bluuebunny/crossref_metadata_2025 huggingface dataset:
TMIC-23. INVESTIGATING THE FUNCTIONAL ROLE OF SECRETED FERRITIN FROM GLIOBLASTOMA CELL TYPES
Authors: Tufano Emily , Palsa Kondaiah , Serpa Rebecka , Connor James
Published: November 2024

Glioblastoma (GBM) is the most common primary malignant brain tumor in adults with a survival rate ~5%. Our laboratory previously showed the inhibitory effect of iron and ferritin-bound iron on migration and spheroid formation in GBM cells. Furthermore, studies have shown the release of extracellular vesicles (EVs) and their iron-related cargoes are regulated by cellular iron concentrations. Building on these findings, this study aims to investigate the impact of EV-associated ferritin on migration in GBM. To achieve this, we examined the influence of iron on EV release from four GBM cell lines, comprising two cancer stem cells (CSCs) and two non-CSCs: T387, T3691, LN229, and T98G. Using Nanoparticle tracking analysis (NTA), we demonstrated that 100 μM of ferric ammonium citrate (FAC) significantly increases EV release in the CSC line after 24 hours (p-value <0.01). Conversely, there was no significant impact on the non-CSC line under the same conditions. Based on these results, the ability of iron to regulate EV release is dependent on CSC versus non-CSC status. Given the functional role of ferritin on migration potential in GBM, we used immunoblotting to examine ferritin release from cells following iron treatment. We observed a reduction in ferritin release from CSCs following FAC treatment compared to the control, while non-CSCs, in contrast, released more ferritin after treatment. Interestingly, ferritin released from CSCs was predominantly within EVs, whereas ferritin released from non-CSCs was not associated with EVs. To assess the functional impact of ferritin release on migration, we utilized a Transwell migration system, and observed that EVs from iron-treated cells reduced migration in the recipients. Together, these findings expand our understanding of differential responses of ferritin in CSC and non-CSC EVs to iron treatment and will allow us to identify new targets and strategies for managing GBM progression.
TMIC-32. INFIGRATINIB INHIBITION OF THE THBS1 PATHWAY IN MELANOMA BRAIN METASTASIS
Authors: Tran Andrew , Choi John , Bergsneider Brandon , Kim Lily , Schonfeld Ethan , Abikenari Matthew , Verma Rohit , Cho Kwang Bog , Lim Michael
Published: November 2024

INTRODUCTION

Despite promising preclinical studies, BRAF/MEK inhibition with vemurafenib in melanoma brain metastases (MBM) has suboptimal durable response in humans. Thrombospondin 1 (THBS1), a matricellular protein, is upregulated in vemurafenib-resistant metastatic melanoma, and its inhibition reverses BRAF inhibitor chemoresistance. However, the characteristics of THBS1-mediated signaling pathways as well as methods of inhibition require further exploration in the context of MBM.

OBJECTIVE

To characterize THBS1-mediated signaling pathways in MBM and evaluate the therapeutic potential of targeting THBS1 pathways.

METHODS

We analyzed human parenchymal MBM samples using publicly available single-cell RNA sequencing (scRNA-seq) data. Protein-protein interaction networks were assessed using the STRING database. Syngeneic B16 melanoma cells were intracranially implanted in C57BL/6J mice and treated with intraperitoneal injection of FGF7/FGFR2 small-molecule inhibitor Infigratinib or PBS (sham). Mice were immunophenotyped using flow cytometry and monitored for survival.

RESULTS

THBS1 is highly upregulated in immunosuppressive MBM-associated cell populations such as S100A8+ Metastasis-Associated Macrophages and Mesenchymal Stromal Cell-like cells. Gene set enrichment analysis of THBS1-positive cells paradoxically revealed upregulation of inflammation-associated gene sets. Differential expression analysis identified Fibroblast Growth Factor 7 (FGF7) as strongly co-expressed in THBS1-positive MBM cells, and STRING-DB confirmed close protein-protein interactions between FGF7 and THBS1. In vivo inhibition using Infigratinib for FGF7’s receptor, FGFR2, decreased the proportion of monocytic myeloid-derived suppressor cells (M-MDSCs) (P=0.030), increased T-cell activation marker IFN-y (P=0.027) and proliferation marker CD69 (P=0.047), and resulted in improved overall survival (P=0.0001).

CONCLUSION

MBM scRNA-seq data suggests that THBS1 is associated with inflammation-induced immunosuppression. Inhibiting the FGF7/FGFR2-THBS1 axis with Infigratinib modulates the tumor microenvironment by reducing immunosuppressive M-MDSCs and increasing T cell activation and proliferation. This data supports THBS1 inhibition as a viable therapeutic option that in the future can also be combined with other current immunotherapeutic strategies.
TMIC-34. INFILTRATIVE BEHAVIOR OF PATIENT-DERIVED GLIOBLASTOMA CELL LINES IN REGION-SPECIFIC HUMAN BRAIN ORGANOIDS
Authors: Ganta Saisrinidhi , Bhatia Tarun , King Alexia , Sing Anson , Read Renee , Sloan Steven
Published: November 2024

