bluuebunny/crossref_metadata_2025 huggingface dataset:
Naturally occurring androgen excess cows are present in dairy and beef herds and have similar characteristics to women with PCOS
Authors: Abedal-Majed Mohamed A , Springman Shelby A , Jafar Hanan D , Bell Brooke E , Kurz Scott G , Wilson Kyle E , Cupp Andrea S
Published: June 2022

Beef cows with excess androstenedione (A4; High A4) in follicular fluid (FF) and secreted by the ovarian cortex have been reported from the University of Nebraska–Lincoln physiology herd displaying characteristics reminiscent of polycystic ovary syndrome (PCOS). Thus, we hypothesized that naturally occurring High A4 cows were present in other dairy and beef herds. Fourteen Jordan (Amman, Jordon) dairy heifers and 16 U.S. Meat Animal Research Center beef heifers were classified by FF (High A4: A4 > 40 ng/mL and Control: A4 < 20 ng/mL) and/or cortex culture media (High A4 > 1 ng/mL/d or Control < 1 ng/mL/d). High A4 dairy heifers (n = 6) had greater A4 concentrations (7.6-fold) in FF and (98-fold) greater in ovarian cortex culture media with greater numbers of primordial and fewer later-stage follicles than Controls (n = 8) even after 7 d of culture. Also, the ovarian cortex had greater staining for Picro Sirius red in High A4 dairy heifers compared with Controls indicating increased fibrosis. Thecal cells from High A4 dairy heifers had greater STAR, LHCGR, CYP17A, CD68, and PECAM mRNA expression with increased mRNA abundance of CYP17A1 and CD68 in the ovarian cortex cultures compared with Control dairy heifers. Similarly, cortex culture media from High A4 beef heifers (n = 10) had increased A4 (290-fold; P ≤ 0.001), testosterone (1,427-fold; P ≤ 0.001), and progesterone (9-fold; P ≤ 0.01) compared with Control heifers with increased primordial follicles and decreased later-stage follicles even after 7 d of culture, indicating abnormal follicular development. High A4 ovarian cortex cultures from beef heifers also had increased fibrosis markers and greater expression of PECAM (P = 0.01) with a tendency for increased vascular endothelial cadherin compared with Controls (n = 6). These two trials support our hypothesis that naturally occurring androgen excess cows are present in other dairy and beef herds. The ability to identify these females that have excess A4 ovarian microenvironments may allow for their use in understanding factors causing abnormal follicle development linked to androgen excess and inflammation.
Maternal nutrient restriction and over-feeding during gestation alter expression of key factors involved in placental development and vascularization
Authors: Reed Sarah A , Ashley Ryan , Silver Gail , Splaine Caitlyn , Jones Amanda K , Pillai Sambhu M , Peterson Maria L , Zinn Steven A , Govoni Kristen E
Published: June 2022

Poor maternal nutrition can negatively affect fetal and placental growth and development. However, the mechanism(s) that contribute to altered placenta growth and function are not well understood. We hypothesized that poor maternal diet would impact signaling through the C-X-C motif chemokine ligand (CXCL) 12-CXCL4 axis and/or placental expression of the insulin-like growth factor (IGF) axis. Using our established sheep model of poor maternal nutrition, we examined the effects of restricted- and over-feeding on ewe placentome gene and protein expression. Specifically, ewes were fed a control (CON; 100%), restricted (RES; 60%), or over (OVER; 140%) diet beginning at day 30.2 ± 0.02 of gestation, and samples were collected at days 45, 90, and 135 of gestation, representing periods of active placentation, peak placental growth, and near term, respectively. Placentomes were separated into cotyledon and caruncle, and samples snap frozen. Protein was determined by western blot and mRNA expression by real-time PCR. Data were analyzed by ANOVA and significance determined at P ≤ 0.05. Ewes fed a RES diet had decreased CXCL12 and vascular endothelial growth factor (VEGF), and increased tumor necrosis factor (TNF)α protein compared with CON ewes in caruncle at day 45 (P ≤0.05). In day 45 cotyledon, CXCR7 protein was increased and mTOR was decreased in RES relative to CON (P ≤0.05). At day 90, CXCR4 and CXCR7 were reduced in RES caruncle compared with CON, whereas VEGF was reduced and mTOR increased in cotyledon of RES ewes relative to CON (P ≤0.05). In OVER caruncle, at day 45 CXCR4 and VEGF were reduced and at day 90 CXCR4, CXCR7, and TNFα were reduced in caruncle compared with CON (P ≤0.05). There was no observed effect of OVER diet on protein abundance in the cotyledon (P > 0.05). Expression of IGF-II mRNA was increased in OVER at day 45 and IGFBP-3 was reduced in RES at day 90 in caruncle relative to CON (P ≤0.05). Maternal diet did not alter placentome diameter or weight (P > 0.05). These findings suggest that restricted- and over-feeding negatively impact protein and mRNA expression of key chemokines and growth factors implicated in proper placenta development and function.
Constant light exposure in early life induces m6A-mediated inhibition of IGF gene family in the chicken
Authors: Yang Yang , Xu Peirong , Liu Jie , Zhao Mindie , Cong Wei , Han Wanwan , Wang Deyun , Zhao Ruqian
Published: June 2022

