Biology subjects

Deng, L.

Publications and source records attributed to Deng, L..

At least 127 records · Page 7Linked to original sources

bioRxiv2019

Cell motion-coordinated fibrillar assembly of soluble collagen I to promote MDCK cell branching formation

Extracellular Matrix (ECM) assembly and remodeling are critical physiological events in vivo, and abnormal ECM assembly or remodeling is related to pathological conditions such as osteoarthritis, fibrosis, cancers, and genetic diseases. ECM assembly/remodeling driven by cells represents more physiological processes. Collagen I (COL) is very abundant in tissues, which assembly/remodeling is mediated by biochemical and mechanical factors. How cells regulate COL assembly biomechanically still remains to be well understood. Here we used fluorescent COL in the medium to study how cells assembled ECM which represents more physiological structures. The results showed that MDCK cells actively recruited COL from the medium and helped assemble the fibers, which in turn facilitated cell branching morphogenesis, both displaying high spatial associations and mutual dependency. Inhibition of cellular contraction force by ROCK and Myosin II inhibitors attenuated but did not block the COL fiber formation, while cell motion showed high consistency with the fiber assembly. Under ROCK or Myosin II inhibition, further analysis indicated high correlation between local cell movement and COL fiber strength as quantified from different regions of the same groups. Blocking cell motion by actin cytoskeleton disruption completely inhibited the fiber formation. These suggest that cell motion coordinated COL fiber assembly from the medium, possibly through generated strain on deposited COL to facilitate the fiber growth.

biophysics↗

bioRxiv2019

Sensing Traction Force on Matrix Induces Cell-Cell Distant Mechanical Communications for Self-assembly

The long-range biomechanical force propagating across large scale may reserve the capability to trigger coordinative responses within cell population such as during angiogenesis, epithelial tubulogenesis, and cancer metastasis. How cells communicate in a distant manner within the group for self-assembly remains largely unknown. Here we found that airway smooth muscle cells (ASMCs) rapidly self-assembled into well-constructed network on 3D Matrigel containing type I collagen (COL), which relied on long-range biomechanical force across the matrix to direct cell-cell distant interactions. Similar results happened by HUVEC cells to mimic angiogenesis. Interestingly, single ASMCs initiated multiple extended protrusions precisely pointing to neighboring cells in distance, depending on traction force sensing. Separate ASMCs sensed each other to move directionally on both non-fibrous Matrigel and more efficiently when containing fibrous COL, but lost mutual sensing on fixed gel or coated glass due to no long-range force transmission. Beads tracking assay demonstrated distant transmission of traction force, and finite element method modeling confirmed the consistency between maximum strain distribution on matrix and cell directional movements in experiments. Furthermore, ASMCs recruited COL from the hydrogel to build fibrous network to mechanically stabilize cell network. Our results revealed for the first time that cells can sense traction force transmitted through the matrix to initiate cell-cell distant mechanical communications, resulting in cell directional migration and coordinative self-assembly with active matrix remodeling. As an interesting phenomenon, cells sound able to make phone call via long-range biomechanics, which implicates physiological importance such as for tissue pattern formation.

biophysics↗

bioRxiv2019

Age-related compensatory reconfiguration of PFC connections during episodic memory retrieval

During demanding cognitive tasks, older adults (OAs) frequently show greater prefrontal cortex (PFC) activity than younger adults (YAs). This age-related PFC activity increase is often associated with enhanced cognitive performance, suggesting functional compensation. However, the brain is a complex network of interconnected regions, and it is unclear how network connectivity of PFC regions differs for OAs vs. YAs. To investigate this, we examined the age-related difference in functional brain network mediating episodic memory retrieval. YAs and OAs participants encoded and then recalled visual scenes, and age-related differences in network topology during memory retrieval were investigated as a function of memory performance. We measured both quantitative changes in functional integration and qualitative reconfiguration in connectivity patterns. The study yielded three main findings. First, PFC regions were more functionally integrated with the rest of the brain network in OAs. Critically, this age-related increase in PFC integration was associated with better retrieval performance. Second, PFC regions showed stronger performance-related reconfiguration of connectivity patterns in OAs. Finally, the magnitude of PFC reconfiguration increases in OAs tracked reconfiguration reductions in the medial temporal lobe (MTL) - a core episodic memory region, suggesting that PFC connectivity in OAs may be compensating for MTL deficits.

