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Biology subjects

Mishra, S.

Publications and source records attributed to Mishra, S..

At least 109 records · Page 6Linked to original sources

Specific sensory neurons and insulin-like peptides modulate food type-dependent oogenesis and fertilization in Caenorhabditis elegans

AbstractAn animals responses to environmental cues are critical for its reproductive program. Thus, a mechanism that allows the animal to sense and adjust to its environment should make for a more efficient reproductive physiology. Here we demonstrate that in Caenorhabditis elegans specific sensory neurons influence onset of oogenesis through insulin signaling in response to food-derived cues. The chemosensory neurons ASJ modulate oogenesis onset through the insulin-like peptide (ILP) INS-6. In contrast, other sensory neurons, the olfactory neurons AWA, regulate food type-dependent differences in C. elegans fertilization rates, but not onset of oogenesis. AWA modulates fertilization rates at least partly in parallel to insulin receptor signaling, since the insulin receptor DAF-2 regulates fertilization independently of food type, which requires ILPs other than INS-6. Together our findings suggest that optimal reproduction requires the integration of diverse food-derived inputs through multiple neuronal signals acting on the C. elegans germline.

developmental biology↗

Pharmacologic Stabilization of Retromer Rescues Endosomal Pathology Induced by Defects in the Alzheimer's gene SORL1

The Sortilin-related receptor 1 gene (SORL1, SORLA) is strongly associated with risk of developing Alzheimers disease (AD). SORLA is a regulator of endosomal trafficking in neurons and interacts with retromer, a complex that is a master conductor of endosomal trafficking. Pharmacological chaperones stabilize retromer in vitro, enhancing its function. Here we used an isogenic series of human induced pluripotent stem cell (hiPSC) lines with either one or two copies of SORL1 or harboring one copy of a SORL1 variant linked to increased risk for AD. We treated hiPSC-derived cortical neurons with the established retromer chaperone, TPT-260, and tested whether indicators of ADs defining endosomal, amyloid, and Tau pathologies were corrected. We observed that the degree of rescue by TPT-260 treatment varied, depending on the number of copies of functional SORL1 and which SORL1 variant was expressed. Using a disease-relevant preclinical model, our work illuminates how the SORL1-retromer pathway can be therapeutically harnessed.

neuroscience↗

Streptomycin and Pyridomycin drug molecules provide the most potential inhibitors against Rv3871 to cure Tuberculosis

The ESX-1 secretion system of Mycobacterium tuberculosis delivers bacterial virulence factors to host cells during infection. The most abundant factor, the ESAT-6/CFP-10 dimer, is targeted for secretion via a C-terminal signal sequence on CFP-10 that is recognized by the cytosolic ATPase, Rv3871. ATPase that is a component of the ESAT-6/CFP-10 secretion system. ESX locus contains genes encoding conserved secretion machinery components termed EccCb1. these core components are required for ESAT-6/CFP-10 secretion [70, 71]. Rv3871 is a cytoplasmic protein connected with the Rv3870 and encoded by the EccCb1 gene. These proteins supply energy for the secretion process. Each of these ATPases is involved in targeting protein for ESX-1 secretion. EccCb1 binds a seven amino acid C-terminal signal peptide of CFT-10, which is required for secretion of the ESAT-6/CFP-10 complex. In this we target the Rv3871 seven amino acid c-terminal region and block it through multiple drugs so, it cannot be activated by the ATPases and not to supply energy for the secretion protein during infection (Tuberculosis). In the present study we have comparatively studied different Rv3871 inhibitory antagonist drug molecule which could be a potential drug molecule to inhibit ATPases on the Rv3871 molecule that is responsible for the energy supply. We have 16 antagonistic drug molecules as shortlisted based on previous studies, that block the target site on the Rv3871 molecule.

microbiology↗

Raltitrexed and Edatrexate drug molecules against human Proton Coupled Folate Transporter to cure cancer

Proton pump inhibitors (PPIs) (e.g.: - rabeprazole, pantoprazole, omeprazole, lansoprazole, and esomeprazole) are widely used to treat gastroesophageal reflux disease and other acid-related disorders. Folic acid transport is important for proper cell proliferation. In human, folic acid is not synthesized in the body, it is obtained externally. Therefore, specific transporters are involved in absorption of folic acid, which is concentrated in intestine. Malignant cancer cells require this folic acid very frequently in large quantities for the rapid reproduction of cancer cell. In the current study, we compared different types of proton pump inhibitor drug molecules that may be potential candidates for preventing the absorption of folate by (hPCFT), its responsible for unusually large and frequent folate production. Each of these anti-drug candidates has 27 finalists based on previous studies. Among these competitors, leukovorin and noletraxid molecules were found to be particularly associated with the hPCFT transporters key active site loop(G155XXG158).

