bluuebunny/crossref_metadata_2025 huggingface dataset:
Supplementary Figure 8 from SOX10 Loss Sensitizes Melanoma Cells to Cytokine-Mediated Inflammatory Cell Death
Authors: Rosenbaum Sheera R. , Caksa Signe , Stefanski Casey D. , Trachtenberg Isabella V. , Wilson Haley P. , Wilski Nicole A. , Ott Connor A. , Purwin Timothy J. , Haj Jelan I. , Pomante Danielle , Kotas Daniel , Chervoneva Inna , Capparelli Claudia , Aplin Andrew E.
Published: February 2024

Other cytokines have less effect on GSDME cleavage in SOX10/Sox10-knockout cells
Supplementary Figure 7 from SOX10 Loss Sensitizes Melanoma Cells to Cytokine-Mediated Inflammatory Cell Death
Authors: Rosenbaum Sheera R. , Caksa Signe , Stefanski Casey D. , Trachtenberg Isabella V. , Wilson Haley P. , Wilski Nicole A. , Ott Connor A. , Purwin Timothy J. , Haj Jelan I. , Pomante Danielle , Kotas Daniel , Chervoneva Inna , Capparelli Claudia , Aplin Andrew E.
Published: February 2024

Caspase-dependent GSDME cleavage is observed concomitant with cytokine-mediated cell death
Supplementary Figure 4 from SOX10 Loss Sensitizes Melanoma Cells to Cytokine-Mediated Inflammatory Cell Death
Authors: Rosenbaum Sheera R. , Caksa Signe , Stefanski Casey D. , Trachtenberg Isabella V. , Wilson Haley P. , Wilski Nicole A. , Ott Connor A. , Purwin Timothy J. , Haj Jelan I. , Pomante Danielle , Kotas Daniel , Chervoneva Inna , Capparelli Claudia , Aplin Andrew E.
Published: February 2024

SOX10-knockout does not consistently alter cytokine receptor expression
Supplementary Figure 3 from SOX10 Loss Sensitizes Melanoma Cells to Cytokine-Mediated Inflammatory Cell Death
Authors: Rosenbaum Sheera R. , Caksa Signe , Stefanski Casey D. , Trachtenberg Isabella V. , Wilson Haley P. , Wilski Nicole A. , Ott Connor A. , Purwin Timothy J. , Haj Jelan I. , Pomante Danielle , Kotas Daniel , Chervoneva Inna , Capparelli Claudia , Aplin Andrew E.
Published: February 2024

SOX10-knockout does not strongly affect the expression of cytokine signaling mediators
Supplementary Figure 2 from SOX10 Loss Sensitizes Melanoma Cells to Cytokine-Mediated Inflammatory Cell Death
Authors: Rosenbaum Sheera R. , Caksa Signe , Stefanski Casey D. , Trachtenberg Isabella V. , Wilson Haley P. , Wilski Nicole A. , Ott Connor A. , Purwin Timothy J. , Haj Jelan I. , Pomante Danielle , Kotas Daniel , Chervoneva Inna , Capparelli Claudia , Aplin Andrew E.
Published: February 2024

SOX10/Sox10-knockout does not alter sensitivity to cell death mediated by other cytokines
dataset 1 from Integrating Clinical Cancer and PTM Proteomics Data Identifies a Mechanism of ACK1 Kinase Activation
Authors: Balasooriya Eranga R. , Madhusanka Deshan , López-Palacios Tania P. , Eastmond Riley J. , Jayatunge Dasun , Owen Jake J. , Gashler Jack S. , Egbert Christina M. , Bulathsinghalage Chanaka , Liu Lu , Piccolo Stephen R. , Andersen Joshua L.
Published: February 2024

Supplementary dataset 1. Potential driver mutations identified against cancer types.
Data from ORAOV1, CCND1, and MIR548K Are the Driver Oncogenes of the 11q13 Amplicon in Squamous Cell Carcinoma
Authors: Mahieu Céline I. , Mancini Andrew G. , Vikram Ellee P. , Planells-Palop Vicente , Joseph Nancy M. , Tward Aaron D.
Published: February 2024

Abstract
11q13 amplification is a frequent event in human cancer and in particular in squamous cell carcinomas (SCC). Despite almost invariably spanning 10 genes, it is unclear which genetic components of the amplicon are the key driver events in SCC. A combination of computational, in vitro, ex vivo, and in vivo models leveraging efficient primary human keratinocyte genome editing by Cas9-RNP electroporation, identified ORAOV1, CCND1, and MIR548K as the critical drivers of the amplicon in head and neck SCC. CCND1 amplification drives the cell cycle in a CDK4/6/RB1-independent fashion and may confer a novel dependency on RRM2. MIR548K contributes to epithelial–mesenchymal transition. Finally, we identify ORAOV1 as an oncogene that acts likely via its ability to modulate reactive oxygen species. Thus, the 11q13 amplicon drives SCC through at least three independent genetic elements and suggests therapeutic targets for this morbid and lethal disease.
Implications:
This work demonstrates novel mechanisms and ways to target these mechanisms underlying the most common amplification in squamous cell carcinoma, one of the most prevalent and deadly forms of human cancer.
Table S8 from Genome-wide p63-Target Gene Analyses Reveal TAp63/NRF2-Dependent Oxidative Stress Responses
Authors: Napoli Marco , Deshpande Avani A. , Chakravarti Deepavali , Rajapakshe Kimal , Gunaratne Preethi H. , Coarfa Cristian , Flores Elsa R.
Published: February 2024

List of differentially expressed genes in ΔNp63+/+ vs. ΔNp63-/- cells (mouse epidermal, lung basal, and mammary epithelial cells) or in TAp63+/+ vs. TAp63-/- cells (mouse epidermal and mammary epithelial cells).
Table S5 from Genome-wide p63-Target Gene Analyses Reveal TAp63/NRF2-Dependent Oxidative Stress Responses
Authors: Napoli Marco , Deshpande Avani A. , Chakravarti Deepavali , Rajapakshe Kimal , Gunaratne Preethi H. , Coarfa Cristian , Flores Elsa R.
Published: February 2024

