Biology subjects

Chakraborty, S.

Publications and source records attributed to Chakraborty, S..

At least 145 records · Page 8Linked to original sources

bioRxiv2023

A microfluidic model of colonocyte-microbiota interaction mimicking the colorectal cancer microenvironment

Changes in the abundance of certain bacterial species within the colorectal microbiota correlate with colorectal cancer development. While carcinogenic mechanisms of single pathogenic bacteria have been characterized in vitro, limited tools are available to investigate interactions between pathogenic bacteria and both commensal microbiota and colonocytes in a physiologically relevant tumor microenvironment. To address this, we developed a microfluidic device that can be used to co-culture colonocytes and colorectal microbiota. The device was used to explore the effect of Fusobacterium nucleatum, an opportunistic pathogen associated with colorectal cancer development in humans, on colonocyte gene expression and microbiota composition. F. nucleatum altered the transcription of genes involved in cytokine production, epithelial-to-mesenchymal transition, and proliferation in colonocytes in a contact-independent manner; however, most of these effects were diminished by the presence of fecal microbiota. Interestingly, F. nucleatum significantly altered the abundance of multiple bacterial clades associated with mucosal immune responses and cancer development in the colon. Our results highlight the importance of evaluating the potential carcinogenic activity of pathogens in the context of a commensal microbiota, and the potential to discover novel inter-species microbial interactions in the colorectal cancer microenvironment.

bioengineering↗

bioRxiv2023

Rapid mode switching facilitates the growth of Trichodesmium: A model analysis.

Trichodesmium is one of the dominant dinitrogen(N2)-fixers in the ocean, influencing global carbon and nitrogen cycles through its biochemical reactions. Although the photosynthetic activity of Trichodesmium fluctuates rapidly at the cellular level, the physiological or ecological advantage of this characteristic is not clear. Here we develop a metabolic model of Trichodesmium that can perform dinitrogen (N2) fixation during the daytime. We examined (1) the effect of the duration of switches between photosynthetic and non-photosynthetic cellular states and (2) the effect of the presence and absence of N2 fixation in photosynthetic states. Our results show that a rapid switch between photosynthetic and non-photosynthetic states increases Trichodesmium growth rates by improving metabolic efficiencies due to optimized C and N metabolism. Our results show the possibility and advantage of the rapid switch, providing a strategy for previous observations that all Trichodesmium cells can contain nitrogenase, which was previously considered to be a paradox. This study reveals the importance of fluctuating photosynthetic activity and provides a mechanism for daytime N2 fixation that allows Trichodesmium to fix N2 aerobically in the global ocean.

ecology↗

bioRxiv2023

Configurable Full Subtractor and Adder by Lego-like Mixing and Matching of Engineered Bacteria

One of the long-term goals of synthetic bioengineering is to create configurable and programmable biological systems by just mixing and matching "LEGO"-like bio-modules. Here, we introduce a configurable and modular multi-cellular system where, from a small library of nine discrete engineered bacterial cells, a full subtractor and a full adder can be built on demand by just mixing and matching seven appropriate cell types in a culture. Here, each set of engineered bacteria was modelled as an artificial neuro-synapse that, in a co-culture, formed a single layer artificial neural network (ANN) type architecture that worked as a biochemical full subtractor or full adder. The system is configurable with interchangeable cellular modules, whereby through simply interchanging two cell types in the subtractor culture, a full adder can be built and vice versa. This Lego-like mix and match system is mathematically predictive, and provide a flexible and scalable means to build complex cellular functions. This work may have significance in biocomputer technology development, multi-cellular synthetic biology, and cellular hardware for ANN.

synthetic biology↗

bioRxiv2023

The Single-Cell Transcriptome Program of Nodule Development Cellular Lineages in Medicago truncatula

Legumes can establish a symbiotic relationship with nitrogen-fixing rhizobia by developing nodules after root exposure to lipo-chito-oligosaccharides secreted by the bacteria. Nodule development initiates with anticlinal mitotic divisions in the pericycle and endodermal and inner cortical cells, establishing cell lineages that ultimately form each nodule compartment. We characterized these lineages by isolating and sequencing the transcriptome of Medicago truncatula single nuclei derived from uninoculated roots and roots undergoing early nodule development at 24, 48, and 96 hours after inoculation. To enrich samples for cells responding to the rhizobia, we complemented the analysis of the Medicago wild-type genotype A17 with a mutant for the autoregulation of nodulation, sunn-4. Analysis of cell lineage trajectories derived from the cortex indicates that their transcriptome is initially enriched for cytokinin perception and signaling while repressing auxin accumulation. As these cells differentiate to form nodules, expression of genes related to auxin biosynthesis, transport, and signaling was enhanced, while genes involved in cytokinin degradation were activated as lineages bifurcated to form the nodule meristem and infection zones. While the contribution of auxin and cytokinin in nodule development has been recognized, this single-cell resource quantifies the expression of each of their regulators, receptors and targets as cells divide and differentiate to form each nodule compartment.

