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Goswami, M.

Publications and source records attributed to Goswami, M..

13 recordsLinked to original sources

Integrative Multi-Tissue Analysis Identifies Synaptic Gene Networks Specific to Major Depressive Disorder in Women

Major Depressive Disorder (MDD) shows marked gender differences in prevalence and molecular signatures. Transcriptomic studies of post-mortem human brain tissue have reported alterations in the expression of synapse-related genes in MDD, including gender-specific patterns. But it remains unclear whether transcriptional changes observed in the brains of women with MDD are detectable in peripheral blood and conserved in experimental stress models. Whole-blood RNA sequencing was performed in women with MDD (n = 6) and matched healthy controls (n = 4). Differentially expressed genes (DEGs) were compared with previously reported female-specific blood and post-mortem brain transcriptomic datasets where selected overlapping synapse-associated genes were evaluated in the hippocampus and prefrontal cortex of female mice exposed to Chronic Variable Mild Stress (CVMS). Peripheral blood analysis identified DEGs enriched for synaptic organization, neuronal structure, and ion transport pathways. A substantial proportion of DEGs overlapped with previously reported datasets from peripheral blood, female MDD brain transcriptomic studies, and genes showing exclusive/enriched expression in the normal human brain. Network-based prioritization identified seven synapse-associated genes (SHANK2, SHANK3, CACNG8, GPHN, PICK1, NRXN2 and DNM2) for further analysis. In the female CVMS model, several of these genes showed altered expression in the hippocampus and/or prefrontal cortex, alongside behavioural changes and reduced dendritic spine density. These findings highlight shared transcriptional signals across human blood and human brain datasets, as well as in the mouse brain. However, larger studies are required to confirm and validate these observations.

neuroscience↗

Redox imbalance dictates dependence on GOT1 versus GOT2 for rod photoreceptor health during aging and stress

Photoreceptor (PR) loss causes vision loss in many blinding diseases, and effective therapies to prevent this cell loss are lacking. Aspartate aminotransferases (GOTs), located in the cytosol (GOT1) and mitochondria (GOT2), are key components of the malate-aspartate shuttle, which transfers reducing equivalents from cytosol to mitochondria. Previous work has implicated the GOTs as potential modulators of blinding retinal disease. To determine the roles of GOT1 and GOT2 in rod PRs, we generated rod PR-specific Got1 or Got2 conditional knockout mice (Got1 or Got2 cKO). We previously showed that Got1 cKO causes PR degeneration and is accompanied by NADH accumulation and a decreased retinal NAD+/NADH ratio. Here, we show that NADH oxidation via metabolic or genetic means prolongs PR survival in Got1 cKO animals, implicating NADH accumulation, or reductive stress, as a key driver of PR degeneration. In contrast, Got2 cKO causes minimal PR degeneration and alterations in retinal NADH and the NAD+/NADH ratio that oppose reductive stress. Interestingly, GOT2, but not GOT1, is decreased in multiple models of PR degeneration, including retinal detachment (RD) where the NAD+/NADH ratio favors a reductive state. Notably, loss of Got2 in PRs demonstrates a neuroprotective effect after experimental RD suggesting decreased GOT2 expression may be part of a stress response to promote PR survival. Overall, this study illustrates the differential dependence on the GOTs for PR health, provides evidence that an overly reductive environment is detrimental to PR survival, and identifies GOT2 as a novel therapeutic target with potentially broad application in blinding diseases.

neuroscience↗

Systematic Comparative Analysis of Pathway Databases from a User-Perspective

Pathway Databases are essential particularly in the post-genomic era as they provide organized information about biological pathways and processes. However, there are too many pathway databases with varying features. These variations, stemming from multiple factors, create challenges for users in making objective selections among the databases. Hence, there is a need to systematically compare these databases from a users perspective. Indeed, there is a need to compile all such databases first. We first compiled pathway databases through a thorough literature search and shortlisted those suitable for human pathway studies. We designed a query set consisting of genes, pathways, and conditions and performed a quantitative comparison of the output from these shortlisted databases. Furthermore, to assess update frequency among the pathway databases, we curated potential novel genes involved in the MAPK pathway and compared them with the information available in pathway databases. The quantitative comparison of pathway databases showed that PathDIP yielded the highest number of pathways when queried with gene and condition names, followed by other databases. However, when queried for pathway names, Reactome yielded the highest number of pathways, followed by PathDIP, KEGG, and others. The results also support the notion that most scientists may have been making very arbitrary choice of pathway databases.

