bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A051: Mitochondrial metabolism and fission as dynamic drivers of breast cancer metastasis
Authors: Savage Hannah , Ma Dennis , Lawson Devon
Published: February 2024

Approximately 30% of women with early-stage breast cancer will develop metastatic disease, making efforts to define drivers of metastasis critical. Our group found that metastatic breast cancer cells express an oxidative phosphorylation (OXPHOS) gene signature. Further, inhibiting OXPHOS reduced metastatic seeding, thus revealing the functional role of mitochondrial metabolism in breast cancer metastasis. It is understood that mitochondria are highly dynamic organelles that are regulated by a balance of fission and fusion proteins; however, it remains poorly understood how mitochondrial metabolism and their dynamics promote cancer spreading. To interrogate mitochondrial heterogeneity within primary tumors, we sorted tumor cells from patient derived xenograft (HCI010) and murine (4T1) breast tumors stratified by low or high uptake of tetramethylrhodamine methyl ester (TMRM, a dye that accumulates in active mitochondria undergoing OXPHOS). Low or high cell fractions were re-injected intravenously in new cohorts of mice. We found greater metastatic burden in the lungs of mice injected with the TMRM high fraction. Furthermore, we uncovered that the TMRM high cells had greater mitochondria abundance than TMRM low cells. We hypothesized the differences in mitochondrial content we observed in primary tumor cells is caused by shifts in cellular expression of mitochondria fission and fusion proteins. Consistent with this hypothesis, we found that TMRM high, metastatic tumor cells had greater expression of mitochondria fission gene, DRP1, and lower expression of fusion gene, MFN2. Collectively, our results revealed heterogeneity in tumor cell mitochondrial content within a primary tumor that appears to influence the fitness of a cell to metastasize. Lastly, our data supports a role for mitochondrial fission in breast cancer metastasis. Future directions for our studies will include genetic and pharmacological modulation of mitochondrial fission and fusion to better understand why it influences metastasis and at what point during the metastatic cascade.

Citation Format: Hannah Savage, Dennis Ma, Devon Lawson. Mitochondrial metabolism and fission as dynamic drivers of breast cancer metastasis [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr A051.
Abstract IA05: Leveraging preclinical models of triple negative breast cancer for translational research
Authors: Rosen Jeffrey M , Pedroza Diego , Zhao Na , Zhang Xiang H-F , Perou Charles M
Published: February 2024

We have developed multiple novel preclinical syngeneic triple negative breast cancer(TNBC) genetically engineered mouse (GEM) models, which have been characterized genomically and with respect to their immune microenvironments. By integrating the immunological characterization of murine syngeneic mammary tumor models with analyses of human breast cancer datasets, we have demonstrated a relationship between EMT and myeloid cells, specifically tumor-associated macrophages (TAMs). We also have leveraged our syngeneic GEM models to define the response to immune checkpoint blockade therapy (ICBT) with emphasis on the myeloid cell environment. We have used our GEM models to test single agent drugs along with standard-of-care therapies for appropriate durations. Finally, we have tested these treatment regimens on established metastases with sufficient tumor burdens. Specifically, we will present results illustrating how CSF-1R targeting therapy combined with metronomic chemotherapy enhances a B and T cell response for the treatment of metastatic TNBC. In additional we well discuss how targeting the RNA helicase eIF4A that unwinds the secondary structures in the 5’UTR of mRNAs to facilitate translation initiation triggers an interferon response to suppress TNBC.

Citation Format: Jeffrey M Rosen, Diego Pedroza, Na Zhao, Xiang H-F Zhang, Charles M Perou. Leveraging preclinical models of triple negative breast cancer for translational research [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr IA05.
Abstract A029: Merlin/NF2 inhibits pancreatic cancer metastasis by activation of AMPK-dependent autophagic degradation of p62
Authors: Zhang Yuxia , Ojeda Virginia , Wang Xiaobo , Dunner Ken , Giancotti Filippo , Zhou Hao , Hernandez Jonathan , Xu Steven Q.
Published: February 2024

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR004) of the Conference Program/Proceedings.

Citation Format: Yuxia Zhang, Virginia Ojeda, Xiaobo Wang, Ken Dunner, Filippo Giancotti, Hao Zhou, Jonathan Hernandez, Steven Q. Xu. Merlin/NF2 inhibits pancreatic cancer metastasis by activation of AMPK-dependent autophagic degradation of p62 [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr A029.
Abstract A032: Cellos: High-throughput deconvolution of 3D organoid dynamics at cellular resolution for cancer pharmacology
Authors: Mukashyaka Patience , Kumar Pooja , Mellert David J. , Nicholas Shadae , Noorbakhsh Javad , Brugiolo Mattia , Anczukow Olga , Liu Edison T. , Chuang Jeffrey H.
Published: February 2024

Three-dimensional (3D) culture models, such as organoids, are flexible systems to interrogate cellular evolution, cellular growth and morphology, multicellular spatial architecture, and cell interactions in response to drug treatment. However, new computational methods to segment and analyze 3D models at cellular resolution with sufficiently high throughput are needed to realize these possibilities. Here we report Cellos (Cell and Organoid Segmentation), an accurate, high throughput image analysis pipeline for 3D organoid and nuclear segmentation analysis. Cellos segments organoids in 3D using classical algorithms and segments nuclei using a Stardist-3D convolutional neural network which we trained on manually annotated dataset of 3,862 cells. To evaluate the capabilities of Cellos we then analyzed 74,450 organoids with 1.65 million cells, from multiple experiments on triple negative breast cancer organoids containing clonal mixtures with complex cisplatin sensitivies. Cellos was able to accurately distinguish ratios of distinct fluorescently labeled cell populations in organoids, with < 3% deviation from seeding ratios in each well and was effective for both fluorescently labelled nuclei and independent Hoechst stained datasets. Cellos was able to recapitulate traditional luminescence-based drug response quantification by analyzing 3D images, including parallel analysis of multiple cancer clones in the same well. Moreover, Cellos was able to identify organoid and nuclei morphology features associated with treatment and unique to each of the clones. Finally, Cellos enables 3D analysis of cell spatial relationships, which we used to detect ecological affinity between cancer clones beyond what arises from local cell division and organoid composition. Cellos provides powerful tools to perform high throughput analysis for pharmacological testing and biological investigation of organoids based on 3D imaging.