Glioblastoma (GBM) is the most common malignant brain tumor in adults. GBMs are known for their highly invasive and proliferative properties and are associated with a dismal prognosis (~12-18 months post-diagnosis). Neuroanatomical disparities in the occurrence of GBMs suggest that in addition to microenvironmental impacts, tumor cells are impacted by the cellular and molecular diversity across varying regions of this nervous system. This idea has been difficult to study due to the lack of human-specific models that effectively recapitulate the biological behaviors of GBMs in region-specific microenvironments. To address these challenges, we engrafted GFP-tagged patient-derived glioma cell lines into human induced pluripotent stem cell (hiPSC)-derived organoids patterned to mimic the most forebrain (dorsal cortex) and hindbrain (spinal cord) regions of the nervous system. Immunostaining assays at 2, 7, 14, and 21 days post-engraftment compared the extent of infiltration, as measured by the distance traveled by glioma cells, in both regions. We found that glioma cells infiltrated a significantly greater distance in the dorsal cortical organoids compared to the spinal cord organoids. This suggests a more conducive environment for glioma infiltration in the forebrain vs. hindbrain at baseline. We are currently determining if these regional biases in infiltration capacity are modified with either optogenetic stimulation of the host brain organoids or in the presence of interregional interactions in an assembloid system. Thus, using this fully human-specific platform, we aim to gain insights into the infiltrative behaviors of GBM cells, which will advance our understanding of this devastating disease.
TMIC-35. MITOCHONDRIAL TRANSFER PROMOTES ASTROCYTE REACTIVITY IN GLIOBLASTOMA
Authors: Mitchell Jonathan , Bayik Defne , Watson Dionysios
Published: November 2024

Astrocytes are the most abundant glial cell in the brain and have been shown to adopt reactive phenotypes upon interaction with glioma cells. While tumor-associated astrocyte (TAA) reactivity is linked to increased inflammation, proliferation, invasion, and treatment resistance of glioblastoma (GBM), the mechanisms by which cancer cells reprogram TAA are poorly defined. We have previously shown that mitochondrial transfer from astrocytes to GBM cells via GAP43-dependent microtubes alters GBM cellular metabolism and increases tumorigenicity. However, the potential role of this communication mechanism in TAA function remains unknown. Patient-derived GBM cells (PDCs) were transduced with a mito-GFP lentiviral vector and were co-cultured with hTERT-immortalized, mito-mCherry human astrocytes. Using flow cytometry, we observed that the frequency of mitochondrial transfer to hTERT astrocytes was dependent on GBM cell type and noted transfer rates as high as ~40% in co-cultures with L1 PDCs. Utilizing high-resolution confocal microscopy, we subsequently confirmed co-localization of fluorescent protein signal with MitoTracker Deep Red and observed increases in mitochondrial biomass in transfer-positive astrocytes. To evaluate whether mitochondrial transfer occurs in the tumor microenvironment, we implanted immunocompromised mice with L1 mito-mCherry PDCs. Approximately 17.7% of mouse TAAs in the tumor core were positive for GBM mitochondria, whereas long-distance transfer was not observed in the contralateral hemisphere. Functionally, acquisition of tumor mitochondria promoted astrocyte proliferation as measured by cells in G2/M phase and significantly increased total cellular reactive oxygen species (ROS) levels. These findings highlight a process by which astrocytes may become reactive in response to increased ROS from GBM mitochondrial transfer. Future studies will investigate the cellular response to increased ROS in astrocytes and investigate potential links to inflammatory and immunosuppressive signaling. Elucidating how GBM-to-astrocyte mitochondrial transfer reprograms TAA would help identify therapeutic vulnerabilities that can be exploited by blocking mitochondrial transfer.
TMIC-39. SPATIAL MULTIOMICS PROFILING REVEALS HETEROGENEITY OF B:T CELL INTERACTIONS AND PLASMA CELL FORMATION IN TERTIARY LYMPHOID STRUCTURES IN HUMAN GLIOMAS
Authors: Cakmak Pinar , Lun Jennifer H , Singh Aakanksha , Macas Jadranka , Burger Michael , Hasse Lucie Marie , Hattingen Elke , Köhler Miriam , Schupp Jonathan , Starzetz Tatjana , Steidl Eike , Plate Karl H , Reiss Yvonne , Imkeller Katharina
Published: November 2024

Adult-type diffuse gliomas, the most common primary brain tumours, remain a clinical challenge in oncology with limited treatment options, restricted anti-tumour immune response and a dismal patient prognosis. Despite their immunosuppressive microenvironment, the formation of lymphoid aggregates containing adaptive immune cells has been reported in gliomas. However, the cellular compositions, immunological function, and relevance of lymphoid aggregates for adaptive anti-tumour immunity is not well understood. Therefore, we performed a comprehensive, unbiased analysis of lymphoid aggregation in 642 adult-type diffuse gliomas using a multi-modal approach, combining DNA methylation and RNA sequencing with spatial transcriptome and proteome profiling. Overall, B cell aggregates and tertiary lymphoid structures (TLS) were observed in 15% of tumours, with the highest prevalence in primary glioma localised to the temporoparietal lobe. The presence of TLS was associated with an improved overall survival. Gliomas containing TLS displayed a remodelled perivascular space, characterised by transcriptional upregulation and spatial redistribution of collagens associated with barrier functions. Spatial transcriptome and proteome profiling revealed heterogenous B:T cell interactions that were associated with elevated CD8 T-cell numbers and differences in IgA and IgG plasma cell forming capacity, suggestive of dynamic adaptive immune response.
TMIC-40. TARGETING GLIOMA TUMOR MICROENVIRONMENT BY SPECIFIC NEUTRALIZATION OF EXTRACELLULAR NICOTINAMIDE PHOSPHORIBOSYLTRANSFERASE (ENAMPT) USING ALT-100, A NOVEL HUMANIZED MONOCLONAL ANTIBODY
Authors: Sharma Pratibha , Saindane Madhuri , Zhang Katherine , Xu Jihong , Huo Longfei , Ju Zhenlin , Miele Stan , Garcia Skip , Puduvalli Vinay
Published: November 2024

BACKGROUND

Intracellular nicotinamide phosphoribosyltransferase (iNAMPT is the rate-limiting enzyme of NAD salvage pathway and is preferentially used by glioma cells for regulating energy metabolism and homeostasis. When secreted, extracellular NAMPT (eNAMPT) exerts cytokine-like functions and is associated with poor prognosis in cancer patients. We have previously reported an essential role of iNAMPT for glioma cell proliferation and survival. Here, we examined whether eNAMPT influences glioma biology and tumor microenvironment by neutralizing eNAMPT using ALT-100, a novel humanized monoclonal antibody.