neuroscience↗

bioRxiv2019

TGS-GapCloser: fast and accurately passing through the Bermuda in large genome using error-prone third-generation long reads

The completeness and accuracy of genome assemblies determine the quality of subsequent bioinformatics analysis. Despite benefiting from the medium/long-range information of third-generation sequencing techniques, current gap-closing tools to enhance assemblies suffer multi-alignments and high error rates, resulting in huge time and money costs.\n\nWe developed a software tool, TGS-GapCloser that uses the low depth (>=10X) single molecule sequencing long reads without any error correction to close gaps. The algorithm distinguishes gap regions from the alignments of long reads against original scaffolds, corrects only the candidate regions, and assigns the best sequences to each gap. We demonstrate that TGS-GapCloser improves the contig N50 value of draft assembly by 25-fold on average, updating above 90% gaps with 93.96% positive predictive value. Despite of high error rate of raw long reads, improved assemblies archive Q50 (99.999%) single-base accuracy with only 11.8% decrement to inputs. Besides it could complete more gaps, and is also [~]29-fold faster than mainstream gap-closing tools. BUSCO analysis revealed that 3.4%-13.1% more expected genes were complete. TGS-GapCloser also shows its power to fill gaps for ultra large genome assembly of ginkgo ([~]12Gb) with 71.6% of gaps closed. The validation of inserted or merged gap sequences was conducted with NGS reads and reference genomes, respectively. The updated genome assemblies may promote the gene annotation, structure variant calling and thus improving the downstream analysis of ontogeny, phylogeny, and evolution.

bioinformatics↗

bioRxiv2019

Structural controllability predicts functional patterns and neuromodulatory benefits associated with working memory

The brain is an inherently dynamic system, and much work has focused on the ability to modify neural activity through both local perturbations and changes in the function of global network ensembles. Network controllability is a recent concept in network science that purports to predict the influence of individual cortical sites on global network states and state changes, thereby creating a unifying account of local influences on global brain dynamics. Here, we present an integrated set of multimodal brain-behavior relationships, acquired from functional magnetic resonance imaging during a transcranial magnetic stimulation intervention, that demonstrate how network controllability influences network function, as well as behavior. This work helps to outline a clear technique for integrating structural network topology and functional activity to predict the influence of a potential stimulation target on subsequent behaviors and prescribes next steps towards predicting neuromodulatory and behavioral responses after brain stimulation.\n\nHighlights- This study tested the strength of network controllability using fMRI and rTMS\n- Controllability correlates with functional modulation of working memory demand load\n- Controllability is also correlated with the memory improvement from applied rTMS\n- These findings link network control theory with physiology and behavior.\n\n\nIn briefBeynel et al. show that the benefits of functionally targeted brain stimulation on working memory performance can be predicted by network control properties at the stimulated site. Structural controllability and functional activity independently predict this cognitive benefit.\n\nAuthor ContributionsConceptualization & Methodology: L.B, S.W.D., B.L., R.C., L.G.A.; Investigation: L.B., L.D., S.W.D., C.A.C., M.D., H.P., S.H.; Writing--Original Draft: L.B., L.D., S.W.D.; Writing--Review & Editing: L.B., L.D., S.W.D., L.G.A., A.V.P.; Funding Acquisition: S.W.D., R.C., B.L., S.H.L., A.V.P.; Resources: L.G.A., B.L., R.C.; Supervision: L.G.A., S.W.D.

