Modulating microglial polarization is a potential strategy to assuage secondary brain injury caused by intracranial hemorrhage (ICH). However, despite decades of effort, effective therapies targeting ...microglia for ICH are still lacking. Here, a nanorobotic, tetrahedral framework nucleic acid (tFNA), is successfully synthesized and designed to carry C‐C chemokine receptor 2 (siCCR2) for use in in vitro hemin‐induced and in vivo collagenase‐induced ICH models. This nanoscale complex (tFNA‐siCCR2), which possesses biocompatibility, editability, and structural stability, exhibits a favorable effect in inhibiting the expression of CCR2. After treatment with tFNA‐siCCR2, hematoma absorption is accelerated, and neurological inflammation is mitigated by decreasing levels of proinflammatory cytokines, while increasing the release of anti‐inflammatory factors. Consequently, the neurological deficits of mice with ICH improve. These results indicate that inhibiting CCR2 expression during the acute phase of ICH polarizes microglia towards a therapeutic subtype, and restores neurological function, which demonstrates that tFNA has a promising ability to transfer siCCR2 for treating ICH.
Tetrahedral framework nucleic acid (tFNA)‐C‐C chemokine receptor 2 (siCCR2) is investigated as therapy for intracranial hemorrhage (ICH) mice. Four specific designed DNA single strands are selfassembled to construct a tetrahedron and carry siCCR2 to inhibit the expression of CCR2 in microglia. By regulating the polarization of microglia from M1 to M2, tFNA‐siCCR2 accelerates absorption of hematoma, thereby reducing motor nerve function impairment in mice with ICH.
Human amniotic membrane mesenchymal stem cells (hAMSCs) have recently been suggested as ideal candidate stem cells for cell-based therapy. Many studies have reported the therapeutic effects of hAMSCs ...in numerous disease models. However, no studies have used hAMSCs to treat intracerebral hemorrhage (ICH). In the present study, we examined the therapeutic potential of hAMSCs in a rat model of ICH, and characterized the possible mechanisms of action. Adult male Wistar rats were subjected to ICH by intrastriatal injection of VII collagenase, and then were intracerebrally administered hAMSCs, fibroblasts, or phosphate-buffered saline (PBS) at 24 h after ICH. Compared with the fibroblasts and the PBS control, hAMSCs treatment significantly promoted neurological recovery, and reduced the numbers of ED1+ activated microglia, as well as myeloperoxidase (MPO+), and caspase-3+ cells in the brain injury model. In addition, hAMSCs treatment significantly increased the expression of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) in the injured brain, and promoted neurogenesis and angiogenesis, compared with the fibroblasts and the PBS control. The transplanted hAMSCs survived for at least 27 days and were negative for β-tubulin III and glial fibrillary acidic protein (GFAP). Taken together, the results suggest that hAMSCs treatment significantly promotes neurological recovery in rats after ICH. The mechanism of action could be mediated by inhibition of inflammation and apoptosis, increasing neurotrophic factor expression, and promotion of neurogenesis and angiogenesis. Thus, hAMSCs are candidate stem cells for the treatment of ICH.
•hAMSCs treatment promoted neurological recovery in rats after ICH.•hAMSCs treatment reduced inflammatory response and apoptosis.•hAMSCs treatment increased the expression of BDNF and VEGF.•hAMSCs treatment enhanced endogenous neurogenesis and angiogenesis.