Citation Format: Patience Mukashyaka, Pooja Kumar, David J. Mellert, Shadae Nicholas, Javad Noorbakhsh, Mattia Brugiolo, Olga Anczukow, Edison T. Liu, Jeffrey H. Chuang. Cellos: High-throughput deconvolution of 3D organoid dynamics at cellular resolution for cancer pharmacology [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr A032.
Abstract IA021: Digital patient avatars predict cancer evolution and personalised therapeutic strategies
Authors: Fisher Jasmin
Published: February 2024

Tumour heterogeneity manifested by genetic diversity and interchangeable cell states poses considerable challenges for the design of therapeutic strategies to combat cancer. While many strategies have been proposed to address this problem, using combination treatments and sequential delivery of therapies, the scope of combinations and sequencing order of therapies is enormous and is beyond the capacity of exhaustive testing through experimental models. We therefore propose using computational models to rapidly identify effective therapeutic interventions supported by convincing mechanistic explanations. Digital cancer avatars are computational programs representing oncogenic signalling networks and mapping molecular interactions to cell phenotype, which can be interrogated in silico. These computational programs specify how molecular interactions and perturbations produce cellular responses, such as cell proliferation, cell survival or cell death, and as such are well-suited to provide novel mechanistic insights into cellular behaviours as well as to identify therapeutic strategies for targeting them. In this talk, I will demonstrate the utility of bespoke computational models for triple-negative breast cancer, non-small cell lung cancer and melanoma to study how genomic changes drive tumour evolution and therapeutic response, and how these models are used to identify personalised combination treatments to overcome resistance. I will also present an in silico method applied to these models, to screen tens of millions combinations for effective sequential treatments by mapping the evolution of a tumour in response to a single-agent treatment and determining the optimal second-line treatment that will overcome resistance. Together these studies demonstrate the utility of digital avatars to improve our understanding of the mechanisms driving tumour progression and pave the way for personalised therapeutic strategies for improved cancer patient outcomes.

Citation Format: Jasmin Fisher. Digital patient avatars predict cancer evolution and personalised therapeutic strategies [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr IA021.
Abstract PR018: Adaptive local fitness landscapes for aneuploid karyotypes
Authors: Beck Richard J. , Andor Noemi
Published: February 2024

Aneuploidy, which occurs in most solid tumors, dramatically alters the expression of many genes which in turn has broad phenotypic effects. Recent evidence suggests that specific aneuploid karyotypes confer a selective advantage. However, various questions remain unanswered regarding the magnitude of these selective advantages, as well as the repeatability and predictability of karyotype evolution. We developed ALFA-K (Adaptive Local Fitness landscapes for Aneuploid Karotypes) – the first approach to infer karyotype fitness landscapes in a context-dependent manner. Based on the dynamics of just a few subclones derived from longitudinal single cell sequencing data, our method can extrapolate the fitness of thousands of karyotypes. We validate the predictive power of our method on data from several P53 deficient cell lines which exhibited substantial subclonal evolution across multiple passages in vivo or in vitro. Based on topological analyses of the inferred fitness landscapes we investigated how mis-segregation rate itself shapes the stringency of karyotype evolution and whether karyotype evolution is subject to diminishing returns epistasis, sign epistasis or reciprocal sign epistasis. We adapted metrics from geo-statistics to study the fitness landscape of a P53 deficient 184-hTERT diploid breast epithelial cell line that underwent whole genome doubling (WGD) and found support for the hypothesis that WGD benefits tumor cells by enhancing their robustness against chromosomal instability. The stark phenotypic consequences of karyotype changes emphasize the value of predicting the evolutionary trajectory of karyotype composition when exposing cells to a new environment. Our work contributes to accumulating evidence that karyotype evolution is predictable.

Citation Format: Richard J. Beck, Noemi Andor. Adaptive local fitness landscapes for aneuploid karyotypes [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr PR018.
Supplementary Table 1 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

CRISPR sequences.
Supplementary Figure 4 from Spatial Transcriptomics Suggests That Alterations Occur in the Preneoplastic Breast Microenvironment of BRCA1/2 Mutation Carriers
Authors: Caputo Anthony , Vipparthi Kavya , Bazeley Peter , Downs-Kelly Erinn , McIntire Patrick , Duckworth Lauren A. , Ni Ying , Hu Bo , Keri Ruth A. , Karaayvaz Mihriban
Published: February 2024

BRCA2 deficiency increases beta-1 integrin-mediated mechano-signaling compared to BRCA1 deficiency in ATCC's HMEC cells
Supplementary Figure 2 from Exosomal lncRNA NEAT1 Inhibits NK-Cell Activity to Promote Multiple Myeloma Cell Immune Escape via an EZH2/PBX1 Axis
Authors: Wang Qing-Ming , Lian Guang-Yu , Sheng Su-Mei , Xu Jing , Ye Long-Long , Min Chao , Guo Shu-Fang
Published: February 2024