plant biology↗

bioRxiv2023

Single-cell RNA sequencing identifies response of renal lymphatic endothelial cells to acute kidney injury

The inflammatory response to acute kidney injury (AKI) likely dictates future renal health. Lymphatic vessels are responsible for maintaining tissue homeostasis through transport and immunomodulatory roles. Due to the relative sparsity of lymphatic endothelial cells (LECs) in the kidney, past sequencing efforts have not characterized these cells and their response to AKI. Here we characterized murine renal LEC subpopulations by single-cell RNA sequencing and investigated their changes in cisplatin AKI. We validated our findings by qPCR in LECs isolated from both cisplatin-injured and ischemia reperfusion injury, by immunofluorescence, and confirmation in in vitro human LECs. We have identified renal LECs and their lymphatic vascular roles that have yet to be characterized in previous studies. We report unique gene changes mapped across control and cisplatin injured conditions. Following AKI, renal LECs alter genes involved endothelial cell apoptosis and vasculogenic processes as well as immunoregulatory signaling and metabolism. Differences between injury models are also identified with renal LECs further demonstrating changed gene expression between cisplatin and ischemia reperfusion injury models, indicating the renal LEC response is both specific to where they lie in the lymphatic vasculature and the renal injury type. How LECs respond to AKI may therefore be key in regulating future kidney disease progression.

pathology↗

bioRxiv2023

Multimodal gradients of human basal forebrain connectivity

The cholinergic innervation of the cortex originates almost entirely from populations of neurons in the basal forebrain (BF). Structurally, the ascending BF cholinergic projections are highly branched, with individual cells targeting multiple different cortical regions. However, it is not known whether the structural organization of basal forebrain projections reflects their functional integration with the cortex. We therefore used high-resolution 7T diffusion and resting state functional MRI in humans to examine multimodal gradients of BF cholinergic connectivity with the cortex. Moving from anteromedial to posterolateral BF, we observed reduced tethering between structural and functional connectivity gradients, with the most pronounced dissimilarity localized in the nucleus basalis of Meynert (NbM). The cortical expression of this structure-function gradient revealed progressively weaker tethering moving from unimodal to transmodal cortex, with the lowest tethering in midcingulo-insular cortex. We used human [18F] fluoroethoxy-benzovesamicol (FEOBV) PET to demonstrate that cortical areas with higher concentrations of cholinergic innervation tend to exhibit lower tethering between BF structural and functional connectivity, suggesting a pattern of increasingly diffuse axonal arborization. Anterograde viral tracing of cholinergic projections and [18F] FEOBV PET in mice confirmed a gradient of axonal arborization across individual BF cholinergic neurons. Like humans, cholinergic neurons with the highest arborization project to cingulo-insular areas of the mouse isocortex. Altogether, our findings reveal that BF cholinergic neurons vary in their branch complexity, with certain subpopulations exhibiting greater modularity and others greater diffusivity in the functional integration of their cortical targets.

neuroscience↗

bioRxiv2023

Regulation of shelterin proteins TERF2IP and TRF2 by H3K4me3-p65 axis drives hyperglycemia dependent endothelial senescence