METHODS

Native levels of eNAMPT in blood samples of patients with primary/recurrent gliomas were determined using ELISA under an IRB approved protocol. Next, the effects of ALT-100 on glioma microenvironment were determined by treating viable human glioma tissue slices with ALT-100 and assessing downstream effects on RNA and protein using RNASeq, reverse phase protein array and spatial transcriptomics. Lastly, the in vivo effect of ALT-100-mediated eNAMPT neutralization on survival was determined in a mouse intracranial glioma model.

RESULTS

Measurable levels of eNAMPT were noted in all patient samples with a range of (3.7-71.2 ng/ml). Preliminary results showed a correlation of eNAMPT levels with grade of tumor. Human organotypic glioma slice cultures treated with ALT-100, showed a profound alteration of gene and protein expression particularly related to key energy metabolism and proliferation pathways and the PD-1/PD-L1 axis. Additionally, results of spatial profiling of ALT-100 treated human organotypic glioma slices will be presented. Survival and response data from ALT-100 treatment in ongoing in vivo experiment will also be reported.

CONCLUSIONS

This is the first report of ALT-100-mediated eNAMPT neutralization in gliomas and demonstrates profound effects on the tumor and its microenvironment. Given that ALT-100 is in phase 2 trials (NCT05938036) against ARDS, data from our studies can potentially accelerate translating ALT-100-mediated eNAMPT neutralization to treat gliomas.
TMIC-44. ANDROGEN LOSS WEAKENS ANTI-TUMOR IMMUNITY AND ACCELERATES BRAIN TUMOR GROWTH
Authors: Lee Juyeun , Lathia Justin , Chung Yoon-Mi , Sharifi Nima , Curtin Lee , Swanson Kristin
Published: November 2024

Many cancers, including glioblastoma (GBM), exhibit a male-biased incidence and outcome disparity. The underlying reasons for this sex bias are unclear but likely involve differences in tumor cell state and immune response. This effect is further influenced by sex hormones, including androgens, which have been shown to inhibit anti-tumor T cell immunity in other solid tumors. Using a series of pre-clinical models, we observed that androgens drive anti-tumor immunity in brain tumors, contrasting with their effect in other tumor types. Upon castration, tumor growth was accelerated in GBM and brain tumor models, while the opposite effect was observed when tumors were located outside the brain. This castration effect could be reversed by testosterone administration. Flow cytometry revealed decreased T cell function, indicated by reduced anti-tumor cytokines (IFNγ and TNFα), and increased T cell exhaustion in both tumors and peripheral tissues in castrated mice, suggesting systemic immunosuppression. Indeed, the survival difference was completely abrogated in T cell deficient mice (RAG1 knockout). Mechanistically, increased hypothalamus-pituitary-adrenal (HPA) axis activity in castrated mice was responsible for the observed systemic immunosuppression, particularly in those with brain tumors. Blocking glucocorticoid receptors reversed the accelerated tumor growth in castrated mice, indicating that elevated glucocorticoid signaling mediated the castration effect. This mechanism was brain-specific rather than GBM-specific, as hyperactivation of the HPA axis was also observed with intracranial implantation of non-GBM tumors. Neuroinflammation caused by brain tumors was stronger in castrated mice, and proinflammatory cytokines like IL-1 and TNF induced HPA axis hyperactivation. Together, these findings highlight that brain tumors drive distinct endocrine-mediated mechanisms in the androgen-deprived setting and highlight the importance of organ-specific effects on anti-tumor immunity.
TMIC-47. HUMAN BLOOD-BRAIN TUMOR BARRIER ON A CHIP TO INVESTIGATE PERSONALIZED TREATMENT FOR GLIOBLASTOMA PATIENTS
Authors: Lim Jaejoon , Cho Sujin , Lee Gaeun , Ryoo Minsu , Kim Yu Jin , Sim JeongMin , Kim Dongkil , Kang Youn-Jung , Ahn Song Ih , Ahn Jungho
Published: November 2024

The inherent characteristics of glioblastoma multiforme (GBM), including tumoral heterogeneity and invasive capacity, combined with the presence of the blood-brain tumor barrier (BBTB), present challenges in developing effective treatment for GBM. Especially, the margins of GBM, where GBM cells infiltrate normal brain tissue, exhibit high resistance to therapies. Despite the difficulties in controlling tumor progression from this region, the GBM margin remains a critical area to be studied. Here we report a microengineered model that mimics the BBTB within the GBM margin, incorporating a 3D network of normal astrocytes and GBM cells isolated from patients newly diagnosed with GBM. The interaction between GBM cells and stromal cells results in increased vascular permeability, reactive gliosis, and alterations in astrocyte behavior to foster tumor invasiveness and progression. We compare patient-specific tumor responses to conventional chemotherapy in our BBTB on a chip model with clinical outcomes, demonstrating the capability of the model to predict personalized drug responses. Our BBTB model may serve as a personalized tool to examine the interactions between tumors and normal brain tissue, ultimately facilitating the screening of personalized medicine for GBM treatment.
TMIC-49. TUMOR-DERIVED SPERMIDINE DRIVES IMMUNE SUPPRESSION IN THE GLIOBLASTOMA MICROENVIRONMENT VIA CD8 T CELL DEPLETION AND FUNCTIONAL ATTENUATION
Authors: Lathia Justin , Kay Kristen , Lee Juyeun , Wang Zeneng , Grabowski Matthew , Brown Mark , Hazen Stanley , Bayik Defne
Published: November 2024

