bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract WP346: Pharmacological Inhibition of Transforming Growth Factor Beta Receptor Improves Breathing Function and Cognition in Mice Models of Dementia
Authors: El Hamamy Ahmad , IQBAL ZAHID , Ranjan Arya , Zhang Yuxing , Sumani Destiny , McCullough Louise , Li Jun
Published: January 2025

Dementia is associated with respiratory dysfunction. In mice models of dementia, there is increased astrogliosis in the brain stem retro-trapezoid nucleus (RTN), which is crucial for breathing control. Our previous work using a genetic approach suggested that increased transforming growth factor beta receptor (TGFβR) signaling may be responsible for RTN astrogliosis and contribute to respiratory and cognitive dysfunctions. Here we tested our hypothesis that pharmacological inhibition of TGFβR2 in mice models of dementia may reduce RTN gliosis, breathing disorder and cognitive impairment.

We used male 16-month-old Tg-2576 and Tg-SwDI mice that model Alzheimer’s Disease (AD) and Cerebral Amyloid Angiopathy (CAA), respectively with WT controls. LY2109761, a selective TGF-β receptor inhibitor, was administered orally every day for 6 weeks at a 50 mg/kg dose. Barnes Maze and Novel Object Recognition Test (NORT) were used to measure cognitive strength, and plethysmography was used to measure respiratory metrics such as apnea rate after the 6 weeks of treatment. Immunofluorescence was used to quantify gliosis (GFAP) at RTN. All data is presented as Mean±SEM.

Tg-2576 mice performed significantly worse than the WTs in the Barnes Maze (escape latency 184±20.99 vs. 119.4±12, p=.039, n=8-9/group), in the NORT (percentage time with the novel object 48.3±2.061 vs. 63.3±2.071, p=.0003, n=8-9/group), and showed increased apneas per minute (9±.9 vs 4.33±.81, p=.001, n=8-9/group). Similarly, Tg-SwDI mice showed impaired breathing and cognition function. In Tg-2576 mice, TGFβR2 inhibition improved performances in Barnes Maze (112±18.6 vs. 184±20.9, p= .02 n=8-9/group) and NORT (57.7±2.54 vs. 48.3±2.06, p=.023, n=8-9/group), and reduced apneas per minute (4.88±.058 vs. 9.0±.98, p=.004, n=8-9/group) compared with vehicle treatment. In Tg-SwDI mice, TGFβR2 inhibition also improved performance in Barnes Maze (43.8±2.72 vs. 58.2±5.16, p=.03 n=7/group) and NORT (59.1±3.28 vs. 46.6±3.32, p=.02, n=7/group), and reduced apneas per minute (5±1.06 vs. 9.71±1.71, p=.038 n=7/group) compared with vehicle group. TGFβR2 pharmacological inhibition additionally reduced astrogliosis in the RTN of Tg-2576 mice (drug 53.62±4.8 vs. vehicle 70.9±3.38, p=.018, n= 4/group).

In conclusion, pharmacological inhibition of TGFβR2 improved cognition in AD and CAA mice. Improving breathing control via reducing gliosis at breathing center RTN may be the underlying mechanisms of the improvement.
Abstract WP353: Mesenchymal-derived extracellular vesicles mediate shifts in microglial phenotypes in functional recovery in a monkey model of cortical injury
Authors: Medalla Maria , Zhou Yuxin , Mojica Chromewell , McCann Ryan , Pessina Monica , Zeldich Ella , XIN Hongqi , Rosene Douglas , Moore Tara
Published: January 2025

Mesenchymal stem cells (MSCs) have recently received attention as an intervention to reverse or slow neurodegenerative, stroke and injury-related changes in the aging brain as they suppress inflammation and facilitate tissue repair. In support of this idea, we have shown that extracellular vesicles (EVs) derived from bone marrow MSCs enhance recovery of motor function of the hand following cortical injury in aged female rhesus monkeys. Specifically, EV-treated treated monkeys (n=5) exhibited full recovery of fine motor function by 3-5 weeks post-injury while vehicle monkeys (n=5) reached a plateau short of full recovery by 8-12 weeks post-injury. Post-mortem analyses of perilesional brain tissue from the same monkeys, revealed multifaceted cellular effects of EVs including downregulating inflammatory microglial phenotypes, dampening neuronal hyperexcitability, and enhancing neuronal and myelin plasticity. To explore the relationship between these processes and functional recovery, we utilized multi-labeling immunohistochemistry (IHC) and high-resolution confocal microscopy to assess microglia phenotypes, neuronal synaptic marker expression and microglia-neuronal interactions in perilesional cortex. To assess these markers, semi-quantitative stereology in Neurolucida and particle analysis in ImageJ were utilized. Results show that MSC-EV treatment decreased the densities of pro-inflammatory hypertrophic microglia expressing LN3+, a marker for MHC II receptors, upregulated with immune activation. Conversely, MSC-EV treatment and functional recovery was significantly correlated with increased proportion of hypertrophic microglia expressing the key complement pathway protein C1q, a phenotype associated with enhanced debris-clearance and repair after degeneration. Interestingly this EV-associated increase in C1q+ hypertrophic microglia was correlated with decreased putatively damaged synapses tagged with C1q and greater synaptic marker expression in perilesional cortex. These data suggest that MSC-EVs promote a shift from pro- to anti-inflammatory repair microenvironment, via enhancement of debris clearance after injury. These findings demonstrate the efficacy of MSC-EVs as a therapeutic, likely acting to reduce inflammatory cascades, facilitate repair and rebalance neuronal synaptic connections to support recovery after cortical injury.
Abstract WP354: Neuroinflammatory Factor Response in Acute Ischemic Stroke Correlates with Lesion Volume and Stroke Severity: Circulating Blood from a Mobile Stroke Unit and during Hospitalization
Authors: Kowalski Robert , Thompson John , Ledreux Aurelie , Violette John , Paustian Whitney , Neumann Robert , Ornelas David , Monte Andrew , Dylla Layne , Coughlan Christina , Jones William , Grotta James , Graner Michael
Published: January 2025

Background:
Circulating neuroinflammatory factors change rapidly in acute stroke, but the association with ischemic lesion size and stroke severity is unknown. We used a mobile stroke unit (MSU) to identify inflammatory markers within minutes of symptom onset, and from blood at 24 hours. We compared these findings with stroke size and severity.

