bioRxiv Science⌕ Search

Biology subjects

Bhattacharya, J.

Publications and source records attributed to Bhattacharya, J..

At least 19 recordsLinked to original sources

Cardiac Timing Biases Creative Exploration and Exploitation

Creative ideation involves a dynamic exchange between exploring new idea categories and exploiting familiar ones, reflecting optimal foraging principles. Although interoceptive signals, particularly cardiac activity, are associated with differences in attention and cognitive control, their role in explore-exploit dynamics during creative idea generation remains unknown. Recording both electroencephalography (EEG) and electrocardiography (ECG) data, we used convolution general linear models to examine cardiac-brain interactions underlying semantic exploration and exploitation during creative idea generation, in both spontaneous (self-selected) and directed (externally cued) conditions. Cardiac deceleration predicted ideation time: this relationship scaled nonlinearly during exploration (category switching), but linearly during exploitation (category persistence), in both conditions. Cardiac deceleration did not predict semantic distance (between response and cue word) or response accuracy during the directed condition, suggesting that heart rate slowing reflects cognitive effort allocation rather than ideational content. Cardiac phase systematically biased explore-exploit dynamics: spontaneous switching and accurate directed-switching were preferentially associated with diastolic phases and parieto-occipital alpha (8-12 Hz) desynchronization, whereas category persistence was associated with systolic phase timing and frontal theta (4-6 Hz) synchronization. The former activity was coupled to higher CD, as expected. However, the latter time-frequency activity was linked to responses timed to systole. These findings suggest that creative ideation unfolds through embodied cardiac-cortical coordination, whereby diastolic states are associated with flexible semantic exploration and alpha-related attentional dynamics, whereas systolic states are associated with exploitative persistence and theta-related control processes. This suggests interoceptive rhythms as temporal scaffolding that structures when and how ideas emerge, fundamentally expanding creativity neuroscience beyond purely cortical models.

neuroscience↗

Evolutionary turnover of protein structural disorder drives aberrant proteome remodelling in naked mole rat

The naked mole rat is an evolutionary outlier among mammals, exhibiting extreme longevity, cancer resistance, hypoxia tolerance, pain insensitivity, eusociality, poikilothermy and other distinctive physiological traits, most of which likely resulted from its adaptation to highly adverse subterranean habitat. Despite accumulating data, the genetic and molecular basis underlying these traits remain poorly understood. Through analyses of 18 distinct protein attributes and allied datasets across hystricomorphs, myomorphs, carnivores, and primates, we observed lineage-specific evolutionary divergence in intrinsic protein disorder in the naked mole rat. The disorder turnover exhibited functional dichotomy. The gain of disorder preferentially associated with proteostasis, immune regulation, neurodevelopment, skeletal growth and tumour suppressive properties, while loss of disorder modulated mostly the cardiac development. The proteins that gained disorder in NMR exhibited lower degradation rates, consistent with stabilization through phase-separation, while the proteins losing disorder show pronounced divergence in gene expression. The disorder turnover was primarily driven by indels affecting functional regions including Pfam domains, ANCHOR-predicted binding sites, short linear motifs, stress induced modifications of Tyr, Met, and Cys residues. Notably, the gained disordered regions were inferred to be redox-sensitive, aligning to exceptional stress tolerance in naked mole rats. Collectively, our results highlight an unusual and previously overlooked large-scale proteome remodelling that drives the molecular evolution of extraordinary traits of naked mole rat. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=169 SRC="FIGDIR/small/720964v1_ufig1.gif" ALT="Figure 1"> View larger version (39K): org.highwire.dtl.DTLVardef@644d1dorg.highwire.dtl.DTLVardef@102c9e4org.highwire.dtl.DTLVardef@14c770org.highwire.dtl.DTLVardef@31bc5e_HPS_FORMAT_FIGEXP M_FIG C_FIG

evolutionary biology↗

Proviral dynamics and HIV-1C viral diversity in the context of HIV-TB co-infection