bioinformatics↗

Tracing the function expansion for a primordial protein fold in the era of fold-based function prediction: beta-trefoil

The incredibly narrow protein fold bottleneck, which separates the billions of unique proteins on one side to deliver diverse biological functions on the other, arises from folds that tolerate mutations during evolution. One such fold, called the {beta}-trefoil, is present in functionally diverse proteins including cytokines involved in the immune system such as interleukin-1. The unrecognizable sequence-level diversity, even among paralogs of interleukin-1 within the same chromosomal locus, suggests the resilience of this fold to mutational onslaught. Furthermore, {beta}-trefoil domain containing-proteins are known to coexist with other domains to achieve functional diversity. In this study, we challenge the reach and limitations of function prediction using fold-fold comparison using {beta}-trefoil fold as an example. We identified proteins containing {beta}-trefoil fold belonging to thirty-two distinct functional class based on diverse domain architecture and or functional annotation by mining both the PDB and AlphaFold databases using fold-fold comparison. Among the proteins with novel domain architecture we find {beta}-trefoil along with chitinase, lipase, {beta}-glucosidase, protein kinase, peptidoglycan-binding + peptidase matrixin, glycosyl hydrolases family 3 + PA14 + fibronectin type-III, alpha galactosidase A, PhoD-like phosphatase, insecticidal crystal toxin, trypsin, alginate lyase and two novel structurally uncharacterized domains. We demonstrate that fold-fold comparison can extend function prediction beyond the reach of sequence-based approach and provides an opportunity to discover novel domain architecture associated with known folds. However, since extending fold similarity to functional similarity may be challenged by convergent fold evolution, we explore if {beta}-trefoil may be a convergent evolution and share our hypothesis.

bioinformatics↗

Single Cell Spatial Transcriptomics Reveals Immunotherapy-Driven Bone Marrow Niche Remodeling in AML

Given the successful graft-versus-leukemia cell treatment effect observed with allogeneic hematopoietic stem cell transplant for patients with refractory or relapsed acute myeloid leukemia, immunotherapies have also been investigated in the nontransplant setting. Here, we use a multi-omic approach to investigate spatiotemporal interactions in the bone marrow niche between leukemia cells and immune cells in patients with refractory or relapsed acute myeloid leukemia treated with a combination of the immune checkpoint inhibitor pembrolizumab and hypomethylating agent decitabine. We derived precise segmentation data by extensively training nuclear and membrane cell segmentation models, which enabled accurate transcript assignment and deep learning-feature-based image analysis. To overcome read-depth limitations, we integrated the single-cell RNA sequencing data with single-cell-resolution spatial transcriptomic data from the same sample. Quantifying cell-cell distances between cell edges rather than cell centroids allowed us to conduct a more accurate downstream analysis of the tumor microenvironment, revealing that multiple cell types of interest had global enrichment or local enrichment proximal to leukemia cells after pembrolizumab treatment, which could be associated with their clinical responses. Furthermore, ligand-receptor analysis indicated a potential increase in TWEAK signaling between leukemia cells and immune cells after pembrolizumab treatment. HighlightsO_LISpatial transcriptomic analysis of R-AML bone marrow niches provides detailed information about intercellular interactions in the tumor microenvironment. C_LIO_LIImmunotherapy shifts the cell composition of the leukemia neighborhood. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=199 SRC="FIGDIR/small/634753v1_ufig1.gif" ALT="Figure 1"> View larger version (93K): org.highwire.dtl.DTLVardef@955273org.highwire.dtl.DTLVardef@158234corg.highwire.dtl.DTLVardef@175837borg.highwire.dtl.DTLVardef@b0a38_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