Methods:
The study is a prospective, cohort investigation of ultra-early blood-derived inflammatory factors in acute stroke. Blood was obtained on the MSU, in the hospital, and from healthy adult controls. Plasma was analyzed with electrochemiluminescence for 9 inflammatory factors. Lesion volume was quantified from MRI with computational software. Variables included demographics and NIHSS.

Results:
Seventy-one adults were analyzed, including 46 patients treated on the MSU between August 2021 and May 2024, and 25 controls. Twenty-six (57%) MSU patients were confirmed ischemic stroke (AIS). For AIS patients, median age was 65 (range 36-88) years, 42% were female, and 76% white. Median initial NIHSS score was 6 (range 1-22; median hospital discharge NIHSS score was 1 (range 0-20). Twenty-three (88%) patients received IV thrombolysis and 7 (27%) had mechanical thrombectomy. Median lesion volume on MRI was 1.4 (range 0-171) mL. Blood was obtained a median of 57 (range 26-355) minutes from stroke symptom onset on the MSU, and MRI was obtained a median of 12.5 (range 2.2 to 31.8) hours. Compared with controls, of factors evaluated in blood obtained on the MSU, IL-6 and IL-4 were elevated; IFNγ, IL-12p70 and IL-2 were reduced; and IL-10, IL-17, IL-1β and TNF-α were comparable. Plasma levels of IFNγ, IL-17 and IL-2 at 24 hours were inversely correlated with lesion volume (Spearman’s rho: -0.540, -0.558 and -0.423, respectively). Change in IL-17 and IL-2 between the blood obtained on the MSU and blood 24±6 hours later (median +86 fg/mL and +55 fg/mL, respectively) was inversely correlated with lesion volume (Spearman’s rho: -0.580 and -0.495, respectively. Plasma IFNγ in blood on the MSU correlated with NIHSS score at hospital discharge (Spearman’s rho: 0.485).

Conclusions:
This study found the acute stroke response for circulating inflammatory factors IFNγ, IL-17 and IL-2 correlated with stroke lesion volume and IFNγ correlated with severity on the NIHSS. This suggests these factors may serve as surrogate markers for brain imaging. The findings may aid development of targeted immunomodulatory therapies in acute stroke.
Abstract WP359: Increased CCL4 and CCL5 following Mechanical Thrombectomy: A Plasma Biomarker Study in Ischemic Stroke Patients
Authors: Li Kelly , Finger Carson , Delevati Colpo Gabriela , Couture Lucy , Verma Pritt , Dongarwar Deepa , Manwani Bharti
Published: January 2025

Background:
Mechanical thrombectomy has been associated with improved outcomes in patients with acute ischemic stroke. However, not all patients with successful reperfusion experience good functional outcomes. Investigating the post-reperfusion inflammatory milieu could provide valuable insights into the mechanisms that influence stroke outcomes. We hypothesized that successful reperfusion may cause distinct cytokine/ chemokine profiles.

Methods:
Plasma samples were collected from patients with ischemic stroke that were admitted to Memorial Hermann Hospital, Houston within 24 hours of admission. Multiplex was used to analyze the plasma samples for levels of 48 cytokines and chemokines. The relationship between thrombectomy (TICI 2b and above) and analytes was assessed using ANOVA and then adjusted association was examined using multilinear regression models after adjusting for patient demographics and comorbidities.

Results:
The mean age of patients was 63.3 years, 54% were women, 87% had hypertension, 44% had diabetes mellitus, and 54% had hyperlipidemia. Only two out of the 48 cytokines demonstrated differences with thrombectomy. CCL4 levels were significantly higher in patients that underwent thrombectomy, 55.7±17.4 pg/ml vs. 39.5±20.6 pg/ml in the ones that did not undergo this procedure, p=0.01. Similarly, CCL-5 (RANTES) levels were elevated in patients with acute ischemic stroke that underwent thrombectomy 368.6±275.1 vs. others, 216.5±156.3, p=0.01. These remained significant after adjusting for co-morbidities.

Conclusions:
Our study suggests that MIP-1β (CCL4) and RANTES (CCL5) are significantly elevated in ischemic stroke patients that underwent successful reperfusion by mechanical thrombectomy. CCL4 or MIP-1β (macrophage inflammatory protein -1) is a chemoattractant for monocytes in injured tissue. CCL5 or RANTES (
r
egulated on
a
ctivation,
n
ormal
T
-cell
e
xpressed and
s
ecreted) is also a chemokine secreted by T cells and monocytes and directly interacts with CCL4. CCL4 and CCL5 may contribute to the post-ischemia reperfusion inflammatory response, potentially influencing outcomes after reperfusion therapies such as mechanical thrombectomy. Longitudinal analysis of these chemokines and other cytokines after mechanical thrombectomy, along with their correlation with functional outcomes may provide deeper insights into the underlying mechanisms and also identify potential biomarkers of successful mechanical thrombectomy in stroke patients.
Abstract WP361: MBS-1 enhances brain cytoprotection in ischemic stroke using the Stroke Preclinical Assessment Network (SPAN) paradigm
Authors: Wang Yaping , Pan Yan , Zhang Shiqing , Lu Dan , HU LIJUN , Wang Ying , Shi Lei , Ye Wencai , XU ANDING
Published: January 2025

Background:
The latest Global Burden of Disease study indicates that stroke remains the second major cause of death. Promising protective effects have been shown in most studies of ischemic stroke (IS) protection in preclinical animal models, but have failed to enter clinical trials. The objective of this study is to investigate the brain cytoprotective effects of the natural compound MBS-1 in IS, using a novel, rigorous, and feasible paradigm inspired by Stroke Preclinical Assessment Network (SPAN).