BackgroundART effectively suppresses HIV replication and restores CD4+ T cells; however, long-lived HIV latent reservoirs enable viral persistence. Tuberculosis (TB) co-infection further impacts HIV latency and enhances viral replication. Given the high prevalence of latent TB infection (LTBI) in TB-endemic settings, understanding its impact on HIV biology is critical. Our study aims to investigate the influence of TB co-infection on HIV reservoir dynamics, viral diversity, and drug resistance mutations in ART-naive individuals. MethodologySamples from 90 ART-naive HIV-1C individuals, stratified based on IGRA and TB diagnosis, were used in this study. Plasma and PBMCs were isolated for viral RNA and DNA extraction respectively. Total proviral DNA was quantified using gag PCR. Full-length env and pol genes were amplified, purified and sequenced using ONT and Illumina platforms. Pol sequences were subjected to Drug Resistance Mutation (DRM) analysis via Stanford HIVdb with a minimum threshold mutation frequency of [≥]10%. Full length env sequences were used for phylogenetic analysis by aligning with Indian Subtype C reference sequence and phylogenetic tree was generated using ggplot2. ResultProviral load analysis showed no significant differences across HIV+LTBI-, HIV+LTBI+, and HIV+TB+ groups, although a trend toward higher levels was observed in HIV+TB+ individuals. Correlation analysis revealed distinct immune associations, with HIV+LTBI+ individuals showing positive correlations with activation and PD-1 expression. Longitudinal analysis of proviral loads demonstrated a modest decline in proviral load post-ART but remained persistent for up to 18-20 months following initiation of ART accompanied by low level ongoing viral replication. DRM analysis revealed a 33% prevalence in ART-naive individuals, with higher occurrence in HIV+LTBI+ group. Of the identified DRMs, 38% (5/13) and 71% (5/7) in sequences obtained from PBMC and plasma respectively were attributed to polymorphic mutations associated with Integrase strand transfer inhibitors (INSTIs). DRMs within plasma and PBMC derived viruses showed high concordance. Phylogenetic analysis of env sequences indicated overlapping viral populations between the 3 groups, with greater diversity in PBMCs compared to plasma. ConclusionThe study highlights that HIV reservoir dynamics, drug resistance, and viral diversity are significantly influenced by TB co-infection. While proviral loads were comparable, LTBI-associated immune activation and granuloma niches may have driven viral diversification and DRM emergence. High concordance between compartments and presence of transmitted resistance underscore the need for baseline screening, multi-compartment analysis, and sustained surveillance.

microbiology↗

Restructured genomes may have shaped the evolution of clade-specific traits in mammals

Genomic rearrangements (GRs) are pervasive in mammalian evolution, yet their contribution to adaptive trait divergence remains poorly defined. By examining 71 mammalian genomes across eight major clades, we uncovered 206 clade-specific disruptions of ancestral synteny. These rearrangements consistently localize near genes underpinning clade-specific physiological, metabolic and behavioural traits and coincided with the accelerated expression rates in the affected lineage. The disruption of otherwise syntenic chromatin contacts and the fusion of domains with contrasting epigenetic states implied that the alteration in regulatory adjacency may underlie the expression divergence. Primate-specific rearrangements exhibited signatures of selective constraint, including fixation across species, reduced recombination, elevated linkage disequilibrium, and enhanced co-expression of genes flanking rearranged loci, indicating that these genomic architectures are selectively maintained. Our observations imply that the clade-specific genomic rearrangements repeatedly reconfigured the mammalian regulatory landscape and likely served as an evolutionary substrate for lineage-specific adaptation.

evolutionary biology↗

Loss of conserved noncoding elements likely shaped the evolution of regressed phenotypes in cavefish