Insulin-like growth factor (IGF) family plays important roles in regulating the development of various organ systems through stimulating cell proliferation and differentiation. Photoperiod is an important factor affecting growth and development in the chicken, yet the effect of constant light exposure in early life on IGF1 and IGF2 expression in the chicken remains unclear. In this study, one-day-old chickens were kept in either constant light (24L:0D, LL) or natural photoperiod (12L:12D, LD) for the first week of life and then maintained in constant light from 8 to 21 d of age. Constant light exposure in early life reduced mRNA expression of IGF gene family, including mRNA expression of IGF1, IGF2, and IGF2 binding proteins, in the hippocampus, hypothalamus, and liver of chickens at both 7 and 21 d of age. Moreover, constant light exposure increased mRNA expression of genes involved in RNA methylation N6-methyladenosine (m6A) in a tissue-specific manner. Interestingly, higher m6A on 3ʹUTR of IGF2 mRNA coincides with lower IGF2 mRNA, indicating a possible role of m6A in the post-transcriptional regulation of IGF2 expression in the hippocampus, hypothalamus, and liver of chickens. These findings suggest a m6A-mediated gene regulation of IGF gene family in different organs of chicken and expand our knowledge on mechanism of gene regulation in response to early life experience.
Dose-response relationship between dietary choline and serum lipid profile, energy expenditure, and respiratory quotient in overweight adult cats fed at maintenance energy requirements
Authors: Rankovic Alexandra , Godfrey Hannah , Grant Caitlin E , Shoveller Anna K , Bakovic Marica , Kirby Gordon , Verbrugghe Adronie
Published: June 2022

Choline is an essential nutrient linked to hepatic lipid metabolism in many animal species, including cats. The current study investigated the serum lipid profiles, serum liver enzymes, respiratory quotients, and energy expenditures of overweight cats fed maintenance diets, in response to graded doses of supplemental dietary choline. Overweight (body condition score [BCS]: ≥6/9) adult male neutered cats (n = 14) were supplemented with five choline chloride doses for 3-wk periods, in a 5 × 5 Latin square design. Doses were based on individual body weight (BW) and the daily recommended allowance (RA) for choline (63 mg/kg BW0.67) according to the National Research Council. Doses were control (no additional choline: 1.2 × RA, 77 mg/kg BW0.67), 2 × RA (126 mg/kg BW0.67), 4 × RA (252 mg/kg BW0.67), 6 × RA (378 mg/kg BW0.67), and 8 × RA (504 mg/kg BW0.67). Choline was top-dressed over the commercial extruded cat food (3,620 mg choline/kg diet), fed once a day at maintenance energy requirements (130 kcal/kgBW0.4). Body weight and BCS were assessed weekly. Fasted blood samples were taken and indirect calorimetry was performed at the end of each 3-wk period. Serum was analyzed for cholesterol, high-density lipoprotein cholesterol (HDL-C), triglycerides, non-esterified fatty acids, glucose, creatinine, blood urea nitrogen (BUN), alkaline phosphatase (ALP), and alanine aminotransferase. Very low-density lipoprotein cholesterol (VLDL) and low-density lipoprotein cholesterol were calculated. Data were analyzed via SAS using proc GLIMMIX, with group and period as the random effects, and treatment as the fixed effect. Statistical significance was considered at P < 0.05. Body weight and BCS did not change (P > 0.05). Serum cholesterol, HDL-C, triglycerides, and VLDL increased with 6 × RA (P < 0.05). Serum ALP decreased with 8 × RA (P = 0.004). Choline at 4 × and 6 × RA decreased serum BUN (P = 0.006). Fed or fasted respiratory quotient and energy expenditure did not differ among dietary choline doses (P > 0.05). These results suggest that dietary choline supplementation at 6 × RA may increase hepatic fat mobilization through increased lipoprotein transport and beneficially support hepatic health in overweight cats. Future studies that combine these results with existing knowledge of feline weight loss and hepatic lipidosis are warranted.
Association of a home-based nursing service during chemotherapy with the transfer to home care immediately after the last chemotherapy treatment in patients with advanced cancer
Authors: Higashibata Takahiro , Iwata Naoko , Kazama Ikuko , Irie Yoshiko , Yokosuka Kyoko , Okita Junya , Hamano Jun , Nagaoka Hiroka
Published: June 2022

Background

The appropriate timing of introducing the home-based nursing service to patients with advanced cancer has not been clarified. This study conducted a retrospective cohort study to investigate the associations of the early utilization of the home-based nursing service during chemotherapy with the place of care immediately after the last chemotherapy treatment in patients with advanced cancer.

Methods

Among the patients referred to the palliative care team of the University of Tsukuba Hospital between January 2018 and December 2019, patients with advanced cancer undergoing chemotherapy or before the start of chemotherapy at referral were included. The study patients who utilized the home-based nursing service during chemotherapy were classified into the early utilization group. The primary endpoint was the place of care immediately after the last chemotherapy treatment.

Results

Of the 1154 patients referred to the palliative care team during the study period, 261 were eligible for this study. Of these patients, 15.3% of patients (n = 40) received the home-based nursing service during chemotherapy. The proportion of patients transferred to home care after the last chemotherapy treatment was 17.5% in the early utilization group and 7.2% in the control group. The multivariate analysis revealed that the early utilization of the home-based nursing service during chemotherapy was significantly associated with the transfer to home care (odds ratio = 3.077; 95% confidence interval, 1.113–8.502).

Conclusion

The early utilization of the home-based nursing service during chemotherapy might be associated with the transfer to home care immediately after the last chemotherapy treatment.
Development and Intrapuparial Characterization of Hydrotaea aenescens (Diptera: Muscidae) Raised at Different Temperatures Under Laboratory Conditions
Authors: Duarte Marina Lopes , Queiroz Margareth Maria de Carvalho
Published: June 2022