Methods:
This study performs the SPAN methodology in two centers, and takes including a randomization, blinding, and placebo control approach, utilizing a novel multi-arm (multiple groups) and multi-stage (multiple time points) design along with the establishment of comorbidity models. Specifically, the study uses wild-type (WT) C57BL/6J mice (8~10 weeks, Half male and half female), aged mice, and diabetic mice to establish a middle cerebral artery occlusion (MCAO) model. We assess the recovery of motor and sensory functions using behavioral tests such as the corner test, cylinder test, modified neurological severity score (mNSS), and Longa score. To identify the protein expression and localization, we use Western blot analysis and immunofluorescence staining. The infarct volume is detected using magnetic resonance imaging (MRI) or triphenyltetrazolium chloride (TTC) staining. Transmission electron microscopy (TEM) and Golgi staining are applied to assess neuronal morphology and number, while Nissl and TUNEL staining are used to evaluate neuronal function and apoptosis levels.

Results:
In the permanent MCAO model induced by chloride iron in WT young mice, MBS-1 significantly reduces the infarct volume, improves neurological function, and decreases neuronal apoptosis. In the MCAO/reperfusion (MCAO/R) model conducted by intraluminal filament in WT young mice, MBS-1 also effectively reduces the infarct volume, enhances neurological function, and decreases neuronal apoptosis. While in the MCAO/R model conducted by intraluminal filament in WT aged mice, MBS-1 visibly improves the survival rate of mice. Furthermore, in the MCAO/R model of diabetic mice, MBS-1 significantly reduces the infarct volume and improves neurological function.

Conclusion:
Using SPAN paradigm, MBS-l exhibits brain cytoprotection in various IS models.

Key words:
Stroke Preclinical Assessment Network, MBS-1, ischemic stroke, brain cytoprotection
Abstract WP362: Can Plasma Extracellular Vesicle-Associated Proteins Differentiate Cerebrovascular Disease Pathologies?
Authors: Trout Amanda , Roberts Jill , ODell Christopher , Prince Christiaan , Walker Mayah , Whitnel Laura , Frank Jacqueline , Milson nathan , Pahwa Shivani , Harp Jordan , Dornbos David , Pennypacker Keith , Stowe Ann , Fraser Justin
Published: January 2025

The ability to differentiate cerebrovascular disease (CVD) with plasma biomarkers could be an invaluable tool. CVD encompasses a spectrum of conditions that alter intracranial blood flow and activate endothelial cells (ECs), which results in a dysregulated blood brain barrier (BBB). Activated ECs upregulate and cleave many junctional proteins, including soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intracellular adhesion molecule-1 (sICAM-1), and von Willebrand factor (vWF). These junctional complexes regulate BBB permeability and infiltration of leukocytes that release interferon-gamma (IFN-gamma) for immune cell regulation. Key barriers to providing therapies for CVD are due to lack of biomarkers to differentiate pathologies. Utilizing extracellular vesicles (EVs), which are nanoparticles, released from all cell carrying cargo specific to their parent cells microenvironment, may provide insight into cerebrovascular changes before overt pathological and cognitive changes are present. Therefore, we unbanked plasma from the Blood And Clot Thrombectomy Registry And Collaboration” (BACTRAC; NCT03153683) and “Moyamoya and Stroke Tissue Evaluation and Repository” (MASTER) to measure EV sVCAM-1, sICAM-1, vWF and IFN-γ concentrations in aneurysms, emergent large vessel occlusions (ELVO), and Moyamoya. Moyamoya vasculopathy is a unique CVD that presents with internal carotid artery terminus stenosis and abnormal vascular collaterals. EVs were isolated using size exclusion chromatography, concentrated, and the protein ran on MSD assays. Aneurysms (n=17, 76.5% female), ELVO (n=20, 65% female), and Moyamoya (n=21, 66.7 % female) did not have significant differences in sex, body mass indexes, hypertension, smoking status or diabetes between groups. ELVOs were significantly older (72.75 ± 3.3 years) than Moyamoya (47.2.1 ±2.0, p<0.0001) and aneurysms (50.9 ± 5.5, p=0.01) and had significantly lower EV IFN-gamma (p=0.004). In ELVOs, lower EV IFN-gamma correlated to larger edema volumes (p=0.023, r=-0.821) and higher stroke severity (NIHSS, r=-0.812, p=0.05). Moyamoya EVs showed increased sVCAM-1 (p= 0.069) and sICAM (p=0.057), compared to aneurysm, with significantly lower (p=0.001) vWF, compared to aneurysms and ELVOs. Higher EV sICAM-1 trended (p=0.059, r=0.794) for better cognitive outcomes following ELVO. These data suggest that markers of BBB dysregulation in plasma EVs could provide some insight into type and severity of CVD.
Abstract WP363: Metabolic Dysfunction after Ischemic Stroke: Changes in Insulin Resistance and Glycolysis
Authors: Couture Lucy , Banerjee Anik , Delevati Colpo Gabriela , Lee Juneyoung
Published: January 2025

Introduction:
Acute ischemic stroke (AIS) is a leading cause of mortality and disability worldwide. Social Isolation (SI) has been identified as a modifiable risk factor for stroke and a contributing factor to poorer functional outcomes. SI has been increasingly recognized as a significant stressor that can impact metabolic health through various biological mechanisms. The relationship between SI and metabolic dysfunction is complex and involves interactions among the neuroendocrine, immune, and metabolic systems. However, studies investigating the role of SI in the development of host metabolic dysfunction in the context of stroke are limited.

Hypothesis:
Isolation alters metabolic pathways after stroke.

Methods:
We analyzed untargeted serum metabolic profiles of AIS patients (N=100) 24 hours after last known well, along with questionnaires regarding SI and loneliness, to assess metabolic differences between socially isolated and socially interactive individuals. Samples were collected from patients ranging from 29 to 89 years old, with 66 males and 34 females. SI was measured via the Lubben Social Network Scale collected from patients during their hospitalization.

