bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting MYCN upregulates L1CAM tumor antigen in MYCN-dysregulated neuroblastoma to increase CAR T cell efficacy
Authors: Grunewald Laura , Andersch Lena , Helmsauer Konstantin , Schwiebert Silke , Klaus Anika , Henssen Anton G. , Straka Teresa , Lodrini Marco , Wicha Sebastian G. , Fuchs Steffen , Hertwig Falk , Westermann Frank , Vitali Alice , Caramel Carlotta , Büchel Gabriele , Eilers Martin , Astrahantseff Kathy , Eggert Angelika , Höpken Uta E. , Schulte Johannes H. , Blankenstein Thomas , Anders Kathleen , Künkele Annette 
Published: January 2024

AbstractBackgroundCurrent treatment protocols have only limited success in pediatric patients with neuroblastomas harboring amplifications of the central oncogene,MYCN. Adoptive T cell therapy presents an innovative strategy to improve cure rates. However, L1CAM-targeting CAR T cells achieved only limited response against refractory/relapsed neuroblastoma in an ongoing phase I trial to date. Here, we investigate how oncogenic MYCN levels influence tumor cell response to CAR T cells, as one possible factor limiting success in trials.MethodsHigh MYCN levels were induced in SK-N-AS cells harboring the normal diploidMYCNcomplement using a tetracycline-inducible system. The inducible MYCN cell model orMYCN-amplified neuroblastoma cell lines were cocultured with L1CAM-CAR T cells. CAR T cell effector function was assessed via activation marker expression (flow cytometry), cytokine release and tumor cytotoxicity (biophotonic signal assessment). The cell model was characterized using RNA sequencing, and our data compared to publicly available RNA and proteomic data sets from neuroblastomas. ChIP-sequencing data was used to determine transcriptionalL1CAMregulation by MYCN using public data sets. Synergism between CAR T cells and the MLN8237 AURKA inhibitor, which indirectly inhibits MYCN activity, was assessedin vitrousing the Bliss model andin vivoin an immunocompromised mouse model.ResultsInducing high MYCN levels in the neuroblastoma cell model reduced L1CAM expression and, consequently, L1CAM-CAR T cell effector function (activation, cytokine release and cytotoxicity)in vitro. Primary neuroblastomas possessing highMYCNlevels expressed lower levels of both theL1CAMtranscript and L1CAM tumor antigen. Indirectly inhibiting MYCN via AURKA using MLN8237 treatment restored L1CAM expression on tumor cellsin vitroand restored L1CAM-CAR T cell effector function. Combining MLN8237 and L1CAM-CAR T cell treatment synergistically increased neuroblastoma-directed killing in MYCN-overexpressing cellsin vitroandin vivoconcomitant with severein vivotoxicity.ConclusionWe shed new light on a primary resistance mechanism in MYCN-driven neuroblastoma against L1CAM-CAR T cells via target antigen downregulation. These data suggest that combining L1CAM-CAR T cell therapy with pharmacological MYCN inhibition may benefit patients with high-risk neuroblastomas harboringMYCNamplifications.
Brain Vasculature Accumulates Tau and Is Spatially Related to Tau Tangle Pathology in Alzheimer’s Disease
Authors: Hoglund Zachary , Ruiz-Uribe Nancy , del Sastre Eric , Woost Benjamin , Bailey Joshua , Hyman Bradley T. , Zwang Theodore , Bennett Rachel E. 
Published: January 2024

AbstractInsoluble pathogenic proteins accumulate along blood vessels in conditions of cerebral amyloid angiopathy (CAA), exerting a toxic effect on vascular cells and impacting cerebral homeostasis. In this work we provide new evidence from three-dimensional human brain histology that tau protein, the main component of neurofibrillary tangles, can similarly accumulate along brain vascular segments. We quantitatively assessed n=6 Alzheimer’s disease (AD), and n=6 normal aging control brains and saw that tau-positive blood vessel segments were present in all AD cases. Tau-positive vessels are enriched for tau at levels higher than the surrounding tissue and appear to affect arterioles across cortical layers (I-V). Further, vessels isolated from these AD tissues were enriched for N-terminal tau and tau phosphorylated at T181 and T217. Importantly, tau-positive vessels are associated with local areas of increased tau neurofibrillary tangles. This suggests that accumulation of tau around blood vessels may reflect a local clearance failure. In sum, these data indicate tau, like amyloid beta, accumulates along blood vessels and may exert a significant influence on vasculature in the setting of AD.
Multi-Channel smFRET study reveals a Compact conformation of EF-G on the Ribosome
Authors: Johnson Jordan L. , Steele Jacob H. , Lin Ran , Stepanov Victor G. , Gavriliuc Miriam N. , Wang Yuhong 
Published: January 2024

