BACKGROUND:Although successful treatment of papillary craniopharyngiomas (PCPs) with BRAFv600e inhibitors has been reported in clinical trials, studies have shown that approximately 10% of PCPs lack the BRAFv600e mutation and that BRAFv600e inhibitors may not be significantly effective against these tumors. However, no studies have focused specifically on BRAFv600e- PCPs. METHODS:Spatial transcriptome sequencing was performed on calcified PCP tissue to identify novel subtypes of PCP cells. The findings were validated via pathological methods in 51 PCP samples. Primary PCP cells from BRAFv600e- PCP patients and BRAFv600e+ PCP patients were isolated and then injected into the brains of nude mice via stereotactic surgery to establish a stable mouse model of human-originated PCP. Model mice were treated with vemurafenib, a BRAF inhibitor, and anlotinib, an angiogenesis inhibitor. BRAFv600e-PCP patients were treated with anlotinib in a phase 1 clinical trial. Changes in the tumors of the model mice and patients were monitored via pathological methods, CT and MRI. RESULTS:Most of calcified PCPs were negative for the BRAFv600e mutation, and findings from the mouse model confirmed that vemurafenib may not have a significant therapeutic effect on BRAFv600e- PCPs. However, the mouse model verified that, anlotinib may have a significant therapeutic effect on BRAFv600e- PCPs. Two patients with BRAFv600e- PCPs participated in a phase 1 clinical trial and received anlotinib therapy; their tumors disappeared after 3 months of therapy and did not recur within 24 months follow-up after stopping the treatment. CONCLUSION:BRAFv600e- PCPs are characterized by calcification and do not respond to the BRAF inhibitor vemurafenib, and for which the angiogenesis inhibitor anlotinib may have a significant therapeutic effect.

Adamantinomatous craniopharyngioma (ACP) is a histologically benign yet clinically aggressive intracranial tumor characterized by high recurrence rates. Fibroblast activation protein (FAP), a marker of cancer-associated fibroblasts (CAFs), is implicated in tumor progression and microenvironment remodeling. This study evaluates the prognostic significance of FAP and develops a radiomics-based model for non-invasive preoperative risk stratification. Immunohistochemical analysis was performed on 54 ACP cases to assess FAP expression levels. Transcriptomic data from 110 ACP cases across two external databases were analyzed for differential gene expression and pathway enrichment. Immunofluorescence was conducted to determine the spatial correlation of FAP with angiogenic markers (VEGF, CD31, CD34). A radiomics model was developed using preoperative MRI data to predict FAP expression and validated for predictive performance. High FAP expression was associated with extracellular matrix remodeling, angiogenesis, and inflammatory pathway activation. Clinically, high-FAP tumors exhibited larger volumes (P = 0.044), more severe hypothalamic dysfunction (P = 0.001), and shorter progression-free survival (P = 0.03). Multivariate analysis identified high FAP expression (HR = 9.890, P = 0.0340) as an independent predictor of recurrence. Immunofluorescence confirmed the co-localization of FAP+ CAFs with angiogenic markers, suggesting a role in tumor recurrence. The radiomics model integrating T1/T2-weighted MRI features demonstrated robust performance in predicting FAP expression, with AUCs of 0.742 (training set),0.822 (internal validation set) and 0.686(external validation set). FAP is a prognostic biomarker in ACP, with high expression indicative of increased recurrence risk. The radiomics model offers a non-invasive approach for preoperative risk stratification, potentially guiding surgical decision-making and anti-angiogenic therapeutic strategies.

BACKGROUND AND OBJECTIVES:Growth hormone-secreting pituitary adenomas (GH-PAs) that invade the structures surrounding the pituitary gland are a big problem for clinical professionals working in the field. People have tried to find a reliable biomarker for this. The STAT3 and phosphorylated STAT3 (p-STAT3) proteins are increased in many cancers and help cells move and invade. This study looked at the link between p-STAT3/STAT3 levels and the Knosp grading system for GH-PAs. METHODS:A total of 71 patient GH-PAs were obtained between 22 January 2016 and 23 January 2022. An immunohistochemical analysis was conducted to assess the expression of p-STAT3/STAT3 and keratin CAM5.2 in tumor samples. p-STAT3/STAT3 was also used to evaluate the predictive value of tumor invasion propensity and patient prognosis. RESULTS:Tumor invasion, maximum diameter, macroadenoma, high T2-weighted intensity signal, cystic degeneration, low remission, and resection rates were found to be associated with high p-STAT3 expression. A large adenoma, a high T2-weighted intensity signal, cystic change, high insulin-like growth factor 1 levels, sparsely granular adenomas, high p-STAT3 expression, and a low resection rate of postoperative tumor were all associated with increased tumor aggressiveness. However, there was no link between Ki-67 and STAT3 expression and tumor invasion. Using MRI to diagnose the invasiveness of GH-PAs, it was found that p-STAT3 was more effective than sparsely granulated. CONCLUSION:The findings of this study indicate that the elevated expression of p-STAT3 may serve as a potential biomarker for the prediction of the invasiveness and clinical outcome of GH-PAs.