neuroscience↗

bioRxiv2019

Identification of key determinants of Staphylococcus aureus vaginal colonization

Staphylococcus aureus is an important pathogen responsible for nosocomial and community acquired infections in humans, and methicillin-resistant S. aureus (MRSA) infections have continued to increase despite wide-spread preventative measures. S. aureus can colonize the female vaginal tract and reports have suggested an increase in MRSA infections in pregnant and postpartum women as well as outbreaks in newborn nurseries. Currently, little is known about specific factors that promote MRSA vaginal colonization and subsequent infection. To study S. aureus colonization of the female reproductive tract in a mammalian system, we developed a mouse model of S. aureus vaginal carriage and demonstrated that both hospital-associated and community-associated MRSA isolates can colonize the murine vaginal tract. Immunohistochemical analysis revealed an increase in neutrophils in the vaginal lumen during MRSA colonization. Additionally, we observed that a mutant lacking fibrinogen binding adhesins exhibited decreased persistence within the mouse vagina. To further identify novel factors that promote vaginal colonization, we performed RNA-sequencing to determine the transcriptome of MRSA growing in vivo during vaginal carriage at 5 hours, 1-day, and 3-days post-inoculation. Over 25% of bacterial genes were differentially regulated at all time points during colonization compared to laboratory cultures. The most highly induced genes were those involved in iron acquisition, including the Isd system and siderophore transport systems. Mutants deficient in these pathways did not persist as well during in vivo colonization. These results reveal that fibrinogen binding as well as the capacity to overcome host nutritional limitation are important determinants of MRSA vaginal colonization.\n\nIMPORTANCEStaphylococcus aureus is an opportunistic pathogen able to cause a wide variety of infections in humans. Recent reports have suggested an increasing prevalence of MRSA in pregnant and postpartum women, coinciding with the increased incidence of MRSA infections in the NICU and newborn nurseries. Vertical transmission from mothers to infants at delivery is a likely route of MRSA acquisition by the newborn, however, essentially nothing is known about host and bacterial factors that influence MRSA carriage in the vagina. Here, we established a mouse model of vaginal colonization and observed that multiple MRSA strains can persist in the vaginal tract. Additionally, we determined that MRSA interactions with fibrinogen as well as iron uptake can promote vaginal persistence. This study is the first to identify molecular mechanisms which govern vaginal colonization by MRSA, the critical initial step preceding infection and neonatal transmission.

microbiology↗

bioRxiv2019

SLR-superscaffolder: a de novo scaffolding tool for synthetic long reads using a top-to-bottom scheme

Synthetic long reads (SLR) with long-range co-barcoding information have been recently developed and widely applied in genomics researches. We proposed a scaffolding model of the co-barcoding information and developed a scaffolding tool with adopting a top-to-bottom scheme to make full use of the complementary information in SLR datasets and a screening algorithm to reduce negative effects from misassembled contigs in an input assembly. In comparison with other available SLR scaffolding tools, our tool obtained the best quality improvement for different input assemblies, especially for those assembled by the next-generation sequencing reads, where the improvement of contiguity is about several hundred-folds.

bioinformatics↗

bioRxiv2019

Characterization of the vaginal DNA virome in health and dysbiosis: an opening study in patients with non-female factor infertility

BackgroundBacterial vaginosis (BV) is characterised by a reduction in Lactobacillus spp. abundance and increased abundance of facultative anaerobes, like Gardnerella vaginalis. BV aetiology is not fully understood, but bacteriophages could play a pivotal role causing perturbation of the vaginal bacterial community. Here we investigate the vaginal viral community, including bacteriophages, and its association to the bacterial community and BV-status.\n\nMethodsVaginal samples from 48 patients undergoing IVF treatment for non-female factor infertility were subjected to metagenomic sequencing of purified virus-like particles. The vaginal viral community was characterized and correlated with BV-status, bacterial community structure and presence of key vaginal bacterial species.\n\nResultsThe majority of identified vaginal viruses belonged to the class of double-stranded DNA bacteriophages, with eukaryotic viruses constituting 4% of total reads. Clear links between viral community composition and BV (q = 0.006, R = 0.26) as well as presence of L. crispatus (q = 0.001, R = 0.43), L. iners, Gardnerella vaginalis and Atopobium vaginae were found (q < 0.002, R > 0.15). Interestingly, also the eukaryotic viral community was correlated with BV-status (q = 0.018, R = 0.20).\n\nConclusionsThe vaginal virome is clearly linked with bacterial community structure and BV-status.\n\nClinical Trials RegistrationNCT02042352.

