tbuckley/cabot-search huggingface dataset:
In vivo metabolic labeling and detection of specific glycoprotein subclasses in a mouse breast cancer model
Authors: Brian Agnew, Courtenay Hart, Tamara Nyberg, Rajkumar Lakshmanaswamy
Journal: Cancer Research (2007)

2465  Altered glycosylation patterns are a hallmark of malignant transformation and may include oligosaccharide truncation, aberrant terminal oligosaccharide sequences, abnormal oligosaccharide branching, and increased sialic acid expression. Both N- and O-linked glycoproteins display abnormal glycosylation patterns in malignant cells. Current analytical and cell biological methods for the analysis of glycoproteins from normal or tumor tissues suffer from a lack of sensitivity and selectivity, and often require the use of harsh conditions that are incompatible with protein analysis. Here we describe a metabolic labeling approach for the detection of glycoprotein subclasses in tumor tissues using a breast cancer animal model. The two-step labeling technique involves the metabolic incorporation of unnatural azide-modified sugars into protein glycans and subsequent ligation with fluorescent azide-reactive detection probes utilizing the copper (I)-catalyzed cycloaddition reaction between azides and alkynes, or “click” chemistry. The glycoprotein detection reaction is highly sensitive, selective, and is performed under standard protein biochemistry conditions. Metabolic labeling of O-linked and sialic acid-containing glycoproteins was accomplished by administration of the unnatural tetraacetylated azide-modified sugar precursors, GalNAz or ManNAz, respectively, to activated Her2/Neu transgenic mice over a 7 day regimen. After 7 days the mice were sacrificed and breast tumors, control mammary glands, and various organ tissues were dissected, lysed, and protein extracts were reacted with fluorescent alkyne detection probes. Labeled glycoproteins were then analyzed by gel electrophoresis and detected by fluorescence laser scanning. Our results demonstrate highly-efficient labeling of O-linked glycoproteins and sialic acid containing glycoproteins isolated from both tumor and normal tissues, including blood, as determined by gel electrophoresis and fluorescence imaging. Significant differences were observed between glycoprotein profiles from breast tumors of different stages and those from normal mammary epithelial cells. And, as expected, there were distinct differences in glycoprotein profiles obtained from different organ tissues. The metabolic labeling approach demonstrated here will allow for the comprehensive analyses of specific glycoprotein changes during tumor development in live animals. The described labeling technique is fully compatible with downstream mass spectrometry and should allow for the identification of glycoproteins isolated from polyacrylamide gels. This novel “click”-based glycoprotein detection strategy provides selectivity, sensitivity and versatility that are unachievable with currently available lectin-based and antibody-based methods.
Functional interrogation of a SARS-CoV-2 host protein interactome identifies unique and shared coronavirus host factors
Authors: H.-Heinrich Hoffmann, Francisco J. Sánchez-Rivera, William M. Schneider, Joseph M. Luna, Yadira M. Soto-Feliciano, Alison W. Ashbrook, Jérémie Le Pen, Andrew A. Leal, Inna Ricardo-Lax, Eleftherios Michailidis, Yuan Hao, Ansgar F. Stenzel, Avery Peace, Johannes Zuber, C. David Allis, Scott W. Lowe, Margaret R. MacDonald, John T. Poirier, Charles M. Rice
Journal: Cell Host & Microbe (2020)
STEM-10. LONG-LASTING TUMOR TREATING FIELDS (TTFIELDS) STIMULATION OF GLIOBLASTOMA PRIMARY CULTURE DAMAGES PREFERENTIALLY FULLY DIFFERENTIATED CANCER CELLS
Authors: Stefania Castiglione, Matteo Ranucci, Gaetano Cannavale, Francesca Cianci, Ivan Verduci, Michele Mazzanti
Journal: Neuro-Oncology (2020)

Abstract Glioblastoma (GBM) is the most aggressive among brain tumors. Radiotherapy and chemotherapy as adjuvant procedures after surgical removal of the tumor mass, are often accompanied by undesirable symptoms and result in an extremely poor prognosis. In the last twenty years, it has been demonstrated that the only treatment able to improve the life expectancy of patients with GBM has been Tumor Treating Fields (TTFields) application. However, despite different therapeutic actions, tumor relapse occurs inexorably in most of the patients. There is scientific consensus that tumor recurrence is due to a population of cells named GBM stem cells (GSCs). GSCs represent a restricted