bluuebunny/crossref_metadata_2025 huggingface dataset:
TMIC-78. ENDOTHELIAL REGULATION OF THE IMMUNOSUPPRESSIVE PHENOTYPE OF TUMOR-ASSOCIATED MICROGLIA IN GLIOBLASTOMA
Authors: Muthukrishnan Sree Deepthi , Qi Haocheng , Alvarado Alvaro , Kawaguchi Riki , Kornblum Harley
Published: November 2023

Glioblastoma (GBM) is considered a non-immunogenic “cold tumor” with an immunosuppressive microenvironment that blocks T-cell infiltration and function, limiting the efficacy of targeted immunotherapy. Tumor-associated microglia and macrophages (TAM) constitute approximately 30-50% of the non-neoplastic immune cells, and they are closely associated with blood vessels in GBM. However, the precise mechanism by which TAM acquire the immunosuppressive phenotype remains poorly understood. In this study, we show that tumor endothelial cells (TEC) from primary and recurrent GBM express inflammatory cytokines and chemokines that influence the function and polarization of microglia. We demonstrate that conditioned media (CM) of TEC promotes the immunosuppressive phenotype of normal microglia and patient-derived TAM. RNA-sequencing of TAM treated with TEC-CM shows upregulation of immunosuppressive cytokines, as well as T-cell co-inhibitory molecules such as PD-L1. Culturing CD8 T-cells in CM of TAM-polarized by TEC inhibits their cytotoxic function and cytokine production, and induces expression of PD-1 and CTLA-4. Furthermore, gene set enrichment analysis uncovers alterations in calcium signaling-related pathways in TAM polarized by TEC-CM. Blocking calcium signaling with small molecule inhibitors in microglia reverses the immunosuppressive phenotype induced by TEC. To determine whether the interactions between GBM, endothelial cells and TAM promotes immunosuppression in vivo, we utilize the humanized immune system mouse model (huNOG-EXL) that harbor human lymphocytes and monocyte populations. In preliminary studies, we found that co-transplantation of GBM tumor cells with TAM and TEC significantly promotes tumor growth compared with GBM cells alone. Single-cell mass cytometry analysis of peripheral blood mononuclear cells (PBMC) pre- and post-transplant, and scRNA-sequencing and spatial-transcriptomic studies of these co-transplanted mice will potentially reveal key insights into local- and peripheral immune responses regulated by tumor endothelial-microglia interactions. Together, our findings highlight a key role for endothelial-mediated calcium signaling in regulation of the immunosuppressive phenotype of microglia in GBM.
TMIC-80. PERK CONTRIBUTES TO GLIOBLASTOMA PROGRESSION BY REGULATING GLIOBLASTOMA STEM CELL ADAPTATION TO EXTRA CELLULAR MATRIX STIFFENING THROUGH F-ACTIN REMODELING AND FOCAL ADHESION COMPLEX REGULATION
Authors: Khoonkari Mohammad , Liang Dong , Govaerts Chris , Huurman Romy , Kamperman Marleen , van Rijn Patrick , Kruyt Frank
Published: November 2023

The association between a stiffening extracellular matrix (ECM) and increased aggressiveness of gliomas is well established. Originally recognized as a protective mechanism against endoplasmic reticulum (ER) stress, the unfolded protein response (UPR) is primarily responsible for maintaining cellular protein homeostasis and triggering cell death under severe stress conditions. However, emerging evidence indicates that the ER transmembrane UPR sensors, IRE1 and PERK, possess additional functions beyond proteostasis, such as modulating cancer stem cell differentiation, cell adhesion, and migration, which also involve more recently identified protein scaffold functions. In this study, we uncover a novel role of PERK in cellular adaptation to matrix stiffening. Glioblastoma stem cells (GSCs), when cultured on human blood plasma/alginate hydrogels with tuneable stiffness, exhibited stiffness-dependent cellular responses, including cell flattening, differentiation, and enhanced migration. Through genetic depletion and pharmacological inhibition strategies combined with immunofluorescence microscopy, we identified the involvement of a previously found PERK/FLNA signaling mechanism in the adaptive response to matrix stiffness, where F-actin polymerization emerged as a key mechanism. To explore the possible involvement of PERK in sensing ECM stiffness, the expression of various proteins that constitute the focal adhesion complex were studied. Remarkably, PERK also influenced the maturation of focal adhesion complexes components know to be involved in sensing ECM stiffness. Finally, comparative transcriptional profiling of PERK proficient and deficient GSCs uncovered significant alterations in the expression of genes associated with cell adhesion and extracellular matrix (ECM)-related processes. These findings reveal a novel function of PERK in stiffness-dependent cellular adaptations, with ongoing investigations exploring the underlying molecular mechanisms and therapeutic implications for glioblastoma progression
TMIC-85. TGFB2 AND IFNGR2 MRNA EXPRESSION LEVELS IN THE TUMOR MICROENVIRONMENT (TME) PROGNOSITICALLY IMPACT OS IN PEDIATRIC DIPG PATIENTS
Authors: Qazi SanJive , Trieu Vuong
Published: November 2023