Background: The role of KI-67, a tumor proliferation marker, as a biomarker for progression in pituitary neuroendocrine tumors (PitNETs) remains controversial. The aim of this study was to analyze the relationship between KI-67 expression levels and PitNETs progression based on transcriptional factor lineage classification. Methods: A total of 97 PitNETs patients who underwent endoscopic surgery performed by the same surgeon between February 2022 and March 2023 were included in the study. Immunohistochemistry was used to stain tumor specimens for KI-67. Tumor progression was assessed based on Knosp classification and follow-up results, and separate analyses were performed according to hormone secretion status and transcriptional factor lineage classification. Results: We found no significant differences in preoperative tumor characteristics and various prognostic outcomes between patients with high and low KI-67 expression. When classified according to hormone secretion types, no significant differences were observed in the progression of various PitNETs and KI-67. However, when classified based on tumor transcriptional factor lineages, a significant association with invasiveness was observed in the T-PIT lineage with high KI-67 expression (P = 0.035), but not in the PIT-1 and SF-1 lineages. Conclusion: The research has found that KI-67 as a biomarker for tumor progression in PitNETs is unreliable. emphasizing the importance of comprehensively considering pitNETs subtypes and molecular characteristics in patient assessment.

OBJECTIVE:Understanding the differences of suprasellar papillary and adamantinomatous craniopharyngiomas (PCPs/ACPs) is pivotal for target therapy, surgical strategy or postoperative management. Here, the clinical features, surgical nuances and postoperative hypothalamic outcomes of PCPs were systematically recapitulated. METHODS:24 PCPs and 52 ACPs underwent initial surgery were retrospectively reviewed. Clinical data, quantified third ventricle (3rd V) occupation and optic chiasm distortion were compared, as well as intra-operative findings, operating notes and prognosis. Moreover, analysis of tumor/3rd V relationship and hypothalamic outcomes were also performed. RESULTS:Tumors were more likely to occupies the 3rd V cavity in PCPs. Chiasm distortion of "compressed forward" was the most common pattern (45.8 %) in PCPs, whereas "stretched forward" pattern accounted the highest (42.5 %) in ACPs. Besides, round-shaped with less calcification, duct-like recess, solid consistency, rare subdiaphragmatic invasion, visible lower stalk and improved postoperative visual outcome were more frequently observed in PCPs. The basal membranes of the tumor epithelium and the reactive gliosis were separated by a layer of collagen fibers in most PCPs, which differs from ACPs in the morphological examination of tumor/3rd V floor interface. In daytime sleepiness and memory difficulty, the PCPs showed significantly better outcomes than the ACPs groups, and PCPs suffered less postoperative weight gain (p < 0.05) than ACPs among adult-onset cases. CONCLUSION:PCPs are different from ACPs regards the clinical features, operative techniques and outcomes. If necessary, PCPs are suggested more amenable to total removal since its less invasiveness to the 3rd V floor and better hypothalamic outcomes.

Aims: Adamantinomatous craniopharyngioma (ACP) is a rare benign intracranial tumour that occurs in the sellar region and likely originates from the embryonic craniopharyngeal epithelium (a remnant of the ectoderm, also known as Rathke's pouch). However, progress in ACP research has been slow due to the lack of ACP cell lines. Therefore, in this study, we established an immortalised ACP cell line.Methods: Immortalised ACP cells were established from cultured primary ACP cells by the ectopic expression of human telomerase reverse transcriptase (hTERT). Primary and immortalised cells were compared and characterised by morphological examination, short tandem repeat (STR) profiling, whole exome sequencing (WES) and transcriptome sequencing. The ability of immortalised cells to form tumours was examined using an intracranial tumour formation assay in mice. Immunofluorescence staining was performed to compare the characteristics of human ACP and intracranial xenografts derived from immortalised cells.Results: A novel immortalised ACP cell line, STAM4, was successfully established in a 4-year-old male ACP patient. When STAM4 cells were compared with primary ACP cells in terms of morphological characteristics, genetic variants, STR profiles, biological behaviours and transcriptome differences, STAM4 cells were similar to primary ACP cells. Additionally, the STAM4 cells were able to form intracranial tumours that resemble human ACP.Conclusions: This ACP cell line may provide a cell model for further studies on the molecular mechanisms underlying the occurrence and progression of ACP and for the development of new anticancer drugs for ACP.