A hallmark of glioblastoma (GBM) is an immunosuppressive microenvironment that is partially driven by tumor cell-derived secreted factors, highlighting the need for a more complete understanding of local immune suppressive mechanism. Altered secreted metabolite levels are observed in the GBM microenvironment; specifically, spermidine (a polyamine), which functions to drive cell proliferation, apoptosis, T cell lineage commitment, and myeloid cell-mediated suppression. However, the direct link between GBM cell-derived spermidine and the tumor microenvironment has yet to be determined. In syngeneic mouse GBM models, spermidine levels were increased in the tumor microenvironment and exogenous administration of spermidine accelerated tumor aggressiveness in an immune dependent-manner. Mechanistically, spermidine administration increased apoptosis of CD8+ T-cells and generated an exhausted phenotype. When ornithine decarboxylase (ODC1, the rate limiting enzyme in spermidine synthesis), was knocked down in tumor cells, no direct effect was observed on tumor cell growth. However, there was an increase in time to tumor succumbence and a concomitant increase in CD8+ T-cells and decrease in CD8+ T cell exhaustion. In GBM patients, there was a negative correlation between ODC1 expression and CD8+ T-cells present near the tumor. Additionally, patients with a favorable prognosis had significantly lower intratumoral levels of spermidine compared to patients with a poor prognosis. Taken together, these results demonstrate that spermidine functions as a tumor cell-derived metabolite that drives tumor progression through reducing CD8+T cell number and altering their phenotype. These findings have clinical implications as clinical trials using a polyamine biosynthesis inhibitor alone have not been very effective in GBM patients, suggesting the need for additional combination immunotherapies. Finally, spermidine is also produced at prominent levels by commensal gut microbes as well as absorbed from consumption of foods such as mushrooms; investigation into how these distal spermidine sources impact GBM growth via the gut-brain axis is actively ongoing.
TMIC-50. IMPACT OF ZOTIRACICLIB ON GLIOMA TUMOR MICROENVIRONMENT
Authors: Lu Peng , Li Qi , Kim Olga , Sergi Zach , Gilbert Mark , Zhang Meili , Zhang Wei , Song Hua , Briceno Nicole , Davis Dionne , Wong Zalman , Castro Maria G , Wu Jing
Published: November 2024

BACKGROUND

Zotiraciclib, an orally administered multi-kinase inhibitor, has shown anti-glioma effect via suppression of transcriptional process and mitochondrial function. Here, we investigate the impact of zotiraciclib on the glioma immune microenvironment.

METHODS

Mouse glioma was generated by intracranial injection of mouse syngeneic cells (IDH1-wildtype NPA cells or IDH1R132H-mutant NPAI cells) in C57BL/6 mice. Zotiraciclib was administered (30 mg/kg, intraperitoneal injection, twice per week) starting seven days after tumor implantation. NPA and NPAI tumors were dissected after 10 and 22 days’ treatment, respectively. Immune cells in the tumor tissue were isolated and examined using flow cytometry. Primary human CD8+ cells and CD14+ monocytes isolated from the blood of a healthy donor were used to further investigate the effect of zotiraciclib (15 nM and 50 nM) on human cytotoxic T cell activation and immunosuppressive M2 macrophage polarization, respectively, using flow cytometry.

RESULTS

No significant difference in the abundance of T cells, myeloid cells, NK cells, NKT cells, DCs, and B cells between zotiraciclib-treated and vehicle-treated groups was detected in neither NPA nor NPAI tumors. The treatment-induced change in the abundance of cytotoxic T cells, helper T cells, macrophages, monocytes, neutrophils, M-MDSCs, PMN-MDSCs, Tregs, and M2 macrophages wasn’t observed. Zotiraciclib reduced the expansion of human CD8+ cytotoxic T cells, but did not affect the expression of CD25, HLA-DR, TNFα and IFNγ after activation in vitro. The viability of human M2 macrophages during differentiation and polarization was not affected, while the expression of CD163 and CD206 was reduced by zotiraciclib.

CONCLUSIONS

The immunophenotyping performed for this study demonstrates that zotiraciclib does not alter the mouse glioma immune microenvironment. Complementary studies using human immune cells shows that zotiraciclib does not suppress the activation of cytotoxic T cells but reduces the polarization of immunosuppressive M2 macrophages, supporting combining zotiraciclib and immune stimulatory approach in glioma treatment.
TMIC-55. CHARACTERIZATION OF THERAPEUTICS EFFECTS IN GLIOBLASTOMA THROUGH INTEGRATED SPATIAL SINGLE-CELL MULTIOME PROFILING OF LONGITUDINAL SAMPLES: UNVEILING CLINICALLY RELEVANT SUBTYPES OF PATHOLOGICAL CHANGES
Authors: Vanmechelen Maxime , Pouya Nazari , Beckervordersandforth Jan , Leunissen Daphne , Cole Basiel , Decraene Brecht , De Visser Yanti , Shankar Gautam , Verduin Maikel , Bravo Carmen , Moors Tom , Telang Jayesh , Messiaen Julie , Eekers Danielle , Claeys Annelies , Derweduwe Marleen , Zurhausen Axel , Bosisio Francesca , Weyns Frank , Daenekindt Thomas , Van Eyken Peter , Govers Mieke , Mennens Ferdinand , Hovinga Koos , De Vleeschouwer Steven , Paul Clement , Broen Martijn , Vooijs Marc , Sciot Raf , Antoranz Asier , Pey Jon , Speel Ernst Jan , Hoeben Ann , De Smet Frederik
Published: November 2024