DIPG patients expressed significantly higher levels of TGFB2 and IFNGR2 than control pons samples and the expression profile indicated higher levels of TGFB2 than TGFB1 and TGFB3 in DIPG patients suggesting TGFB2 mRNA level is preferentially increased in DIPG patients. TGFB2 is a prognostic indicator (HR for patients in the TGFB2high group of DIPG patients = 2.88 (1.12-7.39); P = 0.028) for poor overall survival (OS) independent of IFNGR2 levels, the age of the patient, and the significant interaction effect observed between IFNGR2 and TGFB2. Predictive modeling using the Cox regression parameters showed that the context of low expression of IFNGR2, TGFB2low resulted in more favorable OS times compared to TGFB2high whereby the median OS time of 11 months for the TGFB2low group of patients was greater than the upper 95% confidence limit for the TGFB2high group of patients (Median = 7 months; Upper 95% Confidence interval = 10 months). At higher levels of IFNGR2 expression, the TGFB2high group of patients exhibited a median OS time of 15 months that was within the (95% confidence interval for the TGFB2low group of patients (Median = 13 months; 95% CI = 10-17 months). Worse survival outcomes comparing DIPG patients with high versus low TGFB2 levels in the context of low levels of IFNGR2 levels suggest that abrogating the TGFB2 mRNA expression in the immunologically cold tumor microenvironment indicated by low levels of IFNGR2 expression can potentially treat pediatric DIPG patients. Furthermore, DIPG patients with low levels of JAK1 or STAT1 mRNA expression in combination with TGFB2high also exhibited poor OS outcomes suggesting that the inclusion of (Interferon-gamma) IFNγ to stimulate and activate JAK1 and STAT1 in anti-tumor immune cells present the brainstem tumor microenvironment can enhance the effect of TGFB2 blockade.
MODL-05. ELUCIDATING THE EPIGENETIC DRIVEN TRANSCRIPTIONAL PROGRAM UNDERLYING THE INVASIVE PHENOTYPE OF THE NOVEL EGFRA289VARIANT OF GBM
Authors: Lin Benjamin , Ziebro Julia , Shelton Abigail , Skinner Kasey , Bash Ryan , Osuka Satoru , Sudarshan Sunil , Binder Zev , O'Rourke Donald M , Nathanson David , Furnari Frank , Miller Christopher
Published: November 2023

Glioblastoma (GBM) is a devastating malignant brain tumor with a dismal 15-month median survival. Current treatments have not been successful at preventing recurrence, thus novel therapeutic modalities are urgently needed. EGFR is a tyrosine kinase receptor that is commonly mutated in ~60% of GBM and is an attractive target for GBM precision oncology. Aberrant EGFR signaling drives tumor proliferation and maintenance, thus further study of common EGFR alterations in GBM, including interstitial deletions (EGFRvIII) and missense mutations (EGFRA289), is needed to develop effective EGFR-targeted therapies. This is especially important in the context of a retrospective study that showed EGFRA289 driven GBM was more clinically aggressive. EGFRA289 positive GBM had a unique, invasive radiographic signature on magnetic resonance imaging (MRI) and patients had a significantly shorter median overall survival of 6-months. We hypothesize that the malignant clinical phenotypes of EGFRA289 GBM are driven by unique epigenomic and transcriptomic, collectively referred to as (epi)genomic, programs linked to EGFRA289 signaling. Prior studies using established cell lines show that EGFRvIII overexpression drives malignant (epi)genomic programs mediated by transcription factor (TF) activation. To better understand variant specific EGFR biology in GBM, we have developed novel genetically engineered mouse astrocyte (mAc) models that overexpress wtEGFR (MA-WT), vIII (MA-v3), and EGFRA289V (MA-A289V). Phospho-immunoblots show that these mAc models have differential EGFR ligand dependance, which match published studies. Despite this difference in EGFR signaling, all mAc models have similar sensitivities to EGFR tyrosine kinase inhibitor (TKI) treatment. However, MA-A289V is more proliferative and migratory compared to the other mAc variants. Preliminary RNA-Seq on our mAc models has identified similar and unique TFs differentially expressed (DE) between MA-v3 and MA-A289V when compared to parental MA-C. Integration of RNA-Seq and CUT&RUN followed by mechanistic genetic interrogation will elucidate the relationship between EGFR variant-specific (epi)genomic mechanisms and their unique malignant phenotype.
MODL-09. SCAFFOLD-BASED (MATRIGELTM) 3D CULTURE TECHNIQUE OF GLIOBLASTOMA RECOVERS A PATIENT-LIKE IMMUNOSUPPRESSIVE PHENOTYPE
Authors: Braun Frank , Rothhammer-Hampl Tanja , Lorenz Julia , Pohl Sandra , Menevse Ayse-Nur , Vollmann-Zwerenz Arabell , Bumes Elisabeth , Büttner Maren , Zoubaa Saida , Proescholdt Martin , Schmidt Nils , Hau Peter , Beckhove Philipp , Winner Beate , Riemenschneider Markus
Published: November 2023

3D (organoid) models in cancer research are considered the novel gold standard for in vitro experiments. Nevertheless, a variety of different protocols exits and the models are often used uncritically in lack of further validation. In this study, we aimed to characterize a scaffold-based (MatrigelTM) 3D glioma cell culture approach. We compared the 3D technique to conventional 2D culturing in its efficacy of generating models and in respect to its exact modeling properties. Models were generated from 62 patients with CNS tumors, most of them with glioblastomas, IDH-wildtype (GBM, n = 43). Pairs of 11 GBM models (2D and 3D) together with corresponding patient material were further characterized using bulk and single-cell RNA sequencing as well as 850k methylation analyses. Functional aspects of 3D tumor organoids were analyzed using a coculture system (Incucyte® SX5 live cell imager) with tumor infiltrating lymphocytes. Compared to conventional 2D culture the MatrigelTM-based 3D protocol was not superior in terms of the successful generation of in vitro models. However, NGS-based bulk and single-cell sequencing of matched pairs of 2D and 3D models showed an altered transcription of key immune regulatory genes in the 3D models, indicating the reoccurrence of an immunosuppressive phenotype. Changes included the presentation of different HLA surface molecules as well as cellular stressors. In the coculturing approach, lymphocyte-mediated cell killing appeared less effective in clearing 3D models than their 2D counterparts. IFN-γ release as well as live cell staining and proliferation analysis pointed towards an elevated resistance of 3D models. In conclusion, we found that the Scaffold-based (MatrigelTM) 3D culture technique depicts clinically highly important aspects of tumor-immune cell interactions, with the potential to explore immunotherapeutic approaches in an easily accessible in vitro system.
MODL-10. SEVERELY IMMUNODEFICIENT NOG-EXL MICE ALLOW FOR HUMANIZATION AND DEVELOPMENT OF A HUMAN GLIOBLASTOMA-DERIVED TUMOR MICROENVIRONMENT
Authors: Binder Zev , Lamrani Lamia , Secreto Anthony , Skuli Nicholas , Jacobson Moriah , Assenmacher Charles-Antoine , Willis Elinor , Radaelli Enrico , O'Rourke Donald M , Alanio Cecile , Nasrallah MacLean P
Published: November 2023