Supplementary Figure 2 shows that MM exosomes derived from U266 cells or MM exosomes derived from U266 cells that NEAT1 knockdown had no effect on EZH2 expression.
Supplementary Figure 1 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 cells are more sensitive to cytotoxic T cell-secreted cytokines
dataset 3 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 data set 3. Potential dominant and co-drivers identified my the ML approach.
Data from Spatial Transcriptomics Suggests That Alterations Occur in the Preneoplastic Breast Microenvironment of BRCA1/2 Mutation Carriers
Authors: Caputo Anthony , Vipparthi Kavya , Bazeley Peter , Downs-Kelly Erinn , McIntire Patrick , Duckworth Lauren A. , Ni Ying , Hu Bo , Keri Ruth A. , Karaayvaz Mihriban
Published: February 2024

Abstract
Breast cancer is the most common cancer in females, affecting one in every eight women and accounting for the majority of cancer-related deaths in women worldwide. Germline mutations in the BRCA1 and BRCA2 genes are significant risk factors for specific subtypes of breast cancer. BRCA1 mutations are associated with basal-like breast cancers, whereas BRCA2 mutations are associated with luminal-like disease. Defects in mammary epithelial cell differentiation have been previously recognized in germline BRCA1/2 mutation carriers even before cancer incidence. However, the underlying mechanism is largely unknown. Here, we employ spatial transcriptomics to investigate defects in mammary epithelial cell differentiation accompanied by distinct microenvironmental alterations in preneoplastic breast tissues from BRCA1/2 mutation carriers and normal breast tissues from noncarrier controls. We uncovered spatially defined receptor–ligand interactions in these tissues for the investigation of autocrine and paracrine signaling. We discovered that β1-integrin-mediated autocrine signaling in BRCA2-deficient mammary epithelial cells may differ from BRCA1-deficient mammary epithelial cells. In addition, we found that the epithelial-to-stromal paracrine signaling in the breast tissues of BRCA1/2 mutation carriers is greater than in control tissues. More integrin–ligand pairs were differentially correlated in BRCA1/2-mutant breast tissues than noncarrier breast tissues with more integrin receptor-expressing stromal cells.
Implications:
These results suggest alterations in the communication between mammary epithelial cells and the microenvironment in BRCA1 and BRCA2 mutation carriers, laying the foundation for designing innovative breast cancer chemo-prevention strategies for high-risk patients.
Supplementary Figure S6 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 promotes the NRF2-dependent oxidative stress response
Abstract 1: Overcoming Clinical Trial Enrollment Challenges by Monitoring EMS Radio Transmissions: Pre-Hospital Screening of Acute Ischemic Stroke Patients
Authors: Sanchez Oriana N , Grotta James C , Bratina Patti , Stephenson Jonathan , Aiken Benjamin , Bowry Ritvij , Czap Alexandra L , Marella Prasen , Parker Stephanie , Barreto Andrew D , Dongarwar Deepa
Published: February 2024

Background:
Enrollment into hyperacute stroke trials may be constrained since candidates are usually not identified until they arrive in the emergency department. Pre-hospital identification may increase and speed study enrollment.

Methods:
Midway through the Multi-Arm Optimization of Stroke Thrombolysis (MOST) trial, our mobile stroke unit research personnel monitored EMS radio for potential stroke patients en route to our study hospital. Time of onset, symptoms, and expected arrival time were relayed to the MOST research team, who met the patient upon ED arrival. Expected site enrollment was 1 patient per month.

Results:
The rate of trial enrollment before intervention (10/2019 - 11/2021) was 0.8 patients per month compared to 2.4 after intervention (12/2021 - 07/2023), 2.43, 95% CI: 1.48-4.10, p<0.01 - see figure. In addition, there was a 17.5-minute reduction in ED arrival to trial randomization (88 vs. 70.5 minutes).

Conclusion:
Pre-hospital screening tripled the rate of trial recruitment and can help overcome challenges when enrolling participants in hyperacute clinical trials.
Abstract 100: Impact of Dissection Location on Subsequent Stroke Risk: A Nationwide Analysis
Authors: Shu Liqi , Strelecky Lukas , Xiao Han , Salehi Setareh , Goldstein Eric D , Nguyen Thanh N , Siegler James E , Penckofer Mary , Khan Farhan , Stretz Christoph , Furie Karen L , Yaghi Shadi
Published: February 2024

Introduction:
Cervical artery dissection (CAD) is a frequent cause of acute ischemic stroke (AIS) in young adults. We conducted a nationwide study to investigate potential predictors for subsequent AIS following CAD.

Method:
Adult patients diagnosed with non-traumatic CAD, and without any previous or concurrent stroke (2016-2020), were identified from the Nationwide Readmission Database using validated and standard ICD-10 CM codes. Significant demographic and clinical factors associated with 90-day AIS risk were identified through weighted survey univariate logistic regression followed by a multivariate Cox regression model.

Result:
Among 13,905 CAD patients (mean age 54.6±0.2, 54.8% female), 215 (1.5%) had subsequent AIS within 90 days of discharge with the median stroke occurrence on day 8. Patients with AIS were significantly older (58.3±1.3 vs. 54.5±0.2,
P
=0.026), and more commonly had vertebral artery dissection (51.6% vs. 41.0%,
P
=0.046), and coronary artery disease (24.7% vs. 15.6%,
P
=0.019). After including all these significant variables, the vertebral artery location of the dissection emerged as the sole significant predictor of subsequent AIS (adjusted HR 1.64, 95% CI 1.07-2.50,
P
=0.022). A Kaplan-Meier curve validated these findings, showing a higher AIS risk with vertebral versus carotid artery dissections (Figure). Notably, other risk factors such as hyperlipidemia and hypertension lacked significant association with AIS.