Previous studies have shown that growth hormone can regulate hypothalamic energy metabolism, stress, and hormone release. Therefore, growth hormone has great potential for treating hypothalamic injury. In this study, we established a specific hypothalamic axon injury model by inducing hypothalamic pituitary stalk electric lesions in male mice. We then treated mice by intraperitoneal administration of growth hormone. Our results showed that growth hormone increased the expression of insulin-like growth factor 1 and its receptors, and promoted the survival of hypothalamic neurons, axonal regeneration, and vascular reconstruction from the median eminence through the posterior pituitary. Altogether, this alleviated hypothalamic injury-caused central diabetes insipidus and anxiety. These results suggest that growth hormone can promote axonal reconstruction after hypothalamic injury by regulating the growth hormone-insulin-like growth factor 1 axis.

Abstract Purpose This study aims to systematically assess patient characteristics, identify clinical factors guiding treatment decisions, and predict the prognosis of H3 K27-altered Diffuse Midline Glioma (DMG/K27M). Methods A retrospective review of 64 consecutive DMG/K27M patients was conducted. Patient clinical profiles, treatment approaches, and follow-up data were collected from a Chinese tertiary institution between August 2016 and August 2022. Analysis of overall survival (OS) was conducted using Kaplan–Meier modeling and univariate and multivariate Cox regression analyses to identify prognostic factors. Results Among the 64 patients included in the study, the median OS was 9 months. Thalamus was the tumor location for 53.1% (34/64), with 39.1% (25/64) presenting dizziness as their initial symptom. Patients with dizziness were younger (23.04 vs. 30.7 years, p = 0.0328), exhibited larger tumor volume (27.380 vs. 11.680 cm3, p = 0.0359), and displayed extended overall survival (13.00 vs. 8.00 months, p = 0.0017) compared to those without dizziness. In addition, univariate and multivariate analysis showed that tumor location, dizziness, Karnofsky performance status (KPS) score and treatment regimen were prognostic factors for OS (p < 0.05), whereas age was not significantly correlated with OS. Conclusion Patients without dizziness as the initial symptom may face a challenging prognosis compared to the dizziness (+) group. Tumor location, dizziness, KPS, and treatment regimen could serve as independent prognostic indicators. These findings contribute novel insights for evidence-based practices in DMG/K27M research.

Background The incidence of growth hormone deficiency (GHD) after aneurysmal subarachnoid hemorrhage (aSAH) is significantly higher than that of other neuroendocrine disorders, but the mechanism is still elusive. We investigated the intrinsic connection between ADAM9 and GHD while focusing on the biological activities of microglia in GHRH neurons.Methods We used mass spectrometry to identify differentially expressed proteins in cerebrospinal fluid samples from a well-characterized cohort comprising patients with GHD (n = 10) and patients without GHD (n = 10). The candidates were further validated in an independent cohort (n = 20) using ELISA. In vivo, Immunofluorescence, ELISA, the Morris water maze, Western blotting and various bioinformatics analyses were used to elucidate the molecular mechanism by which ADAM9 regulates GHRH neurons.Findings A total of 683 proteins were identified, including 39 upregulated proteins in the GHD group. ADAM9 was most highly associated with GHD (AUC=0.98). In vivo, ADAM9 was colocalized with M1 microglia markers, GH and cognitive ability of mice decreased significantly, and microglia secreted ADAM9 significantly. ADAM9 regulates pyroptosis of GHRH neurons by the MAD2L2-JNK-caspase-1 pathway. Sorafenib inhibits ADAM9 secreted by microglia and improve GH levels and cognitive ability of mice.Interpretation This study found that microglia-derived ADAM9 regulates pyroptosis of GHRH neurons via the MAD2L2-JNK-caspase-1 pathway, leading to a significant reduction in GH and cognitive ability in aSAH.Funding: This study was supported by Chinese National Natural Science Foundation (82200870), President Foundation of Nanfang Hospital, Southern Medical University (2021C011), Funding by Science and Technology Projects in Guangzhou (2023A04J2334).Declaration of Interest: The authors declare no conflict of interestEthical Approval: All patients were treated by endovascular coiling by a single cerebrovascular neurosurgeon. The study was approved by the medical ethics committee of Nanfang Hospital, Southern Medical University, and patients consented to the use of their specimens for the study. All animal procedures were approved by the Animal Care Committee of Southern Medical University in accordance with the UK Animal Scientific Procedures Act 1986, the European Union (EU) Directive 2010/63/EU for animal experiments or the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health (NIH Publication No. 8023, revised in 1978). The study was approved by the Ethics Committee of Nanfang Hospital (ID: NFYY-2016-117). In addition, all efforts were made to minimize the number of animals used in the study and their suffering.