microbiology↗

bioRxiv2019

De novo assembly of a Tibetan genome and identification of novel structural variants associated with high altitude adaptation

Structural variants (SVs) may play important roles in human adaption to extreme environments such as high altitude but have been under-investigated. Here, combining long-read sequencing with multiple scaffolding techniques, we assembled a high-quality Tibetan genome (ZF1), with a contig N50 length of 24.57 mega-base pairs (Mb) and a scaffold N50 length of 58.80 Mb. The ZF1 assembly filled 80 remaining N-gaps (0.25 Mb in total length) in the reference human genome (GRCh38). Markedly, we detected 17,900 SVs, among which the ZF1-specific SVs are enriched in GTPase activity that is required for activation of the hypoxic pathway. Further population analysis uncovered a 163-bp intronic deletion in the MKL1 gene showing large divergence between highland Tibetans and lowland Han Chinese. This deletion is significantly associated with lower systolic pulmonary arterial pressure, one of the key adaptive physiological traits in Tibetans. Moreover, with the use of the high quality de novo assembly, we observed a much higher rate of genome-wide archaic hominid (Altai Neanderthal and Denisovan) shared non-reference sequences in ZF1 (1.32%-1.53%) compared to other East Asian genomes (0.70%-0.98%), reflecting a unique genomic composition of Tibetans. One such archaic-hominid shared sequence, a 662-bp intronic insertion in the SCUBE2 gene, is enriched and associated with better lung function (the FEV1/FVC ratio) in Tibetans. Collectively, we generated the first high-resolution Tibetan reference genome, and the identified SVs may serve as valuable resources for future evolutionary and medical studies.

genomics↗

bioRxiv2019

BiasDB: A Comprehensive Database for Biased GPCR Ligands

G protein-coupled receptors transmit signals across membranes via interaction with intracellular binding partners. While there is an imprinted signaling profile for each receptor, biased ligands are able to shift intracellular pathways resulting in different recruitment profiles. We present the first comprehensive database of all literature-known biased ligands as a resource for medicinal chemistry and pharmacology. In addition to careful manual curation, we provide an analysis of the data. BiasDB is available at https://biasdb.drug-design.de/.

pharmacology and toxicology↗

bioRxiv2019

The effects of biodegradable mulch film on the growth, yield, and water use efficiency of cotton and maize in an arid region

Plastic residual film pollution in China is serious, and the use of degradable mulch film instead of plastic mulch can effectively alleviate this situation. The substitution of common polyethylene plastic mulch film with biodegradable mulch film in the agricultural production of cotton and maize in an arid region was investigated in the present study. Using bare soil as the control, we compared the effects of common polyethylene plastic film and biodegradable mulch film on crop growth, yield, and water use efficiency (WUE) in maize and cotton. The results indicated that: (1) the biodegradable mulch film in this region remained intact for 60 days after being laid down, significantly degrading after 120 days, and was associated with increased soil temperature, moisture conservation, and degradability in comparison to a bare soil control. (2) Both the biodegradable mulch film and the polyethylene plastic film significantly increased various physiological parameters, such as crop height, stalk diameter, and leaf area. (3) The biodegradable mulch film significantly increased maize and cotton crop yield by 69.4-76.2% and 65.2-71.9%, respectively, compared to the bare soil control. (4) Compared to the bare soil control, the biodegradable mulch film effectively increased WUE in the crops by 64.5-73.1%. In summary, biodegradable mulch film had comparable results to the common polyethylene plastic film in increasing soil temperature, moisture conservation, crop growth, yield, and WUE. As the biodegradable mulch film causes no residual pollution, it is thus preferable to common plastic mulch film for agricultural applications in arid regions and supports the sustainable development of agroecosystems.