Conclusion:
Our study identifies vertebral artery dissection as a significant risk factor for 90-day AIS occurrence post CAD. This highlights the need for research into tailored management strategies for these patients to reduce stroke risk.
Abstract 101: Early Onset Hypertension Increases Risk of Midlife Stroke Among Black Women
Authors: Aparicio Hugo J , Lioutas Vasileios , Shulman Julie G , Himali Jayandra J , Cozier Yvette , Sheehy Shanshan
Published: February 2024

Background:
Stroke at midlife contributes to earlier mortality and greater disability. There is a critical lack of data about early life stroke risk factors among Black women, a group with higher rates of hypertension compared to other racial groups. The hypothesis of the study is that early onset hypertension increases risk of midlife stroke among Black women.

Methods:
The Black Women’s Health Study is a prospective study of 59000 Black women from across the US who were followed since 1995. Baseline year for this analysis was the 1999 questionnaire. We included stroke-free participants younger than age 65 (46754 participants, mean age 42±9.6 [range 24-64] years). History of hypertension was determined by self-report, defined as physician-diagnosed hypertension with use of an antihypertensive medication or diuretic, or use of an antihypertensive alone. Ischemic and hemorrhagic stroke were ascertained by self-report at biennial health questionnaires. Using Cox models, we estimated risk of midlife stroke, comparing persons with and without hypertension before age 45, between ages 45-64, and at 10-year age intervals. Follow up was from 1999 until whichever came first: incident stroke, death, loss to follow-up, or end of study follow-up (Dec 31, 2019).

Results:
At baseline, 2980 (10.5%) of participants age <45 and 7147 (38.7%) of participants between ages 45-64 had hypertension. Stroke occurred in 1485 participants (3.2%) during up to 23 years of follow up (mean 17.1±5.0 years). Compared to women with no history of hypertension, Black women with hypertension before age 45 had a higher risk of midlife stroke (HR=2.23, 95% CI=1.79-2.78), after adjustment for age, neighborhood socioeconomic status, residence in Stroke Belt, smoking, body mass index, and diabetes. Risk was also increased with hypertension at midlife age 45-64 (HR=1.69, 95% CI=1.47-1.95) and was highest among those with hypertension at age 24-34 (HR=3.15, 95% CI=1.92-5.16).

Conclusion:
Black women with hypertension before age 45 have more than twice the risk of stroke at midlife and hypertension before age 35 more than triples the risk. The findings are important to inform public health interventions to expand hypertension screening and treatment in this high-risk population.
Abstract 120:
CCR5
Polymorphism - A Possible Protective Factor for Post-Stroke PTSD Symptoms
Authors: Hallevi Hen , Tene Oren , Alpernas Aviva , Molad Jeremy , Ben Assayag Einor
Published: February 2024

Background:
Current data suggest 25-30% of stroke survivors will develop post-traumatic stress disorder (PTSD) symptomatology. This common and disabling disorder is thought to be caused by exposure to a traumatic, life-threatening event. It is unrelated to stroke severity and is one of the barriers to successful recovery from the stroke. We have recently reported on the naturally occurring loss-of-function mutation (LOFM) in the C-C Chemokine receptor 5 gene (CCR5-Δ32) as a protective factor in post-stroke recovery and depression. We sought to examine whether it also prevents the development of PTSD symptoms after stroke or transient ischemic attack (TIA).

Methods:
Participants were survivors of first-ever mild-moderate ischemic stroke or TIA from the TABASCO prospective cohort study, who underwent a 3T MRI, and were examined by a multi-professional team 6, 12, and 24 months after the event, using direct interviews, depression scales, neurological, neuropsychological, and functional evaluations. PTSD symptoms were assessed using the PTSD Checklist (PCL). CCR5-Δ32 status and PTSD Checklist data were available for 432 patients.

Results:
Forty-eight participants (11%) developed probable PTSD (PCL ≥44) during the first year after the stroke/TIA.
CCR5
-Δ32 carriers presented fewer PTSD symptoms (lower PCL scores) 6, 12, and 24 months after the index event, compared with non-carriers (p=0.002, p<0.001, p=0.011, respectively). The association remained significant after adjustment for age, gender, education, depression or administration of anti-depressants during the follow-up period, ethnicity, and new infarct on MRI. Compared to baseline,
CCR5
-Δ32 non-carriers tended to remain stable or develop worse PTSD symptoms over time, while
CCR5
-Δ32 carriers tended to improve.

Conclusions:
We report that carriers of the
CCR5
-Δ32 allele had a lower tendency to develop PTSD symptoms post-stroke. These findings might have clinical significance in stroke recovery. They also suggest a mechanism-based treatment target for post-stroke depression. Drugs mimicking this LOFM exist and could offer a novel treatment for this serious and underdiagnosed complication of stroke.
Abstract 125: Marked Variation in Perfusion Imaging Definitions of No-Reflow Phenomenon After Thrombectomy
Authors: Mutimer Chloe A , Mujanovic Adnan , Kaesmacher Johannes , Churilov Leonid , Kleinig Timothy J , Parsons Mark W , Mitchell Peter J , Campbell Bruce C , Ng Felix
Published: February 2024

Introduction:
The no-reflow phenomenon (persistent microvascular hypoperfusion despite macrovascular angiographic reperfusion) represents an explanation to poor outcome despite successful thrombectomy. There remains no universally-accepted definition to standardise future studies. We aim to compare the clinical features and outcomes of patients identified as having no-reflow using different perfusion MRI/CT definitions.