Glioblastoma (GBM) remains a highly malignant, intrinsically resistant and inevitably recurring brain tumor with dismal prognosis. The aggressiveness and lack of effective GBM treatments can be attributed to the highly heterogeneous and plastic nature of GBM tumor cells, which easily confer resistance to standard-of-care (SOC) therapy. While tumor progression has also been attributed to interactions with the tumor microenvironment, quantitative data describing these interactions are still largely missing. Here, we used an multiome approach combining high-dimensional, multiplexed immunohistochemistry and spatial transcriptomics to map evolutions in the spatial, single-cell tissue architecture of 357 paired adult GBM tumor samples derived from 52 patients at diagnosis (ND) and upon recurrence (REC) following SOC treatment. We mapped the spatial distribution of a multitude of GBM tumoral subtypes across this multicentric cohort, through which we identified a high level of heterogeneity defined by specific tumoral niches within and across patients and which evolved when subjected to SOC therapy. In addition, we describe the relationship of the various tumoral niches with their local immune-infiltrates, highlighting an even more immunosuppressive environment following SOC resistance. Finally, by aligning these findings to the observed genomic aberrations and the clinical data of the patients, we are now able to more precisely describe the heterogeneous landscape of glioblastoma and how it evolves under SOC treatment at spatial, single-cell resolution.
TMIC-85. THE MECHANISM OF COMBINED HYDROGEN-CHEMICAL THERAPY BASED ON ACTIVE MAGNESIUM MICROMOTORS FOR INHIBITING GLIOBLASTOMA RECURRENCE BY MODULATING TUMOR MICROENVIRONMENT
Authors: Zhao Shiguang
Published: November 2024

OBJECTIVE

This study aims to develop an innovative therapeutic strategy to prevent postoperative recurrence of Glioblastoma (GBM), a condition with poor prognosis and high recurrence rates. The focus is on exploring the efficacy of synergistic hydrogen-chemical therapy in inhibiting GBM recurrence, especially by reducing surgery-induced inflammation and enhancing the sensitivity of GBM cells to chemotherapy.

METHODS

The study employed the fabrication of Mg-Motor-DOX using a biodegradable polylactic acid-glycolic acid copolymer (PLGA) and gelatin containing Doxorubicin (DOX). The reaction of Mg with water produces hydrogen (H2), which aids in scavenging reactive oxygen species (ROS) and inflammation, alongside the specific release of chemotherapy drugs at the tumor site. A thermosensitive hydrogel was developed for in situ delivery of Mg-Motor-DOX to the tumor residual cavity. Additionally, RNA sequencing was used to study the transcriptional changes in the GBM tumor microenvironment (TME) induced by this therapy.

RESULTS

The study presents initial evidence of the in vivo efficacy of synergistic hydrogen-chemical therapy against GBM. The Mg-Motor-DOX mediated therapy not only scavenged ROS and reduced inflammation but also specifically released chemotherapy drugs at the tumor site, enhancing their effectiveness. RNA-Seq technology elucidated the molecular mechanisms underlying the therapy’s efficacy, revealing significant transcriptional changes in the GBM TME.

CONCLUSION

Synergistic hydrogen-chemical therapy, facilitated by Mg-Motor-DOX, shows promising potential in preventing the postoperative recurrence of GBM. This innovative approach effectively combines anti-inflammatory benefits and enhanced chemosensitivity, offering new strategies for GBM treatment. The molecular insights provided by RNA-Seq technology contribute to a deeper understanding of tumor treatment mechanisms, paving the way for future clinical applications.
TMOD-03.BRAFV600E MUTATION INDUCES TWIST1 UPREGULATION ASSOCIATED WITH MIGRATION AND CEREBRAL FLUID DISSEMINATION IN GLIOBLASTOMA
Authors: Miyake Yohei , Furnari Frank
Published: November 2024

INTRODUCTION

BRAFV600E-mutant glioblastoma (GBM) with epithelioid features frequently show cerebrospinal fluid dissemination (CSFD), which results in poor prognoses. BRAF mutation is worth investigating since BRAF or downstream MEK inhibitors show dramatic response for BRAFV600E-mutant GBM including CSFD. This suggests BRAF mutation could play a role for inducing the clinical manifestation of CSFD found in newly diagnosed and recurrent BRAFV600E-mutant GBM. Because, BRAFV600E-mutant GBM often recur with CSFD, elucidating the mechanism by which BRAFV600E-mutant GBM exhibits CSFD is important to improve prognosis. (Objective) To elucidate the molecular mechanism of CSFD involved in BRAFV600E-mutant GBM.

METHODS

We introduced the heterozygous BRAFV600E mutation in human induced pluripotent stem cells (iPSC) in the context of GBM genetic alterations, CDKN2A/2B, and PTEN deletion, TERT promoter mutation and with EGFRvIII overexpression. We implanted these iPSC-derived neural progenitor cells into immunodeficient mouse brains to establish a BRAFV600E-mutant GBM model. At the same time, we created a model with the same genetic background without BRAFV600E. We compared their gene expression, pathology, and migration patterns to identify BRAFV600E specific characteristics.

RESULTS

RNA sequencing revealed BRAFV600E-mutant tumors upregulated migration, and downregulated cell-to-cell adhesion genes. TWIST1 was one of the most upregulated genes, and was validated by qPCR. Concomitant with TWIST1 expression we detected downregulation of E-cadherin. The BRAFV600E GBM model histologically showed a cluster-like invasion phenotype, while non-BRAF tumors showed homogenous invasion with typical GBM pathological findings. Immunohistochemical staining showed relatively high TWIST1 expression in cluster-like tumor cells of BRAFV600E tumors. Upon re-engraftment of primary tumor-derived cells, secondary BRAFV600E tumors presented leptomeningeal dissemination. TWIST1 knockdown resulted in a decrease in the migratory ability of our BRAFV600E model in vitro, and is under investigation in vivo.

CONCLUSION

BRAV600E mutation induces TWIST1 upregulation, and might facilitate migration and CSFD. This work nominates TWIST1 and its regulated genes as targets for BRAFV600E-mutant GBM with CSFD.
TMOD-08. DETAILED GENERATION OF AN ORTHOTOPIC INTRACRANIAL GLIOBLASTOMA (GBM) PATIENT-DERIVED XENOGRAFT (PDX) MODEL FOR PRECLINICAL IMMUNOTHERAPEUTIC STUDIES
Authors: Jung Tae-Young , Tran Thi-Anh-Thuy , Marcelina Correia Dircia Canisia , Kim Young-Hee , Park Sue-Jee , Kim Yeong Jin , Moon Kyung-Sub , Kim In-Young , Jung Shin , Lee Kyung-Hwa , Park Ae Kyung
Published: November 2024

PURPOSE

This study aimed to establish an orthotopic intracranial PDX mouse model using cryopreserved GBM tissues expanded in a heterotopic subcutaneous PDX model for immunotherapeutic studies. The model is designed to closely replicate human GBM, enabling the exploration of its pathophysiology and the development of effective immunotherapies for this type of cancer.

