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Chakraborty, S.

Publications and source records attributed to Chakraborty, S..

At least 73 records · Page 4Linked to original sources

Absence of posterior commissure and sub-commissural organ precedes encephalocele development in a new mouse model

Encephalocele is a congenital defect involving herniation of the meninges, with or without brain tissue, outside the skull. Although traditionally considered a neural tube defect (NTD) alongside anencephaly and open spina bifida (myelomeningocele), encephalocele typically shows well-formed brain tissue, and is likely to represent a later-arising, post-neurulation developmental anomaly, with a different pathogenic mechanism from the open NTDs. A detailed understanding of encephalocele pathogenesis requires experimental studies and, recently, we developed a new mouse model in which the genes encoding FGF3, 4 and 15 are over-expressed in the embryonic day (E) 9.5 brain. Encephalocele subsequently develops as a broad forebrain-midbrain swelling, visible from E11.5, that resolves by birth into a focal brain herniation resembling human parieto-occipital encephalocele. A structural analysis of the brain in mutant embryos reveals absence of the posterior commissure and sub-commissural organ, and at later stages the pineal gland. These structures normally develop just rostral to the forebrain-midbrain boundary, and Pax6 immuno-histochemistry demonstrates that this boundary remains intact in the mutant embryos. Histological analysis reveals a more general change in tissue composition of the neural tube roof in the forebrain-midbrain region, with diminished thickness of the neuroepithelium and increased thickness of the overlying layer, including the non-neural ectoderm (future epidermis). We conclude that the posterior commissure and sub-commissural organ, previously implicated in hydrocephalus, may also be fundamental for development of the earlier-arising brain malformation, encephalocele.

developmental biology↗

Over-expression of Cyclic Nucleotide-Gated Ion Channel 2 (CNGC2) triggers 1 hypersensitivity to virulent pathogens and elevated Ca2+

The Arabidopsis Cyclic Nucleotide-Gated Ion Channel 2 (CNGC2), also known as Defense No Death 1 (DND1), is the most extensively studied plant CNGC and has been implicated in diverse physiological processes, including floral transition, responses to heat and humidity, and hormone signaling. Its role in immunity has received particular attention due to the autoimmunity phenotype observed in the cngc2/dnd1 knockout mutants. Interestingly, despite this hyperactivation of immunity, the mutant also exhibits impaired hypersensitive cell death--a hallmark of effector-triggered immunity (ETI)--as well as reduced reactive oxygen species (ROS) production and diminished Ca{superscript 2} influx in response to pathogen-associated molecular patterns (PAMPs) such as the bacterial flagellin peptide flg22. These contradictory phenotypes highlight the complex biological functions of CNGC2. To date, most studies have focused on loss-of-function mutants. In this study, we performed a detailed characterization of CNGC2 overexpression lines to gain deeper insight into its role in immunity. Remarkably, overexpression of CNGC2 led to heightened susceptibility to two taxonomically distinct pathogens, despite the plants displaying wild-type morphology. Overexpression of CNGC2 rescued several cngc2 mutant phenotypes, including morphological defects and delayed flowering, yet these plants were also hypersensitive to elevated external Ca{superscript 2} levels. Furthermore, they exhibited attenuated responses to flg22, suggesting that CNGC2 does not act as a simple positive or negative regulator of immunity. Our findings reveal an essential role for CNGC2 where a balanced expression level is critical for maintaining Ca{superscript 2} homeostasis between the apoplast and cytosol, thereby influencing the generation of Ca{superscript 2} signals essential for immune responses.

plant biology↗

PEG400 regulates Falcipain 2 activity through an unprecedented allosteric mechanism

