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Biology subjects

Ramasamy, R.

Publications and source records attributed to Ramasamy, R..

8 recordsLinked to original sources

CTR1-mediated copper uptake orchestrates metabolic-epigenetic regulation of pathogenic TH17 cells in autoimmune disease

Pathogenic T helper 17 (pTH17) cells are a subset of CD4+ T cells driving autoimmune diseases including multiple sclerosis (MS). Compared to homeostatic TH17 cells and other TH subsets, pTH17 have enhanced mitochondrial function and oxidative phosphorylation (OXPHOS) that supports their differentiation and pathogenic function. Here we identify Copper Transporter 1 (CTR1), encoded by Slc31a1, as essential for copper uptake in CD4+ T cells, OXPHOS and pTH17 cell differentiation and function. While copper levels are known to be higher in the cerebrospinal fluid of patients with MS compared to healthy individuals, and excess copper contributes to oligodendrocyte loss in murine models of MS, the effect of copper on T cell function and pathogenicity in MS are unclear. We demonstrate that deletion of Slc31a1 in CD4+ T cells decreased intracellular copper levels, disrupting mitochondrial respiration and rewiring metabolism. These changes altered the epigenetic landscape of pTH17 cells by impairing DNA demethylation capacity, leading to hypermethylated DNA and altered chromatin accessibility at key binding sites for AP-1 transcription factors essential for pTH17 differentiation. As a result, CTR1-deficient T cells showed defective differentiation into pTH17 cells, with decreased production of IL-17A and expression of TH17 signature genes, while the differentiation of other CD4+ T cell subsets remained largely unaffected. Moreover, T cell-specific deletion of Slc31a1 protected mice from central nervous system (CNS) inflammation in the experimental autoimmune encephalomyelitis (EAE) model of MS by suppressing clonal expansion of autoreactive CD4+ T cells. These findings establish copper as a critical regulator of pTH17 differentiation and function, revealing a previously unknown molecular link between copper homeostasis, metabolism and epigenetic regulation governing pTH17-mediated autoimmunity.

immunology↗

The circadian system is affected by Alzheimers disease independently from amyloid beta deposits

Circadian disruption, notably sleep disturbances, serves as an early indicator of Alzheimers disease (AD), preceding cognitive symptoms like memory loss. The suprachiasmatic nucleus (SCN) governs biological rhythms and receives direct retinal input via melanopsin-expressing retinal ganglion cells (mRGCs) to synchronize with environmental light cycles. The anatomical and functional basis for circadian disruption in AD remains unclear. Here, we explored the multi-level relationships between gene expression, the SCN connectome, and regulations of sleep and circadian rhythms in the APP/PS1 mouse model. The sleep architecture of APP/PS1 mice displayed significantly reduced rapid eye movement sleep (REM), associated with a reduced daily core body temperature amplitude and locomotor hyperactivity. Lastly, APP/PS1 mice showed an impaired response to acute light pulse stimulation and present hyperactivity of mRGCs at a young age and hypoactivity of these cells at older ages. These physiological functions are known to be, at least in part, regulated by the SCN, the main target of mRGCs. We noted several modifications in SCN connectomics using serial blockface electron microscopy (SBEM), including a reduction of the dendro-dendritic chemical synapse (DDCS) network that receives a large part of the retinal input and is thought to be crucial for synchronicity between SCN neurons. In addition, we observed multiple signs of dystrophy, including modifications of the shape of dendrites and cell soma, accumulation of aggregated lysosomes, and swelling of axons. At the same time, we investigated the changes in gene expression using spatial transcriptomics. The SCN presents changes in the expression of genes associated with synapse formation, cell adhesion, and neurite growth. These results suggest that, despite the absence of amyloid plaques in the ventral hypothalamus, the SCN of APP/PS1 mice still undergo profound gene expression changes, impacting connectomics and physiological functions. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=157 SRC="FIGDIR/small/744599v1_ufig1.gif" ALT="Figure 1"> View larger version (39K): org.highwire.dtl.DTLVardef@ceedb0org.highwire.dtl.DTLVardef@156cfaaorg.highwire.dtl.DTLVardef@5bc262org.highwire.dtl.DTLVardef@36df4d_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