Methods:
We performed a pooled analysis of thrombectomy patients who underwent 24-hour follow-up perfusion MRI or CTs in the EXTEND-IA, EXTEND-IA TNK part 1 and 2 RCT. Presence of no-reflow was defined according to four definitions identified from a meta-analysis of 13 studies (Definition A = eTICI2c-3 and >15% asymmetry in CBV or CBF within the infarct on follow-up perfusion MRI/CT; definition B = mTICI2c-3 and >40% CBF asymmetry, definition C = mTICI2b-3 and presence of a Tmax>6s lesion; Definition D = mTICI2b-3 and >90% reduction of baseline Tmax>6s lesion). Receiver Operating Characteristics (ROC) analysis was performed with the outcome variable being poor functional outcome at 90 days (mRS≥3).

Results:
Of 325 patients analysed, the prevalence of no-reflow varied between definitions from 1.9 to 29.3% (p<0.001). There was poor agreement between definitions (kappa 0.062-0.745, 5 out 6 comparisons <0.196). Among patients identified as exhibiting no-reflow by any definition, there were significant differences in the intralesional interside differences in CBF (p=0.006), CBV (p<0.001) and MTT (p=0.005). Definition A yielded the highest Area Under the ROC Curve (AUC=0.679) for discrimination of 90-day functional outcome (Definitions C=0.649, D=0.597, B=0.515; p<0.0001). Sensitivity analyses testing across the eTICI≥2b, eTICI≥2c and eTICI3 strata showed consistent results.

Conclusions:
Existing imaging definitions of no-reflow varied significantly in prevalence and post-treatment perfusion imaging profile, suggesting that patients classified as having no-reflow by various definitions differ in their underlying pathophysiological processes. Definition A (eTICI2c-3 & >15% CBV/CBF asymmetry) discriminated prognostic performance best, supporting its use as the reference no-reflow imaging definition.
Abstract 136: The Incidence of Newly Diagnosed Cancer After a First-Ever Episode of Cerebral Venous Thrombosis: A Registry- and Population-Based Cohort Study
Authors: van de Munckhof Anita , Verhoeven Jamie , Vaartjes Ilonca , van Es Nick , de Leeuw Frank-Erik , Coutinho Jonathan M
Published: February 2024

Introduction:
Active cancer is a known risk factor for cerebral venous thrombosis (CVT), but it is unknown whether CVT is also associated with occult cancer. We evaluated the incidence of newly diagnosed cancer after CVT.

Methods:
We performed a registry- and population-based cohort study using the Dutch hospital discharge registry. Patients admitted to any Dutch hospital between 1997 and 2020 with a first episode of CVT were identified using ICD-9 and ICD-10 codes. Patients with a history of cancer and patients diagnosed with cancer during admission for the index CVT were excluded. We calculated the cumulative incidence of cancer after CVT with death as a competing risk. Standardized incidence ratios (SIRs) were calculated relative to a control group matched by age, sex, and calendar year with data from the Netherlands Cancer Registry.

Results:
We identified 2649 patients, 1856 (70%) were women and the median age was 44 (IQR 31-56) years. Cancer was diagnosed in 119 patients during follow-up (median duration 4.7 [IQR 1.9-8.9] years), of which hematologic (24%) and gastrointestinal tract (20%) cancers were the most common. The cumulative incidence of cancer in the overall cohort was 5.9% (95%CI 4.8-7.2) after 10 years. The highest cumulative incidence after 10 years was observed in patients ≥50 years (10.5% [95%CI 7.9-13.4]) and in men (9.9% [95%CI 7.1-13.1]). Patients with CVT had an increased rate of cancer compared to controls during the entire follow-up, but the difference narrowed over time (SIR 3.4 [95%CI 2.4-4.5] at 1 year and 1.4 [1.1-1.7] at 10 years). The rate was increased in both patients younger than 50 (SIR 6.7 [95%CI 4.0-10.6] at 1 year and 1.7 [1.2-2.3] at 10 years) and older than 50 years (SIR 2.4 [95%CI 1.5-3.6] at 1 year and 1.2 [0.96-1.6] at 10 years), as well as in men (SIR 3.6 [95%CI 2.2-5.6] at 1 year and 1.7 [1.2-2.2] at 10 years) and women (SIR 3.2 [95%CI 2.0-4.8] at 1 year and 1.2 [0.9-1.6] at 10 years).

Conclusion:
Patients with CVT have an increased risk of cancer compared to peers from the general population, regardless of age group or sex. The absolute risk of cancer was highest in older patients and men, while the relative risk was highest in younger patients. These results support the need for further research on screening for occult cancer in patients with CVT.
Abstract 145: Anticoagulation-Related Intracerebral Hemorrhage and Time to Reversal Treatment After Intracerebral Hemorrhage in the United States
Authors: Sheth Kevin N , Solomon Nicole , Alhanti Brooke , Messe Steven , Xian Ying , Bhatt Deepak , Hemphill Claude C , Frontera Jennifer , CHANG RAY , Danelich Ilya , Huang Joanna , Schwamm Lee H , Smith Eric E , Goldstein Joshua N , MacGrory Brian , Fonarow Gregg C , Saver Jeffrey
Published: February 2024

Background:
Anticoagulation-associated intracerebral hemorrhage (AC-ICH) often results in death. Specific reversal agents are available, but it is not clear whether there is a time-dependent treatment effect. We characterized patients with AC-ICH and investigated the relationship between time to treatment and outcome.

Methods:
We analyzed data from 9492 AC-ICH patients who presented within 24 hours of onset across 465 hospitals reporting any anticoagulation reversal treatment in GWTG-Stroke from 2015 to 2021. For patients with available door-to-treatment (DTT) times, outcomes were analyzed using logistic regression models adjusted for demographic, history, baseline, and hospital characteristics. A spline plot for mortality was generated for DTT and in-patient mortality from the adjusted predicted outcome probability.