Species belonging to the Muscidae family are important in the medical-sanitary and forensic fields due to their biology. The intrapuparial stage of Hydrotaea aenescens (Wiedemann, 1830), which represents about 50% of the immature development cycle in muscoid Diptera, has not yet been studied in detail. This study identifies and describes the main temporal morphological changes that occur in the pupal body of H. aenescens during its development at 22, 27, and 32 ± 1°C, RH 60 ± 10% with a 12-hr photoperiod. Ten pupae were collected, sacrificed, and fixed every 3 hr after pupariation up to 24 hr, and every 6 hr thereafter until the first emergence. Emergence took place at 282, 174, and 126 hr at 22, 27, and 32°C, with 520, 340, and 260 pupae, respectively. External morphology was described and recorded at eight development stages: pupariation, pre-pupae, early and late cryptocephalic pupae, phanerocephalic pupae, pupae-adult apolysis, pharate adults, and imago, employing 18 key morphological characters for pupae age. These morphological pupae characters, comprise, therefore an alternative and/or additional method to aid in determining the minimum Postmortem Interval (minPMI).
Lumbar discal cyst and post-operative discal pseudocyst: a case series
Authors: Jadhav Neha , Sivakumar Lawrence , Talibi Sayed Samed , Momoh Pearl , Rasul Fahid , Hussain Rahim , Shad Amjad
Published: June 2022

Four cases of lumbar cyst (2 discal cysts and 2 post-operative discal pseudocysts) who presented predominantly with features of sciatica without any motor, sensory or sphincteric disturbances. The patients were treated conservatively, and the management was aimed to avert any untoward surgical intervention taking into consideration patient safety and care. Two had previous lumbar decompressive discectomy. During the mean follow-up period of 13 months, there was progressive recovery of symptoms in all our 4 patients. All our patients were successfully managed by conservative approach. An intervertebral disc cyst should be considered in young patients in the differential diagnosis of any extradural intraspinal mass ventral to the thecal sac, notwithstanding its rarity. Alongside, conservative management can be offered as first line of management with appropriate patient selection that is absence of any motor/sensory/sphincteric disturbances. Facetal micro-instability could be one of the aetiologies of this pathology which necessitates further study.
Killing the Letter: Alternate Literacies and Orthographic Distortions in Jude the Obscure
Authors: Creechan Louise
Published: June 2022

When Jude the Obscure (1895) was published as a single volume novel, Hardy added the biblical epithet ‘the letter killeth’ to the title page. In Jude and across his works, Hardy revels in moments in which literacy seems to undo itself. This article traces Hardy’s attempts to ‘kill the letter’ through non-standard engagements with orthography as part of a larger proto-modernist approach that destabilizes the fixity of meaning. There are several concerns linked to the growing primacy of literacy that appear time and again in Hardy’s novels, specifically: the alternative literacies of the lesser educated, semiotic multiplicities, and the transformative potential of spelling mistakes. I suggest that Hardy’s treatment of these themes demonstrates a sustained effort to ‘kill the letter’ and challenge the assumption of progress made by the various educational reforms that had taken place in the latter half of the nineteenth century.
Using all Gene Families Vastly Expands Data Available for Phylogenomic Inference
Authors: Smith Megan L. , Vanderpool Dan , Hahn Matthew W.
Published: June 2022

Traditionally, single-copy orthologs have been the gold standard in phylogenomics. Most phylogenomic studies identify putative single-copy orthologs using clustering approaches and retain families with a single sequence per species. This limits the amount of data available by excluding larger families. Recent advances have suggested several ways to include data from larger families. For instance, tree-based decomposition methods facilitate the extraction of orthologs from large families. Additionally, several methods for species tree inference are robust to the inclusion of paralogs and could use all of the data from larger families. Here, we explore the effects of using all families for phylogenetic inference by examining relationships among 26 primate species in detail and by analyzing five additional data sets. We compare single-copy families, orthologs extracted using tree-based decomposition approaches, and all families with all data. We explore several species tree inference methods, finding that identical trees are returned across nearly all subsets of the data and methods for primates. The relationships among Platyrrhini remain contentious; however, the species tree inference method matters more than the subset of data used. Using data from larger gene families drastically increases the number of genes available and leads to consistent estimates of branch lengths, nodal certainty and concordance, and inferences of introgression in primates. For the other data sets, topological inferences are consistent whether single-copy families or orthologs extracted using decomposition approaches are analyzed. Using larger gene families is a promising approach to include more data in phylogenomics without sacrificing accuracy, at least when high-quality genomes are available.
ATRT-05. Infants and newborns with atypical teratoid/rhabdoid tumors (ATRT) and extracranial malignant rhabdoid tumors: a unique and challenging population
Authors: Nemes Karolina , Johann Pascal D , Steinbügl Mona , Gruhle Miriam , Bens Susanne , Kachanov Denis , Teleshova Margarita , Peter Hauser , Simon Thorsten , Tippelt Stephan , Eberl Wolfgang , Chada Martin , Lopez Vicente Santa-Maria , Grigull Lorenz , Hernáiz-Driever Pablo , Eyrich Matthias , Pears Jane , Milde Till , Reinhard Harald , Leipold Alfred , de Wetering Marianne v , João Gil-da-Costa Maria , Ebetsberger-Dachs Georg , Kerl Kornelius , Lemmer Andreas , Boztug Heidrun , Furtwängler Rhoikos , Kordes Uwe , Siebert Reiner , Vokuhl Christian , Hasselblatt Martin , Bison Brigitte , Kröncke Thomas , Melchior Patrick , Timmermann Beate , Gerss Joachim , Frühwald Michael C
Published: June 2022