Caught in the act of triplication: TNF superfamily

In humans, the signaling mechanisms of the 19 paralogs of the tumor necrosis factor superfamily (TNFSF) and the 29 receptor paralogs of the tumor necrosis factor receptor superfamily (TNFRSF) are extensively characterized because of their therapeutic relevance. The functional expansion of TNFSF in vertebrates from a single ancestral gene through successive duplication events is also well established. However, apart from the first identification of a TNFSF homolog, Eiger (dmEiger), in Drosophila melanogaster in 2002, together with its receptor homologs Wengen (dmWgn) and Grindelwald (dmGrnd), this signaling system has remained largely unexplored in invertebrates. More recently, the implication of an Eiger homolog in Plasmodium resistance in malaria vectors has further highlighted the need for a systematic investigation of this pathway in lower invertebrates. Structural comparison of the dmEiger-dmGrnd complex with the canonical 3:3 ligand-receptor configuration observed in human TNFSF-TNFRSF signaling suggests either conservation of this signaling modality since before the bilaterian split or convergent evolution of a similar architecture in both branches. The recent explosion in high-quality proteomes spanning diverse phyla, together with advances in protein-complex prediction using AlphaFold-multimer, now enables large-scale exploration of ligand-receptor evolution across invertebrates. Here, we analyzed 148 near-complete proteomes spanning major invertebrate phyla and identified 290 TNFSF, 336 wengen (wgn), and 115 grindelwald (grnd) homologs, including homologs from lower invertebrates. Structural characterization of 140 selected complexes using AlphaFold and AlphaFold-multimer revealed several key findings: (i) TNFSF and TNFRSF homologs are present in majority of the phyla under invertebrates (ii) the canonical 3:3 ligand-receptor signaling configuration is conserved across invertebrates; (iii) orthologs of 25 out of the 26 genes implicated in TNF signaling pathways are present in lower invertebrates; and (iv) signaling through grnd-like receptors containing a single cysteine-rich domain with CXXCXXXC signature is the predominant signaling mode in invertebrates and becomes highly prevalent in Arthropoda. We also elaborate a hypothesize on the evolutionary trajectories toward a genetically parsimonious signaling by this complex system before functional expansion in vertebrates and species diversification in Arthropoda.

bioinformatics↗

Proteomic analysis of gut in Labeo rohita reveals ECM as Key Player in host's Response to Aeromonas hydrophila Infection

In the aquaculture sector, one of the challenges include disease outbreaks such as bacterial infections, particularly from Aeromonas hydrophila (Ah), impacting both wild and farmed fish. In this study, we conducted a proteomic analysis of the gut tissue in Labeo rohita following Ah infection to elucidate the protein alterations and its implications for immune response. Our findings reveal significant dysregulation in extracellular matrix (ECM) associated proteins during Ah infection, with increased abundance of elastin and Collagen alpha-3(VI) contributing to matrix rigidity. Pathway and enrichment analysis of differentially expressed proteins (DEPs) highlights the involvement of ECM-related pathways, including Focal adhesions, Integrin cell surface interactions, and actin cytoskeleton organization. Focal adhesions, crucial for connecting intracellular actin bundles to the ECM, play a pivotal role in immune response during infections. Increased abundance of integrin alpha 1, integrin beta 1, and Tetraspanin suggests their involvement in the hosts response to Ah infection. Proteins associated with actin cytoskeleton reorganization, such as myosin, tropomyosin, and phosphoglucomutase, exhibit increased abundance, influencing changes in cell behavior. Additionally, upregulated proteins like LTBP1 and Fibrillin-2 contribute to TGF-{beta} signaling and focal adhesion, indicating their role in immune regulation. The study also identifies elevated levels of laminin, galectin 3, and tenascin-C, which interact with integrins and other ECM components, influencing immune cell migration and function. These proteins, along with decorin and lumican, act as immunomodulators, coordinating pro- and anti-inflammatory responses. ECM fragments released during pathogen invasion serve as "danger signals," initiating pathogen clearance and tissue repair through Toll-like receptor signaling. IMPORTANCEThe study underscores the critical role of the extracellular matrix (ECM) and its associated proteins in the immune response of aquatic organisms during bacterial infections like Aeromonas hydrophila (Ah). Understanding the intricate interplay between ECM alterations and immune response pathways provides crucial insights for developing effective disease control strategies in aquaculture. By identifying key proteins and pathways involved in host defense mechanisms, this research lays the groundwork for targeted interventions to mitigate the impact of bacterial infections on fish health and aquaculture production.

microbiology↗

Prolonged in-vivo tracking of vitreous fluid and its early diagnostic imaging biomarkers for cancer growth