Cellular immunotherapy has resulted in tremendous therapeutic advances across liquid tumors. However, for solid tumors, including glioblastoma (GBM), significant limitations still exist in successfully translating immune-based adoptive cell therapeutics from the bench to the clinic. The majority of preclinical in vivo GBM models either use human cell lines grown in immunodeficient animals or syngeneic tumors grown in immunocompetent animals. Both approaches lack critical immune components of the tumor microenvironment (TME), thus inherently limiting the ability to adequately model immunotherapies. Here, we demonstrate the ability of hematopoietic stem cells (HSCs) from a GBM patient to successfully engraft in the severely immunodeficient NOG-EXL mouse strain and generate a patient-derived immune system. Intraosseous injections of a low number (10,000) of CD34+ HSCs, obtained from a bone marrow draw from the patient at the time of tumor resection, resulted in an average of 20% human CD45+ cells in peripheral blood 17 weeks post-transplant. GBM organoids, established from the same patient, were implanted intracranially at Week 18 and allowed to grow for 5 weeks without significant clinical evidence of graft-vs-host disease or macrophage activation syndrome. Immunohistochemical and flow cytometry analyses of mouse brains, with and without tumors, revealed infiltration of human-derived, CD45+/CD33+ cells, in proximity to the tumor engraftment site. These results demonstrate the ability of patient-obtained CD34+ HSCs to engraft and form a TME in severely immunodeficient mice. Subsequent work will focus on the differences between autologous and allogeneic humanized tumor-bearing mice in the context of the originating tumor. Additional future work will focus on testing cellular therapies directed towards the relevant areas of the autologous model.
MODL-11. EXPLORING THE THERAPEUTIC POTENTIAL OF YAP-TEAD INHIBITORS FOR TARGETING GBM CELL MIGRATION AT THE INFILTRATIVE MARGIN
Authors: Hurtado Thenzing Silva , Pai Balagopal , Ramos Susana Isabel , Hambardzumyan Dolores , Yong Raymund , Tsankova Nadejda
Published: November 2023

Glioblastoma (GBM) is an aggressive brain tumor with a median overall survival of only 12-15 months. Despite remarkable cellular and molecular heterogeneity, all GBM subtypes exhibit an infiltrating growth pattern which leads to incomplete surgical resection and inevitable recurrence, representing a major obstacle for effective treatment and an unmet need for therapy. Our research project aims to investigate the efficacy of YAP-TEAD inhibitors, including Verteporfin (VP), on migratory properties of GBM cells at the interface between the tumor core and the infiltrative tumor margin. Recently, several groups have implicated Hippo pathway YAP-TEAD activation in GBM growth and mesenchymal transition, and our laboratory demonstrated anti-migration and anti-invasion efficacy of the YAP-TEAD inhibitor VP across GBM subtypes, in patient-derived cell and xenograft (PDX) mouse models. Here we set to develop an ex vivo model for high-throughput testing of various YAP-TEAD inhibitors using organotypic slice cultures obtained from PDX and primary glioblastoma resection specimens, focusing on tumor areas around the infiltrative margin. Tumor cells within 250µ-thick organotypic slices were infected with GFP labeled retrovirus and subjected to confocal time-lapse imaging 72-96 hours later. Slices were treated first with vehicle (DMSO) for 4 hours, and subsequently with the YAP-TEAD inhibitor VP for 4 hours. To assess drug impact on tumor cell migration, we gathered imaging data on migration properties such as speed and directionality. Analysis of a PDX model generated from a de-novo GBM surgical resection (G-17969) revealed significantly decreased distance traveled in GFP+ cells treated with VP, compared to DMSO (p-value = 0.0002, n=50 cells). These pilot results corroborate other studies demonstrating anti-invasion properties of VP, contributing towards our goal of establishing a preclinical pipeline for rapid, high-throughput screening of anti-invasion drug inhibitors ex vivo. Key words: Glioblastoma, cell migration, YAP-TEAD inhibitors, time-lapse imaging
MODL-15. ESTABLISHMENT OF ORGANOID MODELS OF PATIENT-DERIVED GH-PRODUCING PITUITARY ADENOMA
Authors: Maeda Sachi , Ohka Fumiharu , Kanamori Fumiaki , Okumura Yuka , Takeuchi Kazuhito , Nagata Yuichi , Motomura Kazuya , Yamaguchi Junya , Kibe Yuji , Harada Hideyuki , Shimizu Hiroki , Takido Yuhei , Saito Ryuta
Published: November 2023

BACKGROUND

Growth hormone-producing pituitary adenomas (GHomas) reveal unfavorable clinical course due to systemic complications such as excessive sweating, impaired glucose tolerance, and hypertension, in addition to its characteristic facial appearance. For GHoma, pharmacological treatments including somatostatin analogs and dopamine agonists are widely performed, however, a subset of case reveals dismal prognosis. We need to elucidate molecular mechanisms contributing to GHoma growth, however, experimental models of GHoma such as cell line and animal model are quietly limited.