ABSTRACTWhile elongation factor G (EF-G) is crucial for ribosome translocation, the role of its GTP hydrolysis remains ambiguous. EF-G’s indispensability is further exemplified by the phosphorylation of human eukaryotic elongation factor 2 (eEF2) at Thr56, which inhibits protein synthesis globally, but its exact mechanism is not clear. In this study, we developed a multi-channel single-molecule FRET (smFRET) microscopy methodology to examine the conformational changes ofE. coliEF-G induced by mutations that closely aligned with eEF2’s Thr56 residue. We utilized Alexa 488/594 double-labeled EF-G to catalyze the translocation of fMet-Phe-tRNAPhe-Cy3 inside Cy5-L27 labeled ribosomes, allowing us to probe both processes within the same complex. Our findings indicate that in the presence of either GTP or GDPCP, wild-type EF-G undergoes a conformational extension upon binding to the ribosome to promote normal translocation. On the other hand, T48E and T48V mutations did not affect GTP/GDP binding or GTP hydrolysis, but impeded Poly(Phe) synthesis and caused EF-G to adopt a unique compact conformation, which wasn’t observed when the mutants interact solely with the sarcin/ricin loop. This study provides new insights into EF-G’s adaptability and sheds light on the modification mechanism of human eEF2.
Mapping the developmental structure of stereotyped and individual-unique behavioral spaces inC. elegans
Authors: Harel Yuval , Nasser Reemy Ali , Stern Shay 
Published: January 2024

AbstractDevelopmental patterns of behavior are variably organized in time and among different individuals. However, long-term behavioral diversity was previously studied using pre-defined behavioral parameters, representing a limited fraction of the full individuality structure. Here, we continuously extracted ∼1.2 billion body postures of ∼2,200 singleC. elegansindividuals throughout their full development time, to create a complete developmental atlas of stereotyped and individual-unique behavioral spaces. Unsupervised inference of low-dimensional movement modes of each single individual revealed a dynamic developmental trajectory of stereotyped behavioral spaces and exposed unique behavioral trajectories of individuals that deviate from the stereotyped patterns. Moreover, classification of behavioral spaces within tens of neuromodulatory- and environmentally-perturbed populations efficiently uncovered plasticity in the temporal structures of stereotyped behavior and individuality. These results present a comprehensive atlas of continuous behavioral dynamics across development time and a general framework for unsupervised dissection of shared and unique developmental signatures of behavior.
Autophagy regulator ATG5 preserves cerebellar function by safeguarding its glycolytic activity
Authors: Tutas J. , Tolve M. , Özer-Yildiz E. , Ickert L. , Klein I , Hosseini M , Liebsch F. , Dethloff F. , Gavalisco P. , Endepols H. , Yang M. , Georgomanolis T. , Neumaier B. , Drzezga A. , Daun S. , Schwarz G. , Thorens B. , Gatto G. , Frezza C. , Kononenko NL 
Published: January 2024

