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Joshi, G.

Publications and source records attributed to Joshi, G..

10 recordsLinked to original sources

Cortical Excitation-Inhibition Balance in Autism Varies by Brain Region and Age

Autism spectrum disorder (ASD) is characterized by differences in social communication, interaction, and restricted and repetitive behaviors. The hypothesis that ASD involves altered cortical excitation-inhibition (EI) balance has been extensively investigated, yet evidence remains mixed, partly because EI balance changes substantially during typical maturation. We used resting-state magnetoencephalography (MEG) to examine developmental trajectories of cortical EI alterations with regional specificity in a large cross-sectional cohort (N = 172; 92 typically developing, 80 ASD; ages 6-32). Functional EI (fEI), derived from critical brain dynamics, was estimated across 500 cortical parcellations. Group-averaged fEI maps revealed broadly similar large-scale topographic patterns across groups. Age-stratified analyses revealed increased global fEI in childhood and decreased fEI in adolescence in ASD, with no evidence of a group difference in adulthood. Parcel-wise regressions identified a significant main effect of diagnosis in right dorsolateral prefrontal cortex, where typically developing individuals showed higher fEI across all ages. A significant group-by-age interaction in left inferior parietal cortex indicated diverging developmental trajectories. Finally, fEI in paracentral and precuneus regions was associated with the magnitude of ASD symptoms. These findings indicate that EI imbalance in ASD is neither globally distributed nor static, but expressed through regionally and developmentally specific differences, with relevance for ASD heterogeneity.

neuroscience↗

SMAD4 loss drives cancer chromosomal instability through transcription-replication conflict-induced replication stress

Cancer chromosomal instability (CIN) drives tumor evolution through generating structural and numerical chromosome aberrations. However, its molecular determinants remain poorly characterized. Here, we uncover a yet unrecognized role for the tumor suppressor SMAD4 as a key regulator of genome stability. Mechanistically, loss of SMAD4 induces replication stress by promoting transcription-replication conflicts (TRCs), which induces chromosomal breaks and subsequent mitotic errors, leading to the concomitant emergence of structural and numerical CIN. While the tumor suppressor function of SMAD4 has been predominantly attributed to TGF-{beta} signaling, we find that its role in genome maintenance operates via BMP signaling and its transcriptional target gene ID3. Consequently, inhibition of BMP signaling mimics the loss of SMAD4 whereas re-expression of SMAD4 or ID3 suppresses TRCs, replication stress and mitotic errors in SMAD4-deficient cancer cells. Our findings reveal a TRC- and replication stress-driven mechanism by which loss of BMP-SMAD4 signaling causes cancer chromosomal instability.

cancer biology↗

Coronin1C SUMOylation modulates filopodia formation, neuritogenesis, and neuronal differentiation

Actin dynamics in the cytosol and at the cell periphery are critical for metabolic processes and the formation of cellular projections. Coronin1C, a versatile cytoskeletal regulator, is an actin-binding protein that associates with actin at the leading edge of a cell. Consequences of Coronin1C interactions on actin dynamics enable actin-mediated filopodia and neurite formation. Here, we report that Coronin1C is SUMOylated at multiple lysine residues in its carboxy-terminus, preferentially by SUMO1. SUMOylation of Coronin1C is critically required for efficient neurite formation and extensive cellular projections during neuronal differentiation. Moreover, SUMOylation-deficient Coronin1C mutant forms cytoplasmic aggregates colocalizing with stress granules, a hallmark of neurodegenerative diseases, under neuronal differentiation conditions. We also demonstrate that Coronin1C SUMOylation works in synergy with Cdc42 GTPase towards actin-polymerization and promotes filopodia formation. In conclusion, we report that Coronin1C SUMOylation by SUMO1 in its carboxy-terminus is critical for cellular projections formation and neuronal differentiation.

cell biology↗

Immunogenicity and protection efficacy of self-amplifying and circular mRNA vaccines for SARS-CoV-2

Recent advances in vaccine technology have positioned messenger RNA (mRNA) vaccines as safe and reliable options for human use. Conventionally, mRNA vaccines were designed using linear or self-amplifying mRNA (SAM), the latter considered to be superior. However, limited success was achieved with SAM vaccines during the COVID-19 pandemic. Further, studies on Circular mRNA (Circ-RNA) vaccines against the SARS-CoV-2, Ebola and monkey pox proved their efficacy. Circ-RNAs are highly stable, neither they induce inflammatory response nor require any extracellular protein for their function. Here, we compared the efficacy of SAM- and Circ-RNA vaccines using the SARS-CoV-2-RBD (receptor binding domain) as the antigen. Both SAM-RBD and Circ-RBD induced a comparable anti-RBD IgG titer and virus-neutralizing antibody titer. However, the latter induced a significantly higher memory T-cell response. Immunization with SAM- and Circ-RBD showed no mortality and improved lung pathophysiology against acute SARS-CoV-2 infection in mice. The Circ-RBD vaccine is stable for 4 weeks at 40C. A bivalent vaccine containing Circ-RBD of both delta and omicron SARS-CoV-2 variants potently neutralized these viruses. These findings demonstrate Circ-RNA-RBD as an excellent vaccine candidate against COVID-19 and also provide a platform for developing bivalent Circ-RNA vaccine candidates against SARS-CoV-2 or other viruses with rapidly emerging variants.