ecology↗

bioRxiv2019

Older adults benefit from more widespread brain network integration during working memory

Neuroimaging evidence suggests that the aging brain relies on a more distributed set of cortical regions than younger adults in order to maintain successful levels of performance during demanding cognitive tasks. However, it remains unclear how task demands give rise to this age-related expansion in cortical networks. To investigate this issue, we used functional magnetic resonance imaging to measure univariate activity, network connectivity, and cognitive performance in younger and older adults during a working memory (WM) task. In the WM task investigated, participants hold letters online (maintenance) while reordering them alphabetically (manipulation). WM load was titrated to obtain four individualized difficulty levels. Network integration--defined as the ratio of within-versus between-network connectivity--was linked to individual differences in WM capacity. The study yielded three main findings. First, as task difficulty increased, network integration decreased in younger adults, whereas it increased in older adults. Second, age-related increases in network integration were driven by increases in right hemispheric connectivity to both left and right cortical regions, a finding that helps to reconcile extant theories of compensatory recruitment in aging to address the multivariate dynamics of global network functioning. Lastly, older adults with higher WM capacity demonstrated higher levels of network integration in the most difficult condition. These results shed light on the mechanisms of age-related network reorganization by suggesting that changes in network connectivity may act as an adaptive form of compensation, with older adults recruiting a more distributed cortical network as task demands increase.\n\nSignificance statementOlder adults often activate brain regions not engaged by younger adults, but the circumstances under which this widespread network emerges are unclear. Here, we examined the effects of aging on network connectivity between task regions recruited during a working memory (WM) manipulation task, and the rest of the brain. We found an age-related increase in the more global network integration in older adults, and an association between this integration and working memory capacity in older adults. The findings are generally consistent with the compensatory interpretation of these effects.

neuroscience↗

bioRxiv2019

Branched-Chain Amino Acid Metabolic Reprogramming Orchestrates Drug Resistance to EGFR Tyrosine Kinase Inhibitors

Drug resistance is a significant hindrance to effective cancer treatment. Although resistance mechanisms of epidermal growth factor receptor (EGFR)-mutant cancer cells to lethal EGFR tyrosine kinase inhibitors (TKI) treatment have been investigated intensively, how cancer cells orchestrate adaptive response under sublethal drug challenge remains largely unknown. Here we find that 2-hour sublethal TKI treatment elicits a transient drug-tolerant state in EGFR-mutant lung cancer cells. Continuous sublethal treatment reinforces this tolerance and eventually establishes long-term TKI resistance. This adaptive process involves H3K9 demethylation-mediated epigenetic upregulation of branched-chain amino acid aminotransferase 1 (BCAT1) and subsequent metabolic reprogramming, which promotes TKI resistance through attenuating reactive oxygen species (ROS) accumulation. Combinational treatment with TKI and ROS-inducing reagents overcomes this drug resistance in preclinical mouse models. Clinical information analyses support the correlation of BCAT1 expression with EGFR TKI response. Collectively, our findings reveal the importance of epigenetically regulated BCAT1-engaged metabolism reprogramming in TKI resistance in lung cancer.\n\nHIGHLIGHTSSublethal EGFR TKI treatment induces transient drug-tolerant state and long-term resistance in EGFR-mutant lung cancer cells\n\nEpigenetically regulated BCAT1-mediated metabolic reprogramming orchestrates EGFR TKI-induced drug resistance\n\nCombinational treatment with TKI and ROS-inducing agents overcomes the drug resistance induced by EGFR TKI treatment