AbstractDysfunctions in autophagy, a highly conserved cellular mechanism responsible for the degradation of intracellular components within lysosomes, often result in neurodegeneration. The neuroprotective effect of autophagy varies across neuronal subtypes, and the mechanisms of selective vulnerability of neurons to autophagy dysfunction are currently unknown. Utilizing a mouse model of ATG5 deficiency in inhibitory neurons and a comprehensive approach, including PET imaging, metabolomics, stable-isotope labeling studies, and live cell imaging, we establish that autophagy contributes to the survival of cerebellar Purkinje cells (PCs) by safeguarding their glycolytic activity. We show that the core autophagy protein ATG5 downregulates the levels of the glucose transporter 2 (GLUT2) during brain maturation. Autophagy-deficient PCs exhibit increased glucose uptake, along with elevated levels of glycolytic intermediates and methylglyoxal-modified proteins. We propose lysophosphatidic acid and serine as glycolytic intermediates inducing PC death and demonstrate that deletion of GLUT2 in ATG5-deficient mice mitigates PC neurodegeneration and restores their ataxic gait. Taken together, this study reveals a novel neuroprotective role of autophagy in preventing excessive glycolytic metabolism in the brain.
Visualizing chaperone-mediated multistep assembly of the human 20S proteasome
Authors: Adolf Frank , Du Jiale , Goodall Ellen A. , Walsh Richard M. , Rawson Shaun , von Gronau Susanne , Harper J. Wade , Hanna John , Schulman Brenda A. 
Published: January 2024

AbstractDedicated assembly factors orchestrate stepwise production of many molecular machines, including the 28-subunit proteasome core particle (CP) that mediates protein degradation. Here, we report cryo-EM reconstructions of seven recombinant human subcomplexes that visualize all five chaperones and the three active site propeptides across a wide swath of the assembly pathway. Comparison of these chaperone-bound intermediates and a matching mature CP reveals molecular mechanisms determining the order of successive subunit additions, and how proteasome subcomplexes and assembly factors structurally adapt upon progressive subunit incorporation to stabilize intermediates, facilitate the formation of subsequent intermediates, and ultimately rearrange to coordinate proteolytic activation with gated access to active sites. The structural findings reported here explain many previous biochemical and genetic observations. This work establishes a methodologic approach for structural analysis of multiprotein complex assembly intermediates, illuminates specific functions of assembly factors, and reveals conceptual principles underlying human proteasome biogenesis.
Turnover of RNA-binding Proteins and MicroRNAs by intrinsically disordered region-directed ZSWIM8 ubiquitin ligase during brain development
Authors: Lei Jing , Zhong Siming , Fan Rong , Shu Xin , Wang Guan , Guo Jiansheng , Xue Shuting , Zheng Luqian , Ren Aiming , Ji Junfang , Yang Bing , Duan Shumin , Wang Zhiping , Guo Xing 
Published: January 2024

AbstractWidely present in mammalian proteomes, intrinsically disordered regions (IDRs) in proteins play important biological functions by conferring structural flexibility and mediating biomolecular interactions. IDR-containing proteins, including many RNA-binding proteins (RBPs), are prone to misfolding and aggregation and must be constantly monitored. Here we show that the conserved ZSWIM8-type Cullin-RING ubiquitin ligase (CRLZSWIM8) is a master regulator of such proteins during brain development. ZSWIM8 selects its substrates via an IDR-dependent mechanism, and deletion of ZSWIM8 causes aberrant accumulation of numerous RBPs including AGO2 and ELAV1 in neonatal brains. Furthermore, AGO2 ubiquitination by ZSWIM8 is triggered by microRNA binding, leading to target-directed microRNA degradation (TDMD) of MiR7. Dysregulation of MiR7 in the absence of ZSWIM8 results in defects in oligodendrocyte maturation and functions. Together, our findings have demonstrated that, by utilizing variable target-recognition strategies, ZSWIM8 controls the abundance of conformationally flexible RBPs and miRNA metabolism that are essential for brain development.TeaserA conserved ubiquitin ligase controls the quality of disordered proteins to ensure brain development.
In vitrosurvival and neurogenic potential of central canal-derived neural stem cells depend on spinal cord injury type
Authors: Schiro Lars Erik , Bauer Ulrich Stefan , Bjorkli Christiana , Sandvig Axel , Sandvig Ioanna 
Published: January 2024