Results:
We found that there is a significant correlation between Lubben Social Network score and alterations in genes related to insulin resistance and glycolysis (FDR adjusted p <0.05). Interestingly, acetyl-CoA carboxylase and glycogen phosphorylase were significantly downregulated in patients with lower Lubben Social Network scores, suggesting isolated individuals have lower rates of metabolic turnover. Our results also demonstrated that suppressor of cytokine signaling (SOCS) was shown to be upregulated with an increased Lubben Social Network scores, suggesting that SI patients have systemic cytokine signaling and inflammation compared to non-SI stroke patients.

Conclusion:
In summary, systemic alterations in metabolic activity are pronounced after stroke, correlate with SI status pre-injury and functional outcome post-injury, and have the potential to directly affect cellular function. We demonstrated that socially isolated individuals are more likely to have metabolic dysfunction, which is a significant risk factor for post stroke disability and recurrent stroke. These metabolic factors should be analyzed additionally through the lens of sex, race, and other social determinants of health that affect stroke severity and functional outcomes.
Abstract WP366: Escape of
Kdm6a
from X Chromosome is Detrimental to Ischemic Brains via IRF5 Signaling
Authors: Ngwa Conelius , Misrani Afzal , Manyam Kanaka Valli , Xu Yan , McCullough Louise , Liu Fudong
Published: January 2025

Introduction:
The fundamental roles of chromatin biology and epigenetics in progression of diseases have been increasingly recognized. Some genes escaping from X chromosome inactivation (XCI) impact neuroinflammation through epigenetic regulation. Our previous studies have suggested that the X escapee genes
Kdm6a
and
Kdm5c
were involved in microglial activation after stroke in aged mice. However, the underlying mechanisms remain elusive.

Hypothesis:
We hypothesized that
Kdm6a/5c
demethylate H3K27Me3/H3K4Me3 in microglia respectively, and mediate the transcription of interferon regulatory factor 5 (IRF5) and IRF4, leading to microglial pro-inflammatory responses and exacerbated stroke injury.

Methods:
Aged (17–20 months)
Kdm6a/5c
microglial conditional knockout (CKO) female mice (one allele of the gene) were subjected to a 60-min middle cerebral artery occlusion (MCAO). Gene floxed females (two alleles) and males (one allele) were included as controls. Infarct volume and behavioral deficits were quantified 3 days after stroke. Immune responses including microglial activation and infiltration of peripheral leukocytes in the ischemic brain were assessed by flow cytometry. Epigenetic modification of IRF5/4 by Kdm6a/5c were investigated by CUT&RUN ChIP assay.

Results:
The demethylation of H3K27Me3 by
Kdm6
a, increased IRF5 transcription; meanwhile Kdm5c demethylated H3K4Me3 to repress IRF5 (IRF5 ratio H3K27Me1/H3K27Me3 in fl/y vs. fl/fl, p = 0.0331; fl/fl vs. CKO, p = 0.0082, Figure 1A). Both Kdm6a
fl/fl
and Kdm5c
fl/fl
mice had worse stroke outcomes compared to fl/y and CKO mice (Figure 1B). Gene floxed females showed more robust expression of CD68 in microglia (
Kdm6a
fl/y vs. fl/fl, p < 0.0001), elevated brain and plasma levels of IL-1β or TNF-α, after stroke (Figure 1C, 1D).

Conclusion:
We concluded that IRF5 signaling plays a critical role in mediating the deleterious effect of Kdm6a; whereas Kdm5c’s effect is independent of IRF5.

Keywords:
Stroke, Aging,
Kdm6a/5c
, Microglia, Epigenetics, Ischemia, IRF
Abstract WP372: Edaravone Dexborneol Protects Against Blood-brain Barrier Disruption Following Cerebral Ischemia/reperfusion By Activating The PDGFB/PDGFR-β Signaling Pathway To Inhibit Pericyte Injury
Authors: Sun Zhiyu , Zhao Hanshu , Yang Shanshan , Liu Ruijia , Yi Lian , Gao Jiadi , Liu Sihan , Chen Yinlin , Zhang Zhongling
Published: January 2025

Introduction:
Recent advances in brain cytoprotection therapies following cerebral ischemia-reperfusion (I/R) injury have become an emerging interest. Our previous study has shown the great potency of Edaravone Dexborneol (Eda.B), the latest generation multitarget cytoprotective agent, in alleviating I/R injury, while the mechanism warrants further investigation. Pericytes play important roles in protecting blood-brain barrier (BBB) integrity and regulating haemodynamics. This study aims to explore of Eda.B's protective effect on pericyte injury after I/R and its underlying mechanisms.

Methods:
The rat transient middle artery occlusion (t-MCAO) model was established. Rats were randomly divided into Sham group (Sham, n=24), tMCAO group (tMCAO, n=24), Edaravone group (Eda, n=24), Dexborneol group (Dexborneol, n=24), and Eda.B group (Eda.B, n=24). Neurological function recovery, infarct volume, BBB disruption was assessed using Zea-Longa scoring, TTC staining, and Evans Blue extravasation, respectively. Basement membrane (BM) edema and pericyte coverage were assessed by transmission electron microscopy (TEM). The expression levels of pericyte marker proteins NG2 and PDGFR-β in the ischemic zone, BBB transcellular transport-related proteins VTN, α5, PDGFB were detected by western blot. Furthermore, a high-affinity monoclonal antibody against PDGFB, MOR8457, was used to explore Eda. B’s protectiveness on pericytes injury via PDGFB/PDGFR-β (Eda.B+MOR8457, n=8).