List of the siRNA sequences.
Table S2 from Genome-wide p63-Target Gene Analyses Reveal TAp63/NRF2-Dependent Oxidative Stress Responses
Authors: Napoli Marco , Deshpande Avani A. , Chakravarti Deepavali , Rajapakshe Kimal , Gunaratne Preethi H. , Coarfa Cristian , Flores Elsa R.
Published: February 2024

List of genes differentially expressed between of TAp63-/- or ΔNp63-/- mouse epidermal cells and the WT mouse epidermal cells. For each gene, the count of the TAp63 or ∆Np63 bound peaks within 10 kb from the gene bodies is indicated.
Supplementary Figure S3 from Genome-wide p63-Target Gene Analyses Reveal TAp63/NRF2-Dependent Oxidative Stress Responses
Authors: Napoli Marco , Deshpande Avani A. , Chakravarti Deepavali , Rajapakshe Kimal , Gunaratne Preethi H. , Coarfa Cristian , Flores Elsa R.
Published: February 2024

∆Np63 cooperates with the FOX family members to regulate cell motility genes
FIGURE 6 from Genome-wide p63-Target Gene Analyses Reveal TAp63/NRF2-Dependent Oxidative Stress Responses
Authors: Napoli Marco , Deshpande Avani A. , Chakravarti Deepavali , Rajapakshe Kimal , Gunaratne Preethi H. , Coarfa Cristian , Flores Elsa R.
Published: February 2024

TAp63 and NRF2 coordinately regulate the expression of TAp63-specific target genes. A, qRT-PCR for the indicated TAp63-specific target genes in epidermal cells of the indicated genotype. Data are mean ± SD, n = 3, * versus WT, P < 0.005, two-tailed t test. B, qRT-PCR of TAp63 ChIP assays using epidermal cells of the indicated genotype on the TAp63-specific peaks of the indicated TAp63-specific target genes. Data are mean ± SD, n = 3, * versus WT, P < 0.005, two-tailed t test. C, qRT-PCR of Nrf2 ChIP-re-ChIP assays using the TAp63 chromatin immunoprecipitated genomic regions of the indicated TAp63-specific target genes from epidermal cells of the indicated genotype. Data are mean ± SD, n = 3. * versus WT, P < 0.005, two-tailed t test. D, qRT-PCR of the indicated TAp63-specific target genes in WT, ΔNp63−/−, and TAp63−/− epidermal cells transfected with the indicated siRNAs. Data are mean ± SD, n = 3, * vs. WT siControl, # versus WT siNrf2, P < 0.005, two-tailed t test. E, Model depicting the coordinated regulation of TAp63-specific metabolic genes by TAp63 and NRF2.
FIGURE 5 from Genome-wide p63-Target Gene Analyses Reveal TAp63/NRF2-Dependent Oxidative Stress Responses
Authors: Napoli Marco , Deshpande Avani A. , Chakravarti Deepavali , Rajapakshe Kimal , Gunaratne Preethi H. , Coarfa Cristian , Flores Elsa R.
Published: February 2024

∆Np63 and TAp63 control the expression of their common target genes together with Stat proteins. A, qRT-PCR for the indicated ΔNp63 and TAp63 common target genes in epidermal cells of the indicated genotype. Data are mean ± SD, n = 3, * versus WT, P < 0.005, two-tailed t test. B, qRT-PCR of ∆Np63 ChIP assays using epidermal cells of the indicated genotype on the common peaks of the indicated common target genes. Data are mean ± SD, n = 3, * versus WT, P < 0.005, two-tailed t test. C, qRT-PCR of TAp63 ChIP assays using epidermal cells of the indicated genotype on the same genomic regions as in B. Data are mean ± SD, n = 3, * versus WT, P < 0.005, two-tailed t test. D, qRT-PCR of Stat4 ChIP-re-ChIP assays using the ∆Np63 chromatin immunoprecipitated genomic regions of the indicated common target genes from epidermal cells of the indicated genotype. Data are mean ± SD, n = 3. * versus WT, P < 0.005, two-tailed t test. E, qRT-PCR of Stat4 ChIP-re-ChIP assays using the TAp63 chromatin immunoprecipitated genomic regions of the indicated common target genes from epidermal cells of the indicated genotype. Data are mean ± SD, n = 3. * versus WT, P < 0.005, two-tailed t test. F, qRT-PCR of the indicated common target genes in WT, TAp63−/−, and ΔNp63−/− epidermal cells transfected with the indicated siRNAs. Data are mean ± SD, n = 3, * versus WT siControl, # versus WT siStat4, and $ versus WT siStat5a, P < 0.005, two-tailed t test. G, Model depicting the coordinated regulation of ΔNp63 and TAp63 common target genes with the Stat family members, Stat4 and Stat5a.
Data from Genome-wide p63-Target Gene Analyses Reveal TAp63/NRF2-Dependent Oxidative Stress Responses
Authors: Napoli Marco , Deshpande Avani A. , Chakravarti Deepavali , Rajapakshe Kimal , Gunaratne Preethi H. , Coarfa Cristian , Flores Elsa R.
Published: February 2024