INTRODUCTION: Malignant rhabdoid tumors (MRT) predominantly affect infants. Patients below six months represent a particularly challenging group: intensity of therapy is limited by toxicity to developing organs. Information on prognostic factors, toxicity and long term outcome is sparse. METHODS: Clinical, genetic, and treatment data of 100 patients (less than 6 months at diagnosis) from 13 European countries were analyzed (2005-2020). Tumors and matching blood samples were examined for SMARCB1 mutations using FISH, MLPA and Sanger sequencing. DNA-methylation subgroups (ATRT-TYR, ATRT-SHH, and ATRT-MYC) were determined using DNA methylation arrays. RESULTS: A total of 45 patients presented with ATRT, 29 with extracranial, extrarenal (eMRT) and 9 with renal rhabdoid tumors (RTK). Seventeen patients demonstrated synchronous tumors (SYN). Distant metastases at diagnosis (M+) were present in 27% (26/97). A germline mutation (GLM) was detected in 55% (47/86). Methylation subgroup status was available in 50% (31/62) of ATRT or SYN (SHH=13, TYR=13, MYC=4, SHH+TYR=1). The 5-year overall- (OS) and event free survival (EFS) rates were 23.5±4.6% and 19±4.1%, respectively. Male sex (11±5% vs. 35.8±7.4%), M+ (6.1±5.4% vs. 36.2±7.4%), presence of SYN (7.1±6.9% vs. 26.6±5.3%) and -GLM (7.7±4.2% vs. 45.7±8.6%) were significant prognosticators of 5-year OS, in univariate analysis. Molecular subgroup and survival analyses confirmed the previously described survival advantage of ATRT-TYR. In an adjusted multivariate model clinical factors that influence prognosis were: male sex [HR: 2.1 (1.2 – 3.6)], M+ [3.3 (1.8 – 6)], GLM [HR: 2 (1.1 – 3.6)] and maintenance therapy [HR: 0.3 (0.1 – 0.8)]. CONCLUSION: In this large cohort of homogenously treated infants with MRT, significant predictors of outcome were sex, M+, GLM and maintenance therapy. We confirm the need to stratify which patient group benefits from multimodal treatment, and which patients need novel therapeutic strategies. Biomarker-driven tailored trials may be a key option.
ATRT-06. Atypical Teratoid Rhabdoid Tumors (ATRT): results from a Single Institution in Brazil - São Paulo University
Authors: Barreto Juliana Silveira , Gomez Karen Nirit Melo , Dutra Alvaro Pimenta , De Azambuja Alessandra Milani Prandini , Cristofani Lilian Maria , Sanders Felipe Hada , Petito Carlo , Baraldi Helena Espindola , Weltman Eduardo , Ferraciolli Suely Fazio , Frassetto Fernando Pereira , Lucato Leandro Tavares , Rosemberg Sérgio , Filho Vicente Odone
Published: June 2022

OBJECTIVES: Atypical teratoid/rhabdoid tumor (AT/RT) is a rare, highly malignant tumor of the central nervous system with poor prognosis. Nowadays, multimodal management, including surgery, chemotherapy (CMT), radiation therapy (RT) and Bone Marrow Transplantation (BMT). The aim of this study was to assess the experience and survival in a center of reference of treatment in childhood cancer in Brazil. PATIENTS AND METHODS: Medical records of AT/RT patients who underwent surgery from 2008 to 2020 at a center of childhood cancer treatment (ITACI) in São Paulo University were retrospectively reviewed and statistically analyzed. RESULTS: Eight patients (2 males and 6 females) were presented with AT/RTs. Median age during presentation was 22 months (range, 0 - 6 years). Seven patients (88%) were < 18 months and one patient were >18 months. Tumor location was supratentorial in four patients, infratentorial in 2 patients. Kidney disease as the primary diagnosis in 2 patients (25%). Surgical treatment was performed in 4 patients. Seven children underwent total CMT and 3 children were treated with RT. Only 3 patients underwent Autologous Bone Marrow Transplantation (ABMT). The chemotherapy management protocol of the patients was variable: 2 patients received the EU-RHAB protocol, 2 patients received the HEAD START III protocol, 3 patients received chemotherapy in the ICE regimen (Ifosfamide + Carboplatin + Etoposide) and 1 patient received chemotherapy in the CDDP+CTX+VCR (Cyclophosphamide + Cisplatin + Vincristine) regimen. All patients had episodes of neutropenic fever when they received chemotherapy, requiring hospitalization and use of an antibiotic treatment. Among the 8 patients analyzed, all died. CONCLUSIONS: Despite progress in treatment, AT/RT of the CNS disease or primary kidney disease associated with a lack of standardization in a regimen contributes to the dismal prognosis. There is a high mortality in patients with AT/RT, similar to that found in the literature.
ATRT-15. Primordial germ cells identified as one potential cell of origin of MYC rhabdoid tumors
Authors: Graf Monika , Interlandi Marta , Moreno Natalia , Roy Rajanya , Holdhof Dörthe , Göbel Carolin , Melcher Viktoria , Mertins Julius , Albert Thomas K , Kastrati Dennis , Alfert Amelie , Holsten Till , de Faria Flavia , Meisterernst Michael , Rossig Claudia , Warmuth-Metz Monika , Nowak Johannes , Hörste Gerd Meyer zu , Mayère Chloe , Nef Serge , Johann Pascal , Frühwald Michael C , Dugas Martin , Schüller Ulrich , Kerl Kornelius
Published: June 2022