METHODS

Glioblastoma tissues from 16 fresh and cryopreserved samples were used to establish a heterotopic subcutaneous PDX mouse model for tumor expansion. Successful engraftments were then used to create an orthotopic intracranial PDX mouse model. The tumor formation rates and durations were estimated in both models. Tumor fidelity to the original GBM tumors was confirmed through immunohistology (GFAP, P53, and Ki67) and genetic analysis (whole exome sequencing, RNA sequencing, and methylome analysis). The therapeutic effects of immunotherapy (allogenic NK cells) in combination with Avastin and Irinotecan were tested in the orthotopic intracranial PDX model.

RESULTS

The orthotopic intracranial GBM mouse model achieved 100% engraftment from successful tumor growth in the subcutaneous PDX model. The overall success rate was 81.25% (85.71% for cryopreserved tissues), with a median process duration of 26.5 weeks (19 weeks for the subcutaneous and 7.5 weeks for intracranial establishment). No significant differences in tumor formation were observed within one year for cryopreserved tissue in the subcutaneous model. Molecular and genetic analysis confirmed similar tumor characteristics between the intracranial PDX tumors and the original patient tumors. Additionally, NK cells combined with Avastin and Irinotecan demonstrated efficacy in this model.

CONCLUSIONS

We successfully established an orthotopic intracranial PDX mouse model using cryopreserved GBM tissues, with no observed differences within one year. These models accurately mirror patients’ tumor characteristics, promising advancements in glioblastoma research and treatment, particularly for immunotherapy studies. Keywords: Glioblastoma (GBM), cryopreserved GBM tissues, patient-derived xenograft (PDX) model, NOD/SCID IL-12Rγnull (NSG) mice
TMOD-30. DISRUPTION OF NORMAL RADIAL GLIA DEVELOPMENT IN AN NF2-ALTERED NEUROEPITHELIAL STEM CELL MODEL
Authors: Burket Noah , Dershem Victoria , Dixon Shelley , Cooper Scott , Angus Steve , Clapp Wade , Tailor Jignesh
Published: November 2024

NF2-related schwannomatosis is a tumor predisposition syndrome caused by mutations in the NF2 gene and associated with spinal ependymomas (SP-EPN). SP-EPNs are suspected to originate from aberrations in the radial glia (RG) cell lineage. These tumors are only effectively treated through high-risk surgical resection, emphasizing the critical need for identification of targets for medical therapy. Yet, the role of NF2-dependent disruption in RG cell development is poorly understood. We hypothesize that this mutation may be halting normal RG cell differentiation, resulting in a RG-like stem cell that is driving SP-EPN formation. An NF2-knockout was generated in neuroepithelial stem (NES) cells using CRISPR/Cas9. Knockouts were validated using Western blot and Sanger sequencing. In vitro differentiation was induced with removal of growth factors. NF2-knockout phenotypes were assessed with real-time polymerase chain reaction and compared with wildtype NES cells. Preliminary data has shown that NF2-knockout cells express similar levels of early pan-neural and neural stem cell genes compared to wildtype after CRISPR editing. The NF2-knockout cells retain this stem cell-like gene expression following attempted differentiation, whereas wildtype cells take on a primarily neural phenotype. The knockout cells also form what appears to be pre-neoplastic spheres when allowed to differentiate. NF2-knockout NES cells fail to differentiate normally compared to wildtype NES cells. They retain early RG-like markers, including SOX2, BLBP, and GFAP. Furthermore, formation of spheres when growth factors were removed hints at NF2 loss being relevant to formation of pre-neoplastic growths. Future work will include investigating downstream effects of NF2 loss in this model.
Mimicking osteoid 3D porous dense microfiber silk fibroin embedded poly(vinyl alcohol) scaffold for alveolar ridge preservation
Authors: Sangkert Supaporn , Kannan Perumal Ramesh , Meesane Jirut , Santavalimp Kanokporn , Phongthanawarakun Jutharat , Promkaew Walaiporn , Anupan Wachiratan , Thuaksuban Nuttawut
Published: November 2024

Alveolar ridge loss presents difficulties for implant placement and stability. To address this, alveolar ridge preservation (ARP) is required to maintain bone and avoid the need for ridge augmentation using socket grafting. In this study, a scaffold for ARP was created by fabricating a 3D porous dense microfiber silk fibroin (mSF) embedded in poly(vinyl alcohol) (PVA), which mimics the osteoid template. The research utilized a freeze–thawing technique to create a mimicked osteoid 3D porous scaffold by incorporating different amounts of mSF into the PVA, namely, 1%, 3%, 5% and 7%. Subsequently, a 3D profilometer machine and a scanning electron microscope were employed to examine the morphology and size of the mSF and the mimicked osteoid 3D porous scaffold in all groups. Thermal characteristics and crystalline structure were analyzed before assessing the water contact angle, swelling behavior, degradation and mechanical properties. The experiment evaluated the biological performance of the mimicked osteoid 3D porous scaffold by examining the efficacy of osteoblast cell adhesion, proliferation, viability, protein synthesis, alkaline phosphatase (ALP) activity and calcium synthesis. Finally, the ability of osteoblast cells to regulate the osteoid matrix deposition on the osteoid 3D porous scaffold was assessed by mimicking the dynamic bone environment using rat mesenchymal stem cells. The findings suggest that incorporating mSF into PVA enhances the interconnective pore size, crystalline structure and thermal behavior of the mimicked osteoid 3D porous scaffold. The hydrophilicity of PVA decreased with an increase in the proportion of mSF, while a higher proportion of mSF resulted in increased swelling and mechanical characteristics. Incorporating a greater proportion of mSF, specifically 5% and 7%, led to a reduced rate of degradation. The addition of 5% mSF to the PVA 3D porous scaffold resulted in remarkable biological properties and excellent osteoconductive activity.
The Impact of Complication Timing on the Outcomes of Implant-based Breast Reconstruction
Authors: Alnaseri Tahera , Prabhu Shamit , Anderson Lexy , Kwan Lorna , Demirjian Maral , Kwok Alvin , Reid Christopher , Hollenbeck Scott , DeLong Michael R.
Published: November 2024

