Biology subjects

Gordon, N.

Publications and source records attributed to Gordon, N..

7 recordsLinked to original sources

bioRxiv2025

FOXO1 inhibition and FADD knockdown have opposing effects on anticancer drug-induced cytotoxicity and p21 expression in osteosarcoma

Forkhead box class O1 (FOXO1) and fas-associated death domain (FADD) regulate cell death pathways and homeostatic processes such as cell cycle progression and apoptosis. FADD phosphorylation promotes nuclear localization of FOXO1 and FOXO1 regulates FADD expression. Therefore, it is plausible that FOXO1 and FADD have synergistic or antagonistic effects on cell cycle regulation and the response to anticancer drug treatment in cancer cells. In the present study we report that AS1842856-mediated inhibition of FOXO1 reverses anticancer drug-induced cytotoxicity while FADD knockdown increases anticancer drug-induced cytotoxicity in osteosarcoma (OS). Reversed anticancer drug-induced cytotoxicity was accompanied with G2/M cell cycle arrest and increased expression of p21. The anti-cancer function of FOXO1 was further supported by the observation that OS cells that express higher basal levels of FOXO1 had increased sensitivity to camptothecin-induced cytotoxicity. FADD knockdown reversed FOXO1 inhibition-induced increase in p21 expression. The results presented in this study indicate that FOXO1 has a tumor suppressor function while FADD has a tumor promoting function in OS following anticancer drug treatment. The experimental approach used in this investigation also indicates that FADD antagonists the effect of FOXO1 on p21 expression in OS.

cancer biology↗

bioRxiv2024

Labeling Natural Killer cells with superparamagnetic iron oxide nanoparticles for detection by preclinical and clinical-scale magnetic particle imaging

IntroductionClinical adoption of NK cell immunotherapy is underway for medulloblastoma and osteosarcoma, however there is currently little feedback on cell fate after administration. We propose magnetic particle imaging (MPI) for the detection, localization, and quantification of VivoTrax-labeled NK cells. MethodsHuman-derived NK-92 cells were labeled by co-incubation with VivoTrax for 24 hours then the excess nanoparticles were washed with centrifugation. Cytolytic activity of labeled vs. unlabeled NK-92 cells was assessed after 4 hours of co- incubation with medulloblastoma cells (DAOY) or osteosarcoma cells (LM7 or OS17) using bioluminescent or GFP counts. Labeled NK-92 cells at two different doses (0.5 or 1 x 106) were administered to excised mouse brains (cerebellum), tibias, and lungs then imaged by 3D preclinical MPI (MOMENTUM imager) and localized relative to fiducial markers. NK-92 cells were imaged by clinical-scale MPI under development at Magnetic Insight Inc. ResultsNK-92 cells were labeled with an average of 3.17 pg Fe/cell with no measured effects on cell viability or cytolytic activity against 3 tumor cell lines. MPI signal was directly quantitative with the number of VivoTrax-labeled NK-92 cells, with preclinical limit of detection of 3.1 x 104 cells on MOMENTUM imager. Labeled NK-92 cells could be accurately localized in mouse brains, tibias, and lungs within < 1 mm of stereotactic injection coordinates with preclinical scanner. Feasibility for detection of a clinically relevant dose of 4 x 107 labeled NK-92 cells was demonstrated on clinical-scale MPI. ConclusionMPI can provide sensitive, quantitative, and accurate spatial information on NK cell delivery, showing its potential to resolve a significant unmet clinical need to track NK cell treatments in patients.

biophysics↗

bioRxiv2024

Thermal Characterization and Preclinical Validation of an Accessible, Carbon Dioxide-Based Cryotherapy System

