Quantitative protein profiling and pathway analysis of spinal arteriovenous malformations.

Spinal arteriovenous malformations (sAVM) are rare and heterogeneous group of blood vessel disorders that affect spinal cord function directly or indirectly; however, the pathogenesis of sAVM is still unclear. In this study, we compared four sAVM specimens obtained during surgery and donated control samples in a Tandem Mass Tag (TMT)-labeled proteomic analysis. We identified 3101 proteins, 654 of which were differentially expressed in sAVM samples compared with the controls. Of these, 96 proteins were upregulated and 358 proteins were downregulated. Gene ontology (GO) analysis revealed that extracellular matrix organization in the biological process category and integrin-binding proteins in the molecular function category were the most enriched items. Two significant differentially expressed proteins (MYLK and MMP9) were verified by Western blot analysis. The pathway analysis indicated that the differentially expressed proteins in the pathways of angiogenesis, focal adhesion and cytoplasmic ribosome contributed to sAVM. The changes in protein profiles identified in this proteomic study provide an improved understanding of the pathogenesis of sAVM. The proteomics data are available via ProteomeXchange with identifier PXD007982.

Integrative proteomics and transcriptomics identify novel invasive-related biomarkers of non-functioning pituitary adenomas.

Non-functioning pituitary adenomas (NFPAs) are usually macroadenomas and display invasion into surrounding tissues. The treatment for invasive NFPAs is still challenging. This study describes the differential patterns of gene expression between invasive and non-invasive NFPAs and identifies novel biomarkers involved in invasion of NFPAs for diagnosis and treatment. Using gene microarray technology, we examined the gene expression profile and found 1160 differentially expressed messenger RNA (mRNA) between invasive and non-invasive NFPAs. Then, we examined the protein profile by liquid chromatography tandem mass spectrometry (LC-MS/MS) and found 433 differentially expressed proteins between invasive and non-invasive NFPAs. Subsequently, we integrated the proteomics and transcriptomics datasets and identified 29 common changed molecules. Through bioinformatics analysis using Ingenuity Pathway Analysis (IPA) software, we showed that the 29 molecules were enriched in 25 canonical signaling pathways, 25 molecular and cellular functions, and 2 networks. Eight genes were identified involved in the invasion function by the molecular and cellular functions analysis, including CAT, CLU, CHGA, EZR, KRT8, LIMA1, SH3GLB2 and SLC2A1. Furthermore, we validated the decreased CHGA expression and increased CLU expression in invasive NFPAs by qRT-PCR and Western blot. Our study demonstrated that integration of proteomics and transcriptomics could prove advantageous for accelerating tumor biomarker discovery and CHGA and CLU might be important novel biomarkers and therapeutic targets for invasion of NFPAs.

Low expression of secreted frizzled-related protein 4 in aggressive pituitary adenoma.

OBJECTIVE: The secreted frizzled-related proteins (sFRPs) are reported to be antagonists of a number of tumors. This study was designed to investigate the relationship of sFRP4 with aggressiveness of pituitary adenomas. MATERIAL AND METHOD: Specimens were classified into three groups: normal control (n = 10), non-aggressive group (n = 42) and aggressive group (n = 26) according to preoperative magnetic resonance imaging (MRI)/computed tomography. sFRP4 were investigated by PCR, Western blotting, and immunohistochemistry (IHC). The methylation status of the sFRP4 promoter region was observed by MassArray. Cell culture and 5-aza-2-deoxycytidine treatment was performed to observe the relationship of downregulation of sFRP4 with methylation of the sFRP4 gene. RESULTS: PCR and Western blot results showed that sFRP4 expression was downregulated in aggressive pituitary adenomas, which was confirmed by IHC. Methylation of the sFRP4 promoter was increased in aggressive pituitary adenomas. And methylation of the sFRP4 promoter lead to downregulation of sFRP4 expression. CONCLUSIONS: sFRP4 expression is inversely related to the aggressiveness of pituitary adenomas, and act as a tumor suppressor.

Emotional Problems, Quality of Life and Symptom Burden in Patients with Chordoma.