Background:

The National Surgical Quality Improvement Program (NSQIP) database provides an important resource for determining complication rates and risk factors for surgical procedures. However, NSQIP is limited to 30-day follow-up, and it is unclear whether this is reliable for evaluating prosthetic breast reconstruction outcomes.

Methods:

A single-institution, cross-sectional, retrospective review was performed for patients undergoing mastectomy with immediate, prepectoral tissue expander reconstruction. Timing of complications was stratified as early (within 30 days of operation) versus late (after 30 days). Categorical variables were compared using χ2 (or Fisher exact) tests, and continuous variables were analyzed using Kruskal-Wallis or Wilcoxon rank-sum tests.

Results:

There were 301 patients (509 reconstructed breasts) included with a median follow-up time of 11 months. Of them, 176 patients (58%) experienced a postoperative complication—140 patients (47%) experienced an early complication and 36 patients (12%) experienced a late complication. Patients with late complications had a significantly higher rate of reconstructive failure compared with the early complication group (17% versus 10%; P = 0.001) and were more likely to require a flap (28% versus 7%; P = 0.001) for final reconstruction. Revision surgery rates after final implant placement were higher in the late complication group (36% versus 64%; P = 0.285).

Conclusions:

Late complications after prepectoral breast reconstruction have a more prominent impact on reconstructive failure and revisions than early complications. This finding may inform strategies to revise national databases such as NSQIP to include more detailed information and longer capture periods.
Causal effects of Parkinson’s disease on the risk of osteoporosis: A two-sample Mendelian randomization study
Authors: Huang Yu , Yi Nan , Li Qinglong , Guo Song , Mo Bingfeng , Yin Dong , Li Hongmian
Published: November 2024

Employing a two-sample Mendelian randomization (MR) analysis, we aimed to investigate the potential causal effect of Parkinson disease (PD) on osteoporosis. We conducted an in-depth MR analysis by leveraging extensive genome-wide association study datasets from the International Parkinson Disease Genomics Consortium and the Genetic Factors for Osteoporosis Consortium. We meticulously selected instrumental variables based on strict criteria, including significance thresholds, linkage disequilibrium, and the exclusion of confounding single-nucleotide polymorphisms. Our investigation utilized diverse MR methods, including inverse variance weighted, MR Egger regression, weighted median, and MR-PRESSO, to robustly evaluate the causal relationship. Our comprehensive analysis revealed noteworthy associations between PD and distinct measures of bone mineral density (BMD) (forearm BMD, femoral neck BMD, lumbar spine BMD). Specifically, the inverse variance weighted method underscored potential significant relationships between PD and forearm BMD (P = .037; odds ratio [OR], 1.04; 95% confidence interval [CI], 1.00–1.09), femoral neck BMD (P = .034; OR, 1.02; 95% CI, 1.00–1.05), and lumbar spine BMD (P = .043; OR, 1.03; 95% CI, 1.00–1.06). The consistency of results across various methods and sensitivity analyses indicated both robustness and minimal pleiotropy concerns. Through a two-sample MR approach, this study establishes a plausible causal relationship between PD and decreased BMD. The outcomes underscore the urgency of targeted interventions to mitigate bone loss and manage osteoporosis in individuals with PD.
Exploring causal associations between dietary intake and liver diseases: A bidirectional Mendelian randomization study
Authors: Wang Hong , Yun Zhangjun , Li Liling , Wang Hui , Zeng Haotian , Ran Yun
Published: November 2024

Previous evidence suggests that dietary intake can affect liver diseases; However, the causal relationship between dietary intake and liver diseases remains unclear. To investigate this, we conducted a bidirectional Mendelian randomization (MR) analysis to comprehensively assess the potential causal relationship between dietary intake and liver diseases. Two-sample bidirectional MR was performed based on genome-wide association studies summary data from the UK Biobank and FinnGen database. The primary analysis method for evaluating causal relationships was inverse-variance weighted. Supplementary analyses included MR-Egger and weighted median methods. Subsequently, sensitivity analyses were performed using Cochran Q test, MR-Egger intercept test, MR-PRESSO, RadialMR, and leave-one-out analysis to assess heterogeneity and horizontal pleiotropy. MR evidence indicated that genetically predicted poultry intake (adjusted odds ratio [OR] = 0.04, 95% confidence interval [CI] = 0.00–0.43, P = .007) and salad/raw vegetable intake (adjusted OR = 0.18, 95% CI = 0.04–0.83, P = .028) were directly associated with a reduced risk of cirrhosis. Conversely, there is no causal association between dietary intake and nonalcoholic fatty liver disease, alcoholic liver disease, or hepatocellular carcinoma. This study provides evidence supporting the impact of dietary intake on liver disease. Increased intake of poultry and salad/raw vegetables is associated with a reduced risk of cirrhosis. These findings can inform preventive and therapeutic strategies for cirrhosis.
Global trends in ischemic stroke burden attributable to high BMI
Authors: Gao Qiongya , Chao Wei , Xu Jiali , Yu Wangfang
Published: November 2024