Results:
Eda.B significantly reduced cerebral infarct volume and promoted neurological function recovery, and showed more significant promotion compared to tMCAO group, Eda group and Dexbornrol group. Additionally, Eda.B significantly reduced the BBB leakage, decreased pericyte coverage, the number of endotheliocytes (EC) vesicles, and the extent of BM edema than the other groups. Furthermore, Eda.B inhibited the downregulation of NG2, PDGFB/PDGFR-β, VTN, and the upregulation of α5 protein expression, which were induced after I/R. Blocking PDGFB with MOR8457 after receiving Eda.B indicated that, Eda.B improved pericyte loss and BBB permeability by activating PDGFB/PDGFR-β signaling.

Conclusions:
This study unravels new insights into mechanisms of pericyte as a promising therapeutic target and Eda.B’s protectiveness against pericytes loss and BBB permeability after I/R injury.
Abstract WP375: A New Neurological Assessment Focused on Consciousness and Quantitative Motor Function in a Rat Model of Subarachnoid Hemorrhage
Authors: Kanamaru Hideki , Zhu Shiyi , Han Mingyang , Huang Lei , Sherchan Prativa , Suzuki Hidenori , Tang Jiping , Zhang John
Published: January 2025

Objective:
In clinical practice, initial neurological status such as impaired consciousness is believed to correlate with patient outcomes in subarachnoid hemorrhage (SAH). However, evaluation methods such as the modified Garcia Score (mGS) commonly used in SAH basic research are inadequate for assessing consciousness, lack quantitative measures, and are subjective. This study aimed to develop an evaluation method that overcomes these limitations.

Methods:
A SAH model was created using Sprague-Dawley rats (Male, 280–320 g). The study included a SAH group (n=18) and a sham-operated group (n=4), established via the endovascular perforation method. In addition to conventional neurological assessments (mGS and beam balance test [BB]), consciousness and brainstem function were evaluated using seven parameters: respiration, whisker movement, eye blink, pinna reflex, vision, auditory response, and righting reflex. These evaluations were conducted by three independent raters. A grip strength test was also performed to quantitatively assess motor function. The primary outcome evaluation was conducted 24 hours post-SAH, followed by brain removal and analysis after euthanasia. Brain swelling was assessed by measuring brain volume, and neurodegeneration was evaluated using Fluoro-Jade C staining. Inter-rater reliability (Fleiss Kappa) was calculated for each evaluation item, and correlations with outcomes were analyzed using Pearson's correlation coefficient.

Results:
Two rats in the SAH group died within 24 hours and were excluded from the analysis. The following seven parameters showed significantly high inter-rater agreement and a correlation with primary outcome: spontaneous activity, whisker movement, auditory response, righting reflex, climbing movement, forelimb movement, and BB). These seven parameters plus a grip strength test was included to develop a new scoring system (Figure), and this new method was compared with the conventional mGS method. The new assessment showed improved correlation with outcomes and reduced variability between raters’ scores.

Conclusion:
The new scoring system, incorporating assessments of consciousness and quantitative motor function, provided a more accurate reflection of brain damage in SAH model rats compared to traditional methods, with improved inter-rater agreement.
Abstract WP378: Fingolimod (FTY720) attenuates neuroinflammation, neuronal cell death, and white matter injury in the rodent model of subarachnoid hemorrhage
Authors: Rangasamy Suresh , Geraghty Joseph , Testai Fernando
Published: January 2025

Introduction:
Subarachnoid hemorrhage (SAH) is associated with elevated morbidity and mortality. We previously showed that the immunomodulatory agent FTY720 improves neurological outcome in rats subjected to SAH.

Hypothesis:
FTY720 protects against SAH-associated neuronal cell death and white matter injury (WMI).

Methods:
We used the endovascular perforation model. Animals were divided into Sham, SAH-vehicle, and SAH-FTY720 groups. The SAH-FTY720 group received a single dose of FTY720 (0.5 mg/kg) intraperitoneally 3 hours after surgery. Brains were isolated and analyzed on day 7 post-injury. Neuroinflammation was ascertained by immunostaining for astrocyte (GFAP) and microglia (Iba-1) cell and the expression of proinflammatory mediators inducible nitric-oxide synthase (iNOS) through the NF-κB signaling pathway and TLR4. WMI was determined by quantifying the expression of myelin basic protein (MBP) and oligodendrocyte precursor cells (NG2). Neuronal death was determined by NeuN staining. Statistical analyses were performed with one-way ANOVA followed by Tukey’s post-hoc test using Graph Prism 10. Data are represented as mean±SD. Statistical significance was determined at the level of
p
<0.05.

Results:
5 rats were included per group. Relative to the Sham group, SAH-vehicle animals had a decreased number of NeuN(+) cells but an increased number of Iba-1(+) (
p<
0.001) and GFAP(+) cells (
p<
0.001) in the ipsilateral cortex and CA1 region of the hippocampus. FTY720 after SAH normalized the number of NeuN(+), Iba-1(+), and GFAP(+) cells in both regions (
p<
0.001 vs SAH-FTY720). These findings were also confirmed by Western blot analysis for GFAP and Iba1. The SAH-vehicle group also had an increased expression of iNOS and TLR4 in cortex and hippocampus (
p
<0.001 vs Sham) which colocalized with Iba-1 and GFAP. Furthermore, SAH increased the expression of activated NF-κB (Pp65) but decreased the expression of the NF-kB inhibitor, IkBα in the cortex and hippocampus as compared to the Sham group (
p<
0.001 vs Sham). The expressions of iNOS, TLR4, IkBα, and Pp65 were normalized in the SAH-FTY720 group. The number of MBP(+) and NG2 (+) cells was decreased in the ipsilateral cortex and corpus callosum of SAH-vehicle animals as compared to Sham group (
p<
0.001 vs Sham). Conversely, the expression of both markers in the SAH-FTY720 group was similar to the Sham group in both regions.

