bluuebunny/crossref_metadata_2025_split huggingface dataset:
Shielding of actin by the endoplasmic reticulum impacts nuclear positioning
Authors: Janota Cátia Silva , Pinto Andreia , Pezzarossa Anna , Machado Pedro , Costa Judite , Campinho Pedro , Franco Cláudio A. , Gomes Edgar R. 
Published: May 2022

AbstractNuclear position is central to cell polarization, and its disruption is associated with various pathologies. The nucleus is moved away from the leading edge of migrating cells through its connection to moving dorsal actin cables, and the absence of connections to immobile ventral stress fibers. It is unclear how these asymmetric nucleo-cytoskeleton connections are established. Here, using an in vitro wound assay, we find that remodeling of endoplasmic reticulum (ER) impacts nuclear positioning through the formation of a barrier that shields immobile ventral stress fibers. The remodeling of ER and perinuclear ER accumulation is mediated by the ER shaping protein Climp-63. Furthermore, ectopic recruitment of the ER to stress fibers restores nuclear positioning in the absence of Climp-63. Our findings suggest that the ER mediates asymmetric nucleo-cytoskeleton connections to position the nucleus.
TUBB4A interacts with MYH9 to protect the nucleus during cell migration and promotes prostate cancer via GSK3β/β-catenin signalling
Authors: Gao Song , Wang Shuaibin , Zhao Zhiying , Zhang Chao , Liu Zhicao , Ye Ping , Xu Zhifang , Yi Baozhu , Jiao Kai , Naik Gurudatta A. , Wei Shi , Rais-Bahrami Soroush , Bae Sejong , Yang Wei-Hsiung , Sonpavde Guru , Liu Runhua , Wang Lizhong 
Published: May 2022

AbstractHuman tubulin beta class IVa (TUBB4A) is a member of the β-tubulin family. In most normal tissues, expression of TUBB4A is little to none, but it is highly expressed in human prostate cancer. Here we show that high expression levels of TUBB4A are associated with aggressive prostate cancers and poor patient survival, especially for African-American men. Additionally, in prostate cancer cells, TUBB4A knockout (KO) reduces cell growth and migration but induces DNA damage through increased γH2AX and 53BP1. Furthermore, during constricted cell migration, TUBB4A interacts with MYH9 to protect the nucleus, but either TUBB4A KO or MYH9 knockdown leads to severe DNA damage and reduces the NF-κB signaling response. Also, TUBB4A KO retards tumor growth and metastasis. Functional analysis reveals that TUBB4A/GSK3β binds to the N-terminal of MYH9, and that TUBB4A KO reduces MYH9-mediated GSK3β ubiquitination and degradation, leading to decreased activation of β-catenin signaling and its relevant epithelial-mesenchymal transition. Likewise, prostate-specific deletion of Tubb4a reduces spontaneous tumor growth and metastasis via inhibition of NF-κB, cyclin D1, and c-MYC signaling activation. Our results suggest an oncogenic role of TUBB4A and provide a potentially actionable therapeutic target for prostate cancers with TUBB4A overexpression.
The phosphatase PTEN links platelets with immune regulatory functions of mouse T follicular helper cells
Authors: Chen Xue , Xu Yanyan , Chen Qidi , Zhang Heng , Zeng Yu , Geng Yan , Shen Lei , Li Fubin , Chen Lei , Chen Guo-Qiang , Huang Chuanxin , Liu Junling 
Published: May 2022

AbstractBeyond a function in hemostasis and thrombosis, platelets can regulate innate and adaptive immune responses. Hyperactive platelets are frequently associated with multiple human autoimmune diseases, yet their pathogenic functions in these diseases have not been fully established. Emerging studies show an essential function of the phosphatase and tensin homolog (PTEN) in maintenance of immune homeostasis. Here, we show that mice with platelet-specific deletion of Pten, develop age-related lymphoproliferative diseases and humoral autoimmunity not seen in wildtype animals. Platelet-specific Pten-deficient mice have aberrant T cell activation, excessive T follicular helper (Tfh) cell responses and accumulation of platelet aggregates in lymph nodes. Transferred Pten-deficient platelets are able to infiltrate into the peripheral lymphoid tissues and form more aggregates. Moreover, Pten-deficient platelets are hyperactive and overproduce multiple Tfh-promoting cytokines via activation of the PDK1/mTORC2-AKT-SNAP23 pathway. Pten-deficient platelets show enhanced interaction with CD4+ T cells and promote conversion of CD4+ T cells into Tfh cells. Our results implicate PTEN in platelet-mediated immune homeostasis, and provide evidence that hyperactive platelets function as an important mediator in autoimmune diseases using mouse models.
C. elegans enteric motor neurons fire synchronized action potentials underlying the defecation motor program
Authors: Jiang Jingyuan , Su Yifan , Zhang Ruilin , Li Haiwen , Tao Louis , Liu Qiang 
Published: May 2022