DIANNE: Segmentation-Free Localization of Histology Differential Attributes

Pathologist-guided distinctions within histology and spatial omic images provide insights into health and disease, with digital pathology leveraging artificial intelligence to automate such assessments. To train computational models, current digital pathology methods rely on upfront manual annotations, which are time-consuming to generate. Pre-annotation is poorly suited to investigating novel spatial behaviors--a major need driven by advances in spatial profiling--for which annotation criteria and data needs will be uncertain. To address these challenges, we present DIANNE, a digital pathology annotation support and discovery approach for rapid training and inference of spatial differential attributes, enabled by train-time Positive Class Mixup Augmentation. DIANNE can compute foundation model-derived segmentation-free localization of differential classifiers across whole slide H&E images within seconds on a workstation, enabling interactive investigation of spatial niches. Predictive models can be re-trained in real-time in response to patch or regional annotation changes, clarifying determinative biological attributes across slides from only a few dozen annotated patches. We demonstrate the effectiveness of DIANNE for tumor detection, artifact identification, and exploration of pancreatic, fetal membranes and kidney tissue structures. DIANNE also provides analogous capabilities for IHC, multiplex immunofluorescence, and registered spatial transcriptomic+H&E images. DIANNE is implemented in a Jupyter toolkit, enabling rapid development of high-resolution classifiers from weakly-supervised training. DIANNE provides a practical system to quantitatively understand known and novel spatial phenotypes.

pathology↗

Alternative splicing expands the functional portfolio of a plant virus to control the viral cycle

Viruses frequently have limited coding space, yet efficiently manipulate complex hosts by deploying intricate, often poorly understood strategies. Geminiviruses, devastating plant pathogens with small single-stranded (ss) DNA genomes, replicate in the nucleus aided by the viral replication-associated protein (Rep), the most conserved protein within this family and related ssDNA viruses. Rep initiates viral DNA replication and represses its own promoter to regulate the infection cycle. The molecular mechanisms enabling this dual functionality are so far unknown. Here, we show that the geminivirus tomato yellow leaf curl virus (TYLCV) exploits the host spliceosome to produce novel Rep splice variants. Splicing generates Rep isoforms lacking the central oligomerization domain that cannot initiate replication but strongly repress the Rep promoter. Conversely, Rep mutants deficient in splicing promote replication but fail to repress transcription. Reduced Rep splicing interferes with the viral gene expression hierarchy and decreases infectivity. Our findings therefore reveal a previously unrecognized viral strategy in which alternative splicing produces functionally specialized viral protein isoforms, providing a mechanistic explanation for the dual role of Rep in viral replication and gene regulation. Splicing events with potentially similar functional consequences in related viruses suggest that this strategy may have convergently evolved across diverse viral lineages infecting different domains of life.

plant biology↗

Age-Related Decline in NCKX4-Mediated Calcium Clearance Accelerates Aortic Remodeling and Drives Early Vascular Aging

Aging is the primary nonmodifiable risk factor for cardiovascular diseases (CVDs), with older women facing a greater risk of CVDs than age-matched men. Vascular smooth muscle cells (VSMCs) dysfunction and impaired calcium (Ca2+) handling are recognized as central contributors to arterial stiffening and calcification. However, the molecular and functional determinants of Ca2+ clearance in vascular aging remains a topic of ongoing research. We identify the (Na+)-sodium/Ca2+-calcium (K+)-potassium-dependent exchanger 4 (NCKX4) as a critical functional regulator of VSMCs Ca{superscript 2} clearance and vascular integrity. We demonstrate that NCKX4 (coded by Slc24A4) expression is markedly reduced in aorta of aged (72-78 weeks) mice, with a pronounced decline in females. Functional assays revealed impaired Ca2+ clearance in both aged and Nckx4-/- VSMCs, which was accompanied by increased calcification. Histomorphometric analyses of young Nckx4-/- mice revealed fragmentation of elastic fibers, collagen accumulation, wall thickening, and extracellular matrix (ECM) remodeling, all hallmarks of vascular aging that closely resembled those of aged wild-type mice. Transcriptomic profiling of VSMCs showed that loss of NCKX4 alters pathways linked to Ca2+-integrin signaling, ECM turnover, and mineralization, including dysregulation of protective anchorage integrins, microfibril-stabilizing, osteogenic drivers and pro-fibrotic integrins. These findings support a model in which impaired Ca2+ clearance promotes maladaptive inside-out integrin signaling, disrupting VSMCs anchorage, ECM homeostasis, and mineralization processes. Collectively, our results establish NCKX4 as a previously unrecognized determinant of vascular aging, whose decline accelerates premature arterial remodeling and calcification. This study positions NCKX4 as a potential mechanistic link between age, sex-dependent vulnerability, and vascular stiffening, with implications for novel therapeutic strategies targeting Ca2+ handling in CVDs prevention.