PurposeEstimation of a correlation between cells in vitreous humour and growth in glioblastoma xenografts. MethodsStreams of cells in vitreous humor are observed in optical coherence tomography (OCT) imaging data of animal (NSGS and Athymic Nude-Foxn1nu) eyes (34 in total) subjected to xenograft growth study, in-vivo. The cancer disease model is studied with and without nanodrug-based treatment protocols. ResultsThe presence of CD8+ and CD4+ is reported inside the tumor using the same data earlier. The transition of these cells is shown to take place from the optic nerve via the vitreous into the nerve fiber layer (NFL) at tumor locations and xenograft -related injuries. Functional analysis of dense temporal imaging series (varying from 28 to more than 100 days) reveals a mild correlation between the volumetric growth of the tumor with the density of these cells, quantitatively and qualitatively. The cross-correlation analysis indicates imaging assisted photodynamic treatment protocol perform relatively better if started with certain delay. Doxorubicin treatment to Nu/Nu Male and NSGS female transforms mild weak negative correlation into mild weak positive correlation. ConclusionsThe plots indicate that the mix of the cells in vitreous humor are effectively dominated by immunosuppressor cytotoxic component. Translational RelevanceWe propose that the vitreous cell density can be used as imaging biomarker helpful for clinicians in early diagnosis and treatment planning of similar disease models. The limitation of this work is that high resolution OCT systems and data-dependent image segmentation methods are required.

cancer biology↗

Proteomic analysis of liver tissue reveals Aeromonas hydrophila infection mediated modulation of host metabolic pathways in Labeo rohita

Aeromonas hydrophila (Ah) is an opportunistic Gram-negative bacterium and a serious global pathogen causing Motile Aeromonas Septicaemia (MAS) in fish and many other vertebrates. The pathogenesis of aeromonas septicaemia is complex and involves multiple perturbed pathways. Molecular analysis of host tissues could be a powerful approach to identify mechanistic and diagnostic immune signatures of disease. We performed a deep proteomic analysis of Labeo rohita liver tissue to examine changes in the host proteome during Ah infection. A total of 2525 proteins were identified of which 158 were found differentially expressed during Ah infection. Functional analysis of significant proteins identified the dysregulation of several metabolic enzymes, antioxidative proteins, cytoskeletal proteins and immune related proteins. Proteomic analysis revealed the alterations in the cellular defence mechanisms including phagolysosomal killing and apoptosis during Ah infection. Our systemic approach revealed the protein dynamics in the host cells to explore the putative biological processes underlying the metabolic reprogramming of the host cells during Ah infection. Our findings paved the way for future research into the role of Toll-like receptors (Tlr3), C-type lectins (Clec4e) and metabolic enzymes in Ah pathogenesis leading towards host directed immunotherapies to tackle the Ah infection in fish. IMPORTANCEBacterial disease is one of the most serious problems in aquaculture industry. Aeromonas hydrophila (Ah), a Gram-negative bacterium causes motile aeromonas septicaemia (MAS) in fish. Small molecules that target the metabolism of the host have recently emerged as potential treatment possibilities in infectious diseases. However, the ability to develop new therapies is hampered due to lack of knowledge about pathogenesis mechanisms and host-pathogen interactions. Molecular level analysis of host tissues could be helpful in finding mechanistic immunological markers of diseases. We examined alterations in the host proteome during Ah infection in Labeo rohita liver tissue to find cellular proteins and processes affected by Ah infection. Our systemic approach revealed protein dynamics underlying the host cells metabolic reprogramming during Ah infection. Our work is an important step towards leveraging host metabolism in targeting the disease by providing a bigger picture on proteome pathology correlation during Ah infection.

systems biology↗

Biocompatible soft hydrogel lens as topical implants for diabetic retinopathy

Fashioned contact lenses can be converted into novel drug delivery vehicles. Ocular disease like diabetic retinopathy is a major microvascular complication where its early diagnosis and treatment is still a mystery to clinicians. Delivery of pharmaceuticals to the posterior part of eye is quite difficult, with exemption from the injectable formulations. Drug loaded hydrogel like contact lens implants can be utilized in place of commercial contact lens. Such hydrogel lens implants are developed with precise manner in which they are to be inserted as implants in the mice models. The present work is the preliminary outcome of our ongoing research work where the polymerized lens implant can be used as payload carrier for retinopathy. The aim is to propose such appropriate implants suitable for in vivo mice models.