AbstractC. elegansneurons were thought to be non-spiking until our recent discovery of action potentials in the sensory neuron AWA; however, the extent to which theC. elegansnervous system relies on analog or digital coding is unclear. Here we show that the enteric motor neurons AVL and DVB fire synchronous all-or-none calcium-mediated action potentials following the intestinal pacemaker during the rhythmicC. elegansdefecation behavior. AVL fires unusual compound action potentials with each depolarizing calcium spike mediated by UNC-2 followed by a hyperpolarizing potassium spike mediated by a repolarization-activated potassium channel EXP-2. Simultaneous behavior tracking and imaging in free-moving animals suggest that action potentials initiated in AVL propagate along its axon to activate precisely timed DVB action potentials through the INX-1 gap junction. This work identifies a novel circuit of spiking neurons inC. elegansthat uses digital coding for long-distance communication and temporal synchronization underlying reliable behavioral rhythm.
Neuronal hyperexcitability drives central and peripheral nervous system tumor progression in models of neurofibromatosis-1
Authors: Anastasaki Corina , Mo Juan , Chen Ji-Kang , Chatterjee Jit , Pan Yuan , Scheaffer Suzanne M. , Cobb Olivia , Monje Michelle , Le Lu Q. , Gutmann David H. 
Published: May 2022

AbstractNeuronal activity is emerging as a driver of central and peripheral nervous system cancers. Here, we examined neuronal physiology in mouse models of the tumor predisposition syndrome Neurofibromatosis-1 (NF1), with different propensities to develop nervous system cancers. We show that central and peripheral nervous system neurons from mice with tumor-causing Nf1 gene mutations exhibit hyperexcitability and increased secretion of activity-dependent tumor-promoting paracrine factors. We discovered a neurofibroma mitogen (COL1A2) produced by peripheral neurons in an activity-regulated manner, which increases NF1-deficient Schwann cell proliferation, establishing that neurofibromas are regulated by neuronal activity. In contrast, mice with the Arg1809Cys Nf1 mutation, found in NF1 patients lacking neurofibromas or optic gliomas, do not exhibit neuronal hyperexcitability or develop these NF1-associated tumors. The hyperexcitability of tumor-prone Nf1-mutant neurons results from reduced NF1-regulated hyperpolarization-activated cyclic nucleotide-gated (HCN) channel function, such that neuronal excitability, activity-regulated paracrine factor production, and tumor progression are attenuated by HCN channel activation. Collectively, these findings reveal that NF1 mutations act at the level of neurons to modify tumor predisposition by increasing neuronal excitability and activity-regulated paracrine factor production.
A bio-functional polymer that prevents retinal scarring through modulation of NRF2 signalling pathway
Authors: Parikh Bhav Harshad , Liu Zengping , Blakeley Paul , Lin Qianyu , Singh Malay , Ong Jun Yi , Ho Kim Han , Lai Joel Weijia , Bogireddi Hanumakumar , Tran Kim Chi , Lim Jason Y. C. , Xue Kun , Al-Mubaarak Abdurrahmaan , Yang Binxia , R Sowmiya , Regha Kakkad , Wong Daniel Soo Lin , Tan Queenie Shu Woon , Zhang Zhongxing , Jeyasekharan Anand D. , Barathi Veluchamy Amutha , Yu Weimiao , Cheong Kang Hao , Blenkinsop Timothy A. , Hunziker Walter , Lingam Gopal , Loh Xian Jun , Su Xinyi 
Published: May 2022