The Mexican cavefish, Astyanax mexicanus, is a captivating model for probing cave adaptations, showcasing pronounced divergence in traits like vision, brain morphology, behaviour, pigmentation, and hypoxia tolerance compared to its surface-dwelling counterpart. Very few protein-coding variants are identified in cave-morphs, and the vast phenotypic gap between the two morphs remains inadequately explained. We investigated the noncoding genomes of teleosts and found that 3,343 conserved non-coding elements (CNEs) were independently lost in cave-morphs. These CNEs, confirmed in Zebrafish, displayed enhancer-associated histone modifications, possessed binding sites of neuronal transcription factors and interacted with cognate genes through chromatin loops. Genes crucial for eye and nervous system development were located adjacent to CNEs lost in cave morphs. Notably, these flanking genes were gradually downregulated during embryonic development of cave-morphs, contrasting with surface morphs. These insights underscore how compromised developmental pathways, stemming from the loss of distal regulatory elements, contribute to the regression of phenotypes in cave morphs. Article SummaryDespite availability of genome sequences and allied datasets, the genetic underpinning of regressed traits of cavefish remains enigmatic. By aligning the genome sequences of teleosts, we identified thousands of noncoding elements specifically lost in cavefish, exhibited enhancer-associated hallmarks, and were enriched with the binding sites of neuronal transcription factors. Their cognate genes were associated with eye and nervous system development, and exhibited developmental downregulation in cavefish. This study highlights how the loss of regulatory elements impacted the cavefish evolution and adaptation.

genomics↗

A mitochondria-targeted PPR protein restores cytoplasmic male sterility by post-transcriptional modification of ORF147 in Cajanus cajanifolius

Restoration factors (Rfs) belonging to the pentatricopeptide repeat proteins (PPRs) family play an essential role in plant growth and development including their binding to CMS-associated mitochondrial RNAs leading to fertility restoration. The present study identified 22 mitochondrial-specific PPRs in pigeonpea and explored the underlying mechanisms of restoration of fertility in the A4 CMS system through yeast-three hybrid studies. The identified gene was functionally validated through transgenic expression in Arabidopsis model system and obtained conclusive evidence that the identified Rf-PPR was responsible for fertility restoration. The sub-cellular localization studies implied that the identified Rf-PPR is mitochondrial targeting. The study demonstrated that due to the interaction between mitochondrial CMS mRNA and nuclear Rf-PPR protein, post-transcriptional modification occurred, leading to the inability to translate and accumulate cytotoxic CMS protein resulting in fertility restoration. The study specifically looks into the RNA-protein interaction occurring at the nucleo-cytoplasmic level in the A4 cytoplasm of Cajanus cajanifolius. HighlightsThe study identifies the restoration of fertility genes corresponding to the CMS-causing orf147 gene.

developmental biology↗

Cardiac Cycle Modulates Alpha and Beta Suppression during Motor Imagery.

IntroductionThe baroreceptor hypothesis posits that baroreceptors, located on the cardiac walls, are most active during systole, translating cardiac contraction information to the brain. Studies within this context have suggested that the systolic phase, characterised by increased noise, may compromise the processing of sensory stimuli. Although the effect of systolic and diastolic cardiac cycle phases on cognition, perception, and action has been widely documented, there remains a gap in applying these interoceptive insights to enhance assistive technologies such as brain-computer interfaces (BCIs). In the context of BCIs, motor imagery (MI) --the mental rehearsal of movement--serves as a widely used control paradigm, yet its modulation through the cardiac cycle has not been empirically tested. Bridging this gap, this study examined how the cardiac cycle phases influence MI by assessing their effect on contralateral suppression of alpha (8-13 Hz) and beta (14-30 Hz) activity in primary sensorimotor cortices. Materials & MethodsTwenty-nine participants performed left/right thumb abductions based on the direction of an arrow presented on the screen to get familiarised with kinesthetic sensations. They then completed a MI task of the same movements. We recorded both electroencephalography (EEG) and electrocardiography (ECG), focusing our analysis on data epochs aligned with the experimental cue, based on whether it occurred during the systolic or diastolic phase of the cardiac cycle. Time-frequency analysis of source-reconstructed data assessed cue-induced changes in power spectral density (PSD) within the alpha and beta bands in the postcentral and precentral gyrus. ResultsWe found that alpha and beta suppression in the contralateral primary motor and somatosensory cortex was more pronounced when the cue fell during the diastolic phase of the cardiac cycle than during the systolic phase. Validating the main results, an analysis with circular statistics revealed that trials with particularly pronounced contralateral alpha and beta suppression featured cues with latencies clustering during diastole, the quietest time of the cardiac cycle. Accompanying the EEG effects, EMG activity on the side of the movement was enhanced during diastole. ConclusionThese findings provide evidence that MI performance can be enhanced by considering the cardiac cycle phases, offering promising implications for BCI-based applications. O_LIKey point 1: The phases of the cardiac cycle influence motor imagery performance. C_LIO_LIKey point 2: Alpha and beta contralateral suppression over sensorimotor cortices is more pronounced when movement direction is cued during diastole. C_LIO_LIKey point 3: Contralateral suppression is more likely to cluster during the quietest time of the diastolic phase: between the T-wave and the P-wave. C_LI