Rhabdoid tumors (RT) are embryonal neoplasms occurring most frequently in the central nervous system where they are termed atypical teratoid rhabdoid tumor (ATRT). A common hallmark of RT is homozygous loss of the BAF complex subunit SMARCB1. RT patients have a poor prognosis with an overall survival time of 17 months and >60% of patients suffer from relapses. The lack of an optimal treatment strategy could be attributed to the heterogeneity within and between different subgroups of ATRT. Despite the recent advancements in characterizing RT at a molecular level, the cellular origin of RT remains elusive. Thus, this study focused on the identification of the cellular origin of MYC-RT and underlying epigenetic deregulations which account for the cellular heterogeneity in these tumors. We showed that Smarcb1 abrogation in Sox2-positive progenitor cells at E6.5 give rise to RT of the MYC and SHH subgroup in genetically engineered mouse models (GEMM). To uncover distinct cells of origin (COO) for the SHH and MYC subgroups, unbiased computational approaches were used to compare single-cell transcriptomes of GEMMs with single-cell reference maps of murine early embryogenesis. While SHH tumors arise from mid/hindbrain progenitor cells, primordial germ cells (PGCs) emerge as COO of both intracranial and extracranial MYC tumors. PGCs as COO of MYC-RT were validated in vivo by using PGC-specific Smarcb1 knockout mouse model. We further characterized a deregulated transcriptome in MYC-RT compared to PGCs, which is sustained by a subset of epigenetically driven tumor cells. Deregulated expression of genes driving methylation/demethylation processes in MYC tumors and regression of these tumors upon treatment with decitabine in vitro and in vivo, indicates that DNA methylation plays a key role in cellular transformation and development of MYC-RT.
RARE-08. Profiling of recurrent adamantinomatous cranionpharyngioma confirms the activation of the MAPK pathway and identifies copy number aberrations in relapsed tumours
Authors: Apps John , Pickles Jessica , Stone Thomas , Chalker Jane , Ogunbiyi Olumide , Schwalbe Edward , Brandner Sebastian , Otto Georg , Castellano Sergi , Jacques Thomas , Deutschbein Timo , Martinez-Barbera Juan Pedro
Published: June 2022

Craniopharyngiomas are rare challenging tumours, with around 25% of cases recurring despite surgery and/or radiotherapy. Whilst transcriptomic and proteomic profiling of adamantinomatous craniopharyngioma (ACP) have revealed activation of the MAPK pathway and IL-6 mediated inflammation, relatively little is known about the biological processes involved in tumour recurrence. To address this, we have analysed a cohort of primary and relapsed ACP tumour accessed from tissue banks, local pathology departments and international collaborators. Serial samples of recurrent ACP (n=11), recurrent papillary craniopharyngioma (PCP) (n=4) and 3 non-recurrent ACP were analysed using β-catenin, pERK1/2 immunostaining, DNA methylation array and RNA sequencing. Differential expression and methylation analyses confirmed differences between ACP and PCP, with over representation of WNT pathway genes in ACP and the MAPK pathway genes in PCP. All of the primary and all except for one of the relapsed ACP tumours showed a pERK1/2 expression. Differences in the immune environment were also identified between ACP and PCP, with higher levels of some inflammatory mediators and CD14+ cell signatures in PCP compared with ACP. Whilst differential methylation and expression analysis revealed relatively stable methylomes and transcriptomes between serial samples of cases, segmental chromosomal alterations were identified in recurrence samples from five ACP cases (5/11,45%). One relapsed case showed histological and molecular signs of malignant transformation, including high ki67 and deletion of TP53. Surprisingly, this malignant tumour showed nuclear beta-catenin in all neoplastic epithelial cells and absence of pERK1/2 staining, despite the primary tumour showing the typical beta-catenin and pERK1/2 expression patterns. These results suggest that the molecular landscape of craniopharyngioma remains stable between recurrences in most cases, but, there is evidence of molecular evolution in a subset of cases. Activation of the MAPK pathway in the vast majority of ACP tumours supports the clinical evaluation of MAPK pathway inhibitors in ACP patients.
RARE-18. Pediatric craniopharyngioma; single center experience in 246 cases with different management modalities
Authors: El Beltagy Mohamed , Enayet Abdelrahman , Sameh Rana
Published: June 2022

PURPOSE: To report our experience with different craniopharyngioma management strategies done for 246 patients in a single institution during a period of 14 years. METHODS: The medical records of all children with the diagnosis of craniopharyngioma treated at Children's Cancer Hospital Egypt (CCHE-57357) during the period from July 2007 to December 2021 were retrospectively reviewed. RESULTS : our registry included 246 pediatric craniopharyngiomawith median age of 7.4 years old. The main strategies of management after initial surgery were follow up, radiotherapy or administration of intracystic interferone post ommaya insertion. The number of cases in each group were 92, 147,and three respectively. Three patients were not operated upon because of heavily calcified lesions, two of them received radiotherapy while the third was kept under follow up. Overall gross or near total total excision was achieved in 31.2 % , subtotal resection was the case in 42.1% while Ommaya insertion and biopsy was done in 21.1 % of cases. Total number of patients received radiotherapy initially or on progression was 195 patients( 78.9%). The five-year overall survival was 88% (95% CI 82.9 – 93.1) while 5- year progression free survival (PFS) was 49.2% (95% CI 41.2—57.2). The five-year PFS rates for patients in the follow up group versus radiotherapy group were 25.2% and 68.7% respectively (P< 0.0001). Beta catenin was positive in 76.4% of cases that were available for testing (123/161). CONCLUSION: management of craniopharyngioma should be individualized with the main objective is the quality of life. Conservative surgery which entails gross total safe resection whenever possible or lesser extent of resection followed by radiotherapy is the main strategy followed in our institution.
RARE-22 Characterizing the landscape of structural variants in adamantinomatous craniopharyngioma
Authors: Jomaa Danny , Khadka Prasidda , Novikov Dana , Condurat Alexandra L , Tsai Jessica W , Dubois Frank , Zhang Shu , Zhou Kevin , Gold Rose , Sousa Cecilia , Vogelzang Jayne , Prince Eric , Lu Sophie , Slivova Veronika , Otto Georg W , Hereza Sergi Castellano , Ashley David , Cohen-Gadol Aaron A , Thompson Eric , Beroukhim Rameen , Apps John , Martinez-Barbera Juan Pedro , Hankinson Todd , Bandopadhayay Pratiti
Published: June 2022