AbstractOne common cause of vision loss after retinal detachment surgery is the formation of proliferative and contractile fibrocellular membranes. This aberrant wound healing process is mediated by epithelial-mesenchymal transition (EMT) and hyper-proliferation of retinal pigment epithelial (RPE) cells. Current treatment relies primarily on surgical removal of these membranes. Here, we demonstrate that a bio-functional polymer by itself is able to prevent retinal scarring in an experimental rabbit model of proliferative vitreoretinopathy. This is mediated primarily via clathrin-dependent internalisation of polymeric micelles, downstream suppression of canonical EMT transcription factors, reduction of RPE cell hyper-proliferation and migration. Nuclear factor erythroid 2–related factor 2 signalling pathway was identified in a genome-wide transcriptomic profiling as a key sensor and effector. This study highlights the potential of using synthetic bio-functional polymer to modulate RPE cellular behaviour and offers a potential therapy for retinal scarring prevention.
Topography-induced large-scale antiparallel collective migration in vascular endothelium
Authors: Leclech Claire , Gonzalez-Rodriguez David , Villedieu Aurélien , Lok Thévy , Déplanche Anne-Marie , Barakat Abdul I. 
Published: May 2022

AbstractCollective migration of vascular endothelial cells is central for embryonic development, angiogenesis, and wound closure. Although physical confinement of cell assemblies has been shown to elicit specific patterns of collective movement in various cell types, endothelial migration in vivo often occurs without confinement. Here we show that unconfined endothelial cell monolayers on microgroove substrates that mimic the anisotropic organization of the extracellular matrix exhibit a specific type of collective movement that takes the form of a periodic pattern of antiparallel cell streams. We further establish that the development of these streams requires intact cell-cell junctions and that stream sizes are particularly sensitive to groove depth. Finally, we show that modeling the endothelial cell sheet as an active fluid with the microgrooves acting as constraints on cell orientation predicts the occurrence of the periodic antiparallel cell streams as well as their lengths and widths. We posit that in unconfined cell assemblies, physical factors that constrain or bias cellular orientation such as anisotropic extracellular matrix cues or directed flow-derived shear forces dictate the pattern of collective cell movement.
Multiplex base- and prime-editing with drive-and-process CRISPR arrays
Authors: Yuan Qichen , Gao Xue 
Published: May 2022

AbstractCurrent base- and prime-editing technologies lack efficient strategies to edit multiple genomic loci simultaneously, limiting their applications in complex genomics and polygenic diseases. Here, we describe drive-and-process (DAP) CRISPR array architectures for multiplex base-editing (MBE) and multiplex prime-editing (MPE) in human cells. We leverage tRNA as the RNA polymerase III promoter to drive the expression of tandemly assembled tRNA-guide RNA (gRNA) arrays, of which the individual gRNAs are released by the cellular endogenous tRNA processing machinery. We engineer a 75-nt human cysteine tRNA (hCtRNA) for the DAP array, achieving up to 31-loci MBE and up to 3-loci MPE. By applying MBE or MPE elements for deliveries via adeno-associated virus (AAV) and lentivirus, we demonstrate simultaneous editing of multiple disease-relevant genomic loci. Our work streamlines the expression and processing of gRNAs on a single array and establishes efficient MBE and MPE strategies for biomedical research and therapeutic applications.
Structural mechanism of TRPV3 channel inhibition by the anesthetic dyclonine
Authors: Neuberger Arthur , Nadezhdin Kirill D. , Sobolevsky Alexander I. 
Published: May 2022

AbstractSkin diseases are common human illnesses that occur in all cultures, at all ages, and affect between 30% and 70% of individuals globally. TRPV3 is a cation-permeable TRP channel predominantly expressed in skin keratinocytes, implicated in cutaneous sensation and associated with numerous skin diseases. TRPV3 is inhibited by the local anesthetic dyclonine, traditionally used for topical applications to relieve pain and itch. However, the structural basis of TRPV3 inhibition by dyclonine has remained elusive. Here we present a cryo-EM structure of a TRPV3-dyclonine complex that reveals binding of the inhibitor in the portals which connect the membrane environment surrounding the channel to the central cavity of the channel pore. We propose a mechanism of TRPV3 inhibition in which dyclonine molecules stick out into the channel pore, creating a barrier for ion conductance. The allosteric binding site of dyclonine can serve as a template for the design of new TRPV3-targeting drugs.
Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
Authors: Zeng Leiping , Liu Yanxia , Nguyenla Xammy Huu , Abbott Timothy R. , Han Mengting , Zhu Yanyu , Chemparathy Augustine , Lin Xueqiu , Chen Xinyi , Wang Haifeng , Rane Draven A. , Spatz Jordan M. , Jain Saket , Rustagi Arjun , Pinsky Benjamin , Zepeda Adrianna E. , Kadina Anastasia P. , Walker John A. , Holden Kevin , Temperton Nigel , Cochran Jennifer R. , Barron Annelise E. , Connolly Michael D. , Blish Catherine A. , Lewis David B. , Stanley Sarah A. , La Russa Marie F. , Qi Lei S. 
Published: May 2022