microbiology↗

Hepatitis E Virus-induced antiviral response by plasmacytoid dendritic cells is modulated by the ORF2 protein

Type I and III interferons (IFN-I/III) are critical to protect the host during viral infection. Previous studies have shown that IFN-mediated antiviral responses against hepatitis E virus (HEV) are suppressed and defeated by viral escape mechanisms at play in infected hepatocytes. Here, we studied the anti-HEV function of IFN secreted by plasmacytoid dendritic cells (pDCs), which are specialized producers of IFNs. We showed that pDCs co-cultured with HEV-replicating cells secreted IFN in a cell-to-cell contact-dependent manner. Pharmacological inhibitor and antibodies targeting contact proteins revealed that pDC response against HEV required the endosomal nucleic-acid sensor TLR7 and adhesion molecules, such as ICAM-I and L{beta}2-integrin. IFNs secreted by pDCs reduced viral spread. Intriguingly, ORF2, the capsid protein of HEV, can be produced in various forms by the infected cells. During infection, a fraction of the intracellular ORF2 protein localizes into the nucleus while another ORF2 fraction packages viral genomes to produce infectious virions. In parallel, glycosylated forms of ORF2 are also massively secreted by infected cells. Using viral genome expressing ORF2 mutants, we showed that glycosylated ORF2 forms contribute to better recognition of infected cells by pDCs via regulation of contacts between infected cells and pDCs. ORF2 forms may thus modulate pDC-mediated anti-HEV response. Together, our results suggest that liver-resident pDCs, which exhibit comparable IFN-producing ability as blood-derived pDCs, may be essential to control HEV replication.

immunology↗

Inhibition of BCR::ABL1 tyrosine kinase activity Aids in the Generation of Stable Chronic Myeloid Leukemia Induced Pluripotent Stem Cells

Induced pluripotent stem cells (iPSCs) generated from patients with chronic myeloid leukemia (CML) have the potential for disease modeling to study disease pathogenesis and screening therapeutic interventions. In this study, we aimed to generate iPSCs from CD34+ hematopoietic progenitors of CML patients with varying responses to tyrosine kinase inhibitor (TKI) therapy. The generated CML-CD34-iPSC colonies displayed atypical "dome-shaped" morphology and underwent spontaneous differentiation in a few days. However, supplementation with imatinib (IM), the most widely used TKI to treat CML patients, in the culture medium improved the stability and maintenance of all isolated CML-CD34-iPSC colonies, allowing them to be maintained for more than 20 passages without significant differentiation. In contrast to previous studies, our results indicate that suppressing the BCR::ABL1 oncogenic pathway is essential for efficiently generating stable CML-iPSC colonies. Furthermore, we successfully differentiated these iPSCs to CD34+ hematopoietic progenitors both in the presence and absence of IM. This robust protocol for generating CML-iPSCs provides a valuable resource for disease modelling. The generated iPSCs will be a valuable tool for investigating CML pathophysiology, drug resistance mechanisms, and drug screening to identify novel and effective therapies for this disease.

cancer biology↗

Dietary αKG inhibits SARS CoV-2 infection and rescues inflamed lungs to restore normal O2 saturation in animals

Our recent works described the rescue effect of -ketoglutarate (KG, a metabolite of Krebs cycle) on thrombosis and inflammation in animals. KG augments activity of prolyl hydroxylase 2 (PHD2), which in turn degrades proline residues of substrates like phosphorylated Akt (pAkt) and hypoxia inducible factor (HIF). Here we describe the inhibitory effect of octyl KG on pAkt as well as on HIF1/HIF2, and in turn decreasing SARS CoV-2 replication in Vero E6 cells. KG failed to inhibit the viral replication and Akt phosphorylation in PHD2-knockdown U937 cells transiently expressing ACE2. Contrastingly, triciribine (TCN, an Akt-inhibitor) inhibited viral replication alongside a downmodulation of pAkt in PHD2-KD cells. Dietary KG significantly inhibited viral infection and rescued hamsters from thrombus formation and inflammation in lungs, the known causes of acute respiratory distress syndrome (ARDS) in COVID-19. KG supplementation also reduced the apoptotic death of lung tissues in infected animals, alongside a downmodulation of pAkt and HIF2. KG supplementation neither affected IgG levels against SARS CoV-2 RBD protein nor altered the neutralization antibody response against SARS CoV-2. It did not interfere with the percentage of interferon-{gamma} positive (IFN{gamma}+) CD4+ and IFN{gamma}+CD8+ T cells in infected animals. The extended work in balb/c mice transiently expressing ACE2 showed a similar effect of KG in reducing accumulation of inflammatory immune cells and cytokines, including IL6, IL1{beta} and TNF, in lungs as well as in circulation of infected animals. Pro-thrombotic markers like platelet microparticles and platelet-leukocyte aggregates were reduced significantly in infected mice after KG supplementation. Importantly, KG supplementation restored the O2 saturation (SpO2) in circulation of SARS CoV-2 infected hamsters and mice, suggesting a potential therapeutic role of this metabolite in COVID-19 treatment.

