ABSTRACT
ATRX, a chromatin remodeler protein, is recurrently mutated in H3F3A-mutant pediatric glioblastoma (GBM) and isocitrate dehydrogenase (IDH)-mutant grade 2/3 adult glioma. Previous work has shown that ATRX-deficient GBM cells show enhanced sensitivity to irradiation, but the etiology remains unclear. We find that ATRX binds the regulatory elements of cell-cycle phase transition genes in GBM cells, and there is a marked reduction in Checkpoint Kinase 1 (CHEK1) expression with ATRX loss, leading to the early release of G2/M entry after irradiation. ATRX-deficient cells exhibit enhanced activation of master cell-cycle regulator ATM with irradiation. Addition of the ATM inhibitor AZD0156 doubles median survival in mice intracranially implanted with ATRX-deficient GBM cells, which is not seen in ATRX-wild-type controls. This study demonstrates that ATRX-deficient high-grade gliomas (HGGs) display Chk1-mediated dysregulation of cell-cycle phase transitions, which opens a window for therapies targeting this phenotype.
Subject(s)
Checkpoint Kinase 1/metabolism , Glioma/metabolism , X-linked Nuclear Protein/metabolism , Animals , Brain Neoplasms/metabolism , Cell Cycle/genetics , Cell Cycle Checkpoints/genetics , Cell Line, Tumor , Checkpoint Kinase 1/physiology , Female , Histones/metabolism , Humans , Isocitrate Dehydrogenase/genetics , Male , Mice , Mice, Inbred C57BL , Mutation , Neoplasm Recurrence, Local/metabolism , Primary Cell Culture , X-linked Nuclear Protein/geneticsABSTRACT
Ruminants, which have multi-compartmented stomachs, are adapted to digest cellulosic materials, which constitute the primary expense on ranches and dairies. Industrial byproducts can be repurposed for livestock diets to decrease these costs. Therefore, finding alternative feedstuffs may benefit the economics of livestock production. The goal of this project was to evaluate the variation in nutritive value of ruminal waste as a potential feedstuff. Twelve paunch samples were collected from individual cattle across multiple harvest dates at the Tarleton State University Meat Laboratory, Stephenville, TX. Samples were dried and assayed for dry matter (DM), crude protein (CP), sequential neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL), and physically-effective fiber (peNDF). Samples were subjected to batch-culture in vitro digestibility assays for the determination of digestibility coefficients. Mean NDF, ADF, ADL, CP and peNDF concentrations were 681, 399, 109, 150, and 387 g kg-1 DM, respectively. Contribution to variance from sample for NDF, ADF, ADL, CP, and peNDF were 75.3, 41.9, 33.0, 51.2, and 71.3%, respectively. In vitro true digestibility (IVTD) and in vitro NDF digestibility (IVNDFD) were recorded as 462 and 216 g kg-1 DM, respectively. Contribution to variation of sample for IVTD and IVNDFD were 31.0 and 30.7%, respectively. Results indicate that rumen waste harvested from abattoirs may be useful for sustainable livestock production, while reducing environmental threats posed by disposal, but the viability of the product is highly dependent on the source animal. For full viability of application in a sustainable system, a centralized receiving and compositing system may be useful for developing a consistent product.
ABSTRACT
Pediatric and adult high-grade gliomas (HGGs) frequently harbor PDGFRA alterations. We hypothesized that cotreatment with everolimus may improve the efficacy of dasatinib in PDGFRα-driven glioma through combinatorial synergism and increased tumor accumulation of dasatinib. We performed dose-response, synergism, P-glycoprotein inhibition, and pharmacokinetic studies in in vitro and in vivo human and mouse models of HGG. Six patients with recurrent PDGFRα-driven glioma were treated with dasatinib and everolimus. We found that dasatinib effectively inhibited the proliferation of mouse and human primary HGG cells with a variety of PDGFRA alterations. Dasatinib exhibited synergy with everolimus in the treatment of HGG cells at low nanomolar concentrations of both agents, with a reduction in mTOR signaling that persisted after dasatinib treatment alone. Prolonged exposure to everolimus significantly improved the CNS retention of dasatinib and extended the survival of PPK tumor-bearing mice (mutant TP53, mutant PDGFRA, H3K27M). Six pediatric patients with glioma tolerated this combination without significant adverse events, and 4 patients with recurrent disease (n = 4) had a median overall survival of 8.5 months. Our results show that the efficacy of dasatinib treatment of PDGFRα-driven HGG was enhanced with everolimus and suggest a promising route for improving targeted therapy for this patient population.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Brain Neoplasms/drug therapy , Brain Neoplasms/genetics , Dasatinib/administration & dosage , Everolimus/administration & dosage , Glioma/drug therapy , Glioma/genetics , Receptor, Platelet-Derived Growth Factor beta/genetics , ATP Binding Cassette Transporter, Subfamily B/genetics , Adolescent , Animals , Antineoplastic Combined Chemotherapy Protocols/pharmacokinetics , Brain Neoplasms/metabolism , Cell Proliferation/drug effects , Child , Child, Preschool , Dasatinib/pharmacokinetics , Drug Screening Assays, Antitumor , Drug Synergism , Everolimus/pharmacokinetics , Female , Gene Expression , Glioma/metabolism , Humans , Male , Mice , Molecular Targeted Therapy , Pregnancy , Tumor Cells, CulturedABSTRACT
Reinvestigation of mycothiazole (1) revealed picomolar potency (IC50 = 0.00016, 0.00027, 0.00035 µM) against pancreatic, (PANC-1), liver (HepG2), and colon (HCT-116) tumor cell lines. Reevaluation of 1 provided [α]D data indicating Vanuatu specimens of C. mycofijiensis contain the 8S enantiomer of 1 and not the 8R configuration previously reported. Semisynthesis provided 8-O-acetylmycothiazole (2), 8-oxomycothiazole (8), mycothiazole nitrosobenzene derivatives (MND1, MND2: 9a, 9b), and MND3 (10) with IC50 = 0.00129, >1.0, >1.0, >1.0, >1.0 µM, respectively, against PANC-1 cell lines. These results highlight the significance of the penta-2,4-dien-1-ol residue as a key structural feature of 1 required for its cytotoxicty against tumor cell lines.
ABSTRACT
As the field of neuro-oncology makes headway in uncovering the key oncogenic drivers in pediatric glioma, the role of precision diagnostics and therapies continues to rapidly evolve with important implications for the standard of care for clinical management of these patients. Four studies at major academic centers were published in the last year outlining the clinically integrated molecular profiling and targeting of pediatric brain tumors; all 4 demonstrated the feasibility and utility of incorporating sequencing into the care of children with brain tumors, in particular for children and young adults with glioma. Based on synthesis of the data from these studies and others, we provide consensus recommendations for the integration of precision diagnostics and therapeutics into the practice of pediatric neuro-oncology. Our primary consensus recommendation is that next-generation sequencing should be routinely included in the workup of most pediatric gliomas.