OBJECTIVE

In recent years, organoid models using 3D-culture technologies have gained attention as useful research models for various types of cancers. This study aims to establish a novel organoid model derived from GHoma patients. Using established GHoma organoid model, we analyzed their histological characteristics.

METHODS

Surgical specimens from five cases of GHoma were finely dissected to approximately 1mm and embedded in Matrigel basement membrane matrix. The specimens were then cultured under 37°C conditions in a 5% CO2 incubator using a medium supplemented with various growth factors. After 28 days of culture, formalin-fixed paraffin-embedded sections were prepared, and the histological similarity with patient tumors was examined.

RESULTS

Hematoxylin and eosin staining showed that organoid models consisted of acidophilic, granular cytoplasm and small round nuclei, maintaining a cell morphology similar with the original tumor. Immunohistochemical staining revealed positive expression of growth hormone, Pit-1, and CAM5.2, which are specific markers for GHoma.

CONCLUSIONS

We successfully established a patient-derived organoid model of GHoma that retains histological and endocrinological characteristics. We believe that this model could be a valuable research model for elucidating the molecular mechanisms of GHoma and developing novel treatment approaches targeting important molecular abnormalities. Additionally, in clinical settings, this model might be useful for predicting the responsiveness to drug treatment and selecting appropriate therapies for individual patients.
MODL-22. DEVELOPMENT OF A PATIENT-DERIVED MODEL BIOBANK TO PROVIDE AN ACCURATE TOOL DECIPHERING BRAIN TUMOR INITIATION, PROGRESSION, AND THERAPEUTIC RESISTANCE
Authors: Deschuyter Marlène , Wolf Thibaut , Lhermitte Benoit , Reita Damien , Fuchs Quentin , Pierrevelcin Marina , Foppolo Sophie , Dontenwill Monique , Entz-Werlé Natacha
Published: November 2023

Brain cancers are the most incident and deadliest tumor types in pediatric and young adult populations. Among them, several histotypes are characterized by their molecular drivers and co-alterations currently guiding our therapeutic strategies. Due to inter- and intra-tumoral heterogeneity, those tumors are becoming rapidly chemo- and radioresistant. To encompass all their intrinsic and extrinsic mechanisms of therapeutic resistances, there is an urgent need to generate accurate brain tumors’ modeling to predict therapeutic responses. Patient-derived cell lines (PDCL) remain for now the most reliable model to estimate and mimic evolutive situation. Main difficulties rely on the derivation methods themselves (2D vs. 3D, in vitro vs. in vivo), but also on the methods to validate their molecular similarities comparatively to the primary tumors. Here we collected, through the PEDIAMODECAN program, 104 fresh brain tumors (e.g., 23 pilocytic astrocytomas (PA), 3 low-grade gliomas (LGG), 13 supra-tentorial high-grade gliomas and 10 DIPG/DMGs, 19 ependymomas, 19 medulloblastomas, 17 other rare entities) to generate PDCL and patient-derived xenografts (PDX). Using Next-Generation Sequencing (NGS), methylome assay and RNA sequencing, we were in capacity to underline similarities with initiated samples. Looking to derivation itself, we successfully maintained 80% samples in 2D cultures and 54% were derived in PDX. As expected, the PA/LGG group shows the higher percentage of failure of PDX generation (90%) and all established PA/LGG PDCLs were limited to a low number of passages. Nevertheless, for high-grade subtypes, PDX generation rate increased to 85,7%. No correlation was observed between derivation success and time between surgical removal and dissociation process. Whereas PDX histology was reproducing the same microscopic features than the primary tumors, methylome assay proves to be a good method to confirm original histotype group. Furthermore, those primary models afford a large biobank reproducing the diversity of pediatric brain tumors and were easily drug screened.
MODL-24. NEUROIMMUNE-COMPETENT HUMAN BRAIN ORGANOID MODEL FOR STUDYING BRAIN TUMOR INVASION
Authors: Sarnow Katharina , Majercak Emma , Haque Ishraq , Tang Xin
Published: November 2023

INTRODUCTION

Diffuse intrinsic pontine glioma (DIPG) is a highly aggressive pediatric brain tumor with a very poor prognosis (< 12 months survival). There is currently no effective treatment for DIPG. Complete surgical resection of DIPG is difficult due to its skull base location and infiltrative growth pattern. Gaining insights into the invasion process of the DIPG cells and their interaction with brain cells in the tumor microenvironment, such as microglia - the brain's primary immune cells, may guide the development of effective treatment strategies to halt or even reverse disease progression. To build a clinically relevant model for studying the microglia/tumor interaction in a 3D human tissue context, we developed a neuroimmune-competent brain organoid/DIPG fusion model system.

METHODS

Human embryonic stem cell derived-neural progenitor cells (NPCs) and -GFP-labelled human myeloid precursor cells were combined to form microglia-containing organoids (MiCBOs) through self-assembly and co-development over two months. Using two-photon live imaging, we characterized the distribution and motility of microglia in MiCBOs and corroborated these results with immunofluorescence staining and single-cell RNA-Sequencing (scRNA-Seq). Tumor cell invasion and interaction with microglia in the brain organoids were assessed by confocal microscopy as well as immunofluorescence stainings.

RESULTS

The human microglia in MiCBOs are viable, widely distributed, actively motile, and highly resemble those in the human brain. Confrontation experiments of the MiCBO with DIPG spheroids enable quantitative characterization of the invasion patterns for different cell lines under various conditions.