bioinformatics↗

FtsK, a DNA motor protein, coordinates the genome segregation and early cell division processes in Deinococcus radiodurans

FtsK/SpoIIIE protein family are DNA translocases known as the fastest DNA motor proteins that use ATP for their movement on DNA. Most of the studies in single chromosome-containing bacteria have established the role of FtsK in chromosome dimer resolution (CDR) connecting the bacterial chromosome segregation process with cell division. But, only limited reports are available on the interdependent regulation of genome segregation and cell division in multipartite genome harbouring (MGH) bacteria. In this study, for the first time, we report the characterization of FtsK from the radioresistant MGH bacterium Deinococcus radiodurans R1 (drFtsK). drFtsK shows the activity characteristics of a typical FtsK/SpoIIIE/Tra family. It stimulates the site-specific recombination catalyzed by Escherichia coli tyrosine recombinases. drFtsK interacts with various cell division and genome segregation proteins of D. radiodurans. Microscopic examination of different domain deletion mutants of this protein reveals alterations in cellular membrane architecture and nucleoid morphology. In-vivo localization studies of drFtsK-RFP show that it forms multiple foci on nucleoid as well as on the membrane with maximum density on the septum. drFtsK coordinates its movement with nucleoid separation. The alignment of its foci shifts from old to new septum indicating its cellular dynamics with the FtsZ ring during the cell division process. Nearly, similar positional dynamicity of FtsK was observed in cells recovering from gamma radiation exposure. These results suggest that FtsK forms a part of chromosome segregation, cell envelope, and cell division machinery in D. radiodurans. ImportanceDeinococcus radiodurans show extraordinary resistance to {gamma}-radiation. It is polyploid and harbours a multipartite genome comprised of two chromosomes and two plasmids, packaged in a doughnut-shaped toroidal nucleoid. Very little is known about how the tightly packed genome is accurately segregated and the next divisional plane is determined. FtsK, a multifunctional protein, helps in pumping the septum-trapped DNA in several bacteria. Here, we characterized FtsK of D. radiodurans R1 (drFtsK) for the first time and showed it to be an active protein. The absence of drFtsK causes many defects in morphology at both cellular and nucleoid levels. The compact packaging of the deinococcal genome and cell membrane formation is hindered in ftsK mutants. In-vivo drFtsK is dynamic, forms foci on both nucleoid and septum, and coordinates with FtsZ for the next cell division. Thus, drFtsK role in maintaining the normal genome phenotype and cell division in Deinococcus radiodurans is suggested.

molecular biology↗

Mutations in a set of ancient matrisomal glycoprotein genes across neoplasia predispose to disruption of morphogenetic transduction

Misexpression and remodeling of the extracellular matrix is a canonical hallmark of cancer, although the extent of cancer-associated aberrations in the genes coding for ECM proteins and consequences thereof, are not well understood. In this study, we examined the alterations in core matrisomal genes across a set of nine cancers. These genes, especially the ones encoding for ECM glycoproteins, were observed to be more susceptible to mutations than copy number variations across cancers. We classified the glycoprotein genes based on the ubiquity of their mutations across the nine cancer groups and estimated their evolutionary age using phylostratigraphy. To our surprise, the ECM glycoprotein genes commonly mutated across all cancers were predominantly unicellular in origin, whereas those commonly showing mutations in specific cancers evolved mostly during and after the unicellular-multicellular transition. Pathway annotation for biological interactions revealed that the most pervasively mutated glycoprotein set regulated a larger set of inter-protein interactions and constituted more cohesive interaction networks relative to the cancer-specific mutated set. In addition, ontological prediction revealed the pervasively mutated set to be strongly enriched for basement membrane dynamics. Our results suggest that ancient unicellular-origin ECM glycoproteins were canalized into playing critical tissue morphogenetic roles, and when disrupted through matrisomal gene mutations, associate with neoplastic transformation of a wide set of human tissues.

bioinformatics↗

FtsZ phosphorylation brings about growth arrest upon DNA damage in Deinococcus radiodurans