To investigate the potential of an affordable cryotherapy device for accessible treatment of breast cancer, the performance of a novel carbon dioxide-based device was evaluated through both benchtop and in vivo canine models. This novel device was quantitatively compared to a commercial device that utilizes argon gas as the cryogen. The thermal behavior of each device was characterized through calorimetry and by measuring the temperature profiles of iceballs generated in tissue phantoms. A 45-minute treatment from the carbon dioxide device in a tissue phantom produced a 1.67 {+/-} 0.06 cm diameter lethal isotherm that was equivalent to a 7-minute treatment from the commercial argon-based device which produced a 1.53 {+/-} 0.15 cm diameter lethal isotherm. In vivo validation was performed with the carbon dioxide-based device in one spontaneously occurring canine mammary mass with two standard 10-minutes freezes. Following cryotherapy, this mass was surgically resected and analyzed for necrosis margins via histopathology. The histopathology margin of necrosis from the in vivo treatment with the carbon dioxide device at 14 days post cryoablation was 1.57 cm. While carbon dioxide gas has historically been considered an impractical cryogen due to its low working pressure and high boiling point, this study shows that carbon dioxide-based cryotherapy may be equivalent to conventional argon-based cryotherapy in the size of the ablation zone in a standard treatment time. The validation of the carbon dioxide device performed in this study is an important step towards bringing accessible breast cancer treatment to women in low-resource settings.

bioengineering↗

bioRxiv2023

PARP inhibition and pharmacological ascorbate demonstrate synergy in castration-resistant prostate cancer

Prostate cancer (PCa) is the second leading cause of cancer death for men in the United States. While organ-confined disease has reasonable expectation of cure, metastatic PCa is universally fatal upon recurrence during hormone therapy, a stage termed castration-resistant prostate cancer (CRPC). Until such time as molecularly defined subtypes can be identified and targeted using precision medicine, it is necessary to investigate new therapies that may apply to the CRPC population as a whole. The administration of ascorbate, more commonly known as ascorbic acid or Vitamin C, has proved lethal to and highly selective for a variety of cancer cell types. There are several mechanisms currently under investigation to explain how ascorbate exerts anti-cancer effects. A simplified model depicts ascorbate as a pro-drug for reactive oxygen species (ROS), which accumulate intracellularly and generate DNA damage. It was therefore hypothesized that poly(ADP-ribose) polymerase (PARP) inhibitors, by inhibiting DNA damage repair, would augment the toxicity of ascorbate. ResultsTwo distinct CRPC models were found to be sensitive to physiologically relevant doses of ascorbate. Moreover, additional studies indicate that ascorbate inhibits CRPC growth in vitro via multiple mechanisms including disruption of cellular energy dynamics and accumulation of DNA damage. Combination studies were performed in CRPC models with ascorbate in conjunction with escalating doses of three different PARP inhibitors (niraparib, olaparib, and talazoparib). The addition of ascorbate augmented the toxicity of all three PARP inhibitors and proved synergistic with olaparib in both CRPC models. Finally, the combination of olaparib and ascorbate was tested in vivo in both castrated and non-castrated models. In both cohorts, the combination treatment significantly delayed tumor growth compared to monotherapy or untreated control. ConclusionsThese data indicate that pharmacological ascorbate is an effective monotherapy at physiological concentrations and kills CRPC cells. Ascorbate-induced tumor cell death was associated with disruption of cellular energy dynamics and accumulation of DNA damage. The addition of PARP inhibition increased the extent of DNA damage and proved effective at slowing CRPC growth both in vitro and in vivo. These findings nominate ascorbate and PARPi as a novel therapeutic regimen that has the potential to improve CRPC patient outcomes.

cancer biology↗

bioRxiv2022

Dissociation protocols used for sarcoma tissues bias the transcriptome observed in single-cell and single-nucleus RNA sequencing