BackgroundEndothelial senescence has been linked to several cardiovascular diseases. Dysregulation of proteins of the shelterin complex including TRF2 and TERF2IP causes senescence as it hampers DNA repair and cell proliferation. However, whether exposure to hyperglycemia interplays with proteins of the shelterin complex thus further dictates the senescent phenotype of endothelial cells (EC) remain to be explored. Approach and ResultsIn this study, we observed elevated levels of p21 and p53 in endothelial cells upon exposure to intermittent hyperglycemia. We also noted hyperglycemia exposure increased the levels of TERF2IP and TRF2, part of the shelterin complex. No change in the level of TRF1 and TPP1 were detected. Furthermore, a robust induction was detected in p65 level upon intermittent hyperglycemia challenge. ChIP-qPCR analysis revealed enhanced H3K4me3 enrichment in the promoter regions of p65, TERF2IP and TRF2. Inhibition of catalysis of H3K4me3 either by pharmacological inhibitor or siRNA-mediated knockdown of MLL2 attenuated increase in p65, TERF2IP and TRF2 levels including reversal of senescence markers p53 and p21. Interestingly, pharmacological inhibition of NF-{kappa}B signaling also diminished abrupt increase in TERF2IP and TRF2 levels thus further reversed intermittent hyperglycemia-induced p53 and p21 levels. More importantly, co-immunoprecipitation and co-localization analysis revealed an interaction between nuclear p65 and MLL2 in EC stimulated with hyperglycemia. Further knockdown of either TERF2IP or TRF2 impaired intermittent hyperglycemia-induced p53 and p21 expression and associated endothelial senescence. ConclusionOverall, the present study describes an interplay of epigenetics in defining NF-{kappa}B signaling and shelterin proteins expression which further govern the biochemical and functional state of endothelial senescence in hyperglycemic milieu.

biochemistry↗

bioRxiv2023

Dynamics of a Lipid Vesicle across a Microfluidic Constriction: How does the Fluidity of the Encapsulation and its Micro-EnvironmentMatter?

Constriction in the flow passage in the physiological circulatory system is central to the occurrence of several diseased conditions such as thrombosis and is also pivotal towards the understanding of several regulatory processes in the human microvasculature. It is, therefore, imperative to advance a mechanistic insight on the dynamics of the transiting cellular encapsulations in a physiologically-mimicking micro-confinement, with particular focus on deciphering the role of its mechano-physical properties. Here we bring out a quantitative depiction on the role of the membrane fluidity and the initial deflation (shape deviation from sphericity) of a lipid vesicle during its morphological transition from stretching to tumbling via rolling as it migrates across a microfluidic constriction. Based on our experimental observations as well as theoretical insights, we construct a regime map to elucidate the range of the key dimensionless parameters orchestrating the dynamic transition. Our results further bring out the role of the initial position of the lipid vesicle on its subsequent stretching dynamics, exhibiting characteristic nonlinearities and non-monotonic trends. In addition, our observations on the vesicles stretching dynamics emerge from mapping selectively with the viscosity contrast between the encapsulated and the suspending fluid medium, offering potential physiologically relevant cues on the impact of the aging of a cellular moiety on its deformability as it transits through a constricted path. Such mechanistic insights may potentially enable establishing quantitative correlations between the dynamical transition of a cellular encapsulation and its mechano-physical properties, which may in turn, have decisive implications in various states of health and disease while circulating across microvascular fluidic pathways. Impact StatementThis study brings out a quantitative mechanistic insight into the dynamics of migrating lipid vesicles as they migrate through a constricted microfluidic passage. Having a direct similitude with the movement of red blood cells in human microvascular pathways, the resulting mapping between the initial shape and bending properties of the vesicle membrane with three distinct morphological transitions (stretching, rolling, and tumbling) provides cues for understanding the healthy and diseased states of the cells based on their morpho-dynamic features and establish exclusive connectivity of the same with the cell membrane as well as the cytoplasm properties, a paradigm that is currently non-existant. This, in turn, may lead to a novel mechanistic approach of label-free disease detection based on cellular imaging, for which the current understanding is mostly empirical rather than fundamental.

biophysics↗

bioRxiv2023

Biology of Coital Behavior: Looking Through the Lens of Mathematical Genomics

Research has shown that genetics and epigenetics regulate mating behavior across multiple species. Previous studies have generally focused on the signaling pathways involved and spatial distribution of the associated receptors. However a thorough quantitative characterization of the receptors involved may offer deeper insight into mating behavioral patterns. Here oxytocin, arginine-vasopressin 1a, dopamine 1, and dopamine 2 receptors were investigated across 76 vertebrate species. The receptor sequences were characterized by polarity-based randomness, amino acid frequency-based Shannon entropy and Shannon sequence variability, intrinsic protein disorder, binding affinity, stability and pathogenicity of homology-based SNPs, structural and physicochemical features. Hierarchical clustering of species was derived based on structural and physicochemical features of the four receptor sequences separately, which eventually led to proximal relationships among 29 species. Humans were found to be significantly distant phylogenetically from the prairie voles, a representative of monogamous species based on coital behavior. Furthermore, the mouse (polygamous), the prairie deer mouse (polygamous), and the prairie vole (monogamous) although being proximally related (based on quantitative genomics of receptors), differed in their coital behavioral pattern, mostly, due to behavioral epigenetic regulations. This study adds a perspective that receptor genomics does not directly translate to behavioral patterns.