Early brain injury (EBI) is the most important potentially treatable cause of mortality and morbidity following subarachnoid hemorrhage (SAH). Apoptosis is one of the main pathologies of SAH-induced ...EBI. Numerous studies suggest that human umbilical cord derived mesenchymal stem cells (hucMSCs) may exert neuroprotective effect through exosomes instead of transdifferentiation. In addition, microRNA-206 (miR-206) targets BDNF and plays a critical role in brain injury diseases. However, the therapy effect of miR-206 modified exosomes on EBI after SAH and its regulatory mechanism have not been elucidated. Here, to identify whether hucMSCs-derived miR-206-knockdown exosomes have a better neuroprotective effect, we established SAH rat model and treated it with the exosomes to research the mechanism of miR-206 in EBI after SAH. We found that treatment with hucMSCs-derived miR-206-knockdown exosomes has a greater neuroprotective effect on SAH-induced EBI compared to treatment with simple exosomes. The miR-206-knockdown exosomes could significantly improve neurological deficit and brain edema and suppress neuronal apoptosis by targeting BDNF. Moreover, the BDNF/TrkB/CREB pathway was activated following treatment with miR-206 modified exosomes in vivo. In summary, these findings indicate that the hucMSCs-derived miR-206-knockdown exosomes prevent early brain injury by inhibiting apoptosis via BDNF/TrkB/CREB signaling. This may serve as a novel therapeutic target for treatment of SAH-induced EBI.
•Treatment with hucMSCs-derived miR-206-knockdown exosomes attenuates EBI after SAH.•BDNF is a target of miR-206.•HucMSCs-derived miR-206-knockdown exosomes attenuate EBI after SAH by targeting BDNF in vivo.•HucMSCs-derived miR-206-knockdown exosomes have influence on the BDNF/TrkB/CREB signaling.
Abstract Neuronal injuries have been a challenging problem for treatment, especially in the case of complete and chronic cervical spinal cord injury (SCI). Recently, particular attention is paid to ...the potential of stem cell in treating SCI, but there are only few clinical studies and insufficient data. This study explored the efficacy of autologous bone marrow mesenchymal stem cells (BMMSCs) transplantation in the treatment of SCI. Forty patients with complete and chronic cervical SCI were selected and randomly assigned to one of the two experimental groups, treatment group and control group. The treatment group received BMMSCs transplantation to the area surrounding injury, while the control group was not treated with any cell transplantation. Both the transplant recipients and the control group were followed up to 6 months, postoperatively. Preoperative and postoperative neurological functions were evaluated with AIS grading, ASIA score, residual urine volume and neurophysiological examination. Results showed that in the treatment group 10 patients had a significant clinical improvement in terms of motor, light touch, pin prick sensory and residual urine volume, while nine patients showed changes in AIS grade. Neurophysiological examination was consistent with clinical observations. No sign of tumor was evident until 6 months postoperatively. In the control group, no improvement was observed in any of the neurological functions specified above. BMMSCs transplantation improves neurological function in patients with complete and chronic cervical SCI, providing valuable information on applications of BMMSCs for the treatment of SCI.
Disorders of consciousness are a heterogeneous mixture of different diseases or injuries. Although some indicators and models have been proposed for prognostication, any single method when used alone ...carries a high risk of false prediction. This study aimed to develop a multidomain prognostic model that combines resting state functional MRI with three clinical characteristics to predict one year-outcomes at the single-subject level. The model discriminated between patients who would later recover consciousness and those who would not with an accuracy of around 88% on three datasets from two medical centers. It was also able to identify the prognostic importance of different predictors, including brain functions and clinical characteristics. To our knowledge, this is the first reported implementation of a multidomain prognostic model that is based on resting state functional MRI and clinical characteristics in chronic disorders of consciousness, which we suggest is accurate, robust, and interpretable.
To confirm whether machine learning algorithms (MLA) can achieve an effective risk stratification of dying within 7 days after basal ganglia hemorrhage (BGH). We collected patients with BGH admitted ...to Sichuan Provincial People's Hospital between August 2005 and August 2021. We developed standard ML-supervised models and fusion models to assess the prognostic risk of patients with BGH and compared them with the classical logistic regression model. We also use the SHAP algorithm to provide clinical interpretability. 1383 patients with BGH were included and divided into the conservative treatment group (CTG) and surgical treatment group (STG). In CTG, the Stack model has the highest sensitivity (78.5%). In STG, Weight-Stack model achieves 58.6% sensitivity and 85.1% specificity, and XGBoost achieves 61.4% sensitivity and 82.4% specificity. The SHAP algorithm shows that the predicted preferred characteristics of the CTG are consciousness, hemorrhage volume, prehospital time, break into ventricles, brain herniation, intraoperative blood loss, and hsCRP were also added to the STG. XGBoost, Stack, and Weight-Stack models combined with easily available clinical data enable risk stratification of BGH patients with high performance. These ML classifiers could assist clinicians and families to identify risk states timely when emergency admission and offer medical care and nursing information.
