Hypoxic microenvironment-induced exosomes confer temozolomide resistance in glioma through transfer of pyruvate kinase M2.

OBJECTIVE: Glioma, a malignant primary brain tumor, is notorious for its high incidence rate. However, the clinical application of temozolomide (TMZ) as a treatment option for glioma is often limited due to resistance, which has been linked to hypoxic glioma cell-released exosomes. In light of this, the present study aimed to investigate the role of exosomal pyruvate kinase M2 (PKM2) in glioma cells that exhibit resistance to TMZ. METHODS: Sensitive and TMZ-resistant glioma cells were subjected to either a normoxic or hypoxic environment, and the growth patterns and enzymatic activity of glycolysis enzymes were subsequently measured. From these cells, exosomal PKM2 was isolated and the subsequent effect on TMZ resistance was examined and characterized, with a particular focus on understanding the relevant mechanisms. Furthermore, the intercellular communication between hypoxic resistant cells and tumor-associated macrophages (TAMs) via exosomal PKM2 was also assessed. RESULTS: The adverse impact of hypoxic microenvironments on TMZ resistance in glioma cells was identified and characterized. Among the three glycolysis enzymes that were examined, PKM2 was found to be a critical mediator in hypoxia-triggered TMZ resistance. Upregulation of PKM2 was found to exacerbate the hypoxia-mediated TMZ resistance. Exosomal PKM2 were identified and isolated from hypoxic TMZ-resistant glioma cells, and were found to be responsible for transmitting TMZ resistance to sensitive glioma cells. The exosomal PKM2 also contributed towards mitigating TMZ-induced apoptosis in sensitive glioma cells, while also causing intracellular ROS accumulation. Additionally, hypoxic resistant cells also released exosomal PKM2, which facilitated TMZ resistance in tumor-associated macrophages. CONCLUSION: In the hypoxic microenvironment, glioma cells become resistant to TMZ due to the delivery of PKM2 by exosomes. Targeted modulation of exosomal PKM2 may be a promising strategy for overcoming TMZ resistance in glioma.

A multidimensional pre-operative planning method of unruptured vertebral artery dissecting aneurysms using three-dimensional AWE mapping and hemodynamic simulation.

OBJECTIVE: High-resolution magnetic resonance imaging (HR-MRI) can provide valuable insights into the evaluation of vascular pathological conditions, and 3D digital subtraction angiography (3D-DSA) offers clear visualization of the vascular morphology and hemodynamics. This study aimed to investigate the potential of a multimodal method to treat unruptured vertebral artery dissection aneurysms (u-VADAs) by fusing image data from HR-MRI and 3D-DSA. METHODS: This observational study enrolled 5 patients diagnosed with u-VADAs, who were scheduled for interventional treatment. The image data of HR-MRI and 3D-DSA were merged by geometry software, resulting in a multimodal model. Quantified values of aneurysm wall enhancement (AWE), wall shear stress (WSS), neck velocity, inflow volume, intra-stent flow velocity (ISvelocity), and intra-aneurysmal velocity (IAvelocity) were calculated from the multimodal method. RESULTS: We found the actual lengths of u-VADAs in the multimodal model were longer than the 3D-DSA model. We formulated surgical plannings based on the WSS, IA velocity, and neck velocity. The post-operative value of IAvelocity, neck velocity, and follow-up quantified values of AWE were decreased compared with the pre-operative condition. After that, u-VADAs were complete occlusion in four patients and near-complete occlusion in one patient during the 6th-month follow-up after surgery. CONCLUSION: The multidimensional method combining HR-MRI with 3D-DSA may provide more valuable information for treating VADAs, with the potential to develop effective surgical planning.

Cerebral cavernous malformation development in chronic mouse models driven by dual recombinases induced gene deletion in brain endothelial cells.