genomics↗

bioRxiv2023

Methotrexate-modulated talin-dynamics drives cellular mechanical phenotypes via YAP signalling

Methotrexate is a well-known antineoplastic drug used to prevent cancer aggravation. Despite being a targeted therapeutic approach, its administration comes with the risk of cancer recurrence, plausibly through its proven off-target effect on focal adhesions. Since FA dynamics is dependent on force transmission through its constituent proteins, including talin, methotrexate might affect the mechanical activity of these proteins. Here we have combined single-molecule studies, computational dynamics, cell-based assays, and genomic analysis to unveil the focal adhesion-regulating role of methotrexate central to its effect on talin dynamics and downstream pathways. Interestingly, our single-molecule force spectroscopic study shows that methotrexate modulates the bimodal force distribution of talin in a concentration-dependent manner. Steered molecular dynamics reveal that methotrexate-talin interactions alter talin mechanical stability exposing their vinculin binding sites. Finally, we found that methotrexate-regulated talin-dynamics remodel cancer cell mechanical phenotypes like cell polarity, adhesion, and migration by regulating talin-vinculin association-mediated YAP signaling. These results further correlate with genomic analysis of methotrexate-treated patients, demonstrating its clinical importance. Taken together, these findings disseminate the effects of methotrexate-modulated mechanosensitivity of adhesion proteins on cellular events.

cell biology↗

bioRxiv2023

Semiconducting carbon nanotubes decrease neuronal bursting in a network of rat hippocampal neurons in vitro while increasing intrinsic excitability of single neurons

The diverse electrical, chemical and structural properties of the functional derivatives of carbon nanotubes (CNTs) have shown biomedical possibilities for neuroprosthesis or neural interfaces. However, the studies have been generally confined to metallic CNTs that affect cell viability unless chemically functionalized for biocompatibility. Here, we explored the effects of semiconducting single-walled carbon nanotubes (ssw-CNT), on the active electrical properties of dissociated hippocampal neurons in-vitro using multielectrode array, calcium imaging and whole-cell patch clamp recordings. The findings show that ssw-CNT treatment regulates neural network excitability from burst to tonic firing by changing the calcium dynamics. However, at a single neuronal level, ssw-CNT increases neuronal excitability.

neuroscience↗

bioRxiv2023

Role of GD2 and its biosynthetic enzyme GD3 synthase in prostate cancer tumorigenesis

While better management of loco-regional prostate cancer (PC) has greatly improved survival, advanced PC remains a major cause of cancer deaths. Identification of novel targetable pathways that contribute to tumor progression in PC could open new therapeutic options. The di-ganglioside GD2 is a target of FDA-approved antibody therapies in neuroblastoma, but the role of GD2 in PC is unexplored. Here, we show that GD2 is expressed in a small subpopulation of PC cells in a subset of patients and a higher proportion of metastatic tumors. Variable levels of cell surface GD2 expression were seen on many PC cell lines, and the expression was highly upregulated by experimental induction of lineage progression or enzalutamide resistance in CRPC cell models. GD2high cell fraction was enriched upon growth of PC cells as tumorspheres and GD2high fraction was enriched in tumorsphere-forming ability. CRISPR-Cas9 knockout (KO) of the rate-limiting GD2 biosynthetic enzyme GD3 Synthase (GD3S) in GD2high CRPC cell models markedly impaired the in vitro oncogenic traits and growth as bone-implanted xenograft tumors and reduced the cancer stem cell (CSC) and epithelial-mesenchymal transition (EMT) marker expression. Our results support the potential role of GD3S and its product GD2 in promoting PC tumorigenesis by maintaining cancer stem cells and suggest the potential for GD2 targeting in advanced PC.

cancer biology↗

bioRxiv2023

Force-regulated chaperone activity of BiP/ERdj3 is opposite to their homologs DnaK/DnaJ: explained by strain energy