neuroscience↗

SARS-CoV-2 antibodies cross-react and enhance dengue infection

The presence of non-neutralizing antibodies of any dengue serotype, increase the severity of subsequent infection by other dengue serotypes. During SARS-CoV-2 pandemic, the number of symptomatic dengue cases increased in India. We describe that antibodies isolated from convalescent plasma from COVID-19 patients enhances DENV2 infection in vitro. CR3022, one antibody against SARS-CoV-2 spike protein, also showed elevated DENV2 infection in vitro. In silico protein-protein interactions between the spike antibodies and the DENV2 E-protein revealed significant interactions. Likewise, few monoclonal/polyclonal antibodies against SARS-CoV-2 showed increased dengue infection in vitro. Importantly, the AG129 mice infected with SARS-CoV-2 three-weeks prior to DENV2 infection, showed elevated dengue pathogenesis. Thus, highlighting the possibilities of elevated infection and symptomatic dengue disease in COVID-19 survivors.

immunology↗

Sessile alveolar macrophage connexin-43 determines mechano-immunity in the lung

The lungs mechanosensitive immune response, which occurs when pulmonary alveoli are overstretched, is a major impediment to ventilation therapy for hypoxemic respiratory failure. The cause is not known. We tested the hypothesis that alveolar stretch causes stretch of alveolar macrophages (AMs), leading to the immune response. In lungs viewed by optical imaging, sessile AMs expressed gap junctional protein connexin-43 (Cx43), and they communicated with the alveolar epithelium through gap junctions. Alveolar hyperinflation increased Ca2+ in the AMs but did not stretch the AMs. The Ca2+ response, and concomitant TNF secretion by AMs were blocked in mice with AM-specific deletion of Cx43. The AM responses, as also lung injury due to mechanical ventilation at high tidal volume, were inhibited by AM-specific delivery of lipid nanoparticles containing Xestospongin C, which blocked the induced Ca2+ increases. We conclude, Cx43- and Ca2+-dependent AM-epithelial interactions determine the lungs mechanosensitive immunity, providing a basis for therapy for ventilator- induced lung injury.

physiology↗

High resolution cryo-EM structures of two potently SARS-CoV-2 neutralizing monoclonal antibodies of same donor origin that vary in neutralizing Omicron variants