Cerebral cavernous malformation (CCM) is a brain vascular disease which can cause stroke, cerebral hemorrhage and neurological deficits in affected individuals. Loss-of-function mutations in three genes (CCM1, CCM2 and CCM3) cause CCM disease. Multiple mouse models for CCM disease have been developed although each of them are associated with various limitations. Here, we employed the Dre-Cre dual recombinase system to specifically delete Ccm genes in brain endothelial cells. In this new series of CCM mouse models, robust CCM lesions now develop in the cerebrum. The survival curve and lesion burden analysis revealed that Ccm2 deletion causes modest CCM lesions with a median life expectance of ∼10 months and Ccm3 gene deletion leads to the most severe CCM lesions with median life expectance of ∼2 months. The extended lifespan of these mutant mice enables their utility in behavioral analyses of neurologic deficits in adult mice, and allow the development of methods to quantify lesion burden in mice over time and also permit longitudinal drug testing in live animals.

Role of pericytes in the development of cerebral cavernous malformations.

Cerebral cavernous malformation (CCM) is caused by loss-of-function mutations in CCM1, CCM2, or CCM3 genes of endothelial cells. It is characterized by pericyte deficiency. However, the role of pericytes in CCMs is not yet clarified. We found pericytes in Cdh5Cre ERT2 ;Ccm1 fl/fl (Ccm1 ECKO ) mice had a high expression of PDGFRß. The inhibition of pericyte function by CP-673451 aggravated the CCM lesion development. RNA-sequencing analysis revealed the molecular traits of pericytes, such as highly expressed ECM-related genes, especially Fn1. Furthermore, KLF4 coupled with phosphorylated SMAD3 (pSMAD3) promoted the transcription of fibronectin in the pericytes of CCM lesions. RGDS peptide, an inhibitor of fibronectin, decreased the lesion area in the cerebella and retinas of Ccm1 ECKO mice. Also, human CCM lesions had abundant fibronectin deposition, and pSMAD3- and KLF4-positive pericytes. These findings indicate that pericytes are essential for CCM lesion development, and fibronectin intervention may provide a novel target for therapeutic intervention in such patients.

Efficacy of stereotactic gamma knife surgery and microvascular decompression in the treatment of primary trigeminal neuralgia: a retrospective study of 220 cases from a single center.

OBJECTIVES: A retrospective study was undertaken to compare the efficacy of stereotactic gamma knife surgery (GKS) and microvascular decompression (MVD) in the treatment of primary trigeminal neuralgia (TN) at a single center. The study included the evaluation of clinical outcomes of pain relief and pain recurrence and complications associated with GKS and MVD. METHODS: The study included 202 patients with primary TN and was conducted between January 2013 and December 2014; about 115 patients were treated with GKS and 87 patients were treated with MVD. TN pain was evaluated using the Barrow Neurological Institute and the visual analog scale scoring systems. Preoperative magnetic resonance tomographic angiography was performed for all patients. Microscope-assisted MVD used the suboccipital retrosigmoid sinus approach. GKS targeted the trigeminal nerve root entry zone with a margin radiation dose of 59.5 Gy, and brainstem dose <12 Gy. Posttreatment follow-up was for 2 years. RESULTS: Postoperative Barrow Neurological Institute scores for patients treated with GKS and MVD were significantly improved compared with preoperative scores (P<0.01). Reduction in postoperative pain following MVD (95.4% patients) was significantly greater than that following GKS (88.7% patients) (P<0.01). Postoperative visual analog scale scores of the MVD group were significantly reduced compared with those of patients treated with GKS at the same postoperative time points (P<0.01). Patients treated with GKS had a significantly increased rate of loss of corneal reflex compared with patients treated with MVD (P=0.002). CONCLUSION: Both GKS and MVD are safe and effective first-line and adjunctive treatment options for patients with TN. The clinical outcomes of pain relief and reduction of pain recurrence were better with MVD. For GKS, this study showed that the optimal radiation therapeutic dose range was 70-90 Gy, but brainstem radiation protection is recommended.