AbstractThe central canal (CC) of the spinal cord is a neurogenic niche consisting of quiescent neural stem cells (NSCs) capable of responding to traumatic damage to the spinal cord by increasing their proliferative activity and sending migrating progeny toward the site of injury, where they contribute to the formation of the glial scar. However, CC NSCs have been demonstrated to have the capability to differentiate into all neural lineage cellsin vitro, but alsoin vivo, in response to infusion of specific growth factors that promote neuronal induction after injury, as well as when transplanted into other neurogenic niches, such as the subgranular zone of the hippocampus. This suggests that CC NSCs may represent a recruitable endogenous source of neural lineage cells that could be harnessed to replenish damaged or lost neural tissue after traumatic spinal cord injury (SCI).NSCs isolated from the CC neurogenic niche of uninjured rats and mice have been shown to display limited proliferative capacityin vitro, with significantly greater proliferative activity achieved with NSCs isolated from SCI-lesioned rats and mice indicating an injury-specific activation of the quiescent CC NSC pool. A central question that currently remains unanswered is whether, and to what extent the CC niche can spontaneously generate viable neurons, and act as a potential source of new cells to replace lost neuronal populationsin situ, and whether SCI sequalae impact future NSC neurogenic potential. To address this question, we need to understand whether the nature of the injury plays a role in the CC neurogenic niche response. In this study, we compared the intrinsic proliferative response and neurogenic potential of NSCs harvested from the CC neurogenic niche in adult female Sprague Dawley rats by culturing said NSCs across three conditions; (i) control, i.e., uninjured tissue, (ii) afterin vivocompression injury 3 days before harvesting, and (iii) afterin vivosimulated burst fracture injury 3 days before harvestingin vitro. We found that lacerations of the dura mater surrounding the spinal cord during a compression injury resulted in drastically altered and persistentin vitroNSC behavior encompassing both proliferation and development compared to uninjured control and compression injury with the dura intact.
Functional characterization of CCHamides and deorphanization of their receptors in the yellow fever mosquito,Aedes aegypti
Authors: Tan Jinghan , Neupert Susanne , Paluzzi Jean-Paul 
Published: January 2024

AbstractAs a widely distributed anthropophilic mosquito species and vector of various arboviruses,Aedes aegyptiposes a significant threat to human health on a global scale. Investigating mosquito neuropeptides allows us to better understand their physiology. The neuropeptides CCHamide1 (CCHa-1) and CCHamide-2 (CCHa-2) and their associated G protein-coupled receptors (CCHa-1R and CCHa-2R) were recently identified and studied across insects. However, expression profiles and physiological roles of CCHamides and their receptors in many other insects, includingA. aegypti, remain unclear. This research aimed to quantify and localize the expression of CCHamides along with their receptors and to elucidate their physiological function in the yellow fever mosquito. RT-qPCR analysis revealed transcript abundance of CCHamides and receptors changes over development. Differential expression was also observed in tissues/organs of adult mosquitoes indicatingCCHa-1andCCHa-2transcripts are most highly enriched in the midgut, while receptors are expressed across various tissues. Further, CCHamides were immunolocalized in neurons in the brain and ventral nerve cord along with enteroendocrine cells in the posterior midgut adjacent to the midgut-hindgut junction, corroborating their transcript expression profiles. Using different mass spectrometrical approaches, presence of CCHamides were confirmed in the brain of both sexes, including thepars intercerebralisof female mosquitoes, as well as in the gut of adult mosquitoes. For chemical identification of predicted CCHamides, we analyzed brain and gut extracts by ESI-Q Exactive Orbitrap MS and resulting fragmentations confirmed CCHa1 and CCHa2 in brain and midgut samples of both male and female mosquitoes. A heterologous functional assay was used to confirm the specificity and sensitivity of the two CCHamide receptors by assessing their activation in response to diverse mosquito peptides, which confirmed CCHa-1 and CCHa-2 as the natural ligands. Finally, using a capillary feeder (CAFE) bioassay, our results suggest that CCHa-2 modulates feeding behaviour in female mosquitoes.
Disruption of autism-associatedPcdh9gene leads to transcriptional alterations, synapses overgrowth and aberrant excitatory transmission in the CA1
Authors: Miozzo Federico , Murru Luca , Maiellano Greta , Zippo Antonio , Moretto Edoardo , Avendano Annalaura Zambrano , Metodieva Verjinia D. , Riccardi Sara , D’Aliberti Deborah , Spinelli Silvia , Canu Tamara , Chaabane Linda , Hirano Shinji , Kas Martien J. H. , Francolini Maura , Piazza Rocco , Passafaro Maria 
Published: January 2024