CONCLUSION

Our MiCBOs represent a valuable model for studying tumor cell invasion within a neuroimmune-competent human tissue context. Ongoing experiments with different primary DIPG cell lines as well as genetically engineered microglia will deliver more detailed insights into the invasion patterns of tumor cells into brain tissue and shed light on tumor cell/microglia interplay during the DIPG invasion process.
MODL-28. LIVING EX VIVO ORGANOTYPIC BRAIN SLICE CULTURES AS A COMPREHENSIVE SUBSTRATE FOR TUMOR ENGRAFTMENT AND ANALYSIS
Authors: Adefolaju Adebimpe , Mann Breanna , Zhang Xiaopei , Bell Noah , Thang Morrent , Dasari Rajaneekar , Kanchi Krishna , Valdivia Alain , Yang Yang , Buckley Andrew , Lettry Vivien , Quinsey Carolyn , Rauf Yasmeen , Kram David , Cassidy Noah , Vaziri Cyrus , Corcoran David L , Rego Stephen , Jiang Yuchao , Graves Lee M , Dunn Denise , Floyd Scott , Baldwin Albert , Hingtgen Shawn , Satterlee Andrew B
Published: November 2023

Heterogeneity, drug resistance, tumor infiltration, and invasiveness significantly contribute to the poor prognosis often associated with brain cancers. To enhance drug development, it is crucial to have models that accurately replicate human responses. Standard in vitro methods to culture, treat and analyze brain tumors often lack resemblance to the original tumor and provide limited insight. To address these challenges, we developed an organotypic brain slice culture (OBSC) model that recapitulates in vivo tumor characteristics within a functional tumor microenvironment (TME) and provides rapid results. OBSCs effectively measure engrafted tumor growth, morphology, and invasion rates. Furthermore, engrafted tumors can undergo relevant therapeutic treatments and be analyzed for drug outcomes. In a 4-day assay, we successfully engrafted a diverse range of brain cancer cell lines, including low-passage lines that struggle to establish in vivo. These cell lines maintain tumor-specific invasion patterns, interact with the OBSC microenvironment, and display killing patterns consistent with in vivo tumor responses. Notably, invasive, metastatic, and diffuse tumor cell lines migrate outward on OBSCs, while densely growing lines contract inward. To further validate OBSCs as a model for the TME, we treated MB231BR, LN229, and U373 tumor lines with TR107, an experimental ClpP inhibitor derived from ONC201. This therapeutic family maximizes tumor elimination by targeting normal cells within the TME. In vitro experiments showed TR107 achieving incomplete tumor kill on cell lines grown in vitro, whereas treatment on OBSCs led to complete tumor kill, suggesting OBSCs' contribution to TME-based tumor elimination. Conclusively, OBSCs provide a valuable platform for studying brain cancers and accurately replicating tumor behavior and responses to therapies within the TME. This model has the potential to advance drug development and deepen our understanding of brain cancer biology, ultimately improving prognoses and treatment outcomes for patients.
MODL-39. ELUCIDATING THE TRANSCRIPTOMIC LANDSCAPE OF METASTATIC PEDIATRIC MEDULLOBLASTOMA
Authors: Orozco Ana Isabel Castillo , Aghababazadeh Masoumeh , Khatami Marjan , Danieau Geoffroy , Jawhar Wajih , Khazaei Niusha , Garzia Livia
Published: November 2023

Medulloblastoma (MB) is a highly aggressive and the most common brain tumor in childhood. MB presents a high intertumoral heterogeneity, with at least four molecular subgroups (SHH, WNT, Group 3, and Group 4) identified. MB can metastasize to the leptomeningeal space, known as Leptomeningeal Disease (LMD) and its presence is a universal predictor of poor outcome among MB patients. Metastatic MB is predominantly found in the MB Group 3 type. Although LMD represents a main clinical challenge, its molecular mechanisms remain poorly characterized. Recent research has shown that primary and MB metastasis diverge dramatically. Our work has focused on establishing therapy naïve Group 3 Patient-Derived Xenografts that faithfully replicate the nature of primary and metastatic Medulloblastoma with the aim to perform comparative genomic and transcriptomic analyses between these models to identify genetic drivers/pathways that sustain metastatic MB or leptomeningeal disease. Our results show various signaling pathways enriched across LMD models, such as protein secretion and oxidative phosphorylation. We also have identified differentially expressed genes, where sets of genes have shown to be present in more than one PDX model, such as members of the Solute Carriers family (SLC44A3 and SLC17A9) and FCGBP. Retrieval of short variants from RNAseq data has not revealed thus far mutations enriched in LMD, suggesting these changes could be attributed to epigenetic disruption rather than genetic changes. In conclusion, our results support the notion that primary and LMD are transcriptionally different, with various enriched pathways and sets of DEG among LMD Group 3 PDX models. These findings are in progress for functional validation. Work is currently being done to profile the epigenome of LMD by correlating transcriptomic data with active chromatin markers such as H3K27ac and H3K4me1. Through these approaches, we aim to elucidate the genetic dependencies of metastatic Medulloblastoma that will help for targeted therapies.
Trifluridine/Tipiracil Plus Bevacizumab for Vulnerable Patients With Pretreated Metastatic Colorectal Cancer: A Retrospective Study (WJOG14520G)
Authors: Kito Yosuke , Kawakami Hisato , Mitani Seiichiro , Nishina Shinichi , Matsumoto Toshihiko , Tsuzuki Takao , Shinohara Yudai , Shimokawa Hozumi , Kumanishi Ryosuke , Ohta Takashi , Katsuya Hiroo , Kawakami Takeshi , Nishina Tomohiro , Hasegawa Hiroko , Akiyoshi Kohei , Chiba Yasutaka , Yamazaki Kentaro , Hironaka Shuichi , Muro Kei
Published: November 2023

Background

Trifluridine/tipiracil (FTD/TPI) plus bevacizumab has shown clinical benefit for metastatic colorectal cancer (mCRC) refractory to standard therapy. However, few data have been available for patients with pretreated mCRC who are intolerant of intensive therapy (vulnerable).