Polypeptide chains experiences mechanical tension while translocating through cellular tunnel. In this scenario, interaction of tunnel-associated chaperones with the emerging polypeptide occurs under force; however, this force-regulated chaperone behaviour is not fully understood. We studied the mechanical chaperone activity of two tunnel-associated chaperones BiP and ERdj3 both in the absence and presence of force; and compared to their respective cytoplasmic homologs DnaK and DnaJ. We found that BiP/ERdj3 shows strong foldase activity under force; whereas their cytoplasmic homolog DnaK/DnaJ behave as holdase. Importantly, these tunnel-associated chaperones (BiP/ERdj3) revert to holdase in the absence of force, suggesting that mechanical chaperone activity differs depending on the presence or absence of force. This tunnel-associated chaperone-driven folding event generates additional mechanical energy of up to 54 zJ that could help protein translocation. The mechanical-chaperone behaviour can be explained by strain theory: chaperones with higher intrinsic deformability function as mechanical foldase (BiP, ERdj3), while chaperones with lower intrinsic deformability act as holdase (DnaK and DnaJ). Our study thus unveils the underlying mechanism of mechanically regulated chaperoning activity and provides a novel mechanism of co-translocational protein folding. SignificanceThe mechanical-activity of chaperones, located at the edge of a tunnel, could be different from their cytoplasmic homologs. Translocating substrates within the tunnel are known to experience mechanical constraints, whereas the cytosolic substrates interact with the chaperones in the absence of force. To understand this phenomenon, we investigated two tunnel-associated chaperones BiP/ERdj3 and their cytosolic homologs-DnaK/DnaJ. We observed that BiP/ERdj3 possess strong foldase activity while their substrates are under force; whereas DnaK/DnaJ possess holdase function. Notably all these chaperones function as holdase in the absence of force, which suggest that mechanical chaperone activity is different with and without force. We explained this mechanical behaviour using strain theory, providing a physical mechanism of chaperone-assisted co-translocational protein folding.

biophysics↗

bioRxiv2023

Mechanism of GTPase activation of a prokaryotic small Ras-like GTPase MglA by an asymmetrically interacting MglB dimer

Cell polarity oscillations in Myxococcus xanthus motility are driven by a prokaryotic small Ras-like GTPase, MglA, which switches from one cell pole to the other in response to extracellular signals. MglA dynamics is regulated by MglB, which functions both as a GAP (GTPase activating protein) and a GEF (guanine nucleotide exchange factor) for MglA. With an aim to dissect the role of asymmetry in the dual GAP and GEF activities of MglB, we generated a functional MglAB complex by co-expressing MglB with a linked construct of MglA and MglB. This strategy enabled us to generate mutations of individual MglB protomers (MglB1 linked to MglA or MglB2) and delineate their role in GEF and GAP activities. We establish that the C-terminal helix of MglB1, but not MglB2, stimulates nucleotide exchange through a site away from the nucleotide-binding pocket, confirming an allosteric mechanism. Interaction between the N-terminal {beta}-strand of MglB1 and {beta}0 of MglA is essential for the GEF activity of MglB. Specific residues of MglB2 which interact with Switch-I of MglA partially contribute to its GAP activity. Thus, the role of the MglB2 protomer in the GAP activity of MglB is limited to restricting the conformation of MglA active site loops by steric hindrance. The direct demonstration of the allosteric mechanism of GEF action provides us new insights into the regulation of small Ras-like GTPases, a feature potentially present in many uncharacterized GEFs.

biochemistry↗

bioRxiv2023

Toxoplasma gondii aspartic protease 5 (TgASP5): Understanding structural details and inhibition mechanism.

Toxoplasma gondii, a worldwide prevalent parasite, is responsible for causing toxoplasmosis by infecting almost all warm-blooded animals, including humans. To establish a successful infection, the parasite exports a series of effector proteins which modulates the host immune system; Golgi-resident T. gondii aspartyl protease 5 (TgASP5) plays an essential role in the maturation and export of these effector proteins. This is the first report of the detailed structural investigation of the TgASP5 mature enzyme. Molecular modeling and all-atom simulation provided in-depth knowledge of the active site architecture of TgASP5. The analysis of the binding mode of TEXEL substrate highlighted the importance of the active site residues forming the pocket; the Ser505, Ala776 and Tyr689 in the S2 binding pocket provide the specificity towards Arg at the P2 position. Our study also provides insights into the binding mode of the known inhibitor RRLStatine. Screening the known aspartic protease inhibitors against TgASP5 active site and performing 100 ns all-atom molecular dynamic simulations, MM-PBSA binding energy calculations provided the best nine inhibitor protein complexes. Besides that, Principal Component Analysis (PCA) was employed to identify the change in protein dynamics with respect to the substrate and ligand binding. TgASP5 is essential for the fitness and virulence of the parasite; inhibiting this enzyme can be a possible therapeutic strategy against toxoplasmosis. Our study put forth the inhibitors which can act as initial scaffolds for developing potent mechanistic inhibitors against TgASP5.