Results:
Among 9492 patients admitted within 24h of AC-ICH, the median age was 77 years, 45% female, and NIHSS median 11. 79% (7469) received reversal treatment and 21% (2143) did not. Among reversal patients, pretreatment AC was warfarin in 62.2% and a direct oral anticoagulant in 37.8%. For 5224 patients with documented times, median onset to treatment time was 232 (IQR 142-185) minutes and DTT was 82 (58-117) mins, with DTT ≤60m for 27.7%. DTT ≤60 minutes was associated with lower odds of mortality than a DTT of >61-180 minutes (aOR 0.83 [95% CI: 0.69-1.00]). Factors associated with DTT ≤ 60 minutes include white non-Hispanic race, higher blood pressure, lower NIHSS, and a larger hospital size. Spline plotting showed a steep mortality increase with longer DTT in the first 30-minutes post-arrival (Figure).

Conclusion:
In US hospitals participating in GWTG-Stroke reporting reversal therapy, most patients with AC-ICH received reversal agents. Earlier administration was associated with improved survival. These findings support intensive efforts to accelerate evaluation and treatment for patients with this devastating form of stroke.
Abstract 146: Trained Immunity Induced by Dietary Salt Impedes Long-Term Brain Recovery in Experimental Intracerebral Hemorrhage
Authors: Lin Tze-Yen , Chen Wan-Ru , Zhang Xin-Yu , Lin Jhih Syuan , Hsu Chia-Lang , Chang Che-Feng
Published: February 2024

Introduction:
Excessive sodium intake is the top dietary risk factor for mortality and non-communicable diseases such as stroke. However, whether and how a high salt diet (HSD) impacts long-term stroke outcomes - even after salt reduction/withdrawal prior to a stroke is unknown.

Methods:
Intracerebral hemorrhage (ICH) was modeled by blood or collagenase injections in mice. Bone marrow (BM) progenitor and immune cell compositions as well as monocyte-derived macrophage (MDM) and microglial phenotypes were profiled using flow cytometry. BM chimeras were created to test innate immune memory. Glycolysis and OXPHOS were measured by Seahorse assays. Macrophage phagocytosis was evaluated using
in vivo
and
in vitro
erythrophagocytosis assays. RNA-seq was used to analyze the progenitor transcriptome. Hematoma, neuron loss, and astrogliosis were assessed with immunofluorescence. LysM
Cre/+
: NR4a1
fl/fl
; Nr4a2
fl/fl
; NR4a3
-/-
mice, LysM
Cre/+
: NR4a1
fl/fl
mice, and celastrol treatment were used to elucidate the mechanism underlying HSD-induced trained immunity.

Results:
HSD impaired long-term ICH recovery despite prior salt withdrawal. HSD triggered innate immune training in BM progenitor cells by downregulating the NR4a family and OXPHOS. This training selectively compromised MDM reparative polarization without altering their initial inflammatory response in the ICH brain. Healthy mice transplanted with BM from HSD-fed mice retained training signatures, confirming a persistent form of trained immunity that originates in the BM. Lack of NR4a1 in MDMs recapitulated HSD-induced negative impacts on ICH outcomes. Notably, transplantation of healthy BM and gain of NR4a1 function enabled stroke recovery in the HSD animals.

Conclusions:
These findings provide the first evidence that links HSD-induced trained immunity to the acquisition of persistent dysregulated inflammatory responses and unveil NR4a1 as a potential therapeutic target.
Abstract 15: Pharmacological Activation of Efferocytosis Prevents Intracranial Aneurysm Rupture
Authors: Maeda Takuma , Take Yushiro , Uchikawa Hiroki , Ishiguro Taichi , Ai Jinglu , Eguchi Satoru , Lawton Michael , Hashimoto Tomoki
Published: February 2024

Introduction:
Efferocytosis is the clearance process of dead or dying cells that reduces the tissue-damaging effects of excessive apoptotic cells. The accumulation of apoptotic cells at higher levels in ruptured aneurysms than in unruptured aneurysms suggests impaired efferocytosis. Mer receptor tyrosine kinase (MerTK), a receptor on macrophages, is essential for efferocytosis. Growth arrest-specific gene 6 (Gas6) is a small molecule that binds to and promotes MerTK-mediated efferocytosis. Our previous study suggested that MerTK knockout mice increased the rupture rate of intracranial aneurysms due to impaired efferocytosis. In this study, we investigated the effect of pharmacological activation of MerTK-mediated efferocytosis using the Gas6 in intracranial aneurysms.

Methods:
We used 10-week-old male C57BL/6J mice. We induced intracranial aneurysms by a combination of elastase injection and hypertension. After aneurysm induction, we started treatment with Gas6 or PBS as a vehicle. We compared the aneurysm formation and rupture rates, and the survival curve between the two groups.

Results:
The aneurysm rupture rate was significantly lower in the Gas6 group than in the control group (85.0% versus 50.0%,
P
<0.05, Fig. A and B). There was no statistical difference in the rate of aneurysm formation between the Gas6 and control groups (80.0% versus 100%,
P
=0.07). The Gas6 group had a better survival rate than the control group (
P
<0.05, Fig. C).