AbstractA major challenge in coronavirus vaccination and treatment is to counteract rapid viral evolution and mutations. Here we demonstrate that CRISPR-Cas13d offers a broad-spectrum antiviral (BSA) to inhibit many SARS-CoV-2 variants and diverse human coronavirus strains with >99% reduction of the viral titer. We show that Cas13d-mediated coronavirus inhibition is dependent on the crRNA cellular spatial colocalization with Cas13d and target viral RNA. Cas13d can significantly enhance the therapeutic effects of diverse small molecule drugs against coronaviruses for prophylaxis or treatment purposes, and the best combination reduced viral titer by over four orders of magnitude. Using lipid nanoparticle-mediated RNA delivery, we demonstrate that the Cas13d system can effectively treat infection from multiple variants of coronavirus, including Omicron SARS-CoV-2, in human primary airway epithelium air-liquid interface (ALI) cultures. Our study establishes CRISPR-Cas13 as a BSA which is highly complementary to existing vaccination and antiviral treatment strategies.
Multimodal confined water dynamics in reverse osmosis polyamide membranes
Authors: Foglia Fabrizia , Frick Bernhard , Nania Manuela , Livingston Andrew G. , Cabral João T. 
Published: May 2022

AbstractWhile polyamide (PA) membranes are widespread in water purification and desalination by reverse osmosis, a molecular-level understanding of the dynamics of both confined water and polymer matrix remains elusive. Despite the dense hierarchical structure of PA membranes formed by interfacial polymerization, previous studies suggest that water diffusion remains largely unchanged with respect to bulk water. Here, we employ neutron spectroscopy to investigate PA membranes under precise hydration conditions, and a series of isotopic contrasts, to elucidate water transport and polymer relaxation, spanning ps-ns timescales, and Å-nm lengthscales. We experimentally resolve, for the first time, the multimodal diffusive nature of water in PA membranes: in addition to (slowed down) translational jump-diffusion, we observe a long-range and a localized mode, whose geometry and timescales we quantify. The PA matrix is also found to exhibit rotational relaxations commensurate with the nanoscale confinement observed in water diffusion. This comprehensive ‘diffusion map’ can anchor molecular and nanoscale simulations, and enable the predictive design of PA membranes with tuneable performance.
Multidimensional analysis of adult patients’ care trajectories before a first diagnosis of schizophrenia
Authors: Vanasse Alain , Courteau Josiane , Courteau Mireille , Roy Marc-André , Stip Emmanuel , Fleury Marie-Josée , Lesage Alain , Brodeur Sébastien 
Published: May 2022

AbstractFor patients at high-risk for developing schizophrenia, a delayed diagnosis could be affected, among many reasons, by their patterns of healthcare use. This study aims to describe and generate a typology of patients’ care trajectories (CTs) in the 2 years preceding a first diagnosis of schizophrenia, over a medico-administrative database of 3712 adults with a first diagnosis between April 2014 and March 2015 in Quebec, Canada. This study applied a multidimensional approach of State Sequence Analysis, considering together sequences of patients’ diagnoses, care settings and care providers. Five types of distinct CTs have emerged from this data-driven analysis: The type 1, shared by 77.6% of patients, predominantly younger men, shows that this group sought little healthcare, among which 17.5% had no healthcare contact for mental disorders. These individuals might benefit from improved promotion and prevention of mental healthcare at the community level. The types 2, 3 and 4, with higher occurrence of mental disorder diagnoses, represent together 19.5% of the study cohort, mostly middle-aged and women. These CTs, although displaying roughly similar profiles of mental disorders, revealed very dissimilar sequences and levels of care providers encounters, primary and specialized care use, and hospitalizations. Surprisingly, patients of these CTs had few consultations with general practitioners. An increased attentiveness for middle-aged patients and women with high healthcare use for mental disorders could help to reduce delayed diagnosis of schizophrenia. This calls for further consideration of healthcare services for severe mental illness beyond those offered to young adults.
Proton-gated anion transport governs macropinosome shrinkage
Authors: Zeziulia Mariia , Blin Sandy , Schmitt Franziska W. , Lehmann Martin , Jentsch Thomas J. 
Published: May 2022