Methods

We performed a multicenter retrospective study (WJOG14520G; TWILIGHT) of FTD/TPI plus bevacizumab for vulnerable patients with pretreated mCRC. Eligibility criteria included previous chemotherapy (although patients treated with all key cytotoxic agents, a fluoropyrimidine, oxaliplatin, and irinotecan, were excluded) and intolerance of full-dose combination therapy with oxaliplatin or irinotecan at the start of FTD/TPI plus bevacizumab.

Results

The median age of 93 evaluable patients was 79 years (range, 21-90). Intolerance of intensive therapy was attributable to an older age in 60 (65%) patients, serious concomitant disease in 24 (26%) patients, and a poor performance status in 19 (20%) patients. FTD/TPI plus bevacizumab was administered as second-line treatment in 74 (80%) patients and as third- or fourth-line treatment in 19 (20%) patients. The objective response rate was 4.9% (95% confidence interval [CI], 1.4%-12.2%), and the disease control rate was 67.9% (95% CI, 56.6%-77.8%). With a median follow-up time of 21.6 months, median overall survival and progression-free survival were 18.6 months (95% CI, 12.1-23.2) and 6.3 months (95% CI, 5.0-8.3), respectively. Neutropenia of grade ≥3 developed in 50 (54%) patients, whereas 2 (2%) patients experienced febrile neutropenia, and no treatment-related death was observed.

Conclusion

Our data show the potential efficacy and acceptable safety profile of FTD/TPI plus bevacizumab for vulnerable patients with pretreated mCRC.
early maturity 7 promotes early flowering by controlling the light input into the circadian clock in barley
Authors: Helmsorig Gesa , Walla Agatha , Rütjes Thea , Buchmann Gabriele , Schüller Rebekka , Hensel Götz , von Korff Maria
Published: November 2023

Breeding for variation in photoperiod response is crucial to adapt crop plants to various environments. Plants measure changes in day length by the circadian clock, an endogenous timekeeper that allows plants to anticipate changes in diurnal and seasonal light–dark cycles. Here, we describe the early maturity 7 (eam7) locus in barley (Hordeum vulgare), which interacts with PHOTOPERIOD 1 (Ppd-H1) to cause early flowering under non-inductive short days. We identify LIGHT-REGULATED WD 1 (LWD1) as a putative candidate to underlie the eam7 locus in barley as supported by genetic mapping and CRISPR-Cas9-generated lwd1 mutants. Mutations in eam7 cause a significant phase advance and a misregulation of core clock and clock output genes under diurnal conditions. Early flowering was linked to an upregulation of Ppd-H1 during the night and consequent induction of the florigen FLOWERING LOCUS T1 under short days. We propose that EAM7 controls photoperiodic flowering in barley by controlling the light input into the clock and diurnal expression patterns of the major photoperiod response gene Ppd-H1.
Constraining the cosmological model of modified f(Q) gravity: Phantom dark energy and observational insights
Authors: Koussour M , Myrzakulov N , Alfedeel Alnadhief H A , Abebe Amare
Published: November 2023

Despite the significant accomplishments of general relativity, numerous unresolved issues persist in our understanding of the cosmos. One of the most perplexing challenges is the ongoing accelerated expansion of the Universe, which continues to elude a complete explanation. Consequently, scientists have proposed various alternative theories to general relativity in pursuit of a deeper understanding. In our analysis, we delve into the recently proposed modified f(Q) gravity, where Q represents the nonmetricity scalar responsible for gravitational effects. Specifically, we investigate a cosmological model characterized by the functional form f(Q) = Q + αQn, where α (with α ≠ 0) and n serve as free parameters. Utilizing this functional form, we construct our Hubble rate, incorporating a specific equation of state to describe the cosmic fluid. Furthermore, we leverage a dataset consisting of 31 data points from Hubble measurements and an additional 1048 data points from the Pantheon dataset. These data serve as crucial constraints for our model parameters, and we employ the Markov Chain Monte Carlo (MCMC) method to explore the parameter space and derive meaningful results. With our parameter values constrained, our analysis yields several noteworthy findings. The deceleration parameter suggests a recent accelerated phase in the cosmic expansion. In addition, the EoS parameter paints a portrait of dark energy exhibiting phantom-like characteristics. Furthermore, we delve into the application of cosmological diagnostic tools, specifically the statefinder and the Om(z) diagnostics. Both of these tools align with our previous conclusions, confirming the phantom-like behavior exhibited by our cosmological model. These results collectively contribute to our understanding of the dynamic interplay between gravity, dark energy, and the expanding cosmos.
Association of the triglyceride-glucose index variability with blood pressure and hypertension: a cohort study
Authors: Wang D , Li W , Zhou M , Ma J , Guo Y , Yuan J , He M , Zhang X , Chen W
Published: November 2023

Background

Several studies have indicated that the triglyceride-glucose index (TyG) index is associated with hypertension; however, evidence on the association of change in the TyG index with blood pressure and hypertension is limited.

Aims

To assess the association of the TyG index with blood pressure and hypertension.

Design

A cohort study.

Methods

We included 17 977 individuals with a mean age of 60.5 years from the Dongfeng–Tongji cohort. The TyG index was calculated as ln [fasting triglyceride (mg/dl)×fasting glucose (mg/dl)/2]. Hypertension was defined as blood pressure ≥140/90 mmHg, self-reported current use of antihypertensive medication or self-reported physician diagnosis of hypertension.