The polymerization/depolymerization dynamics of FtsZ plays the pivotal role in cell division in majority bacteria. Deinococcus radiodurans, a radiation resistant bacterium, shows an arrest of growth in response to DNA damage, despite no change in the level of FtsZ. This bacterium does not deploy LexA/RecA type of DNA damage response and cell cycle regulation, and its genome does not encode homologs of E. colis SulA, which attenuate FtsZ functions in response to DNA damage in different bacteria. A radiation responsive Ser/Thr quinoprotein kinase (RqkA) characterized for its role in radiation resistance in this bacterium, could phosphorylate several cognate proteins including FtsZ (drFtsZ) at Serine 235 (S235) and Serine 335 (S335) residues. Here, we report the detailed characterization of S235 and S335 phosphorylation effect in the regulation of drFtsZ functions, and demonstrated that the phospho-mimetic replacements of these residues in drFtsZ had grossly affected its functions that could result in cell cycle arrest in response to DNA damage in D. radiodurans. Interestingly, the phospho-ablative replacements were found to be nearly similar to drFtsZ while phospho-mimetic mutant showed the loss of signatures characteristics to wild type enzyme including the arrest in its dynamics under normal conditions. The post-bleaching recovery kinetics for drFtsZ and phospho-mimetic mutant was nearly similar at 2h post irradiation recovery but found to be different under normal conditions. These results highlighted the role of S/T phosphorylation in the regulation of drFtsZ functions and cell cycle arrest in response to DNA damage and is first time demonstrated in this prokaryote.

molecular biology↗

Constitutive Inhibition of Transient Receptor Potential CanonicalType 6 (TRPC6) by O-GlcNAcylation at Threonine-221

Transient receptor potential canonical type 6 (TRPC6) is a non-voltage gated cation channel that principally conducts calcium to regulate signaling in cardiac, vascular, neuronal and other cells. Abnormally increased TRPC6 expression/conductance and genetic gain of function mutations contribute to fibrosis, hypertrophy, proteinuria, and edema, notably linked to its stimulation of nuclear factor of activated T-cells (NFAT) signaling. Hyperglycemia (HG) also activates TRPC6/NFAT as a cause of diabetic renal disease. While prior work linked HG-TRPC6 activation to oxidant stress, the role of another major HG modification - O-GlcNAcylation, is unknown. Here we show TRPC6 is constitutively O-GlcNAcylated, TRPC6 and O-GlcNAc transferase proteins interact, this modification potently suppresses basal channel conductance and NFAT activity, and it is unaltered by HG. Proteomics identifies O-GlcNAcylation at Ser14, Thr70, and Thr221 in the N-terminus ankyrin-4 (AR4) and neighboring linker (LH1) domains of TRPC6. Of these, T221 is most impactful as a T221A mutation increases basal NFAT activity 11-fold, TRPC6 conductance 75-80% vs wild-type, and when expressed in cardiomyocytes amplifies NFAT-pro-hypertrophic gene expression. T221 is highly conserved and mutating homologs in TRPC3 and TRPC7 also markedly elevates basal NFAT activity. Molecular models predict electrostatic interactions between T221 O-GlcNAc and Ser199, Glu200, and Glu246, and we find similarly elevated NFAT activity from alanine substitutions at these coordinating sites as well. Thus, O-GlcNAcylation at T221 and its interaction with coordinating residues in AR4-LH1 is required for basal TRPC6 channel conductance and regulation of NFAT.

biochemistry↗

Inferring functions of coding and non-coding genes using epigenomic patterns and deciphering the effect of combinatorics of transcription factors binding at promoters

The number of annotated genes in the human genome has increased tremendously, and understanding their biological role is challenging through experimental methods alone. There is a need for a computational approach to infer the function of genes, particularly for non-coding RNAs, with reliable explainability. We have utilized genomic features that are present across both coding and non-coding genes like transcription factor (TF) binding pattern, histone modifications, and DNase hypersensitivity profiles to predict ontology-based functions of genes. Our approach for gene function prediction (GFPred) made reliable predictions (>90% balanced accuracy) for 486 gene-sets. Further analysis revealed that predictability using only TF-binding patterns at promoters is also high, and it paved the way for studying the effect of their combinatorics. The predicted associations between functions and genes were validated for their reliability using PubMed abstract mining. Clustering functions based on shared top predictive TFs revealed many latent groups of gene-sets involved in common major biological processes. Available CRISPR screens also supported the inferred association of genes with the major biological processes of latent groups of gene-sets. For the explainability of our approach, we also made more insights into the effect of combinatorics of TF binding (especially TF-pairs) on association with biological functions.