pathology↗

B.1.617.3 SARS CoV-2 spike E156G/Δ157-158 mutations contribute to reduced neutralization sensitivity and increased infectivity

SARS CoV-2 variants raise significant concerns due to their ability to cause vaccine breakthrough infections. Here, we sequence-characterized the spike gene, isolated from a breakthrough infection, that corresponded to B.1.617.3 lineage. Delineating the functional impact of spike mutations using reporter pseudoviruses (PV) revealed that N-terminal domain (NTD)-specific E156G/{Delta}157-158 contributed to increased infectivity and reduced sensitivity to ChAdOx1 nCoV-19 vaccine (Covishield)-elicited neutralizing antibodies. A six-nucleotide deletion (467-472) in the spike coding region introduced this change in the NTD. We confirmed the presence of E156G/{Delta}157-158 in the RT-PCR-positive cases concurrently screened, in addition to other circulating spike (S1) mutations like T19R, T95I, L452R, E484Q, and D614G. Notably, E156G/{Delta}157-158 was present in more than 85% of the sequences reported from the USA, UK, and India in August 2021. The spike PV bearing combination of E156G/{Delta}157-158 and L452R further promoted infectivity and conferred immune evasion. Additionally, increased cell-to-cell fusion was observed when spike harbored E156G/{Delta}157-158, L452R, and E484Q, suggesting a combinatorial effect of these mutations. Notwithstanding, the plasma from a recovered individual robustly inhibited mutant spike PV, indicating the increased breadth of neutralization post-recovery. Our data highlights the importance of spike NTD-specific changes in determining infectivity and immune escape of variants.

microbiology↗

De novo transcriptome profiling of mustard aphid (Lipaphis erysimi) and differential expression of transcripts associated with developmental stages, feeding and non-feeding conditions

Lipaphis erysimi is a Brassicaceae specialist aphid, which causes significant losses in yield and/or reduction of oil content of vegetable and oilseed brassicas and is a major pest in the Indian subcontinent. This study reports the de novo transcriptome of L. erysimi for the first time. We also present a comparative analysis of nymphs and adult transcriptomes to study the differential expression profiles associated with different developmental stages as well as different feeding conditions. For this, RNA-seq was performed on three different biological samples adults, nymphs (with all nymph stages pooled) and adults starved for 3 hours (referred to as Adult Feeding, AF; Nymphs Feeding, NF, and Adult Starved_3 hr, ANF samples henceforth). A final transcriptome comprising 52,652 transcripts of 1064bp average length and N50 value of 1806 bp was generated. A total of 27,112 transcripts were annotated with insect proteins from SwissProt, of which 4128 transcripts were components of 165 KEGG pathways. A total of 17,296 transcripts were classified based on their Gene Ontology. Potential transcripts for host selection, detoxification, salivary proteins and effectors, molecular chaperones and developmental genes were identified. A total of 23,532 transcripts that remained unannotated were subjected to BLAST against aphid sequences available at AphidBase and a total of 3091 transcripts had hits with sequences of other aphids in the database, out of which 1380 had protein hits. A total of 20441 found to share no homology to any sequence available in the public domain and could therefore represent novel aphid genes or sequences that are unique to L. erysimi. This is an exploratory study with no biological replicates. However, the significant repertoire of feeding- and development-related genes and their differential expression profiles generated in this study adds to the limited data available on L. erysimi and it would facilitate studies on the molecular basis of aphid feeding and development. This could also allow identification of novel target genes for development of RNAi-based aphid control methods.

systems biology↗

Telomere elongation and Telomerase activity in Normal and Cancer cell lines: HEK-293, HeLa and A549

Telomerase, an eukaryotic ribonucleoprotein (RNP) complex, consists of an essential RNA template and a reverse transcriptase termed as human telomerase reverse transcriptase (hTERT). By using this reverse transcriptase enzyme, telomerase RNP maintains its telomeric length in all cancer cells and in few stem cells. As cells age, telomerase level decreases and telomere get shorten, leading to senescence. Presence of telomerase helps delay the senescence due to active production of telomere, leading to cell turning malignant and eventually cancerous. In this experiment, we would investigate the telomerase activity tested using q PCR and telomerase level using western blot in three different cell lines, HEK-293, HeLa and A549.\n\nHypothesisWhere HEK-293 cells have longer telomeres and higher telomerase activity, HeLa and A549 cell lines will have varied length of telomere but higher telomerase activity.

molecular biology↗