Hello, everyone. I'm Xiaofen Jia, professor of the Department of Neurosurgery, University of Maryland School of Medicine. Today, I will introduce human neural stem cell improved functional outcome after cardiac arrest. Annually, around 300,000 patients suffer from cardiac arrest. About 10 to 25 patients survive to hospital discharge. Majority of them, survivors are comatose. Brain injury is a big impediment to functional recovery. Currently, neither pharmacological intervention nor therapeutic hypothermia can resolve the neural injury caused by cardiac arrest. Neural stem cell therapy is a promising treatment for brain injury and has been reported to significantly improve the functional outcomes in stroke, HIE, and TBI. For example, in preclinical studies, it has been reported the ICV delivery or systemically delivery, intracerebral delivery of this human neural stem cell used in the rat stroke model and improves the outcome after that. There's several preclinical and clinical studies have demonstrated that stem cell administration in ischemic brain injury in stroke model is safe and effective. Through multiple possible mechanisms, for example, promoting the growth factor release, preserves the BBB integration protection, increases the exosome release. These mechanism studies have been really limited in the setting of brain injury post-cardiac arrest. We know we have different rules for stem cell administration. They all have their own advantage and disadvantage. For example, intranasal. Intranasal delivery is non-invasive. It directly migrates to the brain. It can easily repeat different doses. However, the clinical evidence is lacking or limited. ICV delivery, which allows for delivery to multiple sites of the CNS. And it seems this is quite fit the global ischemic brain injury after cardiac arrest, though it's invasive. We initially, we started intranasal delivery administration of human neural stem cell in rogen asphyxial cardiac arrest animal model. So this animal, we will subject it to the cardiac arrest. And intranasal is human neural stem cell was delivered and we can see the brain behavioral outcome and histopathological damage score, both are increased. So the intranasal delivery demonstrated as a therapeutic effect on the ischemic brain injury after cardiac arrest. And we then were testing the ICV delivery. And we would like to see the possible mechanism with that ICV administration. So this we tested in both male and female in control and NSC treatment group. This animal was subjected to eight minutes asphyxial cardiac arrest model. The deliver of the HNSC by ICV was at three hours after our SC. So then we were tested by EEG. We were tested neurological recovery by NDS. And we can also test the immunofluorescent staining for the possible, you know, neuron and the core staining with others, for example, human nuclear antibody to identify the survived or differentiated human neural stem cell. And eventually we also tested the patch clamber to see how these transplanted human neural stem cell works. So we can see from the neurodeficit scores, that is, we can see left side is a green, blue one is a control, right side is NSC group. So that is overall NDS scores. And this one is sub NDS score, which is coordinating and the behavioral indicating the high level neural functional activities. So both overall NDS and the sub NDS demonstrated that HNSC group is a significant improvement in NDS compared to control group. And we then do the quantification of ischemic neuron. And by the cross-valid staining and the quantification, we can see, for example, in this CA2, this is a control. The yellow indicated ischemic neuron, and this one is NSC group. Yellow also indicated the ischemic neuron. We quantify that we can conclude it. The ischemic neuron is significantly less by the NSC treatment. We then checking the grafted NSC end cells, and we also checking the neural survival. So we can see from here, this is N, E, U, N, the neural marker. And we can compare in the C1, C2, that's NSC, that's control. Similarly, this is in different C3, and this is DG. We can all quantified them and demonstrated the HNSC significantly improved the neuron survival. And we marked this with QUA6. This is a marker with human nuclear. So indicating this survived the increase, the neural survival with the core staining, the way this KU86 indicating the effect of the human neural stem cell rather than new NSC recruited from local. And we all try to evaluate the possible mechanism. And we tested, we found, noticed that the wing beta-catenin pathway is significantly increased in the NSC together with KI67, which is proliferation marker, so indicated. At four weeks after that, representative animals, which we tested, evaluated this where is the administrated human neural stem cell, we pre-labored with PKH26. This actually, they injected through ICV and migrated to C1. We can see this is co-laboring with PKH26. We pre-labored it before the injection. And we also, at four weeks, we demonstrated with PETR-CLAM that we can show in the spontaneous excitatory synapse current representing the cells receiving synaptic input from other neuron, which indeed is the cells transplanted human neural stem cells. So this study, we also demonstrated that transplanted human neural stem cell via ICV administration markedly improved neurological outcome after cardiorest. And possibly this beneficial mechanism through the neuronal protection and upregulated the proliferation of SVZ cells. So we have a couple clinical trial or clinical studies using stem cells in stroke. They all demonstrated these cells, these treatment are safe, but effects are not clear. So the limitation, possible limitation is, one is lower rates of cell survival and poor outcomes in growth. I mean, they are not differentiated well to the targeted cell type, what we hope they will after the transplantation. Well, they have the multiple strategy for improving stem cell application in ischemic brain injury, like we recently summarized. For example, we can do stem cell pretreatment. We can use biomaterials. We can also use other methods like the one we are using, for example, metabolic glycoengineering. This technology employed a sugar analog to introducing functional group to the human new SVZ surface protons. By that, we will enhance the cell adhesion and the interaction with surrounding cells and consequently improve the survival rates. So we can check the results for neurodefibrillation in vitro, in human neural stem cells. So this is sugar analog, and we can see after the co-culture in vitro with sugar analog, sugar analog, that's in the control. We can see the treated sugar analog treated the human SC. The cell morphology is significantly different from the control group, which shows more mature, longer neurites in this optimized analog treating groups. And then we tested it, the effect of transplanted this sugar analog treated human stem cell in vivo. And using the same asphyxial cardiorest model, we can see here the red is treated the neural stem cell compared to untreated neural stem cell. The NDS, the survival is significantly higher. That's the overall survival rates. That's NDS. They all shows significant difference. And we, again, we going back to check the quantification of ischemic neuron. We can see the treated NSC has a significantly less ischemic neuron compared to the standard NSC, which improved the functional outcome, include the survival rates and the neural deficit scores. So I would like to acknowledge the funding support from NIH and the members working hard in the prior past years in my lab. And thank you for your attention.

neural stem cell therapy

cardiac arrest

brain injury

animal models

stem cell therapies