bioinformatics↗

Evaluation of odorant-binding protein-1 as a molecular marker for identifying biological forms and delimitating sibling species of Anopheles stephensi

BackgroundAnopheles stephensi, an invasive malaria vector, has been reported to have three biological forms identifiable mainly based on the number of ridges present on the eggs floats. Recently, the first intron of the odorant-binding protein-1 (AsteObp1) has been introduced as a molecular marker for the identification of these forms, and based on this marker, the presence of three putative sibling species (designated as species A, B and C) has been proposed. However, there is no data on the association of proposed markers with biological form or putative species on field populations. MethodsField collected and laboratory-reared An. stephensi were characterized for biological forms based on the number of ridges on the egg s float. DNA sequencing of the partial AsteObp1 gene of An. stephensi individuals were performed by Sangers method, either directly or after cloning with a plasmid vector. Additionally, AsteObp1 sequences of various laboratory lines of An. stephensi were retrieved from a public sequence database. ResultsAsteObp1 intron-1 in Indian An. stephensi populations are highly polymorphic with the presence of more than 13 haplotypes exhibiting nucleotides as well as length-polymorphism (90-to-121 bp). No specific haplotype or a group of closely related haplotypes of intron-1 was found associated with any biological form identified morphologically. High heterozygosity for this marker with a low inbreeding coefficient in field and laboratory populations indicates that this marker is not suitable for the delimitation of putative sibling species, at least in Indian populations. ConclusionsAsteObp1 cannot serve as a marker for identifying biological forms of An. stephensi or putative sibling species in Indian populations.

molecular biology↗

Elucidation of sub-cellular H2S metabolism in Solanum lycopersicum L. and its assessment under development and biotic stress

The signalling molecules serve as a fundamental requirement in plants and respond to various internal and external cues. Among several signalling molecules, the significance of gasotransmitters has been realized in several plant developmental and environmental constraints. The hydrogen sulfide (H2S) is a novel signalling molecule in higher plants and is involved in several physiological processes right from seed germination to flowering and fruit ripening. Moreover, H2S also assist plants in managing biotic and abiotic stresses, therefore serves as one of the imperative choice of chemical priming. Yet, the metabolism of H2S is not much explored and only appraisal study is made till date from Arabidopsis thaliana. Therefore, the present investigation explored the elucidation of H2S metabolism in crop plant Solanum lycopersicum L. Through in silico investigations the study demonstrated the participation of 29 proteins involved in H2S metabolism, which are mainly localized in cytosol, chloroplast, and mitochondria. Additionally, the relevant protein-protein interactomes were also inferred for sub-cellular compartments and expression data were explored under development and biotic stresses namely PAMPs treatment and bacterial infection. The information generated here will be of high relevance to better target the H2S metabolism to enhance the tomato prospects and also serve a preliminary investigation to be adopted in other agronomic important crops.

plant biology↗

The degree of context un/familiarity impacts the emotional feeling and preaware cardiac-brain activity: a study with emotionally salient naturalistic paradigm using DENS Dataset

Emotion experiments with naturalistic paradigms are emerging and giving new insights into dynamic brain activity. Context familiarity is considered as an important dimensions of emotion processing by appraisal theorists. However, how the context un/familiarity of the naturalistic stimuli influences the central and autonomic activity is not probed yet [check it]. Hence, we tried to address this issue in this work by breaking it down into three questions. 1) What is the relation between context un/familiarity with the neural correlates of self-assessment affective dimensions viz. valence and arousal; 2) the influence of context un/familiarity in cardiac-brain mutual interaction during emotion processing; 3.) brain network reorganization to accommodate the degree of context familiarity. We found that the less-context familiarity is primarily attributed to negative emotion feeling mediated by lack of predictability of sensory experience. Whereas, with high-context familiarity, both positive and negative emotions are felt. For less-context familiarity, the arousal activity is negatively correlated with EEG power. In addition, the cardiac activity for both high and less context familiarity is modulated before the reported self-awareness of emotional feeling. The correlation of cortical regions with cardiac activity and connectivity patterns reveals that ECG is modulated by salient feature during pre-awareness and correlates with AIC and conceptual hub in high-familiarity. Whereas, for the low familiarity, the cardiac activity is correlated with the exteroceptive sensory regions. In addition, we found that OFC and dmPFC have high connectivity with less-context familiarity, whereas AIC has high connectivity with high-context familiarity. To the best of our knowledge, the context familiarity and its influence on cardiac and brain activity have never been reported with a naturalistic paradigm. Hence, this study significantly contributes to understanding automatic processing of emotions by analyzing the effect of context un/familiarity on affective feelings, the dynamics of cardiac-brain mutual interaction, and the brains effective connectivity during pre-awareness.