Introduction: Insulin-like growth factor type 1 receptor (IGF1R) is overexpressed in various malignant tumors, which relates to their transformation and recurrence. Craniopharyngioma is a benign tumor with malignant results, often accompanied by a severe inflammatory reaction. However, the relationship between IGF1R expression and the inflammatory response of craniopharyngioma is unclear. Methods: We enrolled 85 patients with adamantinomatous craniopharyngioma (ACP) in a study to explore the relationship between IGF1R expression and clinical features of this disease. Results: Patients in the IGF1R high-expression group had a significantly higher incidence of hypopituitarism, higher recurrence rate, and lower progression-free survival. β-Catenin can further regulate expression of the stem cell marker, CD44, by regulating IGF1R. Using immunofluorescence, we found that tumor stem cell-like cells did not express phosphorylated (p)-ERK, although p-ERK activation was evident in the surrounding cells. Picropodophyllin, a specific inhibitor of IGF1R, increased the expression of p-ERK protein and decreased the transcription level of interleukin-6. Conclusions: High expression of IGF1R might promote inflammation of ACP, which might be an unfavorable factor for pituitary function and prognosis. The high expression of IGF1R in tumor stem cell-like cells might inhibit the expression of p-ERK and promote the generation of inflammatory factors. IGF1R plays a stemness maintenance role in ACP and regulates the production of inflammatory factors through a p-ERK pathway, which suggests that targeting IGF1R and p-ERK might provide a new direction for alleviating tumor inflammation.

Some patients with paediatric craniopharyngiomas (PCs) showed normal growth despite growth hormone deficiency, which is known as growth without GH (GWGH); however, its mechanism remains unclear. We aimed to develop a novel clinical score to predict the probability of GWGH in PCs. A total of 708 PC patients were prospectively enrolled from six hospitals, among which 431 patients were finally included. Data from four of the six hospitals (n = 325) were used to develop the innovative clinical score (ICS), which was further validated using the data from the other two hospitals (n = 106). To establish and validate the ICS, sequential logistic regression was used to analyse the clinical characteristics including tumour growth pattern and tumour size and so on. Furthermore, C-statistic was employed to calibrate the discriminatory ability of the established clinical score, while a calibration plot was adopted for further assessment. The overall incidence of GWGH was 16.9% (73/431). The ICS ranged from 2 to 23, with an optimism-corrected C-statistic of 0.820, Furthermore, the optimism-corrected C-statistic of external validation was 0.835, indicating good discriminatory power and robustness of the clinical score. Additionally, no apparent overestimation or underestimation was observed in the calibration plots, which showed excellent calibration power of the clinical score. Based on tumour growth patterns and PC patients’ clinical characteristics, individualized surgical strategies were promising to achieve long-term effective management of PC patients. The ICS is valuable for the evaluation of probability of developing postoperative GWGH. ClinicalTrials.gov ID: NCT00949156.

Craniopharyngiomas (CPs) are usually benign, non-metastasizing embryonic malformations originating from the sellar area. They are, however, locally invasive and generate adherent interfaces with the surrounding brain parenchyma. Previous studies have shown the tumor microenvironment is characterized by a local abundance of adenosine triphosphate (ATP), infiltration of leukocytes and elevated levels of pro-inflammatory cytokines that are thought to be responsible, at least in part, for the local invasion. Here, we examine whether ATP, via the P2X7R, participates in the regulation of cytokine expression in CPs. The expression of P2X7R and pro-inflammatory cytokines were measured at the RNA and protein levels both in tumor samples and in primary cultured tumor cells. Furthermore, cytokine modulation was measured after manipulating P2X7R in cultured tumor cells by siRNA-mediated knockdown, as well as pharmacologically by using selective agonists and antagonists. The following results were observed. A number of cytokines, in particular IL-6, IL-8 and MCP-1, were elevated in patient plasma, tumor tissue and cultured tumor cells. P2X7R was expressed in tumor tissue as well as in cultured tumor cells. RNA expression as measured in 48 resected tumors was positively correlated with the RNA levels of IL-6, IL-8 and MCP-1 in tumors. Furthermore, knockdown of P2X7R in primary tumor cultures reduced, and stimulation of P2XR7 by a specific agonist enhanced the expression of these cytokines. This latter stimulation involved a Ca2+-dependent mechanism and could be counteracted by the addition of an antagonist. In conclusion, the results suggest that P2X7R may promote IL-6, IL-8 and MCP-1 production and secretion and contribute to the invasion and adhesion of CPs to the surrounding tissue.