While vaccines have by large been found to effective against the evolving SARS-CoV-2 variants, the profound and rapid effectivity of monoclonal antibodies (mAbs) in significantly reducing hospitalization to severe disease outcomes have also been demonstrated. In the present study, by high resolution cryo-electron microscopy (cryo-EM), we examined the structural insights of two trimeric spike (S) protein bound mAbs isolated from an Indian convalescent individual infected with ancestral SARS-CoV-2 which we recently reported to potently neutralize SARS-CoV-2 from its ancestral form through highly virulent Delta form however different in their ability to neutralize Omicron variants. Our findings showed binding and conformational heterogeneities of both the mAbs (THSC20.HVTR04 and THSC20.HVTR26) bound to S trimer in its apo and hACE-2 bound forms. Additionally, cryo-EM resolved structure assisted modeling highlighted key residues associated with the ability of these two mAbs to neutralize Omicron variants. Our findings highlighted key interacting features modulating antigen-antibody interacting that can further aid in structure guided antibody engineering to enhance their breadth and potency. HighlightsO_LITwo potent human mAbs obtained from a single donor differ binding to Omicron spikes C_LIO_LIPattern of binding and conformation of these mAbs bound to full length spike differs C_LIO_LIAntibody binding alters the conformational states of S trimer in its apo and hACE-2 bound forms. C_LIO_LICryo-EM structure guided modeling highlighted correlates of interacting residues associated with resistance and sensitivity of BA.1, BA.2, BA.4/BA.5 resistance and sensitivity against these mAbs. C_LI

biophysics↗

Ancestral SARS-CoV-2 driven antibody repertoire diversity in an unvaccinated individual correlates with expanded neutralization breadth

Understanding the quality of immune repertoire triggered during natural infection can provide vital clues that form the basis for development of humoral immune response in some individuals capable of broadly neutralizing pan SARS-CoV-2 variants. We assessed the diversity of neutralizing antibody responses developed in an unvaccinated individual infected with ancestral SARS-CoV-2 by examining the ability of the distinct B cell germline-derived monoclonal antibodies (mAbs) in neutralizing known and currently circulating Omicron variants by pseudovirus and authentic virus neutralization assays. The ability of the antibodies developed post vaccination in neutralizing Omicron variants was compared to that obtained at baseline of the same individual and to those obtained from Omicron breakthrough infected individuals by pseudovirus neutralization assay. Broadly SARS-CoV-2 neutralizing mAbs representing unique B cell lineages with non-overlapping epitope specificities isolated from a single donor varied in their ability to neutralize Omicron variants. Plasma antibodies developed post vaccination from this individual demonstrated neutralization of Omicron BA.1, BA.2 and BA.4 with increased magnitude and found to be comparable with those obtained from other vaccinated individuals who were infected with ancestral SARS-CoV-2. Development of B cell repertoire capable of producing antibodies with distinct affinity and specificities for the antigen immediately after infection capable of eliciting broadly neutralizing antibodies offers highest probability in protecting against evolving SARS-CoV-2 variants. ImportanceDevelopment of robust neutralizing antibodies in SARS-CoV-2 convalescent individuals is known, however varies at population level. We isolated monoclonal antibodies from an individual infected with ancestral SARS-CoV-2 in early 2020 that not only varied in their B cell lineage origin but also varied in their capability and potency to neutralize all the known VOC and currently circulating Omicron variants. This indicated establishment of unique lineages that contributed in forming B cell repertoire in this particular individual immediately following infection giving rise to diverse antibody responses that could compensate each other in providing broadly neutralizing polyclonal antibody response. Individuals who were able to produce such potent polyclonal antibody responses after infection have a higher chance of being protected from evolving SARS-CoV-2 variants.

immunology↗

Rescue of alveolar wall liquid secretion blocks fatal lung injury by influenza-staphylococcal coinfection