Abstract
The p53 family member TP63 encodes two sets of N-terminal isoforms, TAp63 and ΔNp63 isoforms. They each regulate diverse biological functions in epidermal morphogenesis and in cancer. In the skin, where their activities have been extensively characterized, TAp63 prevents premature aging by regulating the quiescence and genomic stability of stem cells required for wound healing and hair regeneration, while ΔNp63 controls maintenance and terminal differentiation of epidermal basal cells. This functional diversity is surprising given that these isoforms share a high degree of similarity, including an identical sequence for a DNA-binding domain. To understand the mechanisms of the transcriptional programs regulated by each p63 isoform and leading to diverse biological functions, we performed genome-wide analyses using p63 isoform-specific chromatin immunoprecipitation, RNA sequencing, and metabolomics of TAp63−/− and ΔNp63−/− mouse epidermal cells. Our data indicate that TAp63 and ΔNp63 physically and functionally interact with distinct transcription factors for the downstream regulation of their target genes, thus ultimately leading to the regulation of unique transcriptional programs and biological processes. Our findings unveil novel transcriptomes regulated by the p63 isoforms to control diverse biological functions, including the cooperation between TAp63 and NRF2 in the modulation of metabolic pathways and response to oxidative stress providing a mechanistic explanation for the TAp63 knock out phenotypes.
Significance:
The p63 isoforms, TAp63 and ΔNp63, control epithelial morphogenesis and tumorigenesis through the interaction with distinct transcription factors and the subsequent regulation of unique transcriptional programs.
Molecular Pathogenic Mechanisms of IgA Nephropathy Secondary to COVID-19 mRNA Vaccination
Authors: Wang Luoyi , Mao Zhaomin , Zhang Lirong , Shao Fengmin
Published: February 2024

Introduction: Accumulating evidence has disclosed that IgA nephropathy (IgAN) could present shortly after the second dose of COVID-19 mRNA vaccine. However, the undying mechanism remains unclear and we aimed to investigate the potential molecular mechanisms.
Methods: We downloaded gene expression datasets of COVID-19 mRNA vaccination (GSE201535) and IgAN (GSE104948). Weighted Gene Co-Expression Network Analysis (WGCNA) was performed to identify co-expression modules related to the second dose of COVID-19 mRNA vaccination and IgAN. Differentially expressed genes (DEGs) were screened, and a transcription factor (TF)-miRNA regulatory network and protein-drug interaction were constructed for the shared genes.
Results: WGCNA identified one module associated with the second dose of COVID-19 mRNA vaccine and four modules associated with IgAN. Gene Ontology (GO) analyses revealed enrichment of cell cycle-related processes for the COVID-19 mRNA vaccine hub genes and immune effector processes for the IgAN hub genes. We identified 74 DEGs for the second dose of COVID-19 mRNA vaccine and 574 DEGs for IgAN. Intersection analysis with COVID-19 vaccine-related genes led to the identification of two shared genes, TOP2A and CEP55. The TF-miRNA network analysis showed that hsa-miR-144 and ATF1 might regulate the shared hub genes.
Conclusions: This study provides insights into the common pathogenesis of COVID-19 mRNA vaccination and IgAN. The identified pivotal genes may offer new directions for further mechanistic studies of IgAN secondary to COVID-19 mRNA vaccination.
Practical requirements for the development of an advanced cardiorenal unit
Authors: Almenar-Bonet Luis , Sánchez-Lázaro Ignacio , Soldevila Amparo , López-Vilella Raquel , Donoso Trenado Víctor , Devesa Ramón , Carmona Paula , Tormo Sergi , Montero Hernández María Jesús , Hernández Julio , Martínez Dolz Luis , Sánchez-Pérez Pilar
Published: February 2024

Background: Heart failure is frequently associated with kidney disease, and patients with kidney disease are at increased risk of heart failure. The co-occurrence of both entities not only significantly increases morbidity and mortality, but also complicates therapy.
Summary: Cardiorenal syndrome often requires a broad, comprehensive, and multidisciplinary approach. As a result, a need has arisen to create specialized cardiorenal units that allow for rigorous and personalized management of this condition. Moreover, in some cases, cardiorenal syndrome is more complex owing to an acute and critical situation that requires the concept of cardiorenal unit to be extended towards advanced diagnostic and therapeutic positions, thus confirming the need for an advanced cardiorenal unit. The creation of these units constitutes a real challenge, necessitating a specific multilevel action plan, covering governance and management, type of patient, personnel requirements, service portfolio, care process, information systems, and other resources. Specific lines of action must be proposed for each of the relevant points in order to facilitate development of these units, together with continuous evaluation of unit activity through specific indicators, and to detect areas for improvement.
Predictors and Prognosis of Early Neurological Outcomes on Patients with Vertebrobasilar Artery Occlusion Undergoing Endovascular Treatment
Authors: Ma Xinan , Li Yajun , Zhang Pan , Yi Jilong , Xu Yingjie , Hu Miaomiao , Wang Jinjing , Lan Wenya , Xu Guoqiang , Lu Yanan , Xu Pengfei , Feng Feng , Sun Wen , Chen Hao , Wu Zongyi
Published: February 2024

Introduction: This research explored the factors influencing early neurological outcomes (ENO) in patients who had vertebrobasilar artery occlusion (VBAO) and received endovascular treatment (EVT), as well as examining the causal influence of ENO on the prognosis of VBAO patients. Methods: A retrospective review was carried out on patients from 65 Chinese stroke centers, all within 24 h of the estimated occlusion time. ENO includes early neurological improvement (ENI) and early neurological deterioration (END), defined as a decrease or an increase of at least 4 points in NIHSS score between baseline and 24 h after EVT. Death within 24 h after EVT was also considered as END. END was further divided into explained END and unexplained END (unEND). Independent predictors of ENO and the association between ENO and outcomes in patients with VBAO were determined using center-adjusted analyses. The study developed a multivariate logistic regression model to examine the comparative risk of unEND versus explained END on the clinical outcomes in VBAO patients. Results: A total of 2,257 patients were included. Glasgow Coma Scale (GCS) (OR: 1.16, 95% CI: 1.03–1.30) and successful reperfusion (OR: 1.15, 95% CI: 1.02–1.30) were associated with ENI. Baseline NIHSS (OR: 0.60, 95% CI: 0.53–0.68), successful reperfusion (OR: 0.79, 95% CI: 0.71–0.89), and puncture to reperfusion time (OR: 1.17, 95% CI: 1.03–1.33) were associated with END. When examining 3-month prognostic indexes, both END and ENI were found to be linked to the 3-month outcomes, but in opposite directions. A subgroup analysis of END suggested that unEND typically demonstrated a more favorable prognosis compared to explained END, although the prognosis remained generally unfavorable. Conclusions: ENO, whether they manifested as early improvement or deterioration, were linked to the prognosis of VBAO patients undergoing EVT. The outcomes after unEND were more favorable than those following explained END.
Outcomes of Post-Immunotherapy Durable Responders of Advanced Hepatocellular Carcinoma– with Emphasis on Locoregional Therapy for Oligoprogression
Authors: Liu Tsung-Hao , Chen San-Chi , Rau Kun-Ming , Lu Li-Chun , Lin Po-Ting , Su Yung-Yeh , Teng Wei , Lai Shiue-Wei , Yeh Ren-Hua , Kao Tsui-Mai , Lee Pei-Chang , Wu Chi-Jung , Chen Chien-Hung , Hsu Chih-Hung , Lin Shi-Ming , Huang Yi-Hsiang , Chen Li-Tzong , Cheng Ann-Lii , Shen Ying-Chun ,   on behalf of Taiwan Liver Cancer Association Research Group (TRG)
Published: February 2024