INTRODUCTION: Adamantinomatous craniopharyngiomas (ACPs) are rare brain tumors that primarily occur in children and impact long-term morbidity and mortality. The canonical driver mutation for ACP growth occurs in CTNNB1 and leads to constitutive activation of the Wnt/β-catenin signaling pathway. In this study, we outline the genomic, transcriptomic, and structural variant (SV) landscape in a cohort of 41 ACP samples. METHODS: We performed whole-genome sequencing (WGS) and RNA-sequencing of 41 ACP samples. Matched normal samples were also characterized by WGS. Mutect2 was used to detect single nucleotide variants (SNVs) and indels, and copy number data was generated using the GATK pipeline. SvABA was used to perform SV analysis and to identify significantly recurrent breakpoints and juxtapositions. DESeq2 was used to perform differential gene expression analysis based on clinical and molecular annotation data. RESULTS: 29/41 (70%) of the ACP samples harbored missense mutations in exon 3 of CTNNB1, all of which have previously been reported in ACP tumors. SV analysis identified a median of 11.5 events per tumor. Overall, 9.7% of events were interchromosomal. Of the remainder, the majority (78.6%) were deletions. No SVs occurred within CTNNB1. A positive correlation (r = 0.533) was observed between the frequency of SVs and SNVs within samples. Analysis of significantly recurring breakpoints (SRBs) did not identify recurrent breakpoint events. Differential gene expression analysis comparing samples with and without CTNNB1 variants identified 2,143 differentially expressed genes with q-value < 0.05. CONCLUSION: This study identifies activating mutations in exon 3 of CTNNB1 in a large cohort of ACP samples. We also integrate SV and transcriptomic data to comprehensively investigate ACP tumor genomes and identify putative novel tumorigenic mechanisms that advance our understanding of ACP biology.
RARE-29. Transcriptome characterization of pediatric adamantinomatous craniopharyngioma at the cellular level
Authors: Prince Eric , Trudeau Tammy , Chatain Oscar , Staulcup Susan , Martinez-Barbera Juan Pedro , Goerg Carsten , Hankinson Todd
Published: June 2022

BACKGROUND: Adamantinomatous Craniopharyngioma (ACP) is a neurologically devastating brain tumor that affects children and adults. It is histologically heterogeneous with epithelial populations that are characterized by the nuclear accumulation of mutated β-catenin, and activated Wnt signaling. Current models suggest that ACP growth is driven through paracine mechanisms characterized by the senescence-associated secretory phenotype (SASP). However, detailed pathogenic mechanisms remain unknown. Improved definition of the various cellular phenotypes that compose ACP will inform and advance our understanding of this disease. METHODS: Single cell RNA-sequencing (scRNA-seq) and multiplex ELISA were performed on pediatric ACP tissue and cyst fluid, respectively. Reference scRNA-seq data was obtained from PanglaoDB. Preprocessing and standard analyses were conducted using Seurat software. Cellular phenotypes were annotated using the Human Primary Cell atlas. Differential expression and functional enrichment analyses were utilized to identify Wnt-signaling activation and epithelial subpopulations. Paracrine signaling was inferred via CellChatDB. SASP Atlas was utilized to query marker gene lists. Pseudotemporal ordering was performed using monocle3. RESULTS: ACP tissue is heterogenous and contains multiple distinct immune signatures. ACP tissue contains 2 unique epithelial subpopulations, which demonstrate canonical Wnt-signaling and SASP, respectively. Pseudotemporal ordering suggests the initial oncogenic event to be of epidermal character, with subsequent aggressive behavior from a separate epithelial cell population. CONCLUSIONS: Based on gene expression, cell populations that correspond to the histologically identifiable epithelial whorls and palisading epithelium can be identified. These subpopulations display unique functional signatures. Simultaneous and synergistic therapeutic targeting of these separate epithelial populations may lead to improved patient care.
DIPG-04. Feasibility and early results of phase 2 open label randomized study of radiotherapy(RT), concomitant nimotuzumab and vinorelbine and re-irradiation at relapse, versus multiple elective radiotherapy courses with concomitant vinorelbine and nimotuzumab for newly diagnosed childhood and adolescence Diffuse intrinsic Pontine Glioma (DIPG)
Authors: Massimino Maura , Biassoni veronica , Mastronuzzi Angela , Schiavello Elisabetta , Barretta Francesco , Quaglietta Lucia , Milanaccio Claudia , Pecori Emilia , Cacchione Antonella , Boschetti Luna , Ruscio Valentina Di , Chiesa Silvia , Scimone Giuseppe , Barra Salvina , De Martino Lucia , Ramaglia Antonia , Picariello Stefania , Verrico Antonio , Alessandro Ombretta , Vennarini Sabina , Podda Marta , Gattuso Giovanna , Cinalli Giuseppe , Antonelli Manila , Modena Piergiorgio , De Cecco Loris , Buttarelli Francesca R , Gandola Lorenza
Published: June 2022

