Biology subjects

Bhattacharya, S.

Publications and source records attributed to Bhattacharya, S..

At least 19 recordsLinked to original sources

bioRxiv2026

A unique intestinal epithelial cell that expresses canonical lymphoid genes

Enterocytes and intestinal secretory cells harbor innate immunity. However, certain transcriptional programs -even beyond rearranged T- and B-cell receptor genes- are considered the exclusive purview of adaptive immune cells, including abundant and diverse intestinal intra-epithelial lymphocytes (IELs). Here, we identify and characterize columnar intestinal secretory cells that co-express classic epithelial and lymphoid genes, including Ptprc, Cd3e, Cd8a, and Gzma/b but not T- or B-cell receptors. These previously unrecognized epithelium-derived lymphoid-like cells (EDLCs) differ from any known resident cell type, including IELs or tuft cells. Rare in healthy mouse and human intestines, their numbers increase upon IL-25 stimulation or IL-10 receptor deficiency in vivo and upon IL-13 stimulation in vitro. EDLC chromatin is uniquely accessible at thousands of cis-elements, including lymphoid and epithelial loci. These findings reveal a transcriptionally and epigenetically distinct intestinal epithelial derivative that expresses prominent lymphoid character.

cell biology↗

bioRxiv2026

SF3B1 mutation generates novel peptides from long non-coding RNAs

While mRNA alternative splicing caused by somatic mutation in SF3B1 is well-characterized, long non-coding RNAs (lncRNAs) impacted by this mutation are completely unknown. Here, we report long-read sequencing of full-length transcripts from cell lines with and without SF3B1 mutation. We present ALLR (Annotation of human lncRNAs using Long-Read sequencing), a computational workflow to identify and quantify lncRNAs, and to annotate their potential roles in impacting cellular functions. Harnessing matched short-read RNA sequencing and proteomic data, we demonstrate that SF3B1 mutation generates thousands of novel lncRNAs with diverse levels of alternative splicing. Moreover, SF3B1 mutation-associated lncRNAs produce translatable peptides with putative functions, immunogenicity, and potential to bind proteins with impact on cellular signalling networks. 47 lncRNAs could also be detected in primary chronic lymphocytic leukemia (CLL) samples and are associated with significant risks towards adverse disease prognosis, in conjunction with other known CLL risk factors. Our work demonstrates the utility of long-read sequencing for lncRNA research and provides a new territory of functional RNA molecules that SF3B1 mutation may use to impact cancer.

genomics↗

bioRxiv2026

Mycobacterium tuberculosis inactivates host oxysterols to impair macrophage cholesterol homeostasis

Host oxysterols coordinate macrophage cholesterol homeostasis and antimicrobial defense, but whether pathogens directly target oxysterol signaling is unknown. Here, we identify pathogen-mediated enzymatic oxysterol inactivation as a mechanism by which Mycobacterium tuberculosis (Mtb) subverts host cholesterol metabolism. Mtb oxidizes the endogenous oxysterols 27-hydroxycholesterol (27-HC) and 3{beta}-hydroxycholest-5-enoic acid (3{beta}-HCA) into 3-oxo-{Delta}4 metabolites. Sputum lipidomics across TB cohorts on three continents revealed an active-disease signature characterized by 27-HC depletion and accumulation of 27-hydroxycholest-4-en-3-one (27-HCO) and 3-oxocholest-4-enoic acid (3O-CA), which normalized with treatment. In infected human macrophages, Mtb 3{beta}-hydroxysteroid dehydrogenase (3{beta}-Hsd) generated 27-HCO. Unlike their parent oxysterols, 27-HCO and 3O-CA did not activate liver X receptor (LXR), and 3O-CA failed to suppress sterol regulatory element-binding protein 2 (SREBP2) target genes. Accordingly, 3{beta}-Hsd-deficient Mtb did not induce macrophage cholesterol retention, a phenotype restored by LXR antagonism. Thus, Mtb enzymatically inactivates host oxysterols, disrupting macrophage cholesterol homeostasis and generating treatment-responsive metabolic signatures in human TB.