Introduction: The progression patterns, dispositions, and outcomes of patients with advanced hepatocellular carcinoma (HCC) who achieved durable responses with immunotherapy remain poorly characterized. Methods: Patients with advanced HCC who received immune checkpoint inhibitor (ICI)-based immunotherapy and achieved durable responses were retrospectively included. A durable response was defined as partial response (PR) or stable disease (SD) per RECIST 1.1 for more than 8 months after initiation of immunotherapy. Oligoprogression and polyprogression were defined as progression at ≤3 and >3 lesions, respectively. Results: A total of 91 durable responders (63 PR and 28 SD) were identified. The majority had chronic viral hepatitis (n = 69, 75.8%). Forty-seven (51.6%) and 44 (48.4%) patients received the index immunotherapy as first-line and second- or beyond-line therapy, respectively. Fifty-four (59.3%) patients subsequently developed progression, with a predominant pattern of oligoprogression (66.7%). The median overall survival (OS) was 46.2 months (95% CI: 34.1–58.3). For patients with subsequent progression, employment of locoregional therapy (LRT) for progression was associated with prolonged OS (univariate analysis: hazard ratio [HR] 0.397, p = 0.009; multivariate analysis: HR 0.363, p = 0.050). Patients with oligoprogression who received LRT showed longer median OS than those who did not (48.4 vs. 20.5 months, p < 0.001). In contrast, the median OS of patients with polyprogression who received LRT was not different from those without LRT (27.7 vs. 25.5 months, p = 0.794). Conclusion: Approximately 60% of the post-immunotherapy durable responders of HCC subsequently develop progression. Proactive LRT may further rescue patients who develop subsequent oligoprogression. Prospective studies are mandatory to clarify the proper management of durable responders with subsequent progression.
Regulation of YAP Promotor Accessibility in Endothelial Mechanotransduction
Authors: Mannion Aarren J. , Zhao Honglei , Zhang Yuanyuan , von Wright Ylva , Bergman Otto , Roy Joy , Saharinen Pipsa , Holmgren Lars
Published: February 2024

BACKGROUND:

Endothelial cells are constantly exposed to mechanical forces in the form of fluid shear stress, extracellular stiffness, and cyclic strain. The mechanoresponsive activity of YAP (Yes-associated protein) and its role in vascular development are well described; however, whether changes to transcription or epigenetic regulation of YAP are involved in these processes remains unanswered. Furthermore, how mechanical forces are transduced to the nucleus to drive transcriptional reprogramming in endothelial cells is poorly understood. The YAP target gene,
AmotL2
(angiomotin-like 2), is a junctional mechanotransducer that connects cell-cell junctions to the nuclear membrane via the actin cytoskeleton.

METHODS:

We applied mechanical manipulations including shear flow, stretching, and substrate stiffness to endothelial cells to investigate the role of mechanical forces in modulating
YAP
transcription. Using in vitro and in vivo endothelial depletion of AmotL2, we assess nuclear morphology, chromatin organization (using transposase-accessible chromatin sequencing), and whole-mount immunofluorescent staining of the aorta to determine the regulation and functionality of YAP. Finally, we use genetic and chemical inhibition to uncouple the nuclear-cytoskeletal connection to investigate the role of this pathway on
YAP
transcription.

RESULTS:

Our results reveal that mechanical forces sensed at cell-cell junctions by the YAP target gene
AmotL2
are directly involved in changes in global chromatin accessibility and activity of the histone methyltransferase EZH2, leading to modulation of
YAP
promotor activity. Functionally, shear stress–induced proliferation of endothelial cells in vivo was reliant on AmotL2 and YAP/TAZ (transcriptional coactivator with PDZ-binding motif) expression. Mechanistically, uncoupling of the nuclear-cytoskeletal connection from junctions and focal adhesions led to altered nuclear morphology, chromatin accessibility, and
YAP
promotor activity.

CONCLUSIONS:

Our findings reveal a role for AmotL2 and nuclear-cytoskeletal force transmission in modulating the epigenetic and transcriptional regulation of YAP to maintain a mechano-enforced positive feedback loop of vascular homeostasis. These findings may offer an explanation as to the proinflammatory phenotype that leads to aneurysm formation observed in AmotL2 endothelial deletion models.
Mechanism of Ventricular Tachycardia Occurring in Chronic Myocardial Infarction Scar
Authors: Donahue J. Kevin , Chrispin Jonathan , Ajijola Olujimi A.
Published: February 2024

