Welcome to the WSC 2022 Interactive Program

Background and Aims

Currently transplantation of many different types of stem and progenitor cells has shown positive therapeutic results in experimental stroke. Each cell type has its own unique properties and mechanisms of action, which potentially can be combined to enhance the effectiveness of therapy. In this work we tested the comination of mesenchymal stem cells (MSCs) and neural progenitor cells (NPCs) in differentent dozes and delivery routs aming to study the cell distribution in ischemic rat brain.

Methods

Male Wistar rats (n=12) were subjected to 90min MCAO. After 24h transplantation of different combinations of SPIO labelled human placenta MSCs and directly reprogrammed NPCs was performed inside 7-MRI scanner: (1) intra-arterial 7x10⁵ drNPCs+ intravenous 20x10⁵ MSCs; (2) intra-arterial 3x10⁵ drNPCs+ intra-arterial 2x10⁵ MSCs; (3) intra-arterial 7x10⁵ drNPCs+ intra-arterial 5x10⁵ MSCs. Cell distribution in rat brain and complications were estimated.

Results

In group (1) drNPCs within 30min after IA administration were detected in the right hemishere, and MSCs were vizualuzed in both hemispheres 15-20 min after injection. In group (2) with IA transplantation of low dozes of drNPCs and MSCs only small amount of labeled cells were detected in the infarct core and peripherally to it. In group (1) and (2) no complications were detected. In group (3) with the IA injection of higher dozes of drNPCs and MSCs cerebral embolism was observed.

Conclusions

It can be concluded that intra-arterial administration of MSCs and drNPCs in low dozes and combination of intra-arterial and intravenous delivery routes are safe and provide adequate cell distibution in the ischemic brain.

Background and Aims

Bone marrow derived mononuclear stem cells (MNC)secrete neural metabolites which help in recovery after stroke. In this study in vitro H-NMR spectroscopy of serum samples in stroke patients were studied after intravenous stem cell transplantation and physiotherapy.

Methods

Fourteen patients were randomized to autologous bone marrow mononuclear stem cells (BM-MNC) infusion (active) and saline (comparator) followed by 8 weeks of physiotherapy (groups 1& 2 undisclosed). Serum venous samples were drawn at baseline and 8 weeks for 1H-NMR spectroscopy. 340μl serum and 30μl TSP (0.5mM) were added in 200ml of D2O using 700 MHz NMR spectrometer (M/s. Agilent Technologies, USA). Proton NMR spectra of serum samples were acquired using 1D CPMG with presaturation using 90˚ pulse sequence, with 64 scans; relaxation delay=70 s; spectral width= 9000 Hz

Results

No adverse reactions were reported. Mean age for all patients was 52.3+4.7 years, 8M/6F. PLS-DA analysis of group 1 patients revealed significant difference in cellular metabolites (fig1). It was also observed that variable projection analysis (VIP) of metabolites showed a trend on an increased choline (3.2ppm), myelene lipids (ML;1.28ppm), lactate (1.3ppm) and glutamate (2.4ppm) in group 2 patients (fig2). Two sample t-test between group 1 & 2 showed moderately significant levels of NAA and mylene lipids (ML) peaks at 8 weeks (95% CI: 3.2 to 12.6, p=0.045; 95 % CI; 6.2-5.4, p=0.05 respectively) as compared to other peaks (p>0.05).

Conclusions

It was observed that choline, myoinositol, lactate, glutamate, histidine, tyrosine, NAA, NAA/cr and myelene lipids were significantly exhibited in both groups 1 & 2 post treatment.

Background and Aims

The tissue plasminogen activator (t-PA) is currently the only drug treatment available for ischemic stroke. We previously found that intravenous administration of human amnion epithelial cells (hAECs) in mice reduces brain injury at 24-72 h following stroke. Here, we assessed the neuroprotective efficacy of hAECs in the presence of t-PA.

Methods

Male C57BL/6 mice (aged 8-12 weeks) were subjected to middle cerebral artery occlusion for 1 h followed by reperfusion. Immediately following reperfusion, vehicle (saline, n=18) or t-PA (Alteplase, 10 mg/kg; n=83) was administered intravenously. After 30 min of reperfusion, t-PA-treated mice were injected intravenously with either hAECs (1x10⁶; n=39) or vehicle (2% human serum albumin; n=44). Mice were randomly assigned to one of the three treatment groups and euthanised at 3, 6 or 24 h (n= 21, 35, and 45, respectively), at which time brains were collected to assess infarct volume progression via analysis of thionin-stained sections.