cell biology↗

Preimaginal development of Aedes aegypti in brackish water produces adult mosquitoes with thicker cuticles and greater insecticide resistance

Aedes aegypti and Aedes albopictus mosquitoes, the principal vectors of many human arboviral diseases, lay eggs and undergo preimaginal development in fresh water habitats. They have also recently been found to develop in brackish water in coastal areas. Adult females emerging from brackish water-developing Ae. aegypti larvae are shown to possess thicker cuticles and greater resistance to common insecticides used against adults (adulticides) compared with fresh water Ae. aegypti. These findings are compatible with previous findings showing that brackish water Ae. aegypti larvae possess thicker cuticles and greater larvicide resistance. Greater resistance of salinity-tolerant Ae. aegypti to adulticides and larvicides is a hitherto unappreciated problem for controlling arboviral diseases and has implications also for other mosquito-borne diseases.

zoology↗

Lectin Microarray Analysis of Salivary Gland Glycoproteins from the Arboviral Vector Aedes aegypti and the Malaria Vector Anopheles stephensi

Salivary gland glycoconjugates in human-feeding insects and arachnids can elicit hypersensitivity reactions. Glycoproteins introduced during blood feeding by arachnid ticks are responsible for red meat allergy or the -gal syndrome. Dengue virus transmitted by Aedes mosquitoes incorporates mosquito glycoproteins into its envelope and malaria parasites transmitted by Anopheles mosquitoes reportedly express -gal epitopes that are potentially derived from mosquito salivary glands. Tick salivary gland glycoproteins possess -gal epitopes. We used microarrays containing forty different lectins to analyse salivary gland protein glycoconjugates from the principal arboviral vector Aedes aegypti and the rapidly spreading malaria vector Anopheles stephensi. Salivary gland glycoproteins of both mosquitoes possessed very similar lectin-binding specificities. Lectin-binding profiles showed the significant presence of oligomannose N-glycans and O-glycans, a limited presence of glycan structures capped with terminal GalNAc, GlcNAc, {beta} linked Gal, 1-6 linked fucose, and no detectable sialic acids or terminal -linked Gal in salivary gland glycoproteins.

zoology↗

S-nitrosoglutathione Inhibits the Growth of Androgen-Receptor Dependent and Castration Resistant Prostate Cancer by Modulating FOXM1 Signaling

Castration resistant prostate cancer (CRPC) is an advanced form of prostate cancer associated with loss of androgen receptor activity and resistance to AR-targeted therapies. CRPC is often associated with higher metastasis, invasion, and proliferation. Thus, theres a strong need for additional molecular markers in CRPC, and therapies that target them directly. Transcriptomic profiling of prostate cancer patients revealed an upregulated signature specific to high-grade tumors in cell cycle progression, E2F targets, G2M checkpoint, DNA repair, Aurora Kinase B, and FOXM1 pathway. Validation in a separate cohort of neuroendocrine prostate cancer patients, the most resistant and invasive form of CRPC which lacks AR expression, showed CCP signature as a dominant driver gene set for NEPC. Moreover, high expression of mRNA targets in each of these pathways is associated with poor overall survival in NEPC patients. Gene set variation analysis revealed a strong inverse correlation between AR signaling and CCP.31, set of 31 markers associated with high proliferation, and a positive correlation between CCP.31 and NEPC signature. FOXM1, a prioritized candidate in CCP is shown to be upregulated in CRPC cell lines and human clinical sections of Gleason 9 patients. GSNO, a nitric oxide donor, known for its inhibitory effect on cell proliferation and an effective inhibitor of cell cycle checkpoints, was able to reverse CCP signature both in-vitro and in-vivo. GSNO exhibits a dependence on AR signaling in-vitro by showing mild effect on CRPC clones, but successfully inhibits tumor growth of AR-null CRPC xenografts. FOXM1 is also targeted by GSNO in-vivo and in-vitro models of both CRPC and AD prostate cancer. This establishes an important link between androgen receptor status in prostate cancer and proposes nitric oxide donors as therapeutic intervention. GSNO is effective against AR-null castration resistant prostate cancer. SignificanceNitric oxide donor, S-glutathione-transferase inhibits tumor growth of castration resistant prostate cancer, regardless of androgen receptor expression status by targeting FOXM1, a critical regulator of cell cycle progression. O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY

cancer biology↗