ Trm cells in the skin of non-GVHD recipients were mostly Foxp3+ Treg cells. Genetic deficiency of T-bet, BHLHE40, STAT3 or BATF in the injected donor T cells markedly ameliorated skin cGVHD, all associated with lower numbers of CD103+ Trm cells and lower production of IFN-g/GM-CSF. Results were similar when skin cGVHD was greatly reduced by post-transplantation treatment with cyclophosphamide (PTCY) in murine models. In addition, scRNA-seq analysis of blood and skin T cells from GVHD patients in a public dataset showed that ~90% blood T cells express stemness related marker Tcf7 and large proportions of skin T cells (>95%) express terminally differentiated markers such as Cxcr6. Taken together, our results suggest that the differentiation of TCF1+ T progenitor cells into Trm cells in the skin is regulated by the T-bet/BHLHE40 axis and the TCR/STAT3/BATF axis. Although Trm cells are differentiated from TCF1+ T progenitor cells in the skin, the TCF1+ T progenitor cells in blood may play an important role in replenishing TCF1+ T progenitor cells in cGVHD target tissues.
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 (>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.
Clonal Hematopoiesis of Indeterminate Potential Does Not Impact Overall Survival in Multiple Myeloma
Authors: Rexha Farah , Gustine Joshua , Han Ryan , Nakamoto-Matsubara Rie , Panaroni Cristina , Shome Rajib , Nardi Valentina , Branagan Andrew R. , Cirstea Diana , Yee Andrew J. , Raje Noopur
Published: December 2024

Introduction:

Clonal hematopoiesis of indeterminate potential (CHIP) is frequently observed in patients with multiple myeloma (MM). A previous study demonstrated that CHIP is associated with shorter progression-free survival in MM patients undergoing autologous stem cell transplantation (ASCT), particularly those who do not receive maintenance therapy with an immunomodulatory agent (Mouhieddine Nat Commun 2020). We sought to further characterize the impact of CHIP on overall survival (OS) in a large cohort of MM patients, as well as the risk of developing a myeloid neoplasm.

Methods:

Study participants include MM patients evaluated at the Massachusetts General Hospital between January 2019 and December 2022 who underwent targeted exome sequencing (i.e., Heme SNaPshot) on DNA extracted from unselected bone marrow for CHIP-associated somatic mutations. CHIP was defined by the presence of a leukemia-associated mutation with a variant allele fraction >2% in unselected bone marrow aspirate samples. Medical records were manually reviewed for all patients to extract data on baseline clinical characteristics and survival outcomes. Time-to-event outcomes were estimated using the Kaplan-Meier method, and comparisons were made with the log-rank test. Cox-proportional hazard regression analyses were performed to fit univariate and multivariate models for OS; the outcome measure was hazard ratio (HR) with 95% confidence interval (CI). OS was defined as the time from CHIP diagnosis to death or last follow-up. Cumulative incidence estimates for myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) were made with the Fine-Gray method to account for death as a competing risk. Only patients who consented to mutational testing were included in this analysis and does not represent all consecutive MM patients seen at our institution.

Results:

A total of 317 patients with MM underwent gene sequencing, of whom 167 (53%) had CHIP. By univariate analysis, patients with CHIP had a significantly shorter median OS (4.2 years versus not reached; p<0.001). However, on multivariate analysis, CHIP was not associated with OS (HR 1.40, 95% CI 0.83-2.38; p=0.21) after adjusting for age, sex, time from MM diagnosis, tumor burden, extramedullary disease, high-risk cytogenetics, R-ISS, prior therapy, albumin, LDH, and beta-2 microglobulin level. Four patients (1.3%) developed MDS/AML in this cohort, of whom 3 out of 4 patients (75%) had CHIP. Between patients with and without CHIP, there was no statistical difference in the risk of developing MDS/AML after both the diagnosis of MM and time of mutational testing when accounting for death as a competing risk (Fine-Gray p=0.49 and p=0.33, respectively).

Conclusion:

The presence of CHIP is not associated with OS and does not appear to be associated with risk of myeloid neoplasms in MM patients.
Cardiovascular Outcome of Heart Failure Patients with Iron Deficiency Anemia Who Received Iron Infusion Vs Red Blood Cell Transfusion: Analysis from a Nationwide Trinetx Database
Authors: Elom Hilary A , Kulasekaran Monika , Adeniran Lanre , Ilonze Onyedika J
Published: December 2024

Background: Heart Failure (HF) is a significant global health burden, and Iron deficiency anemia (IDA) can worsen outcomes in HF. About 30-50% of HF patients are reported to have IDA, commonly due to nutritional deficiencies, gastrointestinal loss, impaired absorption, and chronic inflammatory cytokines present in HF. HF guidelines recommend IV iron to reduce HF hospitalization. This study aims to evaluate the cardiovascular outcome of HF patients with IDA treated with iron infusion vs RBC transfusion.

Methods: A nationwide retrospective cohort study was conducted using the TriNetX platform, analyzing adult HF patients with IDA from 2015 to 2023. The intervention cohort 1 received iron infusions without packed RBCs, while the control cohort 2 received packed RBCs without iron infusions. Cardiovascular and cerebrovascular outcomes, ICU admissions, and mortality were assessed from 1-month post-intervention. Cohorts were matched 1:1 using Propensity Score Matching (PSM) based on demographics, cardiovascular risk factors, chronic medical conditions, and nutritional status to minimize confounding variables in comparing the two treatment modalities.

Results: After propensity score matching, a total of 20,862 study participants were included in each study cohort. The mean age of cohort 1(patients who received iron infusion with no blood transfusion) and cohort 2(those who received RBC transfusion with no iron infusion were 69.2 +/- 15.0 and 69.3 +/-14.5 respectively. Majority of the patients were White (68%) and female (55%) in both cohorts. Patients in cohort 1 were 20 % less likely to have unstable angina, PCI/coronary angioplasty and acute myocardial ischemia compared with cohort 2 (OR 0.80; 95% CI 0.74 - 0.86). Myocardial infarctions were less likely to occur in patients in cohort 1 (OR 0.71; 95% CI 0.66 - 0.76) compared with patients in cohort 2. Additionally, Patients in Cohort 1 also demonstrated a significantly lower odd of composite cerebrovascular disease (stroke, TIA, carotid intervention/angioplasty) (OR 0.45; 95% CI 0.46 - 0.54) compared to those in cohort 2. The analysis further revealed a significant association with ICU admission (aOR 0.011; 95% CI 0.07 - 0.19) and mortality (aOR 0.99; 95% CI 0.287 - 0.312).

Conclusion: HF patients with IDA receiving iron infusion without RBC transfusion exhibited significantly lower risks of composite adverse cardiovascular and cerebrovascular outcomes compared to those receiving RBC transfusion without iron infusion. This supports IV iron's benefit in HF, reducing ICU hospitalization, mortality and adverse composite outcomes.
Mutations and MRD: clinical implications of clonal ontogeny
Authors: Radich Jerald
Published: December 2024

Measurable residual disease (MRD) is a strong but imprecise predictor of relapse in acute myeloid leukemia. Many patients fall into the outlier categories of MRD positivity without relapse or MRD negativity with relapse. Why? We will discuss these states in the context of “clonal ontogeny” examining how mutations, clonal structure, and Darwinian rules impact response, resistance, and relapse.