AbstractIntracellular organelles change their size during trafficking and maturation. This requires the transport of ions and water across their membranes. Macropinocytosis, a ubiquitous form of endocytosis of particular importance for immune and cancer cells, generates large vacuoles that can be followed optically. Shrinkage of macrophage macropinosomes depends on TPC-mediated Na+efflux and Cl−exit through unknown channels. Relieving osmotic pressure facilitates vesicle budding, positioning osmotic shrinkage upstream of vesicular sorting and trafficking. Here we identify the missing macrophage Cl−channel as the proton-activated Cl−channel ASOR/TMEM206. ASOR activation requires Na+-mediated depolarization and luminal acidification by redundant transporters including H+-ATPases and CLC 2Cl−/H+exchangers. As corroborated by mathematical modelling, feedback loops requiring the steep voltage and pH dependencies of ASOR and CLCs render vacuole resolution resilient towards transporter copy numbers.TMEM206disruption increased albumin-dependent survival of cancer cells. Our work suggests a function for the voltage and pH dependence of ASOR and CLCs, provides a comprehensive model for ion-transport-dependent vacuole maturation and reveals biological roles of ASOR.
Rituximab versus tocilizumab in rheumatoid arthritis: synovial biopsy-based biomarker analysis of the phase 4 R4RA randomized trial
Authors: Rivellese Felice , Surace Anna E. A. , Goldmann Katriona , Sciacca Elisabetta , Çubuk Cankut , Giorli Giovanni , John Christopher R. , Nerviani Alessandra , Fossati-Jimack Liliane , Thorborn Georgina , Ahmed Manzoor , Prediletto Edoardo , Church Sarah E. , Hudson Briana M. , Warren Sarah E. , McKeigue Paul M. , Humby Frances , Bombardieri Michele , Barnes Michael R. , Lewis Myles J. , Pitzalis Costantino ,   the R4RA collaborative group, Rivellese Felice , Giorli Giovanni , Nerviani Alessandra , Fossati-Jimack Liliane , Thorborn Georgina , Humby Frances , Bombardieri Michele , Lewis Myles J. , Durez Patrick , Buch Maya H. , Rizvi Hasan , Mahto Arti , Montecucco Carlomaurizio , Lauwerys Bernard , Ng Nora , Ho Pauline , Romão Vasco C. , da Fonseca João Eurico Cabral , Verschueren Patrick , Kelly Stephen , Sainaghi Pier Paolo , Gendi Nagui , Dasgupta Bhaskar , Cauli Alberto , Reynolds Piero , Cañete Juan D. , Ramirez Julio , Celis Raquel , Moots Robert , Taylor Peter C. , Edwards Chris J. , Isaacs John , Sasieni Peter , Choy Ernest , Thompson Charlotte , Bugatti Serena , Bellan Mattia , Congia Mattia , Holroyd Christopher , Pratt Arthur , White Laura , Warren Louise , Peel Joanna , Hands Rebecca , Hadfield Gaye , Pitzalis Costantino 
Published: May 2022

AbstractPatients with rheumatoid arthritis (RA) receive highly targeted biologic therapies without previous knowledge of target expression levels in the diseased tissue. Approximately 40% of patients do not respond to individual biologic therapies and 5–20% are refractory to all. In a biopsy-based, precision-medicine, randomized clinical trial in RA (R4RA; n = 164), patients with low/absent synovial B cell molecular signature had a lower response to rituximab (anti-CD20 monoclonal antibody) compared with that to tocilizumab (anti-IL6R monoclonal antibody) although the exact mechanisms of response/nonresponse remain to be established. Here, in-depth histological/molecular analyses of R4RA synovial biopsies identify humoral immune response gene signatures associated with response to rituximab and tocilizumab, and a stromal/fibroblast signature in patients refractory to all medications. Post-treatment changes in synovial gene expression and cell infiltration highlighted divergent effects of rituximab and tocilizumab relating to differing response/nonresponse mechanisms. Using ten-by-tenfold nested cross-validation, we developed machine learning algorithms predictive of response to rituximab (area under the curve (AUC) = 0.74), tocilizumab (AUC = 0.68) and, notably, multidrug resistance (AUC = 0.69). This study supports the notion that disease endotypes, driven by diverse molecular pathology pathways in the diseased tissue, determine diverse clinical and treatment–response phenotypes. It also highlights the importance of integration of molecular pathology signatures into clinical algorithms to optimize the future use of existing medications and inform the development of new drugs for refractory patients.