microbiology↗

bioRxiv2026

Targeting REV7 exposes a mitotic vulnerability in colorectal cancer cells

Therapeutic resistance remains a barrier to durable treatment of colorectal cancer (CRC), highlighting the need to identify vulnerabilities in resistant tumor cells. REV7 (MAD2L2) is a multifunctional genome-maintenance protein with established roles in REV3-dependent translesion synthesis (TLS) as well as mitotic progression, spindle organization, and chromosome segregation. Although perturbing REV7 dependent DNA damage tolerance has emerged as a strategy to overcome chemoresistance, whether pharmacologic targeting of REV7 exposes vulnerabilities to mitotic disruption remains poorly understood. Here, we investigated the REV7 targeting small molecule inhibitor C10, which disrupts the REV7 and REV3 interaction, and found that C10 sensitized HCT116 and HT29 CRC cells to the microtubule-disrupting agent colchicine, resulting in markedly reduced clonogenic survival. In contrast, C10 did not substantially enhance sensitivity to cisplatin. Genetic loss of REV7 similarly increased sensitivity to colchicine, independently supporting a REV7 dependent vulnerability to microtubule disruption. Combined C10 and colchicine treatment was associated with pronounced alterations in mitotic and post-mitotic nuclear morphology, including condensed/mitotic-like and multinucleated configurations, and increased micronucleus formation. In HT29 cells, combined treatment also increased the frequency of cells containing multiple gamma tubulin positive foci, consistent with disrupted centrosome-associated organization. Together, these findings support a model in which REV7 dependent functions contribute to cellular tolerance and recovery from microtubule mediated mitotic stress, whereas REV7 and REV3 disruption renders CRC cells vulnerable to mitotic perturbation. This study broadens the potential consequences of targeting REV7 beyond its canonical role in TLS and suggests that combining REV7 directed inhibition with mitotic perturbation warrants further investigation as a therapeutic strategy in CRC and potentially other cancers.

cancer biology↗

bioRxiv2026

Amaranthus viridis-derived phytopriming reprograms redox homeostasis and limits arsenic accumulation in rice

Arsenic (As) poses a major threat to rice productivity and food safety due to its high bioaccumulation potential and subsequent entry into the human food chain. In rice, As impairs seed germination, disrupts morpho-anatomical development, and induces oxidative stress. This study evaluates seed priming with an aqueous extract of the agricultural weed Amaranthus viridis (AvE) as a sustainable strategy to alleviate As-induced phytotoxicity. AvE priming significantly improved germination (71-75%) and morpho-physiological performance under As stress. It reduced oxidative stress markers, including H2O2 (21-38%), malondialdehyde (13-26%), and proline (18.9-44.7%), while increasing antioxidant metabolites, polyphenols and glutathione by up to 2.34-fold and 41%, respectively. Microscopy confirmed restoration of cellular integrity and anatomical organisation in primed seedlings. ICP-OES analysis showed that AvE priming reduced root As uptake by up to 39%, root-to-shoot translocation by up to 58%, and grain As accumulation by up to 95% compared with unprimed plants. qRT-PCR revealed modulation of genes involved in As homeostasis, indicating coordinated physiological and transcriptional responses. Importantly, improved agronomic performance further demonstrated the translational potential of this approach. This study provides the first evidence that A. viridis extract is a cost-effective, sustainable biostimulant for producing low-As rice in contaminated regions.

plant biology↗

bioRxiv2026

Assessing the physiological S nitrosoproteome reveals nitric oxide mediated regulatory networks in rice