neuroscience↗

ATR facilitates the degradation of Api5 through the ubiquitin-proteasome pathway via FBXW2 to regulate apoptosis upon DNA damage

Apoptosis inhibitor 5 (Api5) is an inhibitor of apoptosis, which is found to be upregulated in several cancers and promotes invasion as well as metastasis. Over-expression of Api5 is positively co-related with poor survival of cancers and inhibition of DNA damage induced apoptosis in cancerous cells. Acetylation at lysine 251 (K251) on Api5 facilitates the stability of the protein and thus functionally provides resistance to cancer cells against chemotherapeutic or anti-cancerous agents. However, the regulation of Api5 upon DNA damage is not yet known. In this study, we demonstrate that Api5 undergoes degradation following DNA damage via the ubiquitin-proteasome system. Upon DNA damage, ATR was observed to phosphorylate Api5 at serine 138 which led to the cytoplasmic localisation of Api5. The E3-ubiquitin ligase, SCF-FBXW2 ubiquitinates Api5 leading to its proteasomal degradation.

molecular biology↗

Dataset on Emotions using Naturalistic Stimuli (DENS)

Emotions are constructed and emerge through the dynamic interaction of multiple components. It is difficult to capture the dynamics using static or artificial stimuli. Hence, there is a need for an experiment paradigm using ecologically valid film stimuli. The data set described in this work results from an attempt to capture felt emotional experience at a particular point in time using physiological measures like EEG, ECG and EMG as well as self-reported scales. Sixteen emotional film stimuli were used from the film stimuli dataset validated in the Indian population. Participants self-reported the felt emotional category. Both the raw and pre-processed data are provided along with the pre-processing pipeline. The paradigm we have adopted is new which we have termed as Emotional Event Marker Paradigm (EEMP). Hence, the dataset has unique information about temporal markers of emotional experiences while watching the film stimuli, which is not available with any data to date. It is the first EEG data with emotional film stimuli on the Indian population. This data can be utilized to study dynamic activation and connectivity in a whole-brain source localization study, understand the mutual interactions between the central and autonomic nervous system, understand temporal hierarchy using multi-resolution tools, and perform machine learning-based classification and complex networks analysis associated with emotions.

neuroscience↗

A study of spatio-temporal dynamics of emotion processing usingDENS dataset

The emotion research with artificial stimuli does not represent the dynamic processing of emotions in real-life situations. The lack of data on emotion with the ecologically valid naturalistic paradigm hinders the knowledge of emotion mechanism in a real-world interaction. To this aim, we collected the emotional multimedia clips, validated them with the university students, recorded the neuro-physiological activities and self-assessment ratings for these stimuli. Participants localized their emotional feelings (in time) and were free to choose the best emotion for describing their feelings with minimum distractions and cognitive load. The obtained electrophysiological and self-assessment responses were analyzed with functional connectivity, machine learning and source localization techniques. We observed that the connectivity patterns in the theta and beta band could differentiate emotions better. Using machine learning, we observed that the classification of affective self-assessment features, namely dominance, familiarity, and self-relevance, involves midline brain regions responsible for mentalization and event construction activity compared to valence and arousal, which were mainly associated with lateral brain regions. This finding advocates the need for more than two dimensions for emotion representation. In addition, the channels with high predictability were source localized to the brain regions in default-mode, sensorimotor and salience networks. Hence, in this naturalistic study, we find that the domain-general systems contribute to emotion construction.

neuroscience↗

Are members of the Anopheles fluviatilis complex conspecific?