Secondary lung infection by inhaled Staphylococcus aureus (SA) is a common and lethal event in individuals infected with influenza A virus (IAV). It is unclear how IAV disrupts host defense to promote SA infection in lung alveoli, where fatal lung injury occurs. We addressed this issue using the first real-time determinations of alveolar responses to IAV in live, intact, perfused lungs. Our findings show IAV infection blocked defensive alveolar wall liquid (AWL) secretion and induced airspace liquid absorption, thereby reversing normal alveolar liquid dynamics and inhibiting alveolar clearance of inhaled bacteria. Loss of AWL secretion resulted from dephosphorylation, hence inhibition of the cystic fibrosis transmembrane conductance regulator (CFTR) ion channel in alveolar epithelium, and airspace liquid absorption was caused by alveolar epithelial stimulation of the epithelial Na+ channel (ENaC). Loss of AWL secretion promoted alveolar stabilization by SA and alveolar damage by the secreted SA toxin, alpha hemolysin, but rescue of AWL secretion protected against fatal SA-induced lung injury in IAV-infected mice. These findings reveal a central role for AWL secretion in alveolar defense against inhaled bacteria and identify AWL inhibition as a critical mechanism of IAV lung pathogenesis. AWL rescue may represent a new therapeutic approach for IAV-SA coinfection. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=184 SRC="FIGDIR/small/499169v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@180caceorg.highwire.dtl.DTLVardef@82ce71org.highwire.dtl.DTLVardef@1bf9358org.highwire.dtl.DTLVardef@628fec_HPS_FORMAT_FIGEXP M_FIG C_FIG

physiology↗

A combination of potently neutralizing monoclonal antibodies isolated from an Indian convalescent donor protects against the SARS-CoV-2 delta variant

Although efficacious vaccines have significantly reduced the morbidity and mortality due to COVID-19, there remains an unmet medical need for treatment options, which monoclonal antibodies (mAbs) can potentially fill. This unmet need is exacerbated by the emergence and spread of SARS-CoV-2 variants of concern (VOCs) that have shown some resistance to vaccine responses. Here we report the isolation of two highly potently neutralizing mAbs (THSC20.HVTR04 and THSC20.HVTR26) from an Indian convalescent donor, that neutralize SARS-CoV-2 VOCs at picomolar concentrations including the delta variant (B.1.617.2). These two mAbs target non-overlapping epitopes on the receptor-binding domain (RBD) of the spike protein thereby preventing the virus attachment to its host receptor, human angiotensin converting enzyme-2 (hACE2). Furthermore, the mAb cocktail demonstrated protection against the Delta variant at low antibody doses when passively administered in the K18 hACE2 transgenic mice model, highlighting their potential as cocktail for prophylactic and therapeutic applications. Developing the capacity to rapidly discover and develop mAbs effective against highly transmissible pathogens like coronaviruses at a local level, especially in a low- and middle-income country (LMIC) such as India, will enable prompt responses to future pandemics as an important component of global pandemic preparedness. HighlightsO_LIIdentification of an Indian convalescent donor prior to emergence of SARS-CoV-2 Delta variant whose plasma demonstrated neutralization breadth across SARS-CoV-2 variants of concern (VOCs). C_LIO_LITwo (THSC20.HVTR04 and THSC20.HVTR26) monoclonal antibodies isolated from peripheral memory B cells potently neutralize SARS-CoV-2 VOCs: Alpha, Beta, Gamma, Delta and VOIs: Kappa and Delta Plus. C_LIO_LITHSC20.HVTR04 and THSC20.HVTR26 target non-competing epitopes on the receptor binding domain (RBD) and represent distinct germline lineages. C_LIO_LIPassive transfer of THSC20.HVTR04 and THSC20.HVTR26 mAbs demonstrated protection against Delta virus challenge in K18-hACE2 mice at low antibody doses. C_LI Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/474152v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@1f1b55corg.highwire.dtl.DTLVardef@1b9b438org.highwire.dtl.DTLVardef@e6d2a6org.highwire.dtl.DTLVardef@f92cd_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

Neural markers for Musical Creativity in Jazz Improvisation and Classical Interpretation