Conclusions:
Pharmacological activation of MerTK-mediated efferocytosis prevents intracranial aneurysm rupture. Our findings suggest that efferocytosis and MerTK may serve as novel therapeutic targets for the prevention of aneurysm rupture.
Abstract 156: Cerebral Edema in Patients Presenting With Large Ischemic Core Undergoing Thrombectomy versus Medical Therapy: Secondary Analysis of SELECT 2 Trial
Authors: Ng Felix , Johns Hannah Tabitha , churilov leonid , Yogendrakumar Vignan , Hassan Ameer E , Abraham Michael G , Ortega-Gutierrez Santiago , Hussain M. Shazam , Chen Michael , Wechsler Lawrence , Nguyen Thanh , Johanna Fifi , Ribo Marc , sitton clark , Mendes Pereira Vitor , Kasner Scott , Lansberg Maarten G , Pujara Deep K , Sharma Gaganjyoti , Guha Prodipta , Hill Michael D , Parsons Mark W , Davis Stephen M , Grotta James C , Albers Gregory W , Campbell Bruce , Sarraj Amrou
Published: February 2024

Background:
Endovascular thrombectomy (EVT) improves long-term outcome in patients presenting with large ischemic core but is associated with early neurological worsening (ENW), potentially related to cerebral edema post-reperfusion. We investigated whether EVT increases edema development, and the relationship between edema and ENW in a secondary analysis of the SELECT2 trial.

Methods:
SELECT 2 tested the efficacy and safety of EVT versus medical therapy in patients with large ischemic core (ASPECTS 3-5 or core volume ≥50ml on CT Perfusion[CTP]/MR diffusion). Core volume was defined as the larger of the CTP-CBF relative cerebral blood flow <30% or the visible hypodensity on non-contrast CT. Cerebral edema was measured as the maximum midline shift (MLS) on 24h follow-up MRI or CT. ENW was defined as >=4 point worsening in NIHSS at 24h compared to baseline. Probabilistic index model was used in multivariable analyses to assess ≥1 point improvement on modified Rankin Scale at 90-days.

Results:
After exclusion of 10 patients with hemicraniectomy, the median MLS in 327 patients was 2.75mm (IQR 0-5.85) in EVT and 0mm (IQR 0-4.02) in control patients (p=0.005). EVT was independently associated with greater MLS (adjusted odds ratio, aOR 1.50, 95%CI 1.17-1.92, p=0.0012) after adjusting for age and core volume. There was no interaction between EVT and core volume on the association with MLS (p>0.79). Patient with ENW had greater MLS (5.05mm [IQR2.06-10.2] vs 0mm [IQR0-3.94, p<0.001]) MLS was associated with development of ENW (aOR 1.22, 95%CI 1.13-1.32, p<0.001), and lower likelihood of long-term functional improvement (adjusted generalized OR 0.96, 95%CI 0.92-0.97, p<0.001, adjusted for core volume, EVT, age and baseline NIHSS). Sensitivity analysis testing alternative definitions of core volume (CTP core volume only and Hypodensity on non-contrast only) showed concordant results.

Conclusion:
In patients presenting with large ischemic core, EVT was associated with increased cerebral edema at 24h which, in turn, was associated with ENW. Despite overall benefit of EVT, EVT-related edema is independently associated with neurological deterioration and long-term disability and is therefore a potential treatment target to improve EVT outcomes.
Abstract 157: Endovascular Treatment of Acute Ischemic Stroke Patients With Tandem Lesions Presenting With Low Alberta Stroke Program Early Computed Tomography Score
Authors: Farooqui Mudassir , Galecio Castillo Milagros , Hassan Ameer E , Jumaa Mouhammad A , Divani Afshin A , Ribo Marc , Abraham Michael G , Petersen Nils H , Johanna Fifi , Guerrero Waldo , Malik Amer M , Siegler James E , Nguyen Thanh , Sheth Sunil , Yoo Albert J , Linares Guillermo , Janjua Nazli A , Quispe-Orozco Darko , TEKLE WONDWOSSEN , Zaidi Syed F , Sabbagh Sara , Zevallos Cynthia , Taborda Maria Belen , Barkley Tiffany , Majidi Shahram , Ackerman Jiyoun , Abdalkader Mohamad , Soomro Jazba , Gordon Weston , Salazar-Marioni Sergio , Rodriguez Aaron , Vivanco-Suarez Juan , Yavagal Dileep R , Mokin Maxim , Ortega-Gutierrez Santiago
Published: February 2024

Background and objective:
The effect of endovascular therapy (EVT) in tandem lesion (TL) patients with baseline large core infarct is unclear. We investigated the safety and efficacy of EVT in TL patients with low ASPECTS (0-5). We also compared the safety of acute carotid stenting (CAS) in TLs with low ASPECTS.

Methods:
This prospective multicenter study from 16 centers included patients with anterior circulation TL from 2015-2020. Inclusion criteria were age ≥18 years, EVT for intracranial occlusion, and underwent treatment for extracranial ICA lesions with >70% stenosis. Patients were divided into ASPECTS 0-5 and 6-10 groups. Inverse Probability of Treatment Weighting (IPTW) was used to assess the association of outcomes.

Results:
Of the 691 patients, 44 had an ASPECTS 0-5 and 505 had ASPECTS 6-10. After matching, there were decreased odds of favorable shift in mRS at 90 days (30.7% vs 48%, OR: 0.52, CI:0.29-0.97; p=0.002), higher rates of sICH (11.4% vs 3.6%, OR: 3.18, CI: 1.07-9.49; p=0.038), 90-day mortality (35.9% vs 16.3%; OR: 2.35, CI: 1.16-4.74; p=0.017) and comparable rates of successful reperfusion (79.5% vs 88.7%; OR: 0.73, CI: 0.33-1.63, p=0.45), excellent reperfusion (50% vs 49.1%; OR: 1.35, CI: 0.72-2.53; p=0.35) excellent reperfusion (50% vs 49.1%; OR: 1.35, CI: 0.72-2.53; p=0.35) in the ASPECTS 0-5 compared with 6-10. Among patients with ASPECTS 0-5, no significant differences were noted for sICH (OR: 0.31, CI: 0.03-3.05; p=0.32), PH2 (OR: 1.14, CI: 0.26-5.02; p=0.86), petechial hemorrhages (OR: 0.79, CI: 0.10-6.05; p=0.82), and 90-day mortality (OR: 0.68, CI: 0.14-3.45; p=0.6) in patients with stenting (18/44) versus no-stenting (25/44).