bioinformatics↗

bioRxiv2023

Toxic metal uptake by oyster mushrooms grown in sugarcane bagasse

Fungiculture or Mushroom Cultivation is rapidly expanding throughout the world. It is further catalyzed by its high nutritional value and increased market demand. Additionally, the method of cultivation does not require much space and the per unit production is very high compared to other crops. Bagasse is the byproduct left after extracting the juice from sugarcane. It has been speculated that bagasse can be an excellent substrate due to its high carbohydrate and mineral content (Hoa et al., 2015). In the experiment spawns of Oyster Mushroom were inoculated in bagasse collected from industries. Remarkable growth of mushrooms in short period of time was observed. However, the metal toxicity being our primary concern in this experiment, AAS (Atomic Absorption Spectrometry) Test was used to detect the heavy metal ion concentrations in both the bagasse and the fruiting bodies of mushrooms. The concentrations of metals calculated are as follows, in bagasse (mg/kg): Cadmium - 0.015, Arsenic - 8.56, Copper-12.47, Chromium-9.17, Mercury - less than 0.01, and in mushroom (mg/kg): Cd - 0.022, As - 10.12, Cu - 14.29, Cr - 7.59, Hg - less than 0.01. These results indicated biomagnification of metal ion concentration in the mushrooms up taken from bagasse. These metals, in such high concentrations are lethal and causes many diseases in humans and other animals. Hence, from this experiment it can be deduced, that though bagasse is an excellent substrate for mushroom cultivation, the toxicity of metal ions present in it in such high concentrations overshadow the earlier benefits of bagasse as a good substrate. However, it has been proposed that if such metals are removed or their concentrations are lowered then the earlier mentioned benefits of bagasse are restored.

ecology↗

bioRxiv2023

Development and qualification of a high-yield recombinant human Erythropoietin biosimilar

Recombinant human erythropoietin (rhEPO) has been saving millions of lives worldwide as a potent and safe treatment for the lack of erythrocyte, which is caused by chronic kidney disease (CKD) and other issues. Several biosimilars of rhEPO have been approved since the expiry of the relevant patents to provide cost-effective options but the price of rhEPO is still high for the affordability of global community. Therefore, development of biosimilar of rhEPO at a lower price is highly necessary. Here we report the development and characterization of a biosimilar of rhEPO with high-yield satisfying regulatory requirements. The hEPO-expressing cDNA was stably expressed in CHO cells with successive transfection. The master cell bank (MCB) and working cell bank (WCB) were established from the best selected clone and characterized for 50 passages. The rhEPO was expressed from the WCB in single-use suspension culture system with a high-titer (1.24{+/-}0.16 g/L). To the best of our knowledge this is the highest reported rhEPO titer to date. The rhEPO was purified using a series of validated chromatography unit processes including virus inactivation and filtration. The purified EPO was formulated in serum-free buffer, sterile filtered, and analyzed as the biosimilar of reference product Eprex(R). Physicochemical analysis strongly suggested similarities between the developed rhEPO (GBPD002) and the reference. The in vitro and in vivo functional assays confirmed the similar biofunctionality of the GBPD002 and Eprex(R). GBPD002 could provide a less-expensive solution to the needful communities as an effective and safe biosimilar where rhEPO treatment is necessary.

bioengineering↗

bioRxiv2023

EHD1-dependent traffic of IGF-1 receptor to the cell surface is essential for Ewing sarcoma tumorigenesis and metastasis

Overexpression of EPS15 Homology Domain containing 1 (EHD1) has been linked to tumorigenesis but whether its core function as a regulator of intracellular traffic of cell surface receptors plays a role in oncogenesis remains unknown. We establish that EHD1 is overexpressed in Ewing sarcoma (EWS), with high EHD mRNA expression specifying shorter patient survival. ShRNA and CRISPR-knockout with mouse Ehd1 rescue established a requirement of EHD1 for tumorigenesis and metastasis. RTK antibody arrays identified the IGF-1R as a target of EHD1 regulation in EWS. Mechanistically, we demonstrate a requirement of EHD1 for endocytic recycling and Golgi to plasma membrane traffic of IGF-1R to maintain its surface expression and downstream signaling. Conversely, EHD1 overexpression-dependent exaggerated oncogenic traits require IGF-1R expression and kinase activity. Our findings define the RTK traffic regulation as a proximal mechanism of EHD1 overexpression-dependent oncogenesis that impinges on IGF-1R in EWS, supporting the potential of IGF-1R and EHD1 co-targeting.

cancer biology↗