Anopheles fluviatilis sensu lato, a primary malaria vector in India, was identified to be comprised of four cryptic species, provisionally designated as species S, T, U and V. However, Kumar et al. (Mol Ecol Resour, 2013;13:354-61) considered all of the then known three members of this species complex (S, T and U) conspecific. The specific status of species S and T was refuted based on the lack of sufficient barcode gap in mitochondrial-CO1 and the perceived presence of heterozygotes in populations as detected through one of the two species-specific PCR assays employed for the cryptic species identification. The existence of species U was refuted claiming that earlier investigations have already refuted their existence. This conclusion is concerning because of the differential public health implications of members of the Fluviatilis Complex. Here we discuss problems associated with the CO1-based barcode approach for delimitation of cryptic species, the perceived heterozygosity between species S and T based on a species-specific PCR assay, and interpretation of published reports. We demonstrated that fixed differences do exist in the ITS2-rDNA sequence of species S and T with no evidence of heterozygotes in sympatric populations and, that the observed heterozygosity by Kumar et al. in the ITS2-based species diagnostic PCR is due to the high mispriming tendency of the T-specific primer with species S. We infer that mitochondrial DNA-based barcoding gap, an arbitrary threshold recommended for species delimitation, alone, is inadequate to delimit the members of An. fluviatilis complex.

molecular biology↗

The Alzheimer's gene SORL1 is a key regulator of endosomal recycling in human neurons

BackgroundLoss of the Sortilin-related receptor 1 (SORL1) gene seems to act as a causal event for Alzheimers disease (AD). Recent studies have established that loss of SORL1, as well as mutations in autosomal dominant AD genes APP and PSEN1/2, pathogenically converge by swelling early endosomes, ADs cytopathological hallmark. Acting together with the retromer trafficking complex, SORL1 has been shown to regulate the recycling of the amyloid precursor protein (APP) out of the endosome, contributing to endosomal swelling and to APP misprocessing. We hypothesized that SORL1 plays a broader role in neuronal endosomal recycling and used human induced pluripotent stem cell derived neurons (hiPSC-Ns) to test this hypothesis. We examined endosomal recycling of three transmembrane proteins linked to AD pathophysiology: APP, the BDNF receptor Tropomyosin-related kinase B (TRKB), and the glutamate receptor subunit AMPA1 (GLUA1). MethodsWe used isogenic hiPSCs engineered to have SORL1 depleted or to have enhanced SORL1 expression. We differentiated neurons from these cell lines and mapped the trafficking of APP, TRKB and GLUA1 within the endosomal network using confocal microscopy. We also performed cell surface recycling and lysosomal degradation assays to assess the functionality of the endosomal network in both SORL1 depleted and overexpressing neurons. Finally, we analyzed alterations in gene expression in SORL1 depleted neurons using RNA-sequencing. ResultsWe find that as with APP, endosomal trafficking of GLUA1 and TRKB is impaired by loss of SORL1. We show that trafficking of all three cargo to late endosomes and lysosomes is affected by manipulating SORL1 expression. We also show that depletion of SORL1 significantly impacts the endosomal recycling pathway for APP and GLUA1 at the level of the recycling endosome and trafficking to the cell surface. This has a functional effect on neuronal activity as shown by multi-electrode array (MEA). Conversely, increased SORL1 expression enhances endosomal recycling for APP and GLUA1. Our unbiased transcriptomic data further support SORL1s role in endosomal recycling. We observe altered expression networks that regulate cell surface trafficking and neurotrophic signaling in SORL1 depleted neurons. ConclusionCollectively, and together with other recent observations, these findings suggest that SORL1 is a broad regulator of retromer-dependent endosomal recycling in neurons, a conclusion that has both pathogenic and therapeutic implications for Alzheimers disease.

cell biology↗

TGF--dependent Lymphoid Tissue Residency of Stem-like T cells Limits the Response to Tumor Vaccine

Stem-like CD8+ T cells represent the key subset responding to multiple tumor immunotherapies, including tumor vaccination. However, the signals that control the differentiation of stem-like T cells are not entirely known. Most previous investigations on stem-like T cells are focused on tumor infiltrating T cells (TIL). The behavior of stem-like T cells in other tissues remains to be elucidated. Tissue-resident memory T cells (TRM) are often defined as a non-circulating T cell population residing in non-lymphoid tissues. TILs carrying TRM features are associated with better tumor control. Here, we found that stem-like CD8+ T cells differentiated into TRMs in a TGF-{beta} and tumor antigen dependent manner almost exclusively in tumor draining lymph node (TDLN). TDLN-resident stemlike T cells were negatively associated with the response to tumor vaccine. In other words, after tumor vaccine, TDLN stem-like T cells transiently lost TRM features, differentiated into migratory effectors and exerted tumor control.

immunology↗