Nitric oxide metabolism-based protein post-translational modifications, such as reversible S- nitrosylation, have been at the pinnacle of plant redox research owing to their significant correlation with seed dormancy, interaction with other signaling molecules, plant development and metabolism, biotic and abiotic plant stress responses, immune defense responses against plant pathogens, and senescence. The rapid interconversion of reactive nitrogen species, the abrogation of nitric oxide homeostasis by exogenous supplementation of NO donors and scavengers, the lack of spatio-temporal specificity of NO signaling, and the limited bioavailability or assay sensitivity for detection often limit the effectiveness of identifying and characterizing S-nitrosothiols in plants. Hitherto unknown, we report the first experimental evidence of the total in vivo S-nitrosoproteome in Oryza sativa L. subsp. indica, comprising 134 PSNOs, enriched with 169 putative sites susceptible to S-nitrosylation, without any exogenous supplementation of NO donors. In the present study, mercuric salt- driven facile decomposition of S-nitrosoproteins in the presence of nitrone spin trap 5,5- dimethyl-1-pyrroline N-oxide, resulting in the synthesis of DMPO-nitrone adducts with PSNO-derived protein thiyl radicals in O. sativa, has been demonstrated as an efficient and novel strategy for characterizing the PSNOs using mass spectrometry analysis. The evidence of physiological levels of PSNOs was further re-examined in a bi-directional qualitative and quantitative approach involving the 2,3-diaminonaphthalene assay in tandem with fluorescence-based visualization and fluorometric quantification. In silico analyses, involving both functional enrichment and pathway prediction analyses, have furthermore revealed unique protein-protein interaction networks and signaling pathways among the S- nitrosoproteome candidates and their predictable physiological roles in O. sativa indica, awaiting further in vitro validation for their functional correlation in response to S- nitrosylation. In conclusion, the present study provides novel evidence of nitric oxide signaling in rice cultivars under physiological conditions, bringing new insights into the potential in vivo transnitrosylation of regulatory or active-site cysteine thiols.

biochemistry↗

bioRxiv2026

Evaluating Protein Language Model Embeddingsfor Structural Similarity in the Protein-Sequence Twilight Zone

Evaluating protein sequence similarity remains challenging in the protein-sequence twilight zone (20-35% sequence identity), where traditional methods often fail. In this study, we evaluate whether mean-pooled embeddings from four protein language models: ESM-1b, ESM-2, ProtT5, and ProstT5 can estimate pairwise structural similarity without performing sequence alignment. The benchmark dataset includes 20,445 PISCES protein pairs with sequence identity [≤]30%, representing the protein-sequence twilight zone, with TM-align-derived TMmin used as the structural ground truth. Protein embeddings are compared using cosine similarity, Euclidean- and Manhattan-derived similarities, an RBF kernel, and dot product. Among these similarity metrics, cosine similarity performs best across all four models. Moreover, ProstT5 achieves the highest Spearman correlation with TMmin, followed by ESM-2, ProtT5, and ESM-1b, while all four PLMs outperform BLASTP overall. Furthermore, the advantage of PLM embeddings is most pronounced for protein pairs with the lowest sequence identity. ProstT5 also provides the best discrimination between structurally similar and dissimilar protein pairs. Moreover, it offers a favorable balance between similarity performance and the computational requirements of residue-level embedding generation and storage. Overall, these findings support PLM embeddings as an effective alignment-free approach for detecting structural relationships among proteins in the twilight zone.

bioinformatics↗

bioRxiv2026

Bacterial nanotubes extend from a tubeosome organelle to establish an intercellular bridge

Bacterial nanotubes are widespread intercellular membranous bridges mediating molecular exchange among neighboring cells, yet their formation and dynamics remain largely unexplored. By combining live-cell imaging with in situ cryo-electron tomography, we describe nanotube biogenesis in the Gram-positive Bacillus subtilis. Nanotubes form elaborate, highly dynamic networks, with extending nanotubes exhibiting multidirectional movement toward nearby cells, repeatedly scanning their surface. Ultimately, nanotubes penetrate through the recipient cell wall to establish an intercellular bridge. Strikingly, nanotubes emerge within milliseconds from lipid-enriched sites representing a previously unrecognized membranous organelle, termed tubeosome, located in an unusual pseudo-periplasmic space beneath the cell wall. We present evidence that tubeosomes comprise proto-nanotube reservoirs that unfold to project nanotubes. Our study uncovers the tubeosome as a novel bacterial organelle and provides the first real-time visualization of intercellular bridge formation. As connecting nanotubes are found across bacteria, archaea, and eukaryotes, this mechanism may reflect a broadly conserved strategy for cellular communication.