ABSTRACTProtocadherins are cell adhesion molecules with crucial role in cell-cell contacts, whose mutations or altered expression have been implicated in multiple brain disorders. In particular, growing evidence links genetic alterations inProtocadherin 9(PCDH9) gene with Autism Spectrum Disorder (ASD) and Major Depression Disorder (MDD). Furthermore,Pcdh9deletion induces neuronal defects in the mouse somatosensory cortex, accompanied by sensorimotor and memory impairment. However, the synaptic and molecular mechanisms underlyingPcdh9physiological function and its involvement in brain pathology remain largely unknown. To this aim, we conducted a comprehensive investigation of PCDH9 role in the mouse hippocampus at the ultrastructural, biochemical, transcriptomic, electrophysiological and network level. We show that PCDH9 mainly localizes at glutamatergic synapses and its expression peaks in the first week after birth, a crucial time window for synaptogenesis. Strikingly,Pcdh9KO neurons exhibit oversized presynaptic terminal and postsynaptic density (PSD) in the CA1. Synapse overgrowth is sustained by the broad up-regulation of synaptic genes and the dysregulation of key drivers of synapse morphogenesis, as revealed by single-nucleus RNAseq. Synaptic and transcriptional defects are accompanied by increased EPSC frequency and disturbances in the hippocampal network activity ofPcdh9KO mice. In conclusion, our work indicates thatPcdh9regulates the morphology and function of excitatory synapses in the CA1, thereby affecting glutamatergic transmission in hippocampal circuitries.
Analyzing the Usage of the GIM-COVID-19 Long-Term Sequelae Rapid Access to Consultative Expertise (RACE) Line
Authors: Kaushal Saniya , Birks Peter , Greiner Jesse , Levin Adeera , Malbeuf Michelle , Schwartz Zachary 
Published: January 2024

AbstractReal time access to guidance for physicians has been offered through Rapid Access to Consultative Expertise (RACE) in British Columbia (BC) for the past 12 years.1In the context of the novel coronavirus (COVID-19), the service for RACE was expanded to include a Long-COVID RACE line.1We report here the types and frequencies of questions asked to General Internal Medicine (GIM) experts in Long-COVID, by general practitioners in BC. 149 calls over an 11-month period were tracked by GIM experts, and analysis of the call themes was undertaken. These calls mainly involved consults regarding the post-infection COVID-19 symptoms being experienced by Primary Care Practitioners’ patients. Respiratory symptoms were the leading type of symptoms reported, with shortness of breath, cough, fatigue, and fevers being the most common, respectively. This data will be used to inform future resource utilization and provide insights on the usage of the Long-COVID RACE line.
Practical Challenges for Commercial Enterprises in the Ethics Review Process for Digital Health Research: Document Analysis and Interview Study
Authors: Yang Katherine , Potts Henry W W 
Published: January 2024

AbstractIntroductionThe rapid evolution of digital health interventions has created challenges in navigating the ethics approval process for commercial enterprises. Recognising the need for processes that balance ethical considerations with the specifics of digital health research, this study aimed to describe what happens when enterprises seek ethical review in the UK and propose strategies for a smoother process.MethodsInductive thematic analysis was conducted on thirty-two ethics review documents (29 to an NHS Research Ethics Committee, 3 to an ethics committee at a higher education institution) submitted by digital health developers with commercial sponsors and ten semi-structured interviews with digital health enterprise representatives.ResultsEthics committees raised an average of 4.3 action points per submission. We identified five broad themes around committees’ concerns: ethical commitments in care; study design; digital health research peculiarities; data governance; document quality and completeness. Interviewees reported a range of experiences. Here, we identified six broad themes: submission and protocol revisions; the dynamic between parties; application time and procedures; acumen and practicality in digital health; support and guidance from RECs; enterprise expertise and resources.ConclusionWe suggest strategies for applicants to achieve a favourable decision, such as evidence-based study designs and participant support for better inclusion and equity, and identified specific pitfalls to avoid, such as lack of justification for data governance procedures. We recommend that UK research ethics committees provide adapted guidance and foster collaboration through open communication and mutual understanding, to facilitate a smoother approval process in digital health research.