cancer biology↗

bioRxiv2019

Base editing in Streptomyces with Cas9-deaminase fusions

Conventional CRISPR/Cas genetic manipulation has been profitably applied to the genus Streptomyces, the most prolific bacterial producers of antibiotics. However, its reliance on DNA double-strand break (DSB) formation leads to unacceptably low yields of desired recombinants. We have adapted for Streptomyces recently-introduced cytidine base editors (CBEs) and adenine base editors (ABEs) which enable targeted C-to-T or A-to-G nucleotide substitutions, respectively, bypassing DSB and the need for a repair template. We report successful genome editing in Streptomyces at frequencies of around 50% using defective Cas9-guided base editors and up to 100% by using nicked Cas9-guided base editors. Furthermore, we demonstrate the multiplex genome editing potential of the nicked Cas9-guided base editor BE3 by programmed mutation of nine target genes simultaneously. Use of the high-fidelity version of BE3 (HF-BE3) essentially improved editing specificity. Collectively, this work provides a powerful new tool for genome editing in Streptomyces.

microbiology↗

bioRxiv2019

Deconvolute individual genomes from metagenome sequences through short read clustering

MotivationMetagenome assembly from short next-generation sequencing data is a challenging process due to its large scale and computational complexity. Clustering short reads before assembly offers a unique opportunity for parallel downstream assembly of genomes with individualized optimization. However, current read clustering methods suffer either false negative (under-clustering) or false positive (over-clustering) problems.\n\nResultsBased on a previously developed scalable read clustering method on Apache Spark, SpaRC, that has very low false positives, here we extended its capability by adding a new method to further cluster small clusters. This method exploits statistics derived from multiple samples in a dataset to reduce the under-clustering problem. Using a synthetic dataset from mouse gut microbiomes we show that this method has the potential to cluster almost all of the reads from genomes with sufficient sequencing coverage. We also explored several clustering parameters that deferentially affect genomes with various sequencing coverage.\n\nAvailabilityhttps://bitbucket.org/berkeleylab/jgi-sparc/.\n\nContactzhongwang@lbl.gov

bioinformatics↗

bioRxiv2019

Engineering acetyl-CoA metabolic shortcut for eco-friendly production of polyketides triacetic acid lactone in Yarrowia lipolytica

Acetyl-CoA is the central metabolic node connecting glycolysis, Krebs cycle and fatty acids synthase. Plant-derived polyketides, are assembled from acetyl-CoA and malonyl-CoA, represent a large family of biological compounds with diversified bioactivity. Harnessing microbial bioconversion is considered as a feasible approach to large-scale production of polyketides from renewable feedstocks. Most of the current polyketide production platform relied on the lengthy glycolytic steps to provide acetyl-CoA, which inherently suffers from complex regulation with metabolically-costly cofactor/ATP requirements. Using the simplest polyketide triacetic acid lactone (TAL) as a target molecule, we demonstrate that acetate uptake pathway in oleaginous yeast (Yarrowia lipolytica) could function as an acetyl-CoA shortcut to achieve metabolic optimality in producing polyketides. We identified the metabolic bottlenecks to rewire acetate utilization for efficient TAL production in Y. lipolytica, including generation of the driving force for acetyl-CoA, malonyl-CoA and NADPH. The engineered strain, with the overexpression of endogenous acetyl-CoA carboxylase (ACC1), malic enzyme (MAE1) and a bacteria-derived cytosolic pyruvate dehydrogenase (PDH), affords robust TAL production with titer up to 4.76 g/L from industrial glacier acetic acid in shake flasks, representing 8.5-times improvement over the parental strain. The acetate-to-TAL conversion ratio (0.149 g/g) reaches 31.9% of the theoretical maximum yield. The carbon flux through this acetyl-CoA metabolic shortcut exceeds the carbon flux afforded by the native acetyl-CoA pathways. Potentially, acetic acid could be manufactured in large-quantity at low-cost from Syngas fermentation or heterogenous catalysis (methanol carbonylation). This alternative carbon sources present a metabolic advantage over glucose to unleash intrinsic pathway limitations and achieve high carbon conversion efficiency and cost-efficiency. This work also highlights that low-cost acetic acid could be sustainably upgraded to high-value polyketides by oleaginous yeast species in an eco-friendly and cost-efficient manner.