Cardiac arrest is the leading cause of death in the more economically developed countries. Ventricular tachycardia associated with myocardial infarct is a prominent cause of cardiac arrest. Ventricular arrhythmias occur in 3 phases of infarction: during the ischemic event, during the healing phase, and after the scar matures. Mechanisms of arrhythmias in these phases are distinct. This review focuses on arrhythmia mechanisms for ventricular tachycardia in mature myocardial scar. Available data have shown that postinfarct ventricular tachycardia is a reentrant arrhythmia occurring in circuits found in the surviving myocardial strands that traverse the scar. Electrical conduction follows a zigzag course through that area. Conduction velocity is impaired by decreased gap junction density and impaired myocyte excitability. Enhanced sympathetic tone decreases action potential duration and increases sarcoplasmic reticular calcium leak and triggered activity. These elements of the ventricular tachycardia mechanism are found diffusely throughout scar. A distinct myocyte repolarization pattern is unique to the ventricular tachycardia circuit, setting up conditions for classical reentry. Our understanding of ventricular tachycardia mechanisms continues to evolve as new data become available. The ultimate use of this information would be the development of novel diagnostics and therapeutics to reliably identify at-risk patients and prevent their ventricular arrhythmias.
Reversal of Pulmonary Hypertension in a Human-Like Model: Therapeutic Targeting of Endothelial DHFR
Authors: Murugesan Priya , Zhang Yixuan , Huang Yuanli , Chenggong Zong Nobel , Youn Ji Youn , Chen Wenhui , Wang Chen , Loscalzo Joseph , Cai Hua
Published: February 2024

BACKGROUND:

Pulmonary hypertension (PH) is a progressive disorder characterized by remodeling of the pulmonary vasculature and elevated mean pulmonary arterial pressure, resulting in right heart failure.

METHODS:

Here, we show that direct targeting of the endothelium to uncouple eNOS (endothelial nitric oxide synthase) with DAHP (2,4-diamino 6-hydroxypyrimidine; an inhibitor of GTP cyclohydrolase 1, the rate-limiting synthetic enzyme for the critical eNOS cofactor tetrahydrobiopterin) induces human-like, time-dependent progression of PH phenotypes in mice.

RESULTS:

Critical phenotypic features include progressive elevation in mean pulmonary arterial pressure, right ventricular systolic blood pressure, and right ventricle (RV)/left ventricle plus septum (LV+S) weight ratio; extensive vascular remodeling of pulmonary arterioles with increased medial thickness/perivascular collagen deposition and increased expression of PCNA (proliferative cell nuclear antigen) and alpha-actin; markedly increased total and mitochondrial superoxide production, substantially reduced tetrahydrobiopterin and nitric oxide bioavailabilities; and formation of an array of human-like vascular lesions. Intriguingly, novel in-house generated endothelial-specific dihydrofolate reductase (DHFR) transgenic mice (tg-EC-DHFR) were completely protected from the pathophysiological and molecular features of PH upon DAHP treatment or hypoxia exposure. Furthermore, DHFR overexpression with a pCMV-DHFR plasmid transfection in mice after initiation of DAHP treatment completely reversed PH phenotypes. DHFR knockout mice spontaneously developed PH at baseline and had no additional deterioration in response to hypoxia, indicating an intrinsic role of DHFR deficiency in causing PH. RNA-sequencing experiments indicated great similarity in gene regulation profiles between the DAHP model and human patients with PH.

CONCLUSIONS:

Taken together, these results establish a novel human-like murine model of PH that has long been lacking in the field, which can be broadly used for future mechanistic and translational studies. These data also indicate that targeting endothelial DHFR deficiency represents a novel and robust therapeutic strategy for the treatment of PH.
Abstract 103: Acute Ischemia Induces Spatially and Transcriptionally Distinct Microglial Subclusters
Authors: Li Huiya , Liu Pinyi , Zou Xinxin , Zhu Liwen , Hu Yujie , Deng Shiji , Zhang Siyuan , Li Qi , Zhu Xuan , Gao Shenghan , Xu Yun
Published: February 2024

Introduction:
Damage in the ischemic core and penumbra after stroke affects patient prognosis. Microglia immediately respond to ischemic insult after stroke. However, the microglial heterogeneity and the underlying mechanisms remain unclear. In this study, we sought to uncover stroke-associated microglia and identify their heterogenous features.

Methods:
We performed single-cell RNA-sequencing and spatial transcriptomics on middle cerebral artery occlusion (MCAO) mice to determine stroke-associated microglial clusters. The expression of marker genes and the localization of different microglial subclusters were verified through RNAscope and immunofluorescence. GSVA was employed to reveal functional characteristics of microglial subclusters. IPA was used to explore upstream regulators of microglial clusters, which was confirmed by immunofluorescence, RT-qPCR, and targeted metabolomics. Finally, the infarct size and neurological deficits were evaluated in MCAO mice after pharmacological manipulation of specific microglial subcluster.

Results:
We discovered stroke-associated microglial subclusters in MCAO mice. We also identified novel marker genes of these microglial subclusters and defined these cells as ischemic core-associated (ICAM) and ischemic penumbra-associated (IPAM) microglia, according to their spatial distribution. ICAM, induced by damage-associated molecular patterns, are fueled by glycolysis, and exhibit increased pro-inflammatory cytokines and chemokines production. BACH1 is a key transcription factor driving ICAM generation. In contrast, glucocorticoids enriched in the penumbra triggers IPAM formation, which are powered by citrate cycle and oxidative phosphorylation and are characterized by moderate pro-inflammatory responses, inflammation-alleviating metabolic features, and myelinotrophic properties.

Conclusions:
ICAM induce excessive neuroinflammation, aggravating brain injury, whereas IPAM could be essential for the homeostasis and survival of cells in the penumbra. Our findings provide a biological basis for targeting specific microglial subtypes as a potential therapeutic strategy for ischemic stroke.
Abstract 107: Exploring Non-Human DNA Content in Cerebral Thrombi Using Shotgun Metagenomic Sequencing
Authors: Walker Melanie , Tierney Braden T , Espinoza Josh L , Federico Emma , Gonzalez Alexandra , Mason Christopher E , Dupont Christopher L
Published: February 2024

Introduction:
Prior studies that use 16S rRNA amplicon sequencing to profile bacterial components of cerebral thrombi do not provide species-level taxonomic information or functional gene content, which are critical for the development of subsequent clinical applications. To determine the amount and taxonomic nature of non-human DNA in cerebral thrombi, shotgun metagenomic sequencing was performed on specimens retrieved from ischemic stroke patients.