Results

In the longitudinal analyses, we found a linear dose–response relationship between changes in the TyG index and change in blood pressure. Each one-unit change in the TyG index was associated with a 1.93 (1.23–2.63) mmHg increase in systolic blood pressure (SBP) and a 1.78 (1.42–2.16) mmHg increase in diastolic blood pressure (DBP). During a median follow-up of 9.37 years, a total of 3594 individuals were newly diagnosed with hypertension. We also found a linear dose–response relationship between the TyG index and the incidence of hypertension. The hazard ratio (HR) of hypertension for each one-unit increase in the TyG index was 1.21 (1.13–1.29). In addition, the best cut-off point of TyG for predicting hypertension was 8.4797, with sensitivity, and specificity of 57.85% and 55.40%, respectively.

Conclusions

The TyG index had a positive dose–response relationship with blood pressure and could be used to predict the risk of hypertension.
Survey on reference dosimetry practice in Hokuriku region, Japan
Authors: Kinoshita Naoki , Takemura Akihiro , Oosaka Akitane , Shirosaki Nobuyuki , Tsujikawa Tetsuya
Published: November 2023

This study aimed to obtain reference dosimetry practices for flattened photon and electron beams. A questionnaire survey on these practices was sent to 37 facilities that performed radiotherapy using these beams in Fukui, Ishikawa, Niigata and Toyama, Japan. The survey comprised equipment (dosemeters, water phantoms, thermometers, barometers and hygrometers) and procedures for reference dosimetry (water used as the phantoms, verification of chamber placement, periodic checks of ion recombination- and polarity-corrections, pre-irradiation and relative humidity monitoring). Responses were received from 19 institutions. In the 19 institutions, a reference-class dosemeter was used for the reference dosimetry. In 6 of the 19 institutions, a 10-year-old or older dosemeter was used. Any barometers, thermometers and hygrometers were not recalibrated. Dosimetry equipment necessitates regular maintenance. Moreover, it is necessary to have a backup dosimetry system in clinics to account for potential malfunctions or instances when the primary system is sent for calibration.
Corollary Discharge Dysfunction as a Possible Substrate of Anomalous Self-experiences in Schizophrenia
Authors: Beño-Ruiz-de-la-Sierra Rosa M , Arjona-Valladares Antonio , Hernández-García Marta , Fernández-Linsenbarth Inés , Díez Álvaro , Fondevila Estevez Sabela , Castaño Carolina , Muñoz Francisco , Sanz-Fuentenebro Javier , Roig-Herrero Alejandro , Molina Vicente
Published: November 2023

Background and Hypothesis

Corollary discharge mechanism suppresses the conscious auditory sensory perception of self-generated speech and attenuates electrophysiological markers such as the auditory N1 Event-Related Potential (ERP) during Electroencephalographic (EEG) recordings. This phenomenon contributes to self-identification and seems to be altered in people with schizophrenia. Therefore, its alteration could be related to the anomalous self-experiences (ASEs) frequently found in these patients.

Study Design

To analyze corollary discharge dysfunction as a possible substrate of ASEs, we recorded EEG ERP from 43 participants with schizophrenia and 43 healthy controls and scored ASEs with the ‘Inventory of Psychotic-Like Anomalous Self-Experiences’ (IPASE). Positive and negative symptoms were also scored with the ‘Positive and Negative Syndrome Scale for Schizophrenia’ (PANSS) and with the ‘Brief Negative Symptom Scale’ (BNSS) respectively. The N1 components were elicited by two task conditions: (1) concurrent listening to self-pronounced vowels (talk condition) and (2) subsequent non-concurrent listening to the same previously self-uttered vowels (listen condition).

Study Results

The amplitude of the N1 component elicited by the talk condition was lower compared to the listen condition in people with schizophrenia and healthy controls. However, the difference in N1 amplitude between both conditions was significantly higher in controls than in schizophrenia patients. The values of these differences in patients correlated significantly and negatively with the IPASE, PANSS, and BNSS scores.

Conclusions

These results corroborate previous data relating auditory N1 ERP amplitude with altered corollary discharge mechanisms in schizophrenia and support corollary discharge dysfunction as a possible underpinning of ASEs in this illness.
Linking brain structure, cognition, and sleep: insights from clinical data
Authors: Wei Ruoqi , Ganglberger Wolfgang , Sun Haoqi , Hadar Peter  N , Gollub Randy  L , Pieper Steve , Billot Benjamin , Au Rhoda , Eugenio Iglesias Juan , Cash Sydney S , Kim Soriul , Shin Chol , Westover M Brandon , Joseph Thomas Robert
Published: November 2023

Study Objectives

To use relatively noisy routinely collected clinical data (brain magnetic resonance imaging (MRI) data, clinical polysomnography (PSG) recordings, and neuropsychological testing), to investigate hypothesis-driven and data-driven relationships between brain physiology, structure, and cognition.

Methods

We analyzed data from patients with clinical PSG, brain MRI, and neuropsychological evaluations. SynthSeg, a neural network-based tool, provided high-quality segmentations despite noise. A priori hypotheses explored associations between brain function (measured by PSG) and brain structure (measured by MRI). Associations with cognitive scores and dementia status were studied. An exploratory data-driven approach investigated age-structure-physiology-cognition links.

Results

Six hundred and twenty-three patients with sleep PSG and brain MRI data were included in this study; 160 with cognitive evaluations. Three hundred and forty-two participants (55%) were female, and age interquartile range was 52 to 69 years. Thirty-six individuals were diagnosed with dementia, 71 with mild cognitive impairment, and 326 with major depression. One hundred and fifteen individuals were evaluated for insomnia and 138 participants had an apnea–hypopnea index equal to or greater than 15. Total PSG delta power correlated positively with frontal lobe/thalamic volumes, and sleep spindle density with thalamic volume. rapid eye movement (REM) duration and amygdala volume were positively associated with cognition. Patients with dementia showed significant differences in five brain structure volumes. REM duration, spindle, and slow-oscillation features had strong associations with cognition and brain structure volumes. PSG and MRI features in combination predicted chronological age (R2 = 0.67) and cognition (R2 = 0.40).