Two main types of musical creativity in the western canon are improvisation and interpretation. With improvisation, the fundamental structure of the melody, chords, rhythm and tempo of a piece can be modified, while with interpretation, the focus is on the emotional dynamics. Here we characterise electrical brain activity from professional jazz and classical pianists, whilst they were engaged in these different creative tasks with musical excerpts from both genres. Multivariate EEG was recorded during two phases of each task, mental planning and actual performance. Subsequently neuronal activity patterns were source localised with standardised low resolution electromagnetic tomography (sLORETA). For each musical performance, we obtained both subjective (self-rated) and objective (blind, expert-rated) measures of musical creativity. Across both tasks and genre backgrounds, within the first and middle 4 second segments of the performance phase, for musical performances that were judged highly creative objectively by external expert music assessors, we observed an increased activation in the anterior cingulate and medial prefrontal cortex (Brodmann area, BA32), suggesting a maintenance of executive control, and integrating motoric and emotional communication during creativity. Across genre backgrounds, within the performance phase for the interpretation task compared to the improvisation task, there was an increased activity in the insula (BA 13), suggesting a convergent creative task from the linked goal-orientated conscious error-monitoring and audio-visual integration functions. Genre profession also gave rise to differences across phases; jazz pianists presented a decreased parietal (BA7) activity during improvisation tasks suggesting a role for defocussed attention and for classical pianists, both tasks were associated with occipitotemporal (BA 37) activity which is involved in semantic/ metaphorical processing suggesting a close adherence to the visual score. These 3 areas relate the cognitive demands of the creative musical task to the demands of the corresponding genre of music. HighlightsO_LIEEG activity associated to musical creativity types: Improvisation and Interpretation C_LIO_LIIncreased activity in Insula (BA 13) for Interpretation suggest convergent creativity C_LIO_LIDecreased Precuneus (BA7) activity for Improvisation suggest defocussed attention C_LIO_LIIncreased activity in medial prefrontal cortex (BA32) in highly creative performance C_LI

neuroscience↗

Actin fence therapy with exogenous V12Rac1 protects against Acute Lung Injury

High mortality in Acute Lung Injury (ALI) results from sustained proinflammatory signaling by alveolar receptors, such as TNF receptor type 1 (TNFR1). Factors that determine the sustained signaling are not known. Unexpectedly, optical imaging of live alveoli revealed a major TNF-induced surge of alveolar TNFR1 due to a Ca2+-dependent mechanism that decreased the cortical actin fence. Mouse mortality due to inhaled LPS was associated with cofilin activation, actin loss and the TNFR1 surge. The constitutively active form of the GTPase, Rac1 (V12Rac1), given intranasally as a non-covalent construct with a cell-permeable peptide, enhanced alveolar F-actin and blocked the TNFR1 surge. V12Rac1 also protected against ALI-induced mortality resulting from intranasal (i.n.) instillation of LPS, or of Pseudomonas aeruginosa. We propose a new therapeutic paradigm in which actin enhancement by exogenous Rac1 strengthens the alveolar actin fence, protecting against proinflammatory receptor hyperexpression, hence blocking ALI.

physiology↗

Molecular programs of fibrotic change in aging human lung

Aging is associated with both overt and subclinical lung fibrosis, which increases risk for mortality from viruses and other respiratory pathogens. The molecular programs that induce fibrosis in the aging lung are not well understood. To overcome this knowledge gap, we undertook multimodal profiling of distal lung samples from healthy human donors across the lifespan. Telomere shortening, a cause of cell senescence and fibrosis, was progressive with age in a sample of 86 lungs and was associated with foci of DNA damage. Bulk RNA sequencing confirmed activation of cellular senescence and pro-fibrotic pathways as well as genes necessary for collagen processing with increasing age. These findings were validated in independent datasets for lung and sun-exposed skin, but not other organs including heart, liver and kidney. Cell type deconvolution analysis revealed a progressive loss of lung epithelial cells and an increasing proportion of fibroblasts. Consistent with the observed pro-fibrotic transcriptional profile, second harmonic imaging demonstrated increased density of interstitial collagen in aged human lungs. Furthermore, regions of parenchymal fibrosis were associated with decreased alveolar expansion and surfactant secretion. These findings reveal the transcriptional and structural features of fibrosis and associated physiologic impairments in normal lung aging.

genomics↗

The Mitochondrial Calcium Uniporter of Pulmonary Type 2 Cells Determines Severity of ARDS