The malarial parasite Plasmodium falciparum cleaves host hemoglobin by cascade of proteolytic enzymes. The cysteine protease, Falcipain-2 (FP2) plays an essential role in the process and important for parasite survival, making it a potential drug target. However, similarities with host cysteine cathepsins hamper selective inhibition, thus necessitates detailed structural and functional characterizations of FP2. The present study uncovers a novel regulatory role of polyethylene glycol 400 (PEG400) on FP2 activity. PEG400 inhibits FP2 activity on small peptide substrate and azo-casein, while enhancing hemoglobin degradation, rendering a dual effect on FP2 catalysis. A mixed-type of inhibition has been observed for PEG400 against small peptide substrate of FP2, consistent with binding of PEG400 to catalytic cleft, confirmed by fluorescence quenching and docking studies. Unlike typical nonspecific PEG-protein interactions, PEG400 adopts a fit within catalytic region of FP2 and partially overlaps with leupeptin binding sites, albeit with lower affinity. Computational analysis further identifies a novel allosteric binding pocket of PEG400, supported by in-silico mutagenesis and molecular dynamics simulation. This pocket exhibits minimal conservation in human cathepsins, suggesting selective potential. In contrast to this inhibitory role, biochemical assay reveals that PEG400 promotes haemoglobin proteolysis. Spectroscopic analyse suggests PEG400 alter hemoglobin structural dynamics to favour proteolysis. ENM based normal mode analysis reveals upon haemoglobin binding, PEG400 restricts FP2 hinge-bending motion, improves FP2-hemoglobin proximity, and simultaneously PEG400 is dislodged from the active site, thereby promoting proteolysis. The combined experimental and computational findings reveal a novel mechanism of FP2 regulation, opening new therapeutic avenues.

biochemistry↗

Persistence of florpyrauxifen-benzyl in sediments following application to a large oligotrophic lake to control Eurasian watermilfoil

Florpyrauxifen-benzyl (FPB) is a recently registered arylpicolinate herbicide used to control dicots including Eurasian watermilfoil (EWM). Laboratory studies indicate rapid degradation of FPB in water, but sediment persistence in aquatic systems remains poorly understood, creating uncertainty in ecological risk assessments. We evaluated the environmental fate and transport of FPB and its degradants in water, plants, and sediment following application of the herbicide to two bays of Lake George, NY. FPB was rapidly lost from the water column within 72 hours, consistent with prior studies, but persisted in sediments for at least one year. Sediment cores revealed vertical migration of FPB, which may contribute to frequent non-detects in surface sediments and potentially explain why US EPA was unable to establish sediment half-lives in aquatic systems. Measured concentrations exceeded the 28-day no observable adverse effect concentration (NOAEC) for chironomids for at least a year, highlighting the potential for long-term chronic exposure to benthic organisms. Modeled sorption and dilution scenarios aligned with field observations and indicated that circulation dynamics expanded the affected area beyond the direct treatment zones. These results demonstrate that FPB and its degradants persist longer in sediments than suggested by laboratory studies, raising important questions about long-term ecological risks, effects of repeated applications, and underestimation of sediment persistence. Long-term studies such as this are essential for understanding the fate of these compounds and for guiding informed decisions about future herbicide applications.

pharmacology and toxicology↗

Design, Synthesis, Biological Evaluation and Molecular Docking of New Acid-Functionalized Carbazole Derivatives as Potential Antibiotic Agents

The emergence of antibiotic resistance necessitates the discovery of new scaffolds with improved efficacy and drug-like properties. In this work, a new series of acid-functionalized carbazole derivatives was designed, synthesized, and comprehensively evaluated for their antibacterial potential. The incorporation of acidic functionalities into the carbazole framework enhanced physicochemical properties and biological interactions, yielding distinct strain-specific antibacterial activities. Minimum inhibitory concentration (MIC) studies demonstrated that 3-methyl-1,4-dioxo-4,9-dihydro-1H-carbazole-6-carboxylic acid (2) exhibited broad-spectrum potency, particularly against S. aureus and E. coli, while 6-methyl-9H-carbazole-3-carboxylic acid (3) showed selectivity against B. cereus and (E)-3-methyl-1-(2-tosylhydrazono)-2,3,4,9-tetrahydro-1H-carbazole-6-carboxylic acid (1) displayed strong activity toward S. typhimurium. Molecular docking studies revealed favourable binding affinities of all derivatives toward bacterial dihydrofolate reductase (DHFR), with compound 1 showing the highest docking scores. Molecular dynamics simulations further confirmed the broad conformational adaptability of compound 1, the target-specific stability of compound 3, and protein-dependent binding behaviour of compound 2. Complementary ADMET predictions indicated that all compounds adhered to Lipinskis rules, with compound 3 displaying the most favourable pharmacokinetic profile, including high oral bioavailability and low toxicity risk. Together, these experimental and computational findings establish acid-functionalized carbazole scaffolds as promising antibacterial candidates.