Chordomas are very rare malignant bone tumors. Following surgery, their effects on neurological, physical, psychological, social, and emotional functioning are substantial and can have a major impact on a patients' quality of life (QOL). In this survey, we aimed to characterize the postoperation health-related QOL and emotional problem in patients with chordoma using the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire 30 (EORTC QLQ-C30) and Hamilton Depression Rating Scale (HAMD). The cohort included 100 patients who underwent resection surgery between 2014 and 2020. Being single or divorced, living in a rural area, receiving a diagnosis of sacrococcygeal chordoma, Karnofsky performance status (KPS) ≤ 70, and weight loss were associated with increased likelihood of depression (p < 0.05). Patients who were single or divorced, with KPS ≤ 70, and experiencing weight loss had a higher likelihood of a worse QOL (p < 0.05). The uni- and multivariate logistic regression analyses indicated that the KPS level (p = 0.000) and postoperative radiation therapy (p = 0.009) were related to depression; marital status (p = 0.029), KPS level (p = 0.006), and tumor location (p = 0.033) were related to worse QOL. Certain characteristics placed patients with chordoma at increased risk of emotional problems, which are associated with a lowered QOL and a higher symptom burden. Further knowledge regarding emotional problems is key to improving the QOL for patients with chordoma.

Intraspinal clear cell meningioma without dural attachment: A case report and literature review.

RATIONALE: Clear cell meningioma (CCM) is one of the rarest but most aggressive forms of meningioma, with a tendency to occur at a high recurrence rate. Intraspinal CCM, especially the nondura-based type, is even rarer than the intracranial CCM. PATIENT CONCERNS: We report a case of a 45-year-old woman who presented with a 1-month history of episodic pain in the lower back and in both thighs in the front side. Femoral nerve stretch tests were positive on both sides. Magnetic resonance imaging (MRI) demonstrated an intradural tumor at the L3 level, which was isointense on T1- and T2-weighted images (WI) and homogeneously enhanced on gadolinium-contrast T1 WI. DIAGNOSES: The space-occupying lesion was pathologically confirmed as CCM. INTERVENTIONS: During surgery, we found that the tumor adhered to a nerve root, without dural attachment. The nerve root was partially removed to achieve complete resection. OUTCOMES: The pain disappeared after the operation. The 1 year follow-up MRI revealed no evidence of tumor recurrence or metastasis. LESSONS: Nondura-based intraspinal CCM is easier to completely remove, and such complete removal should be achieved during the first operation. Although the recurrence rate of this particular type of meningioma appears to be lower than that of other types, close clinical and radiological follow-up is necessary.

Risk factor analysis of progressive spinal deformity after resection of intramedullary spinal cord tumors in patients who underwent laminoplasty: a report of 105 consecutive cases.

OBJECTIVELaminoplasty has been used in recent years as an alternative approach to laminectomy for preventing spinal deformity after resection of intramedullary spinal cord tumors (IMSCTs). However, controversies exist with regard to its real role in maintaining postoperative spinal alignment. The purpose of this study was to examine the incidence of progressive spinal deformity in patients who underwent laminoplasty for resection of IMSCT and identify risk factors for progressive spinal deformity.METHODSData from IMSCT patients who had undergone laminoplasty at Beijing Tsinghua Changgung Hospital between January 2014 and December 2016 were retrospectively reviewed. Univariate tests and multivariate logistic regression analysis were used to assess the statistical relationship between postoperative spinal deformity and radiographic, clinical, and surgical variables.RESULTSOne hundred five patients (mean age 37.0 ± 14.5 years) met the criteria for inclusion in the study. Gross-total resection (> 95%) was obtained in 79 cases (75.2%). Twenty-seven (25.7%) of the 105 patients were found to have spinal deformity preoperatively, and 10 (9.5%) new cases of postoperative progressive deformity were detected. The mean duration of follow-up was 27.6 months (SD 14.5 months, median 26.3 months, range 6.2-40.7 months). At last follow-up, the median functional scores of the patients who did develop progressive spinal deformity were worse than those of the patients who did not (modified McCormick Scale: 3 vs 2, and p = 0.04). In the univariate analysis, age (p = 0.01), preoperative spinal deformity (p < 0.01), extent of tumor involvement (p < 0.01), extent of abnormal tumor signal (p = 0.02), and extent of laminoplasty (p < 0.01) were identified as factors associated with postoperative progressive spinal deformity. However, in subsequent multivariate logistic regression analysis, only age ≤ 25 years and preoperative spinal deformity emerged as independent risk factors (p < 0.05), increasing the odds of postoperative progressive deformity by 4.1- and 12.4-fold, respectively (p < 0.05).CONCLUSIONSProgressive spinal deformity was identified in 25.7% patients who had undergone laminoplasty for IMSCT resection and was related to decreased functional status. Younger age (≤ 25 years) and preoperative spinal deformity increased the risk of postoperative progressive spinal deformity. The risk of postoperative deformity warrants serious reconsideration of providing concurrent fusion during IMSCT resection or close follow-up after laminoplasty.

Application of Multimodal Image Fusion to Precisely Localize Small Intramedullary Spinal Cord Tumors.