microbiology↗

bioRxiv2026

Microtubule Acetylation Regulates the Malignant Phenotype of Glioblastoma and is a Promising Therapeutic Target

Glioblastoma (GBM) is universally lethal despite decades of research to find effective treatments. This highlights the need to identify druggable targets essential for sustaining the malignant phenotype but dispensable for normal tissue. We propose that the enzyme -tubulin acetyl transferase (ATAT1) meets these criteria. ATAT1 acetylates -tubulin at lysine 40, which increases microtubule stability and promotes microtubule-based transport. While ATAT1 knockout mice have only a very mild phenotype, ATAT1 suppression in GBM has multiple therapeutic effects by reducing tumor invasion, proliferation, and therapeutic resistance. These translate not only into improved survival with ATAT1 targeting by itself, but also into synergy when ATAT1 deletion is combined with FDA approved therapies. This study strongly supports our conclusion that ATAT1 is a promising therapeutic target in GBM.

Cancer Biology↗

bioRxiv2026

Elucidating the Design Space of Generative Models for Single-Cell Perturbation Prediction

Next-token prediction has produced predictable scaling in language, but the recipe presumes a sequence of tokens with a meaningful order. Single-cell RNA-seq counts have no natural gene ordering, so applying the recipe directly to raw expression fails under an ill-suited left-to-right bias. We instead ask whether a learned latent can supply the structure the recipe needs. We introduce ExpressionVAE (eVAE), a discrete-latent perturbation model that compresses each cell into a short sequence of discrete codes through a finite-scalar-quantization (FSQ) bottleneck and trains a perturbation-conditioned discrete prior over those codes. On Replogle and Parse 1M, eVAE sets a new state of the art on every distributional metric and leads on most cell-eval perturbation metrics, with Frechet distance and MMD2 roughly 3 to 20x lower than the strongest continuous-latent baseline. Swapping the prior between autoregressive and masked discrete diffusion leaves performance near-identical, isolating the gain to the discrete latent itself rather than the prior family. A decoder-head ablation then exposes a single design axis, the richness of the predictive distribution at inference, that splits the standard metrics into two groups, variance-sensitive and mean-sensitive, which move in opposite directions along the axis. Finally, on a held-out CRISPRi reversion benchmark of 1,732 perturbations under inflammatory cytokine stress, the frozen eVAE encoder outperforms UMAP and differential expression and matches scGPT on perturbation ranking at a fraction of the data.1 We release our code.2

bioinformatics↗

bioRxiv2026

Epigenome Alterations and 3D Chromatin Architecture Remodeling in Inflammatory Macrophage Activation under Diabetic Conditions

Aberrant monocyte/macrophage activation in diabetes drives chronic inflammation and complications. While epigenetic mechanisms are implicated, the role of 3D-chromatin reorganization remains unclear. Using integrated multi-Omics, we profiled gene expression and 3D-chromatin architecture in human CD14+ monocyte-differentiated macrophages treated with high glucose + TNF- (HT) mimicking the diabetic milieu. HT induced inflammatory programs resembling those in diabetes, and dynamically altered chromatin accessibility, enhancer-promoter loops, transcriptionally active/inactive (A/B) compartments, and topologically associated domains. Inflammatory genes exhibited increased, whereas cell-cycle and metabolic genes showed reduced chromatin accessibility and enhancer-promoter interactions. These architectural changes facilitated the convergence of lineage-specific and signal-dependent transcription factors at enhancer-promoter loops, forming network hubs orchestrating pro-inflammatory responses. Similar enhancer-promoter interactions were observed in monocytes from individuals with diabetes. Perturbing chromatin interactions via CRISPR-interference targeting HT-induced enhancers suppressed inflammatory gene expression. Results from this first comprehensive enhancer connectome maps in macrophages under diabetic conditions reveal disease-associated rewiring of the 3D epigenome, highlighting epigenetic mechanisms as potential therapeutic targets. TeaserDiabetic conditions rewire 3D chromatin organization and enhancer-promoter interactions in human macrophages, promoting chronic inflammation.