Background and Purpose
As a hallmark of glioblastoma multiforme (GBM), CD44 plays a crucial role in promoting glioblastoma stem cell (GSC) stemness phenotypes and multiple drug resistance. The ...therapeutic potential of CD44 has been validated by the clinical successes of several CD44 inhibitors, including antibodies and hyaluronan‐related drugs.
Experimental Approach
We used systemsDock software to predict verbascoside as a candidate CD44 inhibitor. Microscale thermophoresis was used to confirm the interaction between CD44 and verbascoside. Four glioblastoma cell lines and a patient‐derived glioblastoma cell line were used to test the influences of verbascoside on glioblastoma. CD44‐overexpressing and CD44‐knockout cell lines were also used. Real‐time quantitative PCR and western blot analyses were performed. A xenograft mouse model was used to test verbascoside.
Key Results
Verbascoside bound to CD44 and suppressed its dimerization. By inhibiting CD44 dimerization, verbascoside decreased the release of the CD44 intracellular domain (CD44ICD) and suppressed the expression of CD44 downstream genes. Verbascoside treatment suppressed the stemness phenotypes of cells with high CD44 expression. In a mouse model of glioma, verbascoside treatment highly reduced the growth of intracranial tumours and inhibited CD44ICD release. Both stem cell marker and mesenchymal GBM subtype marker genes were down‐regulated in verbascoside‐treated mice.
Conclusion and Implications
Verbascoside suppressed growth of glioblastoma cells by inhibiting CD44 dimerization. Stem cell‐like cell properties and tumour cell growth were also suppressed by verbascoside, both in vitro and in vivo. Verbascoside significantly prolonged survival of xenografted mice.
Glioma is a common primary central nervous system tumor with complex pathogenesis. DNA damage and repair (DDR) is widely involved in regulating cell proliferation and tumorigenesis by correcting and ...repairing DNA damage mechanisms. Recent studies have reported the following properties in cancer cells in glioma, increased DNA damage and reduced DNA repair capacity. However, the relationship between glioma and DDR-related genes was unclear.
DDR-related risk score model was built. The validity of this model was validated in detail through the Kaplan-Meier survival analysis, tumor mutational burden (TMB) analysis, immune cell infiltration, sensitivity to treatment regimens. Moreover, the model's adaptability was validated in different glioma data cohorts and different glioma subgroups. To further investigate the molecular mechanism of one of DDR-related gene (NUDT1) in glioma, U251 cell was used for the knockdown experiment, followed by MTT, wound healing and transwell analysis.
Ten prognostic-related DDR-related signature genes were obtained, including EID3, MGMT, YWHAG, PMS1, SHPRH, HUS1, NUDT1, GADD45G, APEX1 and FAM175A. The RT-qPCR results suggested that the latter five genes were highly expressed in glioma patients. Interestingly, high TMB score had longer survival. In high-risk score groups, reduced immune cell infiltration in the tumor microenvironment lead to poorer patient outcomes. Sensitivity to treatment regimens analysis indicated that low-risk score groups were more sensitive to chemotherapeutics. Moreover, the risk score model had a good prediction effect on different glioma datasets and different glioma subgroups. In vitro mechanism study showed that knockdown of NUDT1 reduced tumorigenesis. Furthermore, knockdown of NUDT1 remarkably reduced the expression level of HIF-1α.
DDR-related risk score model built-in this work has good predictive performance for glioma.
Key messages
Ten prognostic-related DDR-related signature genes were obtained, including EID3, MGMT, YWHAG, PMS1, SHPRH, HUS1, NUDT1, GADD45G, APEX1 and FAM175A.