OBJECTIVE: We sought to study the application of precise intraoperative localization of small intramedullary spinal cord tumors. METHODS: From November 2015 to August 2017, 5 patients with small intramedullary spinal cord tumors were arranged in this group. By using the O-arm image system, we acquired the intraoperative computed tomography images of all patients and sent them to the Stealth navigation system. Medtronic Synergy Cranial software was used to complete the image fusion with preoperative magnetic resonance images, and the fused images were used to localize the intramedullary spinal cord tumors by the navigation system. The navigation errors were evaluated by measuring the maximum distance between the end of the tumor in sagittal magnetic resonance imaging and its real position. RESULTS: Five patients accomplished the multimodal image fusion, and we successfully completed the image-guided surgeries. The mean diameter of tumors was 12.2 ± 3.1 mm in sagittal magnetic resonance imaging, and the mean incision length was 12.7 ± 3.3 mm. The time of image processing was between 13 minutes and 17 minutes, and the mean value was 15 ± 1.6 minutes. The navigation error was between 0.9 mm and 1.5 mm, and the mean value was 1.2 ± 0.2 mm. CONCLUSIONS: The application of the multimodal image fusion combined with intraoperative O-arm image navigation system can be used to localize small intramedullary tumors.

Increased ß­catenin and c-myc expression predict aggressive growth of non-functioning pituitary adenomas: An assessment using a tissue microarray-based approach.

Non-functional pituitary adenomas (NFPAs) account for 80% of pituitary adenomas with the majority of these exhibiting recurrences post-surgery. Overexpression of ß-catenin and c­myc is common in numerous invasive tumors. The present study sought to investigate the correlation of ß­catenin and c­myc expression levels with aggressive growth and recurrence of NFPAs, using immunohistochemical examination of tissue microarrays. Tissue microarrays comprised 212 NFPAs specimens and 10 healthy specimens as controls. NFPAs were categorized as non­aggressive or aggressive. Immunohistochemical examination was performed to determine the expression of ß­catenin and c­myc. Correlation of the expression levels of ß­catenin and c­myc with clinicopathological parameters, including aggressiveness and recurrence, were assessed by univariate, multivariate and logistic regression analysis. Increased expression of ß­catenin and c­myc was detected in the majority of aggressive NFPAs specimens (71.1 and 88.7%, respectively). There was a significant positive correlation between ß­catenin and c­myc expression and aggressiveness [P=0.001, Odds Ratio (OR)=4.011; P<0.001, OR=30.833]. Only ß­catenin expression demonstrated a significant correlation with recurrence in NFPAs (P=0.021, OR=2.571). ß­catenin and c­myc were demonstrated to be potential biomarkers for aggressive NFPAs and in the future, ß-catenin may serve as a marker for aggressive behavior and recurrence in NFPAs.

Low expression of secreted frizzled-related protein 2 and nuclear accumulation of ß-catenin in aggressive nonfunctioning pituitary adenoma.

The identification of a specific molecular marker for aggressiveness of nonfunctioning pituitary adenomas (NFPAs) is urgently required in order to guide the clinical diagnosis and treatment of NFPAs. In the present study, low expression of secreted frizzled-related protein 2 (sFRP2) in NFPAs was demonstrated by reverse transcription-quantitative polymerase chain reaction, western blot and immunohistochemical analyses. The results confirmed an abnormal accumulation of free ß-catenin in the nuclei of NFPAs, which is the core step for the activation of the Wnt canonical signaling pathway. Furthermore, cyclin D1 and c-Myc, the downstream proteins of the Wnt canonical signaling pathway, were overexpressed in aggressive NFPAs. These findings demonstrated the activation of the Wnt canonical signaling pathway in aggressive NFPAs. In addition, sFRP2 expression was observed to be inversely correlated to the aggressiveness of NFPAs. Therefore, sFRP2 may act as a tumor suppressor through modulation of the cellular cytosolic pool of ß-catenin in NFPAs. Furthermore, the expression of sFRP2 may serve as a biomarker for NFPAs aggressiveness and prognosis.

Assessment of sFRP4 as a bio-marker for predicting aggressiveness and recurrence of growth hormone-secreting pituitary adenomas.