genomics↗

bioRxiv2026

Interplay of stability and dynamics in the optimization of a highly proficient de novo enzyme

The development of highly active de novo enzymes that catalyze new-to-nature reactions is becoming increasingly possible. Here, we identify the features underlying the catalytic efficiency of a highly proficient de novo enzyme, from the original computational design to an optimized catalyst obtained through two rounds of directed evolution. Computational, spectroscopic, and biochemical studies reveal successfully designed features, including precise alignment of catalytic residues, transition state stabilization, and environmental tuning. In the most evolved enzyme, binding of a transition state analog also led to widespread increases in backbone conformational stability throughout the protein, except within a helix near the active site entrance, where the introduction of Gly and Pro increased dynamics and catalytic activity. Thus, the entire protein contributes to catalysis in the most optimized enzyme. Also, the initial design considered only the transition state but not substrate binding, leading to a dynamic Michaelis complex prior to optimization. These studies show the multiple features that need to be optimized to achieve high activity in a designed enzyme.

biochemistry↗

bioRxiv2026

LINE-1 transposable element is expressed in Kaposi's sarcoma and regulates gene expression in KSHV-infected cells

Long interspersed nuclear element-1 (LINE-1/L1) retrotransposons are expressed in various human cancers. Kaposis sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposis Sarcoma (KS) and Primary Effusion Lymphoma (PEL), which are characterized by global transcriptional reprogramming. Previously, we have found upregulation of L1 in PEL. Here, we show that knockdown of L1 in PEL cells significantly impaired their proliferation and induced broad transcriptional changes encompassing DNA replication, mitotic progression, and genome maintenance pathways. Analysis of KSHV-infected mouse and human mesenchymal stem cell models revealed that the KS-mimicking microenvironment robustly upregulates L1 expression, with sustained expression observed in KSHV-induced tumor cells even after the loss of the viral episome. Reanalysis of RNA-seq data from KS patient biopsies further showed elevated L1 expression in both endemic and AIDS-associated epidemic KS, with partial reduction following antiretroviral therapy. L1 ORF1p protein expression was confirmed by immunohistochemistry in KS tissues. Reanalysis of the enzymes that can modulate DNA methylation, a process that regulates L1 expression, revealed upregulation of DNMT3A and TET2 in both mouse KSHV tumor models and human KS. Altogether, our findings establish the L1 activation as a hallmark of KSHV-driven oncogenesis.

microbiology↗

bioRxiv2026

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↗

bioRxiv2026

Paralogous guanine deaminases likely acquired from bacteria by horizontal gene transfer promote purine homeostasis in Caenorhabditis elegans

Disruptions in purine metabolism contribute to a range of human diseases, from rare genetic disorders such as Lesch-Nyhan syndrome and xanthinuria to common conditions including gout and cancer. To better understand the metabolic networks that regulate purine homeostasis, we developed a Caenorhabditis elegans model of xanthine dehydrogenase (xdh-1) deficiency. Remarkably, xdh-1 mutant animals form rare xanthine stones, recapitulating a hallmark of human xanthinuria. To uncover genetic regulators of purine homeostasis, we performed a forward genetic screen for mutations that exacerbate xanthine stone formation in xdh-1 mutants. This approach identified multiple loss-of-function alleles in a previously uncharacterized gene, which we named gda-1. We show that gda-1 encodes an intestinal guanine deaminase that mediates a key enzymatic step in purine catabolism. The C. elegans genome also encodes a paralog, gda-2, which shares guanine deaminase activity but is expressed in distinct tissues. While gda-2 can compensate for gda-1 loss in guanine metabolism, the two genes exhibit non-redundant roles in regulating xanthine accumulation and stone formation. Interestingly, our evolutionary analyses suggest that gda-2 was acquired by nematodes via horizontal gene transfer from bacteria. These findings reveal a spatially regulated purine catabolism pathway in C. elegans and suggest that acquisition of bacterial genes has shaped a core nematode metabolic network.