bioengineering↗

Molecular measurable residual disease testing of blood during AML cytotoxic therapy for early prediction of clinical response

AbstractO_ST_ABSPURPOSEC_ST_ABSMeasurable residual disease (MRD) testing after initial chemotherapy treatment can predict relapse and survival in AML. However, it has not been established if repeat molecular or genetic testing during chemotherapy can offer information regarding the chemotherapy sensitivity of the leukemic clone.\n\nPATIENTS AND METHODSBlood from 45 adult AML patients at day 1 and 4 of induction (n = 35) or salvage (n = 10) cytotoxic chemotherapy was collected for both quantitative real-time PCR (qPCR) assessment (WT1) and next generation sequencing (NGS, >500x depth) of 49 gene regions recurrently mutated in MDS/AML.\n\nRESULTSThe median age was 62 (23-78); 42% achieved a complete response. WT1 was overexpressed in most patients tested but was uninformative for very early MRD assessment. A median of 4 non-synonymous variants (range 0-7) were detected by DNA sequencing of blood on day 1 of therapy (median VAF: 29%). Only two patients had no variants detectable. All mutations remained detectable in blood on day 4 of intensive chemotherapy and remarkably the ratio of mutated to wild-type sequence was often maintained. This phenomenon was not limited to variants in DNMT3A, TET2 and ASXL1. The kinetics of NPM1 and TP53 variant burden early during chemotherapy appeared to be exceptions and exhibited consistent trends in this cohort.\n\nCONCLUSIONSMolecular testing of blood on day 4 of chemotherapy is not predictive of clinical response in AML. The observed stability in variant allele frequency suggests that cytotoxic therapy may have a limited therapeutic index for clones circulating in blood containing these mutations. Further validation is required to confirm the utility of monitoring NPM1 and TP53 kinetics in blood during cytotoxic therapy.

cancer biology↗

Human Bone Marrow Assessment by Single Cell RNA Sequencing, Mass Cytometry and Flow Cytometry

New techniques for single-cell analysis have led to insights into hematopoiesis and the immune system, but the ability of these techniques to cross-validate and reproducibly identify the biological variation in diverse human samples is currently unproven. We therefore performed a comprehensive assessment of human bone marrow cells using both single-cell RNA sequencing and multiparameter flow cytometry from twenty healthy adult human donors across a broad age range. These data characterize variation between healthy donors as well as age-associated changes in cell population frequencies. Direct comparison of techniques revealed discrepancy in the quantification of T lymphocyte and natural killer cell populations. Orthogonal validation of immunophenotyping using mass cytometry demonstrated good correlation with flow cytometry. Technical replicates using single-cell RNA sequencing matched robustly, while biological replicates showed variation. Given the increasing use of single-cell technologies in translational research, this resource serves as an important reference dataset and highlights opportunities for further refinement.

immunology↗

Theaflavins, polyphenols of black tea, inhibit entry of hepatitis C virus

The treatment of hepatitis C virus (HCV) infection by combination of direct acting antivirals (DAA), with different mode of action, has made substantial progress in the past few years. However, appearance of resistance and high cost of the therapy is still an obstacle in the achievement of the therapy, more specifically in developing countries. In this context, search for affordable antivirals with new mechanisms of action is still needed. Tea, after water, is the most popular drink worldwide. Polyphenols extracted from green tea have already shown anti-HCV activity as entry inhibitors. Here, three different theaflavins, theaflavin (TF1), theaflavin-3-monogallate (TF2), and theaflavin-3-3-digallate (TF3), which are major polyphenols from black tea, were tested against HCV in cell culture. The results showed that all theaflavins inhibit HCV infection in a dose-dependent manner in an early step of infection. Results obtained with HCV pseudotyped virions confirmed their activity on HCV entry and demonstrated their pan-genotypic action. No effect on HCV replication was observed by using HCV replicon. Investigation on the mechanism of action of black tea theaflavins showed that they act directly on the virus particle and are able to inhibit cell-to-cell spread. Combination study with inhibitors most widely used in anti-HCV treatment regimen demonstrated that TF3 exerts additive effect. In conclusion, theaflavins, that are present in high quantity in black tea, are new inhibitors of HCV entry and hold promise for developing in therapeutic arsenal for HCV infection.

microbiology↗