We aimed to assess the worldwide burden of ischemic stroke (IS) associated with high body mass index (BMI) using data from the Global Burden of Disease 2019. This study examined the impact of high BMI on IS-related age-standardized death rates (ASDR) and disability-adjusted life years (DALYs). Estimated annual percentage changes (EAPC) is estimated annual percentage change. Trends were assessed using EAPCs. Over the past 3 decades, there has been a declining trend in the global burden of IS associated with high BMI, especially in Western Europe (EAPC = −3.09 for DALYs) and high-income Asia Pacific (EAPC = −4.6 for ASDR). However, certain developing regions, notably Southeast Asia, have experienced significant increases in ASDR (EAPC = 3.7) and age-standardized DALY rates (EAPC = 3.64). The most substantial increase in burden was observed in Southeast Asia for both males (EAPC = 3.85) and females (EAPC = 3.53). Importantly, the burden was most pronounced in regions with low to middle sociodemographic index. The rising disease burden linked to high BMI is largely due to insufficient medical infrastructure and ineffective public health policies in the region. Urgent action is needed from decision-makers to improve these areas and implement effective interventions. This study reveals a consistent global decline in IS related to high BMI, with a more significant burden observed in males under the age of 65, particularly in Southeast Asia, where increases in IS associated with high BMI are notable. Public health officials and global policymakers need timely and reliable quantitative data. This information is essential for implementing effective behavioral interventions, such as monitoring diet and physical activity, to address identified risk factors and reduce the burden of high BMI.
Association between hemoglobin-to-red blood cell distribution width ratio and chronic kidney disease: A cross sectional study
Authors: Ning Lin , Tang Junping , Chen Zhiqiang , Zeng Xiaolin , Liu Quan , Tan Liming , He Min
Published: November 2024

The hemoglobin-to-red blood cell distribution width ratio (HRR) is recognized as a novel prognostic biomarker; however, studies exploring its relationship with chronic kidney disease (CKD) are scarce. This study used data from the National Health and National Health and Nutrition Examination Survey database from 2005 to 2018. The analysis included individuals aged ≥ 20 years who had complete HRR and CKD data. Weighted univariate and multivariate logistic regression analyses were used to assess the association between the HRR and CKD prevalence. Additionally, restricted cubic spline and subgroup analyses were conducted for further validation. Ultimately, 19,426 participants were included in this study. After adjusting for confounders, multivariate logistic regression analysis revealed a negative association between HRR and CKD (OR = 0.35, 95% CI = 0.22–0.56). In addition, restricted cubic spline regression analysis revealed a negative linear association between HRR and CKD, with higher levels of HRR associated with a lower prevalence of CKD. The subgroup analysis revealed that the negative association between HRR and CKD was stronger in the male population. HRR is negatively associated with the prevalence of CKD in the adult population of the US. HRR is a potential indicator for assessing the prevalence of CKD and provides a rationale for personalized management.
Case report: Peritonitis caused by Pantoea agglomerans from pet dog exposure in peritoneal dialysis
Authors: Kim Ji Yeon , Cho Nam-Jun , Park Samel , Gil Hyo-Wook , Lee Eun Young
Published: November 2024

Rationale:

Peritonitis caused by Pantoea agglomerans is a rare occurrence in patients undergoing peritoneal dialysis. Cases potentially linked to pet dogs are even rarer, and there is limited literature available.

Patient concerns:

A patient undergoing peritoneal dialysis presented with symptoms of peritonitis, including abdominal pain and cloudy dialysis fluid.

Diagnoses:

Microbiological analysis identified P agglomerans as the causative organism.

Interventions:

The patient was treated with targeted antibiotic therapy and showed a positive response.

Outcomes:

During a subsequent medical interview, it was revealed that the patient had close contact with their pet dog, raising the possibility that the infection may have been associated with this exposure.

Lessons:

This case highlights the importance of considering zoonotic transmission as a potential source of infection in peritoneal dialysis patients, particularly when there is close contact with pets. Healthcare providers should educate patients about the potential risks posed by pets and implement preventive strategies to mitigate such risks.
Efficacy of four internal fixation devices on femoral neck fractures in young adults: A systematic review and network meta-analysis
Authors: Yuan Daotong , Zhang Zhimeng , Wang Xu , Chang Wenjie , Xie Wenpeng , Zhang Yongkui
Published: November 2024

Background:

The primary treatment of femoral neck fracture in young adults is internal fixation. The high complication rate after femoral neck fracture greatly affects the life of patients. There are many internal fixation devices for femoral neck fracture, but each has its advantages and disadvantages. Our aim was to determine the best internal fixation for young people with femoral neck fractures.

Methods:

We searched 5 databases from January, 2016 to December, 2023. Randomized controlled trials and cohort studies that met the inclusion criteria were assessed for quality using the RoB.2 and ROBINS-I scales, respectively. The network meta-analysis was conducted within a Bayesian framework utilizing a random effect model. Data analysis was performed using the “multinma” package within the R 4.2.0 software.

Results:

A network meta-analysis of 34 studies involving 2291 patients was conducted. Results indicated that the inverted triangular cannulated screws demonstrated the lowest intraoperative bleeding volume (surface under the cumulative ranking curve [SUCRA] = 0.8732) based on the SUCRA. The medial buttress plate (MBP) exhibited superior efficacy in improving the Harris hip score (SUCRA = 0.8465), reducing complications (SUCRA = 0.9251), and accelerating fracture healing time (SUCRA = 0.8111). Additionally, the femoral neck system was ranked highest in terms of operation time (SUCRA = 0.7749) and femoral neck shortening (SUCRA = 0.7933).

Conclusion:

This network meta-analysis findings indicated that MBP resulted in superior postoperative hip function, reduced complication rate, faster fracture healing time. Considering the good physical condition of young adults, surgeon may consider utilizing MBP to achieve improved postoperative outcomes.