subpopulation in the tumor mass, which is mainly composed of bulk cells. The main characteristic of GSCs is their tumorigenic potential even if a small number of cells are left after surgery. Our working hypothesis is based on the evidence that TTFields destabilize the mitotic spindle of dividing cells. Thus, TTFields stimulation should preferentially target bulk cells because of their higher proliferation rate compared to GSCs. Preliminary experiments performed using GBM primary cultures obtained from patients’ biopsies confirmed the presence of two different cell populations. In GBM, primary cultures maintained in stem-cell permissive medium, more than 70% of cells do not show stemness properties. After 72 hours of TTFields stimulation, almost 50% of the cells die. The remaining cells were more than 50% positive for CD44, SOX2, OCT4, and Nanog stemness markers, suggesting an enrichment of GSCs within the population. Following long lasting stimulation (up to 288 hours), GSCs percentage further increased and, more importantly, cells restored their ability to proliferate. In conclusion, TTFields treatment demonstrated efficacy by delaying GBM expansion. However, to prevent GBM relapse, it is imperative to increase the inhibitory efficiency of TTFields, specifically on GBM stem cell growth.
PATH-14. FHL1 INTERACTS WITH SP1 TO REGULATE EGFR EXPRESSION IN GLIOBLASTOMA
Authors: Lili Sun, Ming Li
Journal: Neuro-Oncology (2020)

Abstract The four and a half LIM domain 1 (FHL1) has been considered as a tumor suppressor protein in multiple cancers. Here, we show that FHL1 plays a tumor-promoting role in glioblastoma, the most common and incurable brain cancer. Overexpression of FHL1 promotes the growth, migration, and invasion of GBM cells in vivo and in vitro. In contrast, FHL1 silencing exhibits the opposite effects. Mechanically, FHL1 upregulates EGFR expression and activates the downstream AKT / ERK1 / 2 / STAT3 signaling pathways. We further demonstrated that SP1 can also be induced by FHL1 expression, and FHL1 interacts with SP1 to upregulate EGFR expression at both mRNA and protein levels, leading to glioblastoma malignancy. Clinically, FHL1 is highly expressed in glioblastoma and shows positive correlation with EGFR and SP1 in GBM specimens. Our results suggest the key role of FHL1 in the expression of EGFR and highlight the translation potential of inhibiting FHL1 as a treatment for glioblastoma.
DDEL-04. ENGINEERED NANOCARRIERS TO ENHANCE DRUG DELIVERY ACROSS THE BLOOD-BRAIN BARRIER
Authors: Joelle P Straehla, Cynthia Hajal, Hannah Safford, Giovanni Offeddu, Jeffrey Wyckoff, Roger D Kamm, Paula T Hammond
Journal: Neuro-Oncology (2020)

Abstract Pediatric central nervous system tumors are the leading cause of cancer death in children. Promising therapeutics have been identified, but the ability to deliver an effective concentration to the tumor without causing excessive systemic toxicity remains a challenge. To address this, we leveraged a tunable nanocarrier platform to design a brain-penetrant nanocarrier with preferential uptake into tumor cells over healthy brain cells. First, we used the layer-by-layer technique to iteratively coat liposomes with nanometers-thick layers of oppositely charged polyelectrolytes. To investigate the influence of surface chemistry on cellular trafficking, a panel of layered liposomes was tested for interactions with cancer cell lines, identifying poly-L-aspartic acid and hyaluronic acid as the highest-performing formulations across brain tumor lines. To facilitate nanocarrier transit across the blood-brain barrier (BBB), we developed a click chemistry platform to functionalize the nanocarrier with BBB shuttle ligands. To investigate trafficking in vitro, we utilized a microfluidic brain microvascular model comprising endothelial cells, astrocytes, pericytes, and glioma cells that self-assemble into a perfusable vascular network. We found that nanocarrier size influenced vascular transport, and the addition of BBB shuttle ligands improved transport in the presence of a glioma spheroid. To investigate in vivo nanocarrier trafficking, we performed intravital imaging through a cranial window in anesthetized mice. After intravenous administration, nanocarrier transit across intact brain capillaries was visualized using two photon microscopy, and vessel permeability was quantified over time. Ongoing studies in mice bearing patient-derived xenograft medulloblastoma and glioma tumors are being conducted to further characterize trafficking across tumor-associated vasculature.