BACKGROUND: The purposes of this trial were to evaluate the feasibility, response, PFS/OS of a randomized study comparing two different RT schedules for DIPG while administering the same systemic treatment. METHODS: Patients: 2-21 years-old with a not-pretreated radiologically verified DIPG (MRI blindly reviewed at diagnosis and every 12 weeks thereafter) and symptoms duration below 6 months. Biopsy was required if suggested by atypical imaging. Vinorelbine 20 mg/m2+nimotuzumab 150 mg/m2 were administered weekly for 12 weeks; thereafter every other week until tumor progression or for up to 2 years. Standard(ST) arm included focal RT at total dose of 54Gy (1.8Gy/day); for local progression re-irradiation was proposed at 19.8Gy, in case of dissemination craniospinal irradiation(CSI) at 36Gy was adopted. Experimental(SP) arm included three elective courses of RT at defined timepoints at 36Gy, 19.8Gy and 19.8Gy with possible reirradiation for relapse at 9 Gy. Incidences of local(L) and distant(D) progression were assessed in a competing risk setting. RESULTS: Aggregated preliminary results are given for 4 Italian centers. 54 pts were screened and 51 included, 27 in ST, 28 males, median age 7 years (range 3-17). Median time of observation was 17.9 months. Twelve patients needed a shunt, 10 during treatment; 20 were biopsied, in 18 cases according local protocols. 19/20 tumors had H3.3 K27 mutation. 41 relapsed, 28 locally, 13 with a component of dissemination. 36 died, one for tracheotomy bleeding. SP irradiation was feasible and never produced significant radionecrosis. Median EFS/OS were 7.3/12.9 months, respectively; EFS/OS at 1 year were 19.0%/57.3%, not differing between patients with local vs. disseminated relapses. Patients submitted to biopsies had more dissemination (P=0.04) and less local progression (P=0.077). CONCLUSIONS: Treatment was feasible and OS confirmed previous results obtained in a single center. Randomization results will be later reported.
DIPG-19. FOXR2 is an oncogenic driver across pediatric and adult cancers
Authors: Tsai Jessica W , Cejas Paloma , Wang Dayle K , Patel Smruti , Wu David W , Arounleut Phonepasong , Wei Xin , Zhou Ningxuan , Syamala Sudeepa , Dubois Frank P B , Crane Alexander , Pelton Kristine , Vogelzang Jayne , Sousa Cecilia , Baguette Audrey , Chen Xiaolong , Condurat Alexandra L , Dixon-Clarke Sarah E , Zhou Kevin N , Lu Sophie D , Gonzalez Elizabeth M , Chacon Madison S , Digiacomo Jeromy J , Kumbhani Rushil , Novikov Dana , Hunter J'Ya , Tsoli Maria , Ziegler David S , Dirksen Uta , Jager Natalie , Balasubramanian Gnana Prakash , Kramm Christof M , Nathrath Michaela , Bielack Stefan , Baker Suzanne J , Zhang Jinghui , McFarland James M , Getz Gad , Aguet Francois , Jabado Nada , Witt Olaf , Pfister Stefan M , Ligon Keith L , Kleinman Claudia L , Long Henry , Jones David T W , Bandopadhayay Pratiti , Phoenix Timothy N
Published: June 2022

BACKGROUND: Understanding how aberrant transcription factors (TFs) hijack normal development to induce oncogenesis is a critical question in oncology. Forkhead box (FOX) proteins are a superfamily of transcriptional regulators characterized by a forkhead DNA-binding domain. Within this family, Forkhead Box R2 (FOXR2) has been identified as a candidate structural variant (SV) driver in a subset of pediatric cancers including CNS embryonal tumors and peripheral neuroblastoma. While FOXR2 has been shown to stabilize MYC isoforms, the mechanistic details through which it enhances tumor formation, other non-SV mechanisms of activating aberrant expression, and the true extent of its role as an oncogene across all cancers have not been systematically evaluated. METHODS: We applied an integrative approach using transcriptomics, epigenetics, in vitro cancer models, and in vivo mouse models to systematically evaluate the mechanisms by which FOXR2 is activated across human cancers. RESULTS: We performed a pan-cancer analysis of FOXR2 activation across over 10,000 adult and pediatric cancer samples, and surprisingly found FOXR2 to be aberrantly upregulated in 70% of all cancer types (including diffuse midline gliomas), and 8% of all individual tumors. FOXR2 expression occurred predominantly in the absence of rearrangement/fusions, single nucleotide variants, or copy number aberrations at the DNA level. Transcriptomic and epigenomic analyses show the vast majority of tumors (78%) aberrantly express FOXR2 through a previously undescribed epigenetic mechanism via hypomethylation of a novel promoter. Using both in vitro and in vivo models, we demonstrate that FOXR2 expression is both sufficient and necessary for transformation across multiple lineages, including DMGs. CONCLUSION: Taken together, this study demonstrates that FOXR2 is a novel and potent oncogene across pediatric and adult cancers, and highlights a new epigenetic mechanism by which its expression is activated.
DIPG-31. Prognostic and predictive biomarkers of response in children and young adults with H3K27M-altered diffuse intrinsic pontine glioma: results from a multi-center, interventional clinical trial (PNOC003)
Authors: Kline Cassie , Jain Payal , Kilburn Lindsay , Bonner Erin , Gupta Nalin , Crawford John , Banerjee Anu , Packer Roger , Villanueva-Meyer Javier , Luks Tracy , Zhang Yalan , Kambhampati Madhuri , Zhang Jie , Yadavilli Sridevi , Kraya Adam , Kuhn John , Liang Winnie , Byron Sara , Berens Michael , Molinaro Annette , Prados Michael , Resnick Adam , Waszak Sebastian , Nazarian Javad , Mueller Sabine
Published: June 2022