Conclusion:
This multicenter cohort study of AIS patients with TLs demonstrated poor outcomes for functional independence at 90 days and higher rates of sICH and mortality in patients with low ASPECTS (0-5). Among patients with ASPECTS 0-5, rates of sICH and mortality were similar in stenting and non-stenting groups.
Abstract 16: Integrin α9β1 and VCAM-1 Interaction Promotes Neutrophil Hyperactivation and Cognitive Decline Following Subarachnoid Hemorrhage
Authors: Kaur Harpreet , Pandey Nilesh , Zator Jacob , Murnane Kevin , Dhanesha Nirav
Published: February 2024

Background:
Integrin α9β1 is a transmembrane glycoprotein that is highly expressed on neutrophils and mediates transendothelial migration via vascular cell adhesion molecule-1 (VCAM-1). Neutrophils are major contributors to cerebral thrombosis and oxidative stress following subarachnoid hemorrhage (SAH); however, the mechanism by which neutrophil integrin α9 contributes to SAH pathogenesis is unclear. Here, we evaluated the mechanistic role of neutrophil integrin α9 in the post-SAH sensorimotor and cognitive recovery.

Methods:
SAH was induced by filament perforation in neutrophil-specific α9
-/-
(α9
fl/fl
Mrp8Cre
+/-
) mice and littermate controls. Cognitive recovery (Y-maze, Barnes maze, and novel object recognition test) and sensorimotor recovery (modified neurological severity score, corner, and cylinder test) were performed for up to 4-weeks. Unbiased RNA sequencing (RNA-seq) was used to evaluate the potential mechanisms by which neutrophil integrin α9-VCAM-1 interactions promote SAH. Computational modeling, docking, virtual screening, and cell-free assays were used to identify small-molecule inhibitors of the integrin α9-VCAM-1 interactions.

Results:
Experimental SAH resulted in increased levels of neutrophil integrin α9 which was concomitant with neutrophil hyperactivation. Post-SAH neutrophil-specific α9
-/-
mice exhibited improved sensorimotor and cognitive recovery, along with reduced plasma elastase and MPO levels and brain neutrophil infiltration. Mechanistically, integrin α9/VCAM-1 interactions promote neutrophil hyperactivation, extracellular signal-regulated kinase (ERK) phosphorylation, and endothelial cell apoptosis. RNA-seq data revealed that α9/VCAM-1 interactions promotes gene expression related to neutrophil inflammation, exocytosis, NF-κB signaling, and chemotaxis. Virtual screening and cell-free assay identified macitentan as a potent inhibitor of integrin α9-VCAM-1 interactions.
In vitro,
macitentan treatment decreased neutrophil adhesion to VCAM-1 and macitentan treatment at a dose of 5 mg/kg resulted in improved SAH outcomes.

Conclusion:
The results of this study describe a previously unrecognized pathway involving integrin α9 and VCAM-1 in the pathogenesis of SAH.
Abstract 27: A Novel Adjuvant Strategy After Endovascular Therapy: Arterial Glyceryl Trinitrate in Acute Ischemic Stroke for Neuroprotection (AGAIN)
Authors: Cheng Zhe , Geng Xiaokun , Gao Jie , Ding Yuchuan
Published: February 2024

Introduction:
Although mechanical thrombectomy (MT) demonstrates robust clinical efficacy in acute ischemic stroke (AIS), not all stroke patients benefit from successful reperfusion. The protective effect of NO donors has been shown to prevent ischemia-reperfusion injury through reduction of reactive oxygen species formation in cardiovascular studies. Glyceryl Trinitrate (GTN), a Food and Drug Administration-approved vasodilator, is one of the widely used exogenous NO donors used in the clinic and has been trialed for neuroprotection in AIS without successful clinical translation. The recent advances in MT for AIS have paved the way for promising new opportunities to supplementary neuroprotective therapies to patients after reperfusion. This study evaluated the safety, feasibility, and preliminary efficacy of intra-arterial administration of GTN after MT for neuroprotection.

Methods:
This is a prospective randomized controlled study (ID: ChiCTR2100045254). Eligible patients were randomized to receive 800μg GTN or same volume of normal saline through the catheter after recanalization. The primary outcome was symptomatic intracranial hemorrhage (sICH), while secondary outcomes included mortality, functional outcome, infarction volume, complications, and blood nitrate index (NOx).

Results:
A total of 40 patients were enrolled and randomized with no participants being lost to follow-up. There was no significant difference in the proportion of sICH between GTN and control groups. No significant difference was observed in mortality or rates of neurological deterioration and other complications. Favorable trends for both functional independence (mRS 0-2, 75.0% vs. 65.0%) at 90 days and reduction in final infarct volume (33.2 vs. 38.9 ml) have been observed in GTN group, although they did not reach the significant levels. Moreover, the concentration of blood NO
X
was increased (p<0.05) at 2 hours after GTN administration (26.2 vs 18.0 μmol/l).

Conclusion:
The AGAIN study suggests intra-arterial administration of GTN post MT is safe and feasible in AIS patient. The discernible positive trends may be due to the raised NO
X
levels, which potentially highlight the mechanistic reinforcement for these observed benefits.