Methods:
Thrombi were retrieved during standard of care thrombectomy and preserved in DNA/RNA Shield (Zymo), alongside environmental controls. DNA was extracted per manufacturer instructions, using Zymo kits. Quality control was carried out using Bowtie2 and the bbtools suite. To clump reads, bbmap was applied. Sequencing adapter contamination was removed with bbduk. Data was aligned to the human reference genome (hg38) using Bowtie2. Tadpole (mode=correct, ecc=t, ecco=t) was used to correct sequencing errors. MetaSPAdes was used to assemble quality-controlled sequencing reads into contigs. CAT/BAT was then applied to annotate contigs and their Open Reading Frames (ORF) taxonomically using the Lowest Common Ancestor approach.

Results:
Of the 41 specimens, N = 31 met quality control. The preponderance of DNA sequencing reads mapped to the human genome. An average of 7,559,292 +/- 1,615,7061 (9.1%) reads per sample did not align (Fig 1A). Contigs resulting from
de novo
assemblies of these potentially non-human reads contained sequences consistent with bacterial ORFs. The number of these ORFs varied per sample (Fig 1B) and correlated with the number of non-human reads.
Proteobacteria
and
Firmicutes
dominated. (Fig 1C).

Conclusion:
While much work remains to optimize sequencing techniques and controls, the potential to recover near-complete bacterial genomes from thrombus presents an opportunity to further explore the human host-microbiota interactions in acute ischemic stroke.
Abstract 108: Gut Microbiota Composition Modifies Behavioral Phenotypes Into Adulthood Following Neonatal Hypoxic-Ischemia in Mice
Authors: Korf Janelle , Blasco-Conesa Maria , Thompson Hannah , Khan Romeesa , Blixt Frank , McCullough Louise D , Ganesh Bhanu P
Published: February 2024

Neonatal hypoxic ischemic encephalopathy (HIE) increases the risk for attention deficit disorder and autism spectrum disorders in children. Perturbations in the gut microbiome are associated with behavioral changes in pre-clinical models of neurological injury and development. However, the role of the gut microbiome in behavior after HIE has not been investigated. Also, the therapeutic potential of gut microbiota modification after HIE remains unexplored. We hypothesize that altering the gut microbiome after HIE can improve chronic behavioral deficits in mice. The Rice Vannucci Model (RVM) was used to model HIE on 9-day old C57BL/6 mice. 2 months after injury, fecal samples were collected for 16s rRNA sequencing and behavioral tests were performed. We treated HIE and sham mice with fecal microbiota transfers (FMT) from naïve donors. To determine if the microbiota from HIE mice drives behavioral deficits, we also gave FMT from HIE or sham donors into naïve. Tests were repeated 1 and 3 months after FMT, and mice were sacrificed at 5 months of age. 2 months after injury, baseline open field tests revealed a hyperactive phenotype in HIE mice. There was increased mean velocity, distance moved, and cumulative time in border coupled with decreased cumulative time in center compared to shams (P<0.0001, P<0.0001, P=0.0168, P=0.025; T test, two cohorts, n=14-18). PCoA on calculated weighted UniFrac distances reveal a significant difference in β-diversity in males (P=0.015) and females (P=0.033) at 2 months. At 5 months, HIE mice with a naïve FMT had normalization of their hyperactive phenotype, measured by a reduction in mean velocity and distance moved (P=0.028 and P=0.0322; Repeated Measures One-way ANOVA, Dunnett’s test), while sham mice had no change. Surprisingly, naïve mice given a FMT from HIE donors became hypoactive with reduced mean velocity and distance moved (P=0.0004 and P=0.0004; Mixed effects Model, Tukey’s test, 2 cohorts n=6-11), while Naive mice with sham FMT had no change. In conclusion, gut microbiota modification through FMT in adult mice with a history of HIE reduced the severity of their hyperactive phenotype. Conversely, FMT from HIE mice into naïve mice induced mild behavioral changes but did not reproduce the hyperactivity seen in HIE mice.
Abstract 114: LACROSSE Trial: Lumbar Drainage Compared to External Ventricular Drainage in Aneurysmal Subarachnoid Hemorrhage, a Randomized Control Trial
Authors: Rahmani Redi ,   LACROSSE Trial Investigators, Morrow Jay , Bender Matthew T , Khalessi Alexander A
Published: February 2024

Introduction:
Delayed cerebral ischemia (DCI) is a leading cause of morbidity and mortality in aneurysmal subarachnoid hemorrhage (aSAH). External ventricular drainage (EVD) has been regarded as the gold standard, but lumbar drains (LD) may impact the rate of DCI due to preferential clearing of subarachnoid blood. Recent trials have not directly compared these drains therefore our goal was to determine if LD is superior to EVD for prevention of DCI in aSAH.

Methods:
A randomized controlled trial was initiated at UC San Diego, the University of Rochester, and Rochester Regional to randomize drain placement to either EVD or LD in aSAH patients. The primary outcome was DCI, defined as an exclusionary change in GCS or new, none-treatment related infarcts on imaging. Secondary outcomes were ICU measures and functional outcomes.