Author Correction: Senescent cells promote tissue NAD+ decline during ageing via the activation of CD38+ macrophages
Authors: Anthony J. Covarrubias, Abhijit Kale, Rosalba Perrone, Jose Alberto Lopez-Dominguez, Angela Oliveira Pisco, Herbert G. Kasler, Mark S. Schmidt, Indra Heckenbach, Ryan Kwok, Christopher D. Wiley, Hoi-Shan Wong, Eddy Gibbs, Shankar S. Iyer, Nathan Basisty, Qiuxia Wu, Ik-Jung Kim, Elena Silva, Kaitlyn Vitangcol, Kyong-Oh Shin, Yong-Moon Lee, Rebeccah Riley, Issam Ben-Sahra, Melanie Ott, Birgit Schilling, Morten Scheibye-Knudsen, Katsuhiko Ishihara, Stephen R. Quake, John Newman, Charles Brenner, Judith Campisi, Eric Verdin
Journal: Nature Metabolism (2020)
STEM-27. miR-10b-5p MODULATES 5hmC EXPRESSION AND THE STEM-LIKE PHENOTYPE IN GBM
Authors: Harmon Khela, Sweta Sudhir, Maria Lugo-Fagundo, Bachchu Lal, Hernando Lopez-Bertoni, John Laterra
Journal: Neuro-Oncology (2020)

Abstract Epigenetic alterations such as DNA methylation and dysregulation of non-coding RNAs (e.g. miRNAs) are found in all types of cancer and are thought to play important roles in tumorigenesis. GBM is characterized by small subsets of cells, referred to as glioma stem cells (GSCs), that display stem-like properties implicated in tumor initiation, therapeutic resistance, and recurrence. DNA methylation patterns are altered in GBM and GSCs and are thought to play critical roles in tumor initiation and propagation. DNA methylation is a reversible process catalyzed, in part, by the ten-eleven translocation (TET) family of enzymes. These enzymes function as deoxygenases that catalyze the conversion of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC). Multiple studies found negative correlations between 5hmC levels and glioma grade and loss of 5hmC correlates with poor prognosis of GBM patients. However, the mechanisms leading to the loss of 5hmC in glioma and the role this phenomenon plays in gliomagenesis remains poorly understood. We found that Sox2 expression decreases TET2 expression and its product 5hmC in GSCs and identified miR-10b-5p as a molecular intermediary of this process. We show that miR-10b-5p expression is high in GBM compared to non-tumor in clinical specimens and high levels of this miRNA correlate with poor patient outcome. Expression of transgenic miR-10b-5p enhanced sphere formation capacity of GSCs and the expression of stem cell markers and drivers. Additionally, using a combination of molecular and biochemical endpoints, we show that miR-10b-5p modifies 5hmC levels by regulating TET2 in GSCs. Finally, we show that repression of miR-10b-5p increases 5hmC levels and inhibits tumor propagation in GBM xenograft models. Taken together, these results present a new molecular mechanism that controls 5hmC and the tumor propagating capacity of GSCs and suggests that miR-10b-5p inhibition and other strategies for enhancing TET2 function can be developed to treat GBM.