BACKGROUND: Diffuse intrinsic pontine glioma (DIPG) is a fatal brain tumor. Herein, we report on novel prognostic and predictive genomic biomarkers identified in PNOC003, a multi-center precision medicine trial for children and young adults diagnosed with DIPG. METHODS: Patients aged 3-25 years were enrolled on PNOC003 based on radiographic diagnosis of DIPG. Pre-treatment tumor biopsies were analyzed using tumor-normal whole-exome sequencing and mRNA-tumor sequencing to determine biology-informed, multi-agent therapy following radiation therapy (RT). Whole-genome sequencing was performed as an exploratory study aim. Genomic biomarkers were investigated to identify predictors of RT response and overall survival (OS) in patients with confirmed H3K27M-altered DIPG. Prognostic biomarkers were verified in a retrospective, H3K27M-altered diffuse midline glioma cohort (n=22) from the Children’s Brain Tumor Network (CBTN). RESULTS: Thirty patients enrolled on PNOC003 met molecular criteria for H3K27M-altered DIPG. TP53 was the most frequently altered driver gene (73%). Somatic alterations in PTEN>TP53>PDGFRA were independently associated with OS (P<0.05, in order of negative impact on survival). TP53 mutations associated with worse OS (TP53mut 11.1 mo [95% CI 8.7, 14]; TP53wt 13.3 mo [95% CI 11.8, NA]; P=3e-2), chromosomal instability (P=3e-3), and resistance to RT (P=6e-4). Moreover, loss of chromosome 10q, encoding tumor suppressor PTEN, was associated with worse OS, co-occurred with PTEN alterations, biallelic PTEN inactivation and loss of gene expression. The combination of TP53 alterations and loss of 10q/PTEN in H3K27M-altered DIPG was associated with the worst OS in a combined PNOC003 and CBTN cohort (TP53mut/10qdel, n=14, OS 8.4 mo [95% CI 7.4, 15.8]; TP53mut/10qwt, n=20, OS 13.1 mo [95% CI 10.1, 17.2]; TP53wt/10qwt, n=14, OS 15.5 mo [11.8, 29.4]; P=2e-3). CONCLUSION: PNOC003, a tissue-driven clinical trial, provided insights into prognostic and predictive genomic biomarkers and informed a novel molecular tumor classification system for H3K27M-altered DIPGs.
DIPG-46. Radiation induced senescence in diffuse intrinsic pontine glioma cells reveals selective vulnerability to Bcl-XL inhibition
Authors: Vardon Ashley , Guiho Romain , Carvalho Diana , Boult Jessica , Carter Rebecca , Grabovska Yura , Mackay Alan , Zheng Guangrong , Zhou Daohong , Hiley Crispin , Lythgoe Mark , Jones Chris , Hargrave Darren , Martinez- Juan-Pedro
Published: June 2022

Diffuse intrinsic pontine glioma remains a devastating condition with a dismal five year survival rate less than 5%. New approaches for treating this aggressive disease are critical to driving progress. Conventional radiotherapy remains the cornerstone of treatment, with no chemotherapeutic agent found to improve survival. However, radiotherapy is often delivered as a palliative treatment, and disease often recurs 3-6 months after. Radiation causes DNA damage and oxidative stress yielding a senescent state of replicative arrest in susceptible cells. However, increasing evidence demonstrates malignant cells can escape senescence leading to tumour recurrence. Targeted ablation of non-replicating senescent tumour cells following radiation could negate tumour recurrence. It remains unknown whether DIPG undergoes senescence following radiation, and furthermore, whether senolytics can be utilised to target senescent DIPG cells. We employed radiation to induce a senescent state in primary human DIPG cell lines. Senescence was confirmed using SA-β-gal staining, lack of EdU incorporation and qRT-PCR to characterise the SASP in three primary human DIPG cell lines. RNA-sequencing on DIPG cells following radiation revealed senescence and SASP signatures. Likewise, expression of senescence markers has been detected in human tumours. Viable cells that survive radiation were then utilised to screen candidate senolytic drugs, only Bcl-XL inhibitors demonstrated reproducible senolytic activity in radiation treated DIPG cells. In addition, Bcl-XL degradation using PROTACs (proteolysis targeting chimeras) resulted in a significant increase in senolysis of susceptible tumour cells. Conversely, Bcl-2 inhibitors failed to show any consistent senolytic activity. We are currently performing preclinical studies in the mouse to test the efficiency of senolytics against DIPG. These results demonstrate future possibilities of targeting radiation induced senescence in DIPG, using novel senolytic therapies and highlight Bcl-XL dependency as a potential vulnerability of surviving DIPG cells following exposure to radiation.
DIPG-50. Bioinformatic evaluation of genes involved in sphingomyelin biosynthesis in Diffuse Midline Glioma H3K27 altered/DIPG: dysregulation of sphingosine 1-phosphate (SP1)
Authors: Humphreys Aimee , Russell Hanna , Fillmore Helen L
Published: June 2022

Sphingosine 1-phosphate (S1P), a bioactive signalling lipid, interacts with a network of metabolic enzymes, receptors, transporters, and epigenetic partners. This network is well described in many cancers; however, little is known about its potential impact in DIPG. Expression of HDAC1 (binding target of S1P) and genes associated with the sphingomyelin (SM) pathway were examined in datasets identified in the National Centre for Biotechnology Information, Gene Expression Omnibus, and analysed using the R2: Genomics Analysis and Visualization Platform (http://r2.amc.nl). The Paugh-DIPG dataset (27 DIPG samples) and normal samples (20 years and younger - Berchtold dataset) were compared. To avoid issues related to batch effects, expression values for each gene of interest and controls were exported into separate files to determine differentially expressed genes. Internal genes include housekeeping; ACTB, GAPDH, B2M, TBP; downregulated in DIPG; GPR6, NGB, and upregulated in DIPG; MMP16, PDGFRA, TP53, CSPG4. Genes of interest; SPHK1, SPHK2, SGPL1, ACER1, ACER3, KDSR, SMPD1-4, CPTP, GLTP, DEGS1, CERK, CERS1-6, ASAH1, SGPP1, SGPP2 and HDAC1. To test for significance, each dataset was standardised using ACTB housekeeping gene. Values including Log-transformed fold change were analysed using the non-parametric, Mann-Whitney test. 7 of the 16 genes were dysregulated relative to expression in normal brain (p<0.0002). SPHK2 and SMPD3 were downregulated, and HDAC1, SGPL1, DEGS1, CERS4, and ASAH1 were upregulated in DIPG compared to normal. To identify genes more likely associated with DIPG (vs development), we evaluated gene expression in Brainspan dataset (brspv10rs). Validation of SPHK2 and SGPL1 protein expression (responsible for the synthesis and cleavage of SP1) is underway. Current work is focused on the intracellular processing and function (isoform specific inhibitors) of S1P in DIPG cells. Given its reported role in several cancer hallmarks, a better understanding of the sphingomyelin biosynthesis pathway in DMG/DIPG is merited and may lead to novel therapeutic targets.