Acute lung immunity to inhaled pathogens elicits defensive pneumonitis that may convert to the Acute Respiratory Distress Syndrome (ARDS), causing high mortality. Mechanisms underlying the conversion are not understood, but are of intense interest because of the ARDS-induced mortality in the ongoing Covid-19 pandemic. Here, by optical imaging of live lungs we show that key to the lethality is the functional status of mitochondrial Ca2+ buffering across the mitochondrial Ca2+ uniporter (MCU) in the lungs alveolar type 2 cells (AT2), which protect alveolar stability. In mice subjected to ARDS by airway exposure to lipopolysaccharide (LPS), or to Pseudomonas aeruginosa, there was marked loss of MCU expression in AT2. The ability of mice to survive ARDS depended on the extent to which the MCU expression recovered, indicating that the viability of Ca2+ buffering by AT2 mitochondria critically determines ARDS severity. Mitochondrial transfer to enhance AT2 MCU expression might protect against ARDS.

physiology↗

Elicitation of potent serum neutralizing antibody responses in rabbits by immunization with an HIV-1 clade C trimeric Env derived from an Indian elite neutralizer

Evaluating the structure-function relationship of viral envelope (Env) evolution and the development of broadly cross-neutralizing antibodies (bnAbs) in natural infection can inform rational immunogen design. In the present study, we examined the magnitude and specificity of autologous neutralizing antibodies induced in rabbits by a novel HIV-1 clade C Env protein (1PGE-THIVC) vis-a-vis those developed in an elite neutralizer from whom the env sequence was obtained that was used to prepare the soluble Env protein. The thermostable 1PGE-THIVC Env displayed a native like pre-fusion closed conformation in solution as determined by small angle X-ray scattering (SAXS) and negative stain electron microscopy (EM). This closed spike conformation of 1PGE-THIVC Env trimers was correlated with weak or undetectable binding of non-neutralizing monoclonal antibodies (mAbs) compared to neutralizing mAbs. Furthermore, 1PGE-THIVC SOSIP induced potent neutralizing antibodies in rabbits to autologous virus variants. The autologous neutralizing antibody specificity induced in rabbits by 1PGE-THIVC was mapped to the C3/V4 region (T362/P401) of viral Env. This observation agreed with electron microscopy polyclonal epitope mapping (EMPEM) of the Env trimer complexed with IgG Fab prepared from the immunized rabbit sera. While the specificity of antibodies elicited in rabbits associated with neutralizing autologous viruses were distinct to those developed in the elite neutralizer, EMPEM analysis demonstrated significant changes to Env conformations when incubated with polyclonal antibody sera from the elite neutralizer, suggesting these antibodies lead to the destabilization of Env trimers. Our study not only shows distinct mechanisms associated with potent neutralization of sequence matched and unmatched autologous viruses by antibodies induced in rabbits and in the elite neutralizer, but also highlights how neutralizing antibodies developed during the course of natural infection can impact viral Env conformations. Author SummaryThe interplay between circulating virus variants and broadly cross neutralizing polyclonal antibodies developed in a subset of elite neutralizers is widely believed to provide strategies for rational immunogen design. In the present study, we studied the structural, antigenic and immunogenic properties of a thermostable soluble trimeric protein with near native pre-fusion conformation prepared using the primary sequence of an HIV-1 clade C env isolated from the broadly cross neutralizing plasma of an elite neutralizer. This novel SOSIP Env trimer demonstrated comparable antigenic, structural and immunogenic properties that favoured several ongoing subunit vaccine design efforts. The novel clade C SOSIP induced polyclonal neutralizing antibody response developed in rabbits not only differed in its epitope specificity compared to that elicited in natural infection in presence of pool of viral quasispecies but also showed how they differ in their ability to influence Env structure and conformation. A better understanding of how vaccine-induced polyclonal neutralizing antibody responses compares to responses that developed in natural infection will improve our knowledge in designing better vaccine design strategies.

immunology↗