Results

While there was no mortality within 6 h of stroke, high mortality at >6 h in t-PA-treated mice was found compared to t-PA plus hAECs (60% vs 27%). This outcome prevented an unbiased comparison of infarct volume of survivors at 24 h. However, at 6 h, infarcts in the subcortical region (corresponding to the expanding infarct core) were ~50% larger in t-PA- vs vehicle-treated mice (22.1±2.9 mm³ vs 14.7±2.3 mm³) but were ~40% smaller in mice receiving hAECs (13.0±2.2 mm³, p=0.03 vs t-PA alone).

Conclusions

When administered in combination with t-PA, hAECs attenuated infarct growth and mortality following stroke.

Background and Aims

Transient ischemic attack (TIA) presents a high risk for subsequent stroke and Alzheimer’s disease (AD) and related dementia (ADRD). However, the neuropathophysiology of TIA has been rarely studied. By evaluating recurrent TIA-induced neuropathological changes, our study aimed to explore the potential mechanisms underlying the contributions of TIA to ADRD.

Methods

Recurrent TIA was induced by three times 10-min middle cerebral artery occlusion (MCAO) within a week in rats. Neurological symptom and locomotor function were evaluated by Garcia Neurological score and cylinder test. Rats were sacrificed at 1, 3, and 7 days after the last TIA episode and brains were processed for immunohistochemistry and Western blot analysis of AD-related biomarkers, neuronal cytoskeleton, astrogliosis and microgliosis.

Results

Neither neurological deficit nor apoptosis were observed following recurrent TIA. No increase of AD-related biomarkers was indicated after TIA, including tau hyperphosphorylation and β-site APP cleaving enzyme 1. Neuronal cytoskeleton modification and neuroinflammation was found at acute and subacute stage after TIA, evidenced the reduction of microtubule-associated protein 2 (MAP2), elevation of neurofilament-light chain (NFL), and increase of glial fibrillary acidic protein (GFAP)-positive astrocytes and ionized calcium binding adaptor molecule 1 (Iba1)-positive microglia at the affected cerebral cortex and basal ganglion. Similar NFL, GFAP and Iba1 alteration was found in the corpus callosum at the subacute phase after TIA.

Conclusions

Recurrent TIA triggers neuronal cytoskeleton change, astrogliosis and microgliosis at the acute and subacute stage. Our study indicates that the TIA-induced neuronal cytoskeleton modification and neuroinflammation may be involved in the vascular contribution to cognitive impairment and dementia.

Background and Aims

The incidence rate of ischemic stroke over the world at young age is rising of which 10-30% are young adults in the age group between 18-45 years. Whole Exome Sequencing (WES) help in identifying the disease causing rare genetic variants in young Ischemic Stroke. Other contributory genes which were never implicated in the neurological disorders could be uncovered through WES.

To identify the rare or low frequency variants in young Ischemic stroke.

Methods

We executed the WES in 100 young IS cases with the age range of 18-40years. A total of 459 controls were used for frequency and family-based segregation analysis. Identified 41 variations in 22 genes from intra-family and inter-family analysis were further validated in unaffected family members. Confirmed variations were replicated in another cohort of 100 cases and 100 controls with mass array genotyping

Results

A total of 19 variations from 13 genes were identified. Eleven genes (HBB, VWF, HHIPL2, CUL9, KALRN, ZNF318, TGFB2, TTBK1, F13A1, CCND3, and RPL7L1) were from intra-family analysis, and two genes (PDE4DIP and NOS2) from the inter-family analysis. Three genes (TTBK1, CUL9 andRPL7L1) are not reported with any neurological disorder yet. Two variations c.17_18del, p.P6fs in HBB and c.G2561A, p.R854Q in vWF were reported as pathogenic. Two loci rs10482721 in TGFB2 and rs56320013 in KALRN were replicated in another cohort of 100 cases and 100 controls.

Conclusions

Rare or low-frequency variants inclined towards the risk of Ischemic stroke were revealed in this study. The TGFB2 and KALRN could be considered a genetic signature of the stroke.