In high-risk score groups, reduced immune cell infiltration in the tumor microenvironment leads to poorer patient outcomes.
The risk score model had a good prediction effect on different glioma datasets and different glioma subgroups.
Knockdown of NUDT1 reduced tumorigenesis of glioma and remarkably reduced the expression level of HIF-1α.
Inflammation and apoptosis caused by intracerebral hemorrhage (ICH) are two important factors that affect patient prognosis and survival. Toll-like receptor 4 (TLR4) triggers activation of the ...inflammatory pathway, causing synthesis and release of inflammatory factors. The inflammatory environment also causes neuronal apoptosis. However, no studies have reported the role of TLR4 in inflammation and apoptosis.
We performed survival curve analysis and behavioral scores on TLR4 knockout mice and wild-type mice after inducing ICH. We used TLR4 knockout mice and wild-type mice to make ICH models with type VII collagenase and explored the link between TLR4 in inflammation and apoptosis. We used Western blot to detect the expression of apoptosis-related proteins, inflammatory factors, and their receptors at different time points after ICH induction. The effects of TLR4 on apoptosis were observed by TUNEL, Hoechst, and HE staining techniques. The association with TLR4 in inflammation and apoptosis was explored using IL-1β and TNF-α antagonists. Data conforming to a normal distribution are expressed as mean ± standard deviation. Grade and quantitative data were compared with rank sum test and t test between two groups. P < 0.05 was considered statistically significant.
TLR4 knockout significantly increased the survival rate of ICH mice. The scores of TLR4 knockout mice were significantly lower than those of wild-type mice. We found that TLR4 knockout mice significantly inhibited apoptosis and the expression of inflammatory factors after the induction of ICH. The apoptosis of ICH-induced mice was significantly improved after injecting IL-1β and TNF-α antagonists. Moreover, the anti-apoptotic effect of the antagonist in wild-type mice is more pronounced. A single injection of the antagonist failed to improve apoptosis in TLR4 knockout mice.
We conclude that TLR4-induced inflammation after ICH promotes neuronal apoptosis. IL-1β and TNF-α antagonists attenuate this apoptotic effect. Therefore, targeting TLR4 in patients with clinical ICH may attenuate inflammatory response, thereby attenuating apoptosis and improving prognosis.
While repetitive transcranial magnetic stimulation (rTMS) has been applied in treatment of patients with disorders of consciousness (DOC), a standardized stimulation protocol has not been proposed, ...and its therapeutic effects are inconsistently documented.
To assess the efficacy of rTMS in improving consciousness in patients with persistent minimally conscious state (MCS) or unresponsive wakefulness syndrome (UWS), previously known as vegetative state (VS).
A prospective single-blinded study, with selected subjects, was carried out. In total, 16 patients (5 MCS and 11 VS/UWS) with chronic DOC were included. All patients received active 10 Hz rTMS at the left dorsolateral prefrontal cortex (DLPFC), at one session per day, for 20 consecutive days. A single daily session of stimulation consisted of 1,000 pulses (10 s of 10 Hz trains; repeated 10 times with an inter-train interval of 60 s; and 11 min and 40 s for total session). The main outcome measures were changes in the total score on the JFK Coma Recovery Scale-Revised (CRS-R) scale. Additional measures were the impressions of caregivers after the conclusion of the interventions, which were assessed using the Clinical Global Impression-Improvement (CGI-I) scale.
The CRS-R scores were increased in all 5 MCS patients and 4 of 11 VS/UWS patients, while a significant enhancement of CRS-R scores was observed compared to the baseline in all participants (
= 0.007). However, the improvement was more notable in MCS patients (
= 0.042) than their VS/UWS counterparts (
= 0.066). Based on the CGI-I scores, two patients improved considerably, two improved, six minimally improved, six experienced no change, and none deteriorated. Good concordance was seen between the CGI-I result and the increases in CRS-R scores.
Treatment of 10 Hz multisession rTMS applied to the left DLPFC is promising for the rehabilitation of DOC patients, especially those in MCS. Further validation with a cohort of a larger sample size is required.
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