CTNI-41. PHASE I STUDY OF RUXOLITINIB WITH RADIATION AND TEMOZOLOMIDE IN PATIENTS WITH NEWLY DIAGNOSED GRADE III GLIOMAS AND GLIOBLASTOMA
Authors: Yasmeen Rauf, Rachel Hufsey, Kathy Robinson, John Suh, Samuel Chao, Erin Murphy, Jennifer Yu, David Peereboom, Manmeet Ahluwalia
Journal: Neuro-Oncology (2020)

Abstract BACKGROUND Ruxolitinib is a novel, potent, and selective inhibitor of JAK1 (Janus kinase 1) and JAK2 with modest to marked selectivity against TYK2 (tyrosine kinase 2) and JAK3, respectively. Dysregulation of the JAK/STAT pathway has been associated with several types of cancer and increased proliferation and survival of malignant cells. METHODS Newly diagnosed patients with unmethylated MGMT Glioblastoma or grade III glioma were recruited to Arm I. Every patient received ruxolitinib and 60 Gy radiation for 6 weeks over 6 weeks (2Gy x 30). The dose of Ruxolitinib was administered given the 3 + 3 design. Level 1 or starting dose was 10 mg twice daily, level 2 was 15 mg twice daily, level 3 was 20 mg twice daily and level -1 was 5 mg twice daily. Arm two was started once safe dose was established for Arm 1 for each dose level. Patients with methylated MGMT glioblastoma or grade III glioma were eligible for Arm 2. Every patient received ruxolitinib + radiation x 60 Gy + daily temozolomide at 75 mg/m2 for 6 weeks over 6 weeks. RESULTS 30 patients were randomized to Arm I and 16 to arm II. There was one partial response in the methylated Arm and one partial response in the unmethylated arm. The maximum tolerated dose of Ruxolitinib was 20 mg BID in both methylated and unmethylated arm. There were 14 incidences of Grade 3 toxicity including respiratory distress, seizure, gait disturbance, weakness, thrombocytopenia, cognitive disturbance, urinary retention, and bacterial meningitis. There were 4 incidences of Grade 4 toxicity including seizure, respiratory distress, somnolence and thromboembolic events. CONCLUSION This is an ongoing clinical trial and final results will be presented once the trial is complete. We will perform single cell sequencing on good and poor responders, that data will be presented in the meeting.
Intravenous transplantation of olfactory ensheathing cells reduces neuroinflammation after spinal cord injury <i>via</i> interleukin-1 receptor antagonist
Authors: Lijian Zhang, Xiaoqing Zhuang, Päivi Kotitalo, Thomas Keller, Anna Krzyczmonik, Merja Haaparanta-Solin, Olof Solin, Sarita Forsback, Tove J. Grönroos, Chunlei Han, Francisco R. López-Picón, Hechun Xia
Journal: Theranostics (2020)

Rationale: Olfactory ensheathing cell (OEC) transplantation has emerged as a promising therapy for spinal cord injury (SCI) repair.In the present study, we explored the possible mechanisms of OECs transplantation underlying neuroinflammation modulation.Methods: Spinal cord inflammation after intravenous OEC transplantation was detected in vivo and ex vivo by translocator protein PET tracer [ 18 F]F-DPA.To track transplanted cells, OECs were transduced with enhanced green fluorescent protein (eGFP) and HSV1-39tk using lentiviral vector and were monitored by fluorescence imaging and [ 18 F]FHBG study.Protein microarray analysis and ELISA studies were employed to analyze differential proteins in the injured spinal cord after OEC transplantation.The anti-inflammation function of the upregulated protein was also proved by in vitro gene knocking down experiments and OECs/microglia co-culture experiment. Results:The inflammation in the spinal cord was decreased after OEC intravenous transplantation.The HSV1-39tk-eGFP-transduced OECs showed no accumulation in major organs and were found at the injury site.After OEC transplantation, in the spinal cord tissues, the interleukin-1 receptor antagonist (IL-1Ra) was highly upregulated while many chemokines, including pro-inflammatory chemokines IL-1α, IL-1β were downregulated.In vitro studies confirmed that lipopolysaccharide (LPS) stimulus triggered OECs to secrete IL-1Ra.OECs significantly suppressed LPS-stimulated microglial activity, whereas IL-1Ra gene knockdown significantly reduced their ability to modulate microglial activity. Conclusion:The OECs that reached the lesion site were activated by the release of pro-inflammatory cytokines from activated microglia in the lesion site and secreted IL-1Ra to reduce neuroinflammation.Intravenous transplantation of OECs has high therapeutic effectiveness for the treatment of SCI via the secretion of IL-1Ra to reduce neuroinflammation.