Conclusions

Routine clinical data holds extended value in understanding and even clinically using brain-sleep-cognition relationships.
Between Peripherality and Privilege: “Women Wage Peace” as a Case Study of Intersectionality Practices in Women’s Movements
Authors: Lion Veronica
Published: November 2023

This article analyzes Women Wage Peace (WWP), an Israeli grassroots peace movement, as a case study of intersectionality in women’s movements. Using an ethnographic model based on semi-structured interviews with previous and current movement members, I investigate the dilemmas and strategies of WWP in its pursuit of a diverse membership base, a goal considered unique in Israeli peace discourse. This study contributes to understanding intersectional theory and practice, explaining why such an approach is not (always) effective in women’s movements despite the best intentions. Examining WWP through the lens of peripherality and privilege highlights the movement’s hierarchical power structure, identifying the places where intersectional awareness does not translate into political practice. By providing examples of the challenges of intersectionality in women’s movements, these findings offer a response to the need for more nuanced analysis in current feminist and social movement research.
Activation of moderate autophagy promotes survival of fat graft
Authors: Pei Su‐Jun , Zhu Yuan‐Zheng , Yang Juan‐Min , Zhang Min‐Chen , Shi Chen‐Long , Ding Ying , Yi Yang‐Yan
Published: November 2023

Clinically unpredictable retention following fat grafting remains outstanding problems because of the unrevealed mechanism of grafted fat survival. The role of autophagy, a process to maintain cellular homeostasis through recycling cellular debris, has yet been to be reported in fat grafting. This study aims to improve the survival of fat grafting through the autophagy. First, the relationship between cell death and autophagy in the early stage of fat grafting was evaluated through immunostaining, RNA sequencing, and western blot. Next, rapamycin, an autophagic agonist, was used for the culturing of adipose‐derived stem cells and adipocytes during ischemia. Cell death, autophagy, and reactive oxygen species (ROS) were assayed. Finally, rapamycin was used to assist fat grafting in nude mice. The results demonstrated that the peak of cell death at the early stage of fat grafting was accompanied by a decrease in autophagy. In vitro, during ischemia, 25 nM was confirmed as the optimal dose of rapamycin that reduces cell death with enhanced autophagy and mitophagy, improved mitochondrial quality as well as decreased ROS accumulation. In vivo, promoted mitophagy, alleviated oxidative stress, and decreased cell apoptosis of rapamycin‐treated fat grafts were observed in the early stage. In addition, rapamycin increased the survival of fat grafts with increased neovascularization and reduced fibrosis. We suggested that moderate autophagy induced by rapamycin contribute to enhanced ischemic tolerance and long term survival of fat grafts through mitochondrial quality control.
Pediatric Highlights From the Extracorporeal Life Support Organization Registry: 2017–2022
Authors: O’Neil Erika R. , Guner Yigit , Anders Marc M. , Priest John , Friedman Matthew L. , Raman Lakshmi , Di Nardo Matteo , Alexander Peta , Tonna Joseph E. , Rycus Peter , Thiagarajan Ravi R. , Barbaro Ryan , Sandhu Hitesh S.
Published: November 2023

The Extracorporeal Life Support Organization (ELSO) registry which collects data from hundreds of participating centers supports research in ECMO to help improve patient outcomes. The ELSO Scientific Oversight Committee, an international and diverse group of ECMOlogists (https://www.elso.org/registry/socmembers.aspx), selected the most impactful and innovative research articles on pediatric ECMO emerging from ELSO data. Here they present brief highlights of these publications.
Weathering the Storm: Syringe Services Program Laws and Human Immunodeficiency Virus During the COVID-19 Pandemic
Authors: Jackson Hannah , Dunphy Christopher , Grist Mary Blain , Jiang Xinyi , Xu Likang , Guy Gery P. , Salvant-Valentine Sheila
Published: November 2023

Background:

Syringe services programs (SSPs) are community-based prevention programs that provide a range of harm reduction services to persons who inject drugs. Despite their benefits, SSP laws vary across the United States. Little is known regarding how legislation surrounding SSPs may have influenced HIV transmission over the COVID-19 pandemic, a period in which drug use increased. This study examined associations between state SSP laws and HIV transmission among the Medicaid population before and after the COVID-19 pandemic.

Methods:

State-by-month counts of new HIV diagnoses among the Medicaid population were produced using administrative claims data from the Transformed Medicaid Statistical Information System from 2019 to 2020. Data on SSP laws were collected from the Prescription Drug Abuse Policy System. Associations between state SSP laws and HIV transmission before and after the start of the COVID-19 pandemic were evaluated using an event study design, controlling for the implementation of COVID-19 nonpharmaceutical interventions and state and time fixed effects.

Results:

State laws allowing the operation of SSPs were associated with 0.54 (P = 0.044) to 1.18 (P = 0.001) fewer new monthly HIV diagnoses per 100,000 Medicaid enrollees relative to states without such laws in place during the 9 months after the start of the COVID-19 pandemic. The largest effects manifested for population subgroups disproportionately affected by HIV, such as male and non-Hispanic Black Medicaid enrollees.

Conclusion:

Less restrictive laws on SSPs may have helped mitigate HIV transmission among the Medicaid population throughout the COVID-19 pandemic. Policymakers can consider implementing less restrictive SSP laws to mitigate HIV transmission resulting from future increases in injection drug use.

Disclaimer:

The findings and conclusions in this report are those of the author(s) and do not necessarily represent the official position of the Centers for Disease Control and Prevention.