Conclusion:
FTY720 reduces neuroinflammation, neuronal death, and WMI after SAH.
Abstract WP380: Oligodendrocytes Damage in Cerebral Amyloid Angiopathy
Authors: Jiang Danye , Garg Shivi , Khan Romeesa , McCullough Louise
Published: January 2025

Introduction:
Cerebral amyloid angiopathy (CAA) leads to amyloid β (Aβ) deposition in the cortical and leptomeningeal vessels, and it is a largely untreatable cause of intracerebral hemorrhage and dementia. Studies estimate a 10-50% prevalence of CAA in the elderly population, positioning CAA as one of the strongest vascular contributors to age-related cognitive impairment. While reduced white matter (WM) volume and altered WM integrity have been reported in CAA, the response of oligodendrocytes – the specialized glial cells responsible for myelin production – to Aβ deposition remains unclear. Further, the potential role of oligodendrocytes in CAA-induced cognitive decline has yet to be elucidated.

Methods:
TgSwDI mice harboring Swedish, Dutch, and Iowa mutations of human amyloid precursor protein were used as a mouse model of CAA. Motor and cognitive behaviors were assessed in male and female C57BL/6 wildtype (WT) and CAA mice at 3 and 12 months of age using the open field test (OFT) and fear conditioning (FC). At 12 months, oligodendrocyte and microglia phenotypes, vascular structure, and Aβ burden were evaluated using flow cytometry and immunostaining.

Results:
At 12 months, WT and CAA animals exhibited comparable gross motor activity. CAA mice displayed significant cognitive deficits in FC compared to WT mice, which retained intact cognition (n=6-15/grp, p<0.05). Notably, these cognitive deficits were more prominent in female CAA mice. At 12 months, CAA mice also displayed reduced corpus callosum (CC) thickness. Sex differences were observed in CAA mice in oligodendrocyte (Olig2
+
) numbers in the CC and hippocampal CA1 region (n=4/grp, p<0.05), as well as in microglial (Iba1
+
) phagocytosis of oligodendrocytes (n=7/grp, p<0.01) using immunostaining and 3D reconstruction. Additionally, a higher frequency of disease-associated microglia (DAM, Clec7a
+
) was detected near the CC in CAA mice using flow cytometry and immunostaining (n=4/grp, p<0.01).

Conclusion:
CAA mice exhibited cognitive decline without motor impairment, alongside significant changes in oligodendrocyte frequency and increased DAM presence near the corpus callosum. The increase in microglial phagocytosis of oligodendrocytes suggests a potential mechanism underlying oligodendrocyte damage and WM injury in CAA. Further research is needed to clarify whether these changes in oligodendrocytes contribute to cognitive decline in CAA.
Abstract WP381: Spatiotemporal imaging and molecular signatures of stem cell transplant-driven recovery in stroke-injured brains
Authors: Habib Pardes , Liang Xibin , Noristani Rozina , Kim Jieun , Ewbank Sedona , Airan Raag , Bliss Tonya , Steinberg Gary
Published: January 2025

Introduction:
Intraparenchymal transplantation (tx) of human neural stem cells is in clinical trial for chronic ischemic stroke, but the mechanisms of action are unknown. Here we use longitudinal MR imaging and spatial transcriptomics in a rodent stroke model to investigate the regenerative responses of cell therapy on regional and molecular levels.

Methods:
Adult male Sprague Dawley rats were subjected to 30 min of stroke (tMCAo) followed 1 month later by transplantation of embryonic-derived neural stem cells (NR1) or buffer into the ipsilesional striatum. We combined longitudinal MRI scans (T2w, FLAIR, DWI, SWI, DTI) with neurobehavior (whisker paw test, cylinder test, EBST, corner turn test, T-maze) and spatiotemporal transcriptomics.

Results:
NR1 transplantation significantly improved motor, sensorimotor and cognitive performance compared to the vehicle group (p<0.01). Most NR1-treated rats (94%) exhibited a new cortical T2-FLAIR signal, associated with positive outcomes, compared to 33% in the vehicle group. Additionally, the NR1-induced FLAIR signal was more hyperintense and persisted longer than in the vehicle group. Unbiased spatial transcriptomics of the cortical FLAIR region revealed a microglia/inflammation-associated signature including pathways involved in phagocytosis, immune cell response, cytokine signaling, and ER stress. DTI analysis of corticospinal, corticostriatal, nigrostriatal, spinothalamic, and callosal interhemispheric pathways showed that quantitative anisotropy (QA) values in the vehicle group remained stable from pre-transplant to 3 months post-transplant. In contrast, NR1-treated rats had higher pre-transplant QA values, which dropped below vehicle levels by day 7 post-transplant, but then significantly increased by 3 months, exceeding pre-transplant levels. This suggests substantial white matter recovery and fiber tract regeneration in the NR1-treated rats.

Conclusion:
Our study demonstrates T2-FLAIR may be a potential prognostic marker following stem cell transplantation, with further molecular analysis needed to deepen our understanding of brain restoration. Monitoring pre-transplant and early post-intervention QA values may enable patient selection and predict both early and long-term responses to cell therapy.
Abstract WP384: Plasma small extracellular vesicles derived from patients with cerebral aneurysms impair cerebral endothelial permeability and promote angiogenesis
Authors: Chebl Alex , Zhang Jing , Aldilaimi Akram , Teng Hua , Li Yanfeng , Chopp Michael , Zhang Zheng Gang
Published: January 2025

Background:
Small extracellular vesicles (sEVs) from blood samples of patients with cerebral aneurysms (CA) have been used as a biomarker for CA. However, biological function of sEVs in CA has not been studied. We tested the hypothesis that plasma sEVs from patients with CA impair cerebral endothelial cell (CEC) function.

Methods:
Plasma sEVs were isolated using ultracentrifugation from venous and arterial blood of patients with CA undergoing endovascular treatment. Size and marker proteins of sEVs were characterized. Human CECs were treated with sEVs at 1x10
8
particles/ml. The effects of sEVs on CEC permeability and angiogenesis were measured by trans-endothelial and tube formation assays, respectively. One-way ANOVA was used for statistical analysis.