Regulation and targeting of androgen receptor nuclear localization in castration-resistant prostate cancer
Authors: Shidong Lv, Qiong Song, Guang Chen, Erdong Cheng, Wei Chen, Ryan Cole, Zeyu Wu, Laura E. Pascal, Ke Wang, Peter Wipf, Joel B. Nelson, Qiang Wei, Wenhua Huang, Zhou Wang
Journal: Journal of Clinical Investigation (2020)

Nuclear localization of the androgen receptor (AR) is necessary for its activation as a transcription factor. Defining the mechanisms regulating AR nuclear localization in androgen-sensitive cells and how these mechanisms are dysregulated in castration-resistant prostate cancer (CRPC) cells is fundamentally important and clinically relevant. According to the classical model of AR intracellular trafficking, androgens induce AR nuclear import and androgen withdrawal causes AR nuclear export. The present study has led to an updated model that AR could be imported in the absence of androgens, ubiquitinated, and degraded in the nucleus. Androgen withdrawal caused nuclear AR degradation, but not export. In comparison with their parental androgen-sensitive LNCaP prostate cancer cells, castration-resistant C4-2 cells exhibited reduced nuclear AR polyubiquitination and increased nuclear AR level. We previously identified 3-(4-chlorophenyl)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (CPPI) in a high-throughput screen for its inhibition of androgen-independent AR nuclear localization in CRPC cells. The current study shows that CPPI is a competitive AR antagonist capable of enhancing AR interaction with its E3 ligase MDM2 and degradation of AR in the nuclei of CRPC cells. Also, CPPI blocked androgen-independent AR nuclear import. Overall, these findings suggest the feasibility of targeting androgen-independent AR nuclear import and stabilization, two necessary steps leading to AR nuclear localization and activation in CRPC cells, with small molecule inhibitors.
The Pharmacogenetics of Statin Therapy: When the Body Aches, the Mind Will Follow⁎⁎Editorials published in the Journal of the American College of Cardiologyreflect the views of the authors and do not necessarily represent the views of JACCor the American College of Cardiology.
Authors: Joseph S. Rossi, Howard L. McLeod
Journal: Journal of the American College of Cardiology (2009)
PATH-29. COMPARATIVE ANALYSIS OF DNA METHYLATION PROFILES ASSOCIATED WITH IDH-WILDTYPE GLIOMA AND GLIONEURONAL TUMOR SUBTYPES
Authors: Omkar Singh, Kenneth Aldape
Journal: Neuro-Oncology (2020)

Abstract DNA Methylation profiles are highly robust and reproducible as a classification tool. Less is understood regarding the methylation differences that exist among gliomas and glioneuronal tumors. To address this, we analyzed differentially methylated probes (DMPs) of gliomas and glioneuronal tumors compared to normal brain white matter controls. After filtering (Δbeta &amp;gt;0.3, logFC &amp;gt;±1) of significant probes we observed that low grade glioma/glioneuronal tumors (LGGs) had significantly fewer DMPs (Hyper/Hypo) as compared to GBMs. For example, posterior fossa pilocytic astrocytomas (PA’s) showed 2861 DMPs (1916 hypo/945 hyper) versus 9653 for GBM-RTKI ((6563/3090) respectively, while tumors such as PXA and anaplastic PA showed intermediate changes between LGG’s and GBMs. Hypomethylated and hypermethylated probes were analyzed for gene ontology and KEGG pathway enrichment, with LGG subtypes showing hypomethylated probes/genes associated with cell adhesion, blood vessel development and viral infection (P-value = 10-7). In contrast, hypomethylated probes in GBM subtypes were enriched for plasma membrane and cell periphery gene ontologies (P-value = 10-52). With respect to hypermethylated probes, LGG subtypes