Results:
52 patients were enrolled. The rate of DCI was significantly lower in the LD arm (7.4%, 2/27) compared to the EVD arm (36%, 9/25) (p=0.017) with a NNT of 3.5 for LD. There was no significant difference between the two arms with regards to admission characteristics, aneurysm location, treatment type, or total number of vasospasm treatments. LD patients spent fewer days in the ICU (p=0.015), on a ventilator (p=0.004) and had fewer drain days (p=0.003) compared to EVD patients. The proportion of patients with functional independence was significantly higher in the LD group compared to the EVD group 1-month post-hemorrhage (BI p = 0.04; GOS p = 0.04)

Conclusions:
In this trial, we show that LD is superior to EVD in aSAH patients in the reduction of DCI, is associated with significantly improved clinical outcomes, and reduced utilization of ICU resources.
Abstract 115: Presenting Symptoms and Neuroimaging Correlates in North American Moyamoya Vasculopathy
Authors: Mishra Murli , Han Caleb , Garza Maria , Richerson Wesley , Rodeghier Mark , Jordan Lori C , Donahue Manus
Published: February 2024

Introduction:
Moyamoya disease (MMD) is a steno-occlusion of the supraclinoid internal carotid arteries and proximal branches with the development of collateral vessels
1
. Once thought to affect primarily females of East Asian descent, more than 11,000 hospitalizations in patients with MMD of non-Asian ancestry are reported annually in the USA, with the majority presenting with ischemic stroke. Here, we present frequent non-stroke symptomatology in North American MMD along with hemodynamic imaging profiles at presentation.

Methods:
Participants provided informed consent and underwent anatomical MRI with hypercapnic blood oxygenation level-dependent
2
; time regression was applied to quantify cerebrovascular reactivity (CVR) and reactivity delay time (CVR
DELAY
)
3
in right and left MCA territories. A logistic regression analysis based on the Huber-White sandwich estimator compared the CVR metrics between cohorts. Significance criterion: Bonferroni corrected two-sided p<0.05.

Results:
74 patients (age=11-77 years; Asian=6%, White=65%, Black=29%) diagnosed with MMD with symptom durations spanning hours to years prior to presentation were enrolled. Non-stroke symptoms and frequency prior to stroke are listed in Table 1. In more than 1/3
rd
of patients, unprovoked headaches preceded stroke-like symptoms. In patients exhibiting ischemic symptoms within six months of neuroimaging, cerebrovascular reserve capacity was reduced (p=0.007), and cerebrovascular reactivity delay time increased (p<0.001), in the flow territory from which symptoms were derived compared to the contralateral flow territory.

Discussion:
North American MMD is being observed with increasing frequency and increasing the awareness of non-stroke MMD presentations may lead to early diagnosis that can lead to proactive revascularization to minimize the risk of first stroke in these patients.
Abstract 130: Decompression of Corticospinal Tract (CST): An Imaging Biomarker for Post-Stroke Recovery
Authors: Haque Muhammad E , Boren Seth , Vyas Vedang , Steven Emily , Suchting Robert , sitton Clark , Hasan Khader , Savitz Sean I , Aronowski Jaroslaw A
Published: February 2024

Objective:
To investigate the anatomical and physiological impact of intracerebral hemorrhage (ICH) on the mechanical CST distortion measured using serial MRI and its relation to functional recovery.

Introduction:
ICH causes CST (motor pathway for arm movement) deformation/displacement leading to disruption of brain-arm neuronal communication. Limited knowledge exists about the degree and anatomical disruption of CST and long-term recovery of the arm impairment and how temporal resolution of the hematoma/edema volume impacts recovery. Here we applied serial quantitative MRI to establish an association between the CST displacement (CSTD) and arm recovery.

Methods:
We imaged 12 patients with subcortical bleed on day-2 and 90, who underwent routine care. Using DTI, the change in CSTD at the level of posterior limb of internal capsule (PLIC) was calculated by the distance shift between the patient's pre (determined a standard brain template, registered to DTI images) and post-ICH CST location. A centroid (the medians of a volume at 3 intersecting coordinates) at each PLICs was determined and the distance between the two centroids was recorded. ICH severity and arm impairment were measured on NIHSS and Fugl-Meyer assessment for Upper Extremity (FM-UE) scores.

Results:
We enrolled 9M/3F, with an average age of 512.1±8.3y who were serially imaged. The hematoma volume (18.6±28.1 to 4.5±4.9mL) and NIHSS (14.0±5 to 5.0±3) significantly (p<0.05) decreased whereas FM-UE (23.8±19 to 41.5±22) improved. The patient's average CSTD changed from 10.9±4.61 to 5.78±2.2mm over time. The change in CSTD exhibited a negative correlation (R2=0.37) with FM-UE scores. These results are shown in Fig1.

Conclusion:
In conclusion, we present novel quantitative surrogate imaging markers of CST distortion and its association with recovery after ICH.
Abstract 144: Value of CT Perfusion Imaging Among Patients With Minor Stroke
Authors: Zuflacht Jonah , Miller Jenna , Ljungberg Lovisa , Little Jessica , Zhou Sonya , Kasner Scott
Published: February 2024

Introduction:
CTP has increasingly been incorporated into the evaluation of all patients with suspected acute ischemic stroke (AIS), including those with minor symptoms. We aimed to assess the incremental value of CTP in acute treatment decision-making among patients with low NIHSS.

Methods:
We performed a retrospective cohort study of all patients who underwent CTP upon presentation to the ED at three academic, urban hospitals in Philadelphia, PA between January 1, 2022 and December 31, 2022. We collected data on initial NIHSS score, AIS treatment decisions, subsequent neuroimaging, and final diagnosis.

Results:
There were 531 patients with a median age of 64.5 years (IQR 54-73) and 56% were women. 73% were Black or African American, 20% White, and 4% Asian. Frequency of CTP imaging by initial NIHSS score is presented in the Figure. Among 90 patients (16.9%) with NIHSS≤2, CTP imaging was cited as justification for AIS treatment (thrombolysis, thrombectomy, or both) in 0 (0%, 95% CI: 0-4%). Specifically, among 41 patients (7.7%) with NIHSS≤1, 0 received thrombolysis or thrombectomy, and 12 (29.2%) were ultimately diagnosed with AIS on CT/MRI. Among 49 with NIHSS=2 (9.2%), 2 patients received thrombolysis (based on clinical exam, CTP was normal) and one later underwent thrombectomy based on clinical decompensation with repeat NIHSS=7, and 28.6% had AIS on CT/MRI. Meanwhile, among 33 patients with NIHSS=3, acute treatment was given to 3 and CTP influenced AIS treatment decisions in 2 (6.1%). Among 59 patients with NIHSS=4, acute treatment was given to 4 and CTP influenced decision-making in 3 (5.1%). CTP mismatch ratio was > 1.7 in all 5 patients who received acute treatment and had NIHSS of 3 or 4.