BackgroundSingle-cell RNA-seq has emerged as an innovative technology used to study complex tissues and characterize cell types, states, and lineages at a single-cell level. Classification of bulk tumors by their individual cellular constituents has also created new opportunities to generate single-cell atlases for many organs, cancers, and developmental models. Despite the tremendous promise of this technology, recent evidence studying epithelial tissues and diverse carcinomas suggests the methods used for tissue processing, cell disaggregation, and preservation can significantly bias gene expression and alter the observed cell types. To determine whether sarcomas - tumors of mesenchymal origin - are subject to the same technical artifacts, we profiled patient-derived tumor explants (PDXs) propagated from three aggressive subtypes: osteosarcoma, Ewing sarcoma (ES), desmoplastic small round cell tumor (DSRCT). Given the rarity of these sarcoma subtypes, we explored whether single-nuclei RNA-seq from more widely available archival frozen specimens could accurately be identified by gene expression signatures linked to tissue phenotype or pathognomonic fusion proteins. ResultsWe systematically assessed dissociation methods across different sarcoma subtypes. We compared gene expression from single-cell and single-nucleus RNA-sequencing of 125,831 whole-cells and nuclei from ES, DSRCT, and osteosarcoma PDXs. We detected warm dissociation artifacts in single-cell samples and gene length bias in single-nucleus samples. Classic sarcoma gene signatures were observed regardless of dissociation method. In addition, we showed that dissociation method biases can be computationally corrected. ConclusionsWe highlighted transcriptional biases, including warm dissociation and gene-length biases, introduced by the dissociation method for various sarcoma subtypes. This work is the first to characterize how the dissociation methods used for sc/snRNA-seq may affect the interpretation of the molecular features in sarcoma PDXs.

cancer biology↗

bioRxiv2018

Expectation and attention increase the integration of top-down and bottom-up signals in perception through different pathways

Perception results from the integration of incoming sensory information with pre-existing information available in the brain. In this EEG (electroencephalography) study we utilised the Hierarchical Frequency Tagging method to examine how such integration is modulated by expectation and attention. Using intermodulation (IM) components as a measure of non-linear signal integration, we show in three different experiments that both expectation and attention enhance integration between top-down and bottom-up signals. Based on multispectral phase coherence, we present two direct physiological measures to demonstrate the distinct yet related mechanisms of expectation and attention. Specifically, our results link expectation to the modulation of prediction signals and the integration of top-down and bottom-up information at lower levels of the visual hierarchy. Meanwhile, they link attention to the propagation of ascending signals and the integration of information at higher levels of the visual hierarchy. These results are consistent with the predictive coding account of perception.

neuroscience↗

bioRxiv2016

Hierarchical Frequency Tagging reveals neural markers of predictive coding under varying uncertainty

Understanding the integration of top-down and bottom-up signals is essential for the study of perception. Current accounts of predictive coding describe this in terms of interactions between state units encoding expectations or predictions, and error units encoding prediction error. However, direct neural evidence for such interactions has not been well established. To achieve this, we combined EEG methods that preferentially tag different levels in the visual hierarchy: Steady State Visual Evoked Potential (SSVEP at 10Hz, tracking bottom-up signals) and Semantic Wavelet-Induced Frequency Tagging (SWIFT at 1.3Hz tracking top-down signals). Importantly, we examined intermodulation components (IM, e.g., 11.3Hz) as a measure of integration between these signals. To examine the influence of expectation and predictions on the nature of such integration, we constructed 50-second movie streams and modulated expectation levels for upcoming stimuli by varying the proportion of images presented across trials. We found SWIFT, SSVEP and IM signals to differ in important ways. SSVEP was strongest over occipital electrodes and was not modified by certainty. Conversely, SWIFT signals were evident over temporo- and parieto-occipital areas and decreased as a function of increasing certainty levels. Finally, IMs were evident over occipital electrodes and increased as a function of certainty. These results link SSVEP, SWIFT and IM signals to sensory evidence, predictions, prediction errors and hypothesis-testing - the core elements of predictive coding. These findings provide neural evidence for the integration of top-down and bottom-up information in perception, opening new avenues to studying such interactions in perception while constraining neuronal models of predictive coding.\n\nSIGNIFICANCE STATEMENTThere is a growing understanding that both top-down and bottom-up signals underlie perception. But how do these signals interact? And how does this process depend on the signals probabilistic properties? Predictive coding theories of perception describe this in terms how well top-down predictions fit with bottom-up sensory input. Identifying neural markers for such signal integration is therefore essential for the study of perception and predictive coding theories in particular. The novel Hierarchical Frequency Tagging method simultaneously tags top-down and bottom-up signals in EEG recordings, while obtaining a measure for the level of integration between these signals. Our results suggest that top-down predictions indeed integrate with bottom-up signals in a manner that is modulated by the predictability of the sensory input.

neuroscience↗