BACKGROUND: The calcification of adamantinomatous craniopharyngioma (ACP) often creates difficulties for surgical therapy. Nevertheless, the mechanism of ACP calcification is unclear. Our previous studies demonstrated that osteoblastic factors might play important roles in ACP calcification.OBJECTIVE: We examined the effects of recombinant human Bmp2 on ACP cell differentiation by testing osteoblastic proteins and calcium deposition. METHODS: The expression of osteoblastic factors including osteopontin (OPN), Runx2, and osterix in Bmp2-treated ACP cells was examined by western blot and/or real time PCR. ALP activity and calcium deposition after Bmp2 induction were also tested.RESULTS: Bmp2 significantly amplified the expression of Runx2, Osterix and OPN, as well as ALP activity. Both of these effects could be repressed by noggin treatment. Bmp2 also significantly induced the calcification of ACP, and noggin inhibited this calcium deposition.CONCLUSION: Our study demonstrated for the first time that ACP cells could differentiate into an osteoblastic lineage via induction by Bmp2. The mechanism of ACP calcification likely involves osteoblastic differentiation modulated by Bmp2. Further studies targeting Bmp2 cascades could result in novel therapeutic interventions for recurrent ACP.

BACKGROUND:Beta (β)-human chorionic gonadotropin (β-HCG) is used to confirm the diagnosis and plan treatment of intracranial germinomas. However, the cutoff values of serum β-HCG in diagnosis of intracranial germinomas reported in the literature are inconsistent. To establish an appropriate cutoff value of serum β-HCG for diagnosis of intracranial germinomas, we retrospectively reviewed the records of intracranial tumor patients who received serum β-HCG and α-fetoprotein (AFP) tests for diagnostic purposes at our hospital from 2005 to 2014.METHODS:A total of 93 intracranial germinomas and 289 intracranial non-germ cell tumors were included in this study. Receiver operating characteristic (ROC) analysis was used to evaluate the sensitivity and specificity of 3 cutoffs (0.1, 0.4, and 0.5 mIU/mL) for diagnosing intracranial germinomas. The serum β-HCG level of intracranial germinoma patients was further analyzed to investigate the effect of metastasis status and tumor location on serum β-HCG level.RESULTS:The area under the ROC curve was 0.81 (P < .001), suggesting β-HCG is an effective marker. Of the 3 cutoff values, 0.1 mIU/mL possessed a highest sensitivity (66.67%) and good specificity (91%). Although there was no β-HCG level difference between metastatic and non-metastatic intracranial germinoma patients, the diagnostic rate of metastatic neurohypophyseal germinomas was significantly higher than that of its non-metastatic counterpart (P < .05), implying that the location of the germinoma might need to be considered when β-HCG is used as a marker to predict metastasis.CONCLUSIONS:Determining an optimal cutoff of serum β-HCG is helpful for assisting the diagnosis of intracranial germinoma.

Whether a mixed type of craniopharyngioma (CP) exists and whether papillary craniopharyngioma (pCP) is on a histopathological continuum with Rathke's cleft cyst (RCC) remain controversial. Herein, we examined the expression and localization of β-catenin, BRAF p.V600E (V600E), and triggering receptor expressed on myeloid cells-1 (TREM-1) in 58 samples including 20 pCPs, 26 adamantinomatous craniopharyngiomas (aCP), and 12 RCCs. Five aCPs were diagnosed with mixed type CPs and the remaining 21 cases were pure aCPs. Four of the 12 RCCs presented with significant squamous epithelium (SE). V600E immunoreactivity was observed in all pCPs in the cytoplasm, but not in the nuclei. aCPs and RCCs, including mixed type CP, did not express V600E. Nuclear β-catenin translocation was detected exclusively in aCPs. TREM-1 was expressed in pCPs. Additionally, TREM-1 expression was detected in the SE of 5 "mixed type" CPs, while it was absent in pure aCPs. TREM-1 was expressed in 4 RCCs with SE, but not in the remaining 8 RCCs. TREM-1 mRNA levels were compared in cultured pCP and aCP cells. TREM-1 mRNA level was significantly (p < 0.001; up to 4.045 fold) higher in pCPs than in aCPs. Western blotting revealed a significantly (p < 0.001; up to 7.19 fold) lower level of TREM-1 expression in aCP cells compared to that in pCP cells. Our findings further supported that RCC and pCP may represent two ends of a morphological spectrum. A variant showing overlapping histological features of aCP and pCP should not be considered as a mixed type.