genetics↗

bioRxiv2026

Discrete Diffusion for Single-Cell Gene Expression Modeling

AO_SCPLOWBSTRACTC_SCPLOWCurrent generative modeling of single-cell transcriptomics relies on continuous latent representations, transforming inherently discrete and sparse gene counts into continuous space. We propose Discrete Cell Models (DCM), a diffusion-based framework that learns cellular representations directly in the discrete domain. Our framework supports both unconditional and conditional generation, allowing for precise modeling of complex biological scenarios such as cell-type-specific transcriptional responses to genetic perturbations. We demonstrate that DCM scales effectively and achieves strong performance against current state-of-the-art methods, including scVI, CPA, STATE, scGPT, and scLDM. On the Dentate Gyrus benchmark, DCM achieves a 5-fold improvement in MMD2RBF and a nearly 2-fold improvement in W2 distance, over the leading continuous diffusion baseline (scLDM). On the conditional Replogle perturbation benchmark, DCM sets a new state of the art on W2 distance while remaining competitive on MMD2RBF. Together, these results establish discrete diffusion as a promising direction for foundational models of cellular biology.

cell biology↗

bioRxiv2026

The promoter-poised Rpd3 HDAC complex orchestrates global chromatin reprogramming upon nutrient transition

Adaptive transcriptional rewiring underlies the metabolic flexibility of Saccharomyces cerevisiae. We demonstrate that the histone deacetylase Rpd3 mediates nutrient-dependent chromatin reprogramming that coordinates transcriptional shutdown and global acetylation balance during metabolic transitions. Genome-wide analyses reveal that Rpd3 complexes drive rapid, reversible histone deacetylation across promoters and gene bodies, fine-tuning transcriptional output. Rpd3, primarily through the large complex (Rpd3L), localizes at promoters of active genes enriched in H3K9ac and the acetyltransferase Gcn5. Upon nutrient shift, Gcn5 disengages while Rpd3-mediated H3K9 deacetylation enforces repression. Loss of Rpd3 or its Rpd3L-specific subunit, Pho23, disrupts this balance, resulting in the aberrant persistence of growth programs upon starvation and defective activation of respiratory genes in the presence of glucose. HDACs thus can act as metabolic gatekeepers, coupling nutrient cues to chromatin reprogramming and ensuring transcriptional fidelity during metabolic transitions, thereby resolving the long-standing paradox of HDAC enrichment at active promoters.

biochemistry↗

bioRxiv2026

5-Iodotubercidin inhibits Epithelial to Mesenchymal Transition by inhibiting IKK/NFκB-dependent gene expression

Epithelial to mesenchymal transition (EMT) is a process of trans-differentiation important for development, inflammation and cancer. Transforming Growth Factor-{beta} (TGF{beta}) is a physiologically relevant inducer of EMT. We had recently characterized the adenosine analogue, adenosine kinase inhibitor 5-Iodotubercidin (5-ITu), as a compound which preferentially sensitizes MK2-deficient cells to TNF-induced, RIPK1-dependent cell death. Here we investigated the effect of 5-ITu on TGF{beta}-induced EMT. 5-ITu suppressed TGF{beta}-induced morphological changes and migration in A549 (lung cancer) and PANC1 (pancreatic cancer) cell lines. Consistent with these effects, there was significant suppression of EMT markers as indicated by qPCR, immunoblotting and immunofluorescence and confocal microscopy. Mechanistic investigations revealed that 5-ITu-mediated EMT suppression was independent of adenosine kinase inhibition and RIPK1 activation. 5-ITu suppressed NF{kappa}B activity in cells undergoing EMT and IKK inhibition phenocopied the effect of 5-ITu on EMT. Kinase assays revealed IKK{beta} as a potential direct target of 5-ITu. We identified a TGF{beta}-associated, NF{kappa}B-dependent gene signature consisting of 4 genes, which are differentially regulated upon 5-ITu treatment. Interestingly, this 4 gene signature could predict survival in lung and pancreatic cancer. The identification of this role for the multitarget kinase inhibitor 5-ITu in NF{kappa}B activity-dependent EMT, in addition to RIPK1-dependent necroptosis has potential implications in anticancer strategies.

cell biology↗