Results:
Ten patients, 6 with non-ruptured CA and 4 with ruptured CA, ranging in age from 51-88 years old were studied. There were no significant differences in sEV sizes and protein markers between non-ruptured and ruptured CA (141 ± 6 nm vs 144 ± 2 nm). The permeability assay showed that compared to controls, sEVs derived from arterial samples (asEVs) from ruptured and non-ruptured CA significantly increased CEC leakage (116 ± 7 and 119 ± 14 vs 101 ± 6 control, respectively, p<0.05). The angiogenic assay showed that venous sEVs (vsEVs), but not asEVs, of non-ruptured CA, significantly (p<0.05) increased vascular branches (14 ± 6 vs 6 ± 3 control), however, both vsEVs and asEVs of ruptured CA robustly increased the branch numbers (13 ± 7, and 13 ± 5 vs 6 ± 3 control, respectively, p<0.05).

Conclusions:
Increased CEC permeability and vascular branches are highly related to blood brain barrier (BBB) damage and angiogenesis, respectively. Thus, our data suggest that sEVs from CA could damage the BBB and promote angiogenesis, which provides new insights into the role of sEVs in mediating CA progression.
Abstract WP385: Impact of estrous cycle hormones on optogenetic stimulation and stroke recovery dynamics
Authors: Ko Hee Jae , Chiang Terrance , Chen Hansen , Radit Nahin T. , Cheng Michelle , Steinberg Gary
Published: January 2025

Background:
A damaged stroke area can affect both local and connected brain regions, leading to network-wide disruptions in brain functions. Increasing evidence shows that estrogen, particularly 17β-estradiol, plays a protective role after ischemic stroke. Understanding how hormonal levels impact brain stimulation may shed light on varying responses to neuromodulation therapies across estrous cycle phases. Previously, we demonstrated that optogenetic stimulations in the ipsilesional primary motor cortex (iM1) enhance functional recovery in male mice. In this study, we aim to expand upon these findings by exploring functional recovery and post-stroke neuroplasticity changes in in female mice.

Methods:
Female C57BL/6 mice (7-9 weeks old) underwent stereotaxic surgery to express channelrhodopsins-2 and implant an optical fiber in iM1. Four weeks later, transient middle cerebral artery occlusion (MCAO) was induced. Optogenetic stimulation was initiated on day 5 post-stroke and continued for 10 days. Estrous cycles were monitored via vaginal swabbing and estradiol ELISA measurements. Rotating beam tests were performed at pre-stroke, 4, 7, 10, and 14 days post-stroke (PD4, PD7, PD10, and PD14). Infarct size, astrocytic activation and microglia/macrophage activation were assessed.

Results:
Rotating beam tests showed that 71.4% of the stimulated mice significantly recovered by PD10 (p<0.01). We observed irregular hormonal cycles post-stroke, with the diestrus phase noticeably longer in majority of mice. Stimulated mice exhibit a trend toward better recovery when in the diestrus/metestrus phase at the start of stimulation on PD5. Optogenetic stimulation appeared to reset the hormonal cycles, though some irregularities remained during stimulation period. Additionally, recovery varied during estrous cycle stages, with optogenetic stimulation enhancing motor function during specific phases. At PD10, improved recovery was significantly correlated with low estradiol levels (R
2
= 0.61, p<0.05), and most recovered mice were in the estrus phase by PD14.

Conclusions:
These results highlight the importance of considering hormonal cycles in post-stroke brain stimulations. Ongoing studies are investigating the effects of stimulation on hormonal cycles in relation to post-stroke plasticity. Our data suggest that tailoring brain stimulations to specific estrous hormonal phases may optimize recovery outcomes.
Abstract WP386: Juvenile mice have more robust acute inflammation after stroke than adults but less chronic inflammation, suggesting that the juvenile brain terminates the acute inflammatory response more efficiently.
Authors: Vanden Kelly , Buckwalter Marion , Mayne Elizabeth
Published: January 2025

Introduction:
Children typically recover better after stroke than adults. In adults, neuroinflammation exacerbates infarct volume acutely and drives infarct-induced neurodegeneration chronically. The evolution of neuroinflammation after stroke in the juvenile brain is poorly understood compared to perinatal or adult stroke. Here, we quantify the initiation and resolution of inflammation after stroke in juvenile mice.

Methods:
Wild-type male juvenile (p28) and adult (6 months) C57BL/6J mice were randomized to sham surgery or DH (distal hypoxic) stroke, an established model of cortical stroke that produces chronic secondary neurodegeneration in connected gray and white matter regions. We performed immunostaining at 3 days (acute) and 8 weeks (chronic) after stroke to quantify myeloid (CD68, Iba1), astroglial (GFAP), and lymphocytic (B220, CD3) inflammation within the infarct and sites of secondary neurodegeneration.

Results:
Acutely, juvenile mice had 267% more CD68 (p = 0.0023) and 235% more Iba1 (p = 0.0086) staining in the thalamus. Juveniles also had greater myeloid cell activation in connected white matter tracts, with 322% more CD68 (p < 0.001) and 696% more Iba1 (p = 0.0002) staining in the corpus callosum, and 260% more Iba1 staining in the corticospinal tract (p = 0.0059) than adults (n=6-11/group). There was no difference in acute GFAP staining. In contrast, by 8 weeks after stroke, juveniles had less CD68 staining in the thalamus (19% of adult, p <0.0001), corpus callosum (68% of adult, p = 0.0428), and corticospinal tracts (55% of adult, p = 0.0009) (n = 6-10/group). Juveniles and adults had comparable lymphocytic neuroinflammation in the infarct.