The association of sFRP4 expression with aggressiveness and recurrence of growth hormone (GH)-secreting pituitary adenomas was investigated. Ten normal pituitary and 52 GH-secreting pituitary adenoma specimens were classified into three groups: normal pituitary (control) group, non-aggressive group, and aggressive group, according to preoperative evaluation by magnetic resonance imaging (MRI)/computed tomography (CT). Expression of sFRP4 was determined by quantitative real-time polymerase chain reaction (qRT-PCR), western blot analysis, and tissue microarrays, to assess the association between sFRP4 and aggressiveness. Follow-up information of all 52 patients was collected to evaluate the impact of sFRP4 expression on the recurrence/progression of GH-secreting pituitary adenomas. qRT-PCR results showed a lower level of sFRP4 mRNA in the aggressive group, as compared to that in the non-aggressive group (P=0.001). A similar trend was observed on western blot analysis for sFRP4 protein expression (P=0.004). On analysis by tissue microarrays, weak sFRP4 expression was detected in the aggressive group (10/15, 66.7%). Univariate analysis showed a significant relationship between low sFRP4 expression and aggressiveness (P=0.024). On multivariate analysis weak sFRP4 expression was found to be an independent factor of recurrence/progression (odds ratio: 0.063, P=0.026). Methylation of the sFRP4 promoter was increased in low sFRP4 staining group compared to that in the high sFRP4 staining group (P<0.001). In this study, weak sFRP4 expression appeared to predict aggressive behavior, and was associated with recurrence/progression of GH-secreting pituitary adenomas. Methylation of the sFRP4 promoter may account for the low sFRP4 expression.

The role of TGF-ß/Smad signaling in dopamine agonist-resistant prolactinomas.

BACKGROUND: Prolactinomas are the most common secretory pituitary adenomas. The first line of treatment involves dopamine agonists (DAs); however, a subset of patients is resistant to such therapy. Recent studies suggest that dopamine can up-regulate TGF-ß1 synthesis in rat pituitary lactotrophs whereas estradiol down-regulates TGF-ß1. To date, the role of TGF-ß/Smad signaling in DAs-resistant prolactinomas has not been explored. METHODS: High-content screening (HCS) techniques, qRT-PCR, Western blot, immunofluorescence and ELISA, were performed to determine the role of TGF-ß/Smad signaling in DAs-resistant prolactinomas. RESULTS: We reported a significant down-regulation of TGF-ß/Smad signaling cascade in DAs-resistant prolactinomas compared to normal human anterior pituitaries. Following treatment with TGF-ß1, the dopamine agonist, bromocriptine, and the estrogen antagonist (ER), fulvestrant in GH3 cells, we found that TGF-ß1 and fulvestrant caused significant cytotoxicity in a dose- and time-dependent manner and activated Smad3 was detected following exposure to TGF-ß1 and fulvestrant. In addition, treating GH3 cells with fulvestrant increased active TGF-ß1 levels and decreased PRL levels in a dose-dependent manner. CONCLUSION: TGF-ß/Smad signaling pathway may play an important role in DA-resistant prolactinomas and has the potential to be a viable target for the diagnosis and treatment of prolactinomas, particularly in patients who are resistant to DAs.

Overexpression of the cell adhesion molecule claudin-9 is associated with invasion in pituitary oncocytomas.

Pituitary oncocytoma is a subtype of nonfunctioning pituitary adenomas with the potential to be locally invasive. Currently, surgery is the most effective treatment, whereas functional pituitary adenomas can be treated by drugs. We analyzed the invasiveness of pituitary oncocytomas to identify biomarkers that may be useful for guiding future therapeutic decision making. To identify important biomarkers of pituitary oncocytomas, 20 oncocytomas that were negative for hormone-specific immunostaining were confirmed by anti-mitochondria antibody immunostaining and electron microscopy. Our clinical phenotype data showed a prominent male predilection (85%). These tumors were classified as invasive or noninvasive based on preoperative magnetic resonance imaging, intraoperative records, and pathology slide. We observed significantly different expression profiles between pituitary oncocytomas (n = 3) and normal pituitary glands (n = 3). A total of 1937 genes were differentially expressed between the pituitary oncocytomas and normal pituitary glands. Among these 1937 genes, 954 were up-regulated and 983 were down-regulated in pituitary oncocytomas. The most significantly altered gene, claudin-9 (CLDN9), was further confirmed by quantitative real-time polymerase chain reaction, Western blot, and immunohistochemical staining in 10 invasive pituitary oncocytoma samples and 10 noninvasive pituitary oncocytoma samples. High levels of CLDN9 were found in the pituitary oncocytomas, whereas low levels were detected in normal pituitary glands. In addition, the CLDN9 expression level was higher in invasive oncocytomas compared with noninvasive oncocytomas. Bioinformatics Gene Ontology analysis was performed to better understand the critical role of CLDN9 in the development and progression of oncocytomas. Consequently, CLDN9 may be an important biomarker for invasive pituitary oncocytomas.