Conclusions:
CTP is frequently performed in patients with low NIHSS. It had limited impact on acute treatment decisions, notably none among those with NIHSS≤2, suggesting that CTP may be over-utilized in this subset of patients with AIS.
Abstract 150: Regional Molecular Alterations in Cerebral Amyloid Angiopathy-Related Hemorrhagic Lesions
Authors: Gokcal Elif , Becker Alex J , Horn Mitchell J , Das Alvin S , Rosand Jonathan , Viswanathan Anand , Polimeni Jonathan R , Johnson Keith A , Greenberg Steven M , Gurol Edip M
Published: February 2024

Background:
Previous case reports suggested co-localization of tau but not β-Amyloid with cortical superficial siderosis (cSS) in cerebral amyloid angiopathy (CAA). We thus hypothesized that the tau load represents a continuum with amyloid accumulation in the disease cascade and that the tau load would be higher in lobes with cSS and intracerebral hemorrhages (ICH) when compared to the contralateral lobes without hemorrhages in CAA patients.

Methods:
The study included 26 cognitively healthy probable CAA patients (mean age 71±7.5, 53.8% male) who underwent multimodal MRI, Pittsburgh compound B (PiB, for amyloid), and
18
F-flortaucipir (FTP, for tau) PET imaging. Mean global cortical PiB uptake and FTP uptake were calculated in each lobar region. The presence of ICH and cSS were identified in each lobar region independent of molecular imaging. A pairwise comparison of PiB and FTP uptake was done between lobes with any hemorrhagic lesion (ICH/cSS) and their corresponding lobes without hemorrhages.

Results:
The FTP uptake was significantly higher in 8 frontal, 16 temporal, and 11 occipital lobes with hemorrhages compared to their corresponding contralateral lobes without hemorrhages (1.18±0.1 vs. 1.13±0.1; p=0.002, 1.28±0.1 vs. 1.20±0.1; p=0.004, 1.29±0.1 vs. 1.10±0.1; p=0.002, respectively) but not in parietal lobes (p=0.107). The PiB uptake was significantly higher in occipital lobes with hemorrhages compared to their contralateral lobes without hemorrhages (1.41±0.3 vs. 1.35±0.3, p=0.004) but there was no difference in the comparison of amyloid burden for the remaining lobar regions with or without hemorrhage (p>0.2 for all comparisons).

Conclusions:
Reduced PiB uptake at sites of larger hemorrhagic lesions (cSS/ICH) has been observed in the past so the lack of differences in amyloid between most lobes with and without hemorrhage was not surprising in our study. Our results otherwise show significantly increased tau load at most brain lobes with hemorrhagic lesions among CAA patients. Potential clinical effects of the tau burden related to cSS/ICH should be studied in larger CAA cohorts using appropriate clinical testing.
Abstract 153: First Pass Effect in Mechanical Thrombectomy for Anterior Circulation Acute Ischemic Stroke is Modified by Procedure Time: Proposal of a New Measure for Thrombectomy Procedures
Authors: Koo Andrew , Reeves Benjamin , Renedo Daniela , Maier Ilko , Al Kasab Sami , Jabbour Pascal M , Kim Joon-Tae , Quintero Stacey C , rai ansaar , Starke Robert , Psychogios Marios , Shaban Amir , Arthur Adam S , Yoshimura Shinichi , Cuellar Hugo , Grossberg Jonathan A , Alawieh Ali M , Romano Daniele , Tanweer Omar , Mascitelli Justin , Fragata Isabel , Polifka Adam , Osbun Joshua , Crosa Roberto , Park Min , Levitt Michael , Brinjikji Waleed , Moss Mark , Dumont Travis , Williamson Richard , Navia Pedro , Kan Peter , Spiotta Alejandro M , Sheth Kevin N , de Havenon Adam H , Matouk Charles C
Published: February 2024

Objective:
To determine whether first pass effect (FPE) after mechanical thrombectomy (MT) for anterior circulation large vessel occlusion acute ischemic stroke (LVO-AIS) is modified by procedural time (PT).

Methods:
The Stroke Thrombectomy and Aneurysm Registry (STAR), a multi-center international dataset, was retrospectively analyzed for anterior circulation LVO-AIS treated by MT who achieved excellent reperfusion (TICI 2c/3). The primary outcome was good functional outcome as defined by a 90-day modified Rankin Scale (mRS) 0-2. The primary study exposure was first pass success (FPS, 1 pass vs ≥2 passes) and the secondary exposure was PT. Logistic regression models were fit-adjusted and marginal effects used to assess the interaction of PT (≤30 vs >30 minutes) and FPS, adjusting for potential confounders including time from last known well to start of MT.

Results:
A total of 1,310 patients had excellent reperfusion. These patients were divided into two cohorts based on PT: ≤30 minutes (777 patients, 59.3%) and > 30 minutes (533 patients, 40.7%). Good functional outcome was observed in 658 patients (50.2%). The interaction term between FPS and PT was significant (p=0.018). Individuals with FPS in ≤30 minutes had 11.5% higher adjusted predicted probability of good outcome compared with those who required ≥2 passes (58.2% vs. 46.7%, p=0.001). However, there was no significant difference in the adjusted predicted probability of good outcome based on FPS in individuals with PT >30 minutes (p=0.763). This relationship appeared identical in models with PT treated as a continuous variable.

Conclusion:
In a large, real-world, multi-national dataset, we find that FPE is importantly modified by PT. The added clinical benefit of FPE is lost in longer procedures (>30 minutes). These data argue for a new metric for MT procedures, namely, FPE
30
, that better represents the ideal of fast, complete reperfusion with a single pass of a thrombectomy device.