Conclusions:
Acutely, juveniles had widespread myeloid cell activation in gray and white matter sites that are connected to the infarct and undergo secondary neurodegeneration, which, unexpectedly, was markedly greater than adults. Strikingly, this relationship reversed by 8 weeks after stroke, with juveniles demonstrating significantly more efficient resolution of inflammation than adults. These results suggest that children's ability to terminate the acute inflammatory response after stroke may be protective and could contribute to the more favorable outcomes after pediatric stroke. Future studies will test hypothesis that early termination of the acute inflammatory response after stroke is protective and will compare the transcriptomic response to stroke in juvenile vs adult myeloid cells.
Abstract WP39: Pretreatment Factors Associated with Symptomatic Stroke in Moyamoya Disease Patients: Long-Term Multicenter Experience
Authors: Musmar Basel , Roy Joanna , Salim Hamza , Kaul Anand , Atallah Elias , El Naamani Kareem , chen jared , Jabre Roland , Saad Hassan , Grossberg Jonathan , Dmytriw Adam , Patel Aman , Khorasanizadeh Mirhojjat , Ogilvy Christopher , Thomas Ajith , Monteiro Andre , Siddiqui Adnan , Cortez Gustavo , Hanel Ricardo , Porto Guilherme , Spiotta Alejandro , Piscopo Anthony J. , Hasan David , Ghorbani Mohammad , Weinberg Joshua , Nimjee Shahid , Bekelis Kimon , Salem Mohamed , Burkhardt Jan-Karl , Zetchi Akli , Matouk Charles , Howard Brian , Lai Pui Man Rosalind , Du Rose , Abbas Rawad , Sioutas Georgios , Amllay Abdelaziz , Munoz Alfredo , Herial Nabeel , Tjoumakaris Stavropoula , Gooch Michael , Rosenwasser Robert , Jabbour Pascal
Published: January 2025

Background:
Moyamoya disease (MMD) is a cerebrovascular disorder marked by the progressive steno-occlusion of the bilateral internal carotid arteries and the formation of abnormal collateral vessel networks at the base of the brain. Previous studies have attempted to identify risk factors predictive of postoperative complications to improve patient management. This study aims to identify pretreatment factors associated with symptomatic stroke in MMD patients.

Methods:
This study is a multicenter retrospective analysis conducted across 13 academic institutions in North America. A total of 518 patients with MMD were included. Data collected included patient demographics, disease characteristics, and follow-up duration. Stroke-free survival was analyzed using Kaplan-Meier curves. Univariate and multivariable Cox regression analyses were used to identify risk factors for symptomatic stroke.

Results:
The median age of the patients was 43 years (IQR, 34–52 years), and 370 (71%) were females. Hypertension was present in 255 (49%) patients, diabetes mellitus in 144 (28%), and 192 (37%) were smokers. Multivariable Cox regression identified advanced age (HR 1.03, 95% CI 1.01–1.05, p = 0.011), female sex (HR 2.03, 95% CI 1.00–4.11, p = 0.049), diabetes mellitus (HR 2.03, 95% CI 1.14–3.63, p = 0.016), smoking status (HR 2.27, 95% CI 1.27–4.05, p = 0.006), and incidental findings (HR 0.37, 95% CI 0.15–0.93, p = 0.034) as significant factors associated with symptomatic stroke.

Conclusion:
Advanced age, female sex, diabetes mellitus, and smoking status were significant predictors of symptomatic stroke in MMD patients. Patients with incidental findings had a reduced risk of stroke. These findings emphasize the importance of managing modifiable risk factors and the potential benefits of early detection in improving clinical outcomes for MMD patients. Further prospective studies are needed to validate these findings.
Abstract WP391: Preclinical imaging evaluation of protective effect of SEW2871 against blood-brain barrier dysfunction associated with cerebral small vessel disease
Authors: Shal Bushra , Griego Michael , Yang Yirong , Sillerud Laurel , Perez Carlos , McChriston Joanna , Deng Haoran , Chen Haojie , Maez Sean , Akter Halima , Yang Yi
Published: January 2025

Cerebral small vessel disease (SVD) contributes to approximately 50% of human dementias world-wide and pose a burden to the health care system worldwide. Early endothelial/blood-brain barrier (BBB) dysfunction is an important contributor to the pathogenesis of SVD. Sphingosine-1-phosphate (S1P), a membrane-derived signaling sphingolipid, regulates the BBB integrity by binding to its receptor isoform 1 (S1PR1) on endothelial cells. We previously showed that activation of S1PR1 using SEW2871, a selective S1PR1 agonist, reversed the loss of cerebral capillary endothelial S1PR1 and BBB integrity in a rat model of SVD. The current study evaluated multi-doses of SEW2871 as a novel therapy by targeting BBB damage for SVD. Male and female spontaneously hypertensive rat stroke-prone (SHRSPs) were subjected to unilateral carotid artery occlusion (UCAO) followed by Japanese permissive diet (JPD) for 35 days. Three doses of SEW2871 (0.5, 1.0 and 5.0 mg/kg) were tested. Neuropathological outcomes were assessed with MRI, immunohistochemistry, and Morris water maze (MWM). Body weight, oxygen saturation, and heart rate were evaluated weekly. SEW2871 plasma concentrations were measured at 2, 5, and 24 hours after injection. MRI showed increased BBB permeability, white and gray matter lesions (WML and GML) and decreased cerebral blood flow at 2 and 5 weeks after UCAO/JPD onset. T2 and fractional anisotropy images showed that three doses of SEW2871 demonstrated protective effect on WML and GML with most significant improvements in 1.0 mg/kg group. Dynamic contrast-enhanced MRI and arterial spin labeling (ASL) maps showed improved BBB leakage and CBF most remarkable in SEW2871 0.5 and 1.0 mg/kg groups compared to the vehicle rats. Furthermore, MWM test demonstrated that treatment with SEW2871 1.0 mg/kg presented significant improvement in spatial memory compared to UCAO/JPD group. Post-mortem immunohistochemical staining for IgG and Iba-1 showed BBB leakage and active microglia in lesioned WM and GM, which were significantly attenuated by SEW2871 1.0 mg/kg compared to vehicle rats, supporting the results obtained from MRI. Our data suggested that long-term treatment with SEW2871 reduced BBB permeability leading to improvement of brain lesions, CBF reduction, and cognitive impairment. The results also indicated that the effectiveness of SEW2871 in all doses tested, while 1.0 mg/kg showed a most remarkable protection against the neuropathological cascades associated with SVD.