showed enrichment for myelination, glial cell differentiation and sphingolipid metabolism (P-value = 10-5) while DNA-binding transcription factor activity was seen in GBM subtypes (P-value=10-35). Examples of the most significantly hypermethylated genes in GBM included the transcription factors, GATA3 and PAX9. Intermediate-grade gliomas such as anaplastic PA and PXA showed enrichment of hypermethylated genes similar to GBM, but of lower significance (P-values = 10-6 and 10-4). Overall, understanding of cancer-associated DNA methylation changes in glioma subtypes suggests a hierarchy of biological changes that may underlie the pathogenesis of these tumors and interestingly, highlight tumor types such as PXA and anaplastic PA as having intermediate methylation changes, between benign LGG and GBM. Hypermethylation of transcription-factor genes will be investigated in GBM and compared with changes in gene expression to understand possible roles in the pathogenesis.
Модернизация энтерального пищеварения больных хроническим панкреатитом с трофологическими нарушениями после перенесенного острого панкреатита
Authors: L.S. Babinets, B.O. Мigenko
Journal: GASTROENTEROLOGY (2020)

Актуальность. Хронический панкреатит (ХП), по данным многих ученых и практиков, характеризуется прогрессирующей функциональной экзокринной и эндокринной недостаточностью поджелудочной железы (ПЖ). На фоне функциональной несостоятельности ПЖ достаточно часто возникает полинутриентная трофологическая недостаточность (ПТН), которая требует адекватной коррекции. Цель: обосновать целесообразность применения модифицированного метода лаважа кишечника для коррекции мальдигестии и полинутриентной трофологической недостаточности при хроническом панкреатите, возникшей после перенесенного острого панкреатита. Материалы и методы. Исследована эффективность применения модифицированного лаважа кишечника для улучшения энтерального пищеварения у больных ХП с признаками ПТН после перенесенного острого панкреатита (ОП). Обследованы 60 больных ХП после перенесенного в анамнезе ОП. I группа (38 па­циентов) получала общепринятый комплекс лечения ХП (ОЛ). Во II группе (22 больных) комплекс лечения дополнительно к ОЛ включал курс модифицированного лаважа кишечника, 1–5 процедур, в зависимости от тяжести клинической симптоматики, а эффективность и продолжительность определялись по лабораторным, антропометрическим и инструментальным параметрам ХП и ПТН при ХП после перенесенного ОП. Результаты. Предложенная методика кишечного лаважа имеет следующие преимущества: позволяет увеличить время лаважа кишечника за счет «пульсирующей» подачи жидкости, обеспечивает щадящее воздействие промывки на слизистую оболочку кишки, предотвращает дополнительную травматизацию кишечника при необходимости такого введения лекарственных средств. Использование двойного контура позволяет вводить лекарственные средства сразу после лаважа без вынимания зонда, предотвращая таким образом смешивание и возможные побочные эффекты при необходимости одновременного применения нескольких лекарственных средств. Выводы. Проведенный анализ уровня С-реактивного протеина показал, что этот параметр может служить маркером глубины интоксикации и остроты воспаления, а также определяющим для назначения количества процедур кишечного лаважа: при СРП 3,0–7,0 г/мл выполняли одну процедуру лаважа, при СРП 7,0–14,0 мг/л — 3 процедуры, при СРП ≥ 14,0 мг/л — 5 процедур. Применение курса процедур кишечного лаважа в общепринятом лечении пациентов с хроническим панкреатитом после перенесенного острого панкреатита привело к достоверно более эффективному уменьшению болевого синдрома, улучшению антропометрических параметров, показателей копрограммы и нормализации про- и антиоксидантных изменений.