bioengineering↗

bioRxiv2019

Liver X Receptor β Controls Hepatic Stellate Cell Activation via Hedgehog Signaling

Liver X receptors (LXR) and {beta} serve important roles in cholesterol homeostasis, anti-inflammatory processes and the activation of hepatic stellate cells (HSCs). However, the development of therapies for liver fibrosis based on LXR agonists have been hampered due to side-effects such as liver steatosis. In this study, we demonstrated that HSCs expressed high levels of LXR{beta}, but not LXR, and that overexpression of LXR{beta} suppressed fibrosis and HSC activation in a carbon tetrachloride (CCl4)-induced fibrosis mouse model, without resulting in liver steatosis. Furthermore, Hedgehog (Hh)-regulated proteins, markedly increased in the CCl4-affected liver and mainly expressed in activated HSCs, were repressed under conditions of LXR{beta} overexpression. In addition, LXR{beta} knockout led to activation of Hh signaling and triggering of HSC activation, while overexpression of LXR{beta} led to the inhibition of the Hh pathway and suppression of HSC activation. These results suggest that LXR{beta} suppresses the activation mechanism of HSCs by inhibiting Hh signaling. In conclusion, LXR{beta}, by restoring the differentiation of HSCs, may be a promising therapeutic target for liver fibrosis without the adverse side-effects of LXR activation.

cell biology↗

bioRxiv2019

The Group B Streptococcal surface antigen I/II protein, BspC, interacts with host vimentin to promote adherence to brain endothelium and inflammation during the pathogenesis of meningitis

Streptococcus agalactiae (Group B Streptococcus, GBS) normally colonizes healthy adults but can cause invasive disease, such as meningitis, in the newborn. To gain access to the central nervous system, GBS must interact with and penetrate brain or meningeal blood vessels; however, the exact mechanisms are still being elucidated. Here, we investigate the contribution of BspC, an antigen I/II family adhesin, to the pathogenesis of GBS meningitis. Disruption of the bspC gene reduced GBS adherence to human cerebral microvascular endothelial cells (hCMEC), while heterologous expression of BspC in non-adherent Lactococcus lactis conferred bacterial attachment. In a murine model of hematogenous meningitis, mice infected with {Delta}bspC mutants exhibited lower mortality as well as decreased brain bacterial counts and inflammatory infiltrate compared with mice infected with WT GBS strains. Further, BspC was both necessary and sufficient to induce neutrophil chemokine expression. We determined that BspC interacts with the host cytoskeleton component vimentin, and confirmed this interaction using a bacterial two-hybrid assay, immunofluorescent staining, and imaging flow cytometry. Vimentin null mice were protected from WT GBS infection and also exhibited less inflammatory cytokine production in brain tissue. These results suggest that BspC and the vimentin interaction is critical for the pathogenesis of GBS meningitis. AUTHOR SUMMARYGroup B Streptococcus (GBS) typically colonizes healthy adults but can cause severe disease in immune compromised individuals, including newborns. Despite wide-spread intrapartum antibiotic prophylaxis given to pregnant women, GBS remains a leading cause of neonatal meningitis. To cause meningitis, GBS must interact with and penetrate the blood-brain barrier (BBB), which separates bacteria and immune cells in the blood from the brain. In order to develop targeted therapies to treat GBS meningitis, it is important to understand the mechanisms of BBB crossing. Here, we describe the role of the GBS surface factor, BspC, in promoting meningitis and discover the host ligand for BspC, vimentin, which is an intermediate filament protein that is constitutively expressed by endothelial cells. We determined that BspC interacts with the C-terminal domain of cell-surface vimentin to promote bacterial attachment to brain endothelial cells and that purified BspC protein can induce immune signaling pathways. In a mouse model of hematogenous meningitis, we observed that a GBS mutant lacking BspC was less virulent compared to WT GBS and resulted in less inflammatory disease. We also observed that mice lacking vimentin were protected from GBS infection. These results reveal the importance of the BspC-vimentin interaction in the progression of GBS meningitis disease.

microbiology↗