pharmacology and toxicology↗

AKNA drives neural stem cell fate transition through differential localization and coordinating the modulation of chromatin from H3K27me3 to H3K27ac

Neuronal stem cells (NSCs) play pivotal role in adult neurogenesis, however, detail mechanisms of obtaining pluripotency or undergoing differentiation remain unknown. Herein, how AT-hook protein AKNA regulates pluripotency and stemness in neuroblastoma cells is demonstrated by gene knockdown, immunofluorescence, chromatin immunoprecipitation (ChIP), localization of AKNA, signaling interactions and transcriptional activity. AKNA was abundant and mostly nuclear during induction of pluripotency and its knockdown reduced stemness, even in presence of other pluripotency factors, including OCT4 and SOX2. During the course of induction of differentiation, AKNA remain localized in the cytosol, essential for regulated differentiation. Cytosolic retention of AKNA is possibly driven by FAK signaling. In the nucleus, AKNA promotes KDM6B demethylase for H3K27me3 demethylation to H3K27, and subsequently promotes CBP/p300 to acetylate H3K27 to H3K27ac deposition on promoter region of respective genes to trigger their transcription. Data obtained from knockdown and overexpression of AKNA and KDM6B further reinforce its importance that, they physically interact to drive pluripotency and stemness. These findings establish AKNA as a critical regulator for NSCs fate determination in association with epigenetic modifiers and signaling pathway, offering potential targets for neuroblastoma therapies and regenerative medicines for neurodegenerative diseases.

molecular biology↗

Immune cell de novo steroidogenesis regulates inflammation resolution and recovery in acute lung injury

Effective resolution of inflammation following acute lung infection or injury is critical for restoring immune and tissue homeostasis to ensure functional recovery. Prolonged or unresolved inflammation can impair lung repair, promote fibrosis, and contribute to pulmonary dysfunction. While systemic steroid signalling is known to modulate general immune responses, the specific role of immune cell-mediated steroidogenesis in regulating lung inflammation and repair remains unknown. Here, we show that immune cell de novo steroidogenesis is essential for resolving inflammation and promoting recovery in a murine model of acute lung injury. During the resolution phase, steroid-synthesizing immune cells, predominantly basophils, are enriched in the lung. Mice with immune cell-specific ablation of de novo steroidogenesis exhibit exacerbated lung injury, impaired resolution of inflammation, and defective tissue repair. These findings reveal a previously unrecognized immunoregulatory function of immune cell-derived steroids and identify immune cell steroidogenesis as a potential therapeutic target for promoting resolution and recovery in inflammatory lung diseases.

immunology↗

Yeast two-hybrid screening identifies Dad1 as a candidate substrate of Hrr25 kinase in Saccharomyces cerevisiae

In Saccharomyces cerevisiae, sister kinetochores are mono-oriented during meiosis I, ensuring that homologous chromosomes segregate to opposite poles, a process critically dependent on the kinase activity of Hrr25, a casein kinase. However, the direct substrates of Hrr25 involved in this mechanism remain poorly defined. In this study, we used a yeast two-hybrid (Y2H) approach to screen for and identify physical interactors of Hrr25. The HRR25 gene was cloned into a Y2H bait vector, and its functional expression was confirmed by complementation of a temperature-sensitive hrr25-ts mutant. Screening independent Y2H libraries in three reading frames, followed by validation via reporter assays, restriction analysis, and sequencing, we identified six putative interactors: HED1, DAD1, YDR015C (from clone C1-5), REP1 (C2-15), and CYR1 and SYS1 (C3-7). Phosphorylation site prediction and AlphaFold 3.0 structural modeling identified high-confidence Hrr25 target residues, including S70/T73 on Hed1 and S63 on Dad1, S323 on Rep1, and S198/S527 on Cyr1, each located in structurally accessible and potentially functional regions. Plasmid-dependent assays confirmed that reporter activation in C1-5 depended on the presence of the prey plasmid, and restriction mapping demonstrated that C1-5 contained a full-length DAD1 ORF. Given Dad1s known role in DASH/Dam1 kinetochore complex and its function in kinetochore-microtubule attachment, along with previous findings that DAD1 mutations cause meiosis I defects, our data suggest that Dad1 may be a substrate of Hrr25. We propose that Hrr25-mediated phosphorylation of Dad1 could facilitate sister kinetochore co-orientation during meiosis I. These results provide new insights into the molecular mechanisms of chromosome segregation and identify Dad1 as a potential candidate substrate for Hrr25 in meiotic regulation.

cell biology↗

Inhibition of coronaviral exonuclease activity by TRIM-mediated SUMOylation

Members of the TRIM E3 ligase family are effectors of the host innate or intrinsic defense against various viruses; however, how specific TRIM proteins antagonize coronavirus infection is still largely elusive. Through an RNAi screen targeting 71 human TRIM genes, we identified multiple TRIM proteins with antiviral or proviral activity against SARS-CoV-2. TRIM32 potently restricted SARS-CoV-2 replication in a RING E3 ligase-dependent but interferon-independent manner. Mechanistically, TRIM32 binds to and SUMOylates the 3'-to-5' exoribonuclease (ExoN) of NSP14, which is essential for SARS-CoV-2 replication. TRIM32-mediated NSP14 SUMOylation at K9 and K200 inhibits RNA binding and NSP10 cofactor recruitment, respectively, ultimately suppressing ExoN activity. Our study further revealed that NSP14 SUMOylation by TRIM32 and its antiviral activity are broadly conserved for coronaviruses. These results identify the coronaviral NSP14 protein as a direct target of host restriction via SUMOylation, which may uncover novel ways to therapeutically inhibit coronavirus infections in humans.

microbiology↗

Maternal CENP-C restores centromere symmetry in mammalian zygotes to ensure proper chromosome segregation

Across metazoan species, the centromere-specific histone variant CENP-A is essential for accurate chromosome segregation, yet its regulation at the parental-to-zygote transition in mammals is poorly understood. To address this, we developed a CENP-A-mScarlet knock-in mouse model, which revealed sex-specific dynamics: mature sperm retains 10% of the CENP-A levels present in MII-oocytes. However, in zygotes prior to the first mitosis, this difference is resolved, using maternally inherited cytoplasmic-CENP-A. Notably, the increase in CENP-A at paternal centromeres is independent of sensing CENP-A asymmetry or the presence of maternal chromosomes. Instead, CENP-A equalization relies on asymmetric recruitment of maternal CENP-C to paternal centromeres. Depletion of maternal CENP-A decreases total CENP-A in pronuclei without disrupting equalization. In contrast, reducing maternal CENP-C or disruption of its dimerization domains impairs CENP-A equalization and chromosome segregation. Therefore, maternal CENP-C acts a key epigenetic regulator that resets centromeric symmetry at fertilization to preserve genome integrity. Highlights[bullet] CENP-A asymmetry between sperm and oocyte centromeres is a conserved feature from flies to mammals including mice and humans. [bullet]CENP-A asymmetry between parental centromeres is resolved prior to the first zygotic division via maternally inherited, cytoplasmic CENP-A. [bullet]Zygotic CENP-A levels in zygotes are regulated in a pronucleus-autonomous manner. [bullet]CENP-A equalization relies on asymmetric CENP-C recruitment to the paternal pronucleus and requires CENP-C dimerization. Key TermsCentromere; CENP-A; CENP-C; sperm; oocyte; zygote; intergenerational; epigenetics; mouse

genetics↗

Engineering macrophage responses through 3D scaffold microarchitecture

Biomaterial implantation in living organisms triggers a physiological response known as foreign body reaction, leading to the recruitment of macrophages, that can polarize either into a pro-inflammatory (M1) or an anti-inflammatory (M2) phenotype. Currently, there is growing interest in tailoring the physical properties of tissues and biomaterials to promote efficient tissue regeneration. Tridimensionality can profoundly influence macrophage behaviour; however, there is no clear consensus on the underlying mechanisms. 3D microstructures may play a crucial role in modulating immune cells, promoting anti-inflammatory responses, and supporting effective tissue repair and regeneration. In this study, we used two-photon polymerization to fabricate 3D scaffolds with large pores, measuring 50x50x20 m3, and small pores, measuring 15x15x15 m3. Both microstructures effectively influenced macrophage cytoskeletal organization and cellular metabolic activity. Notably, they were not sufficient to induce spontaneous macrophage polarization, indicating that they are intrinsically immunologically inert. When combined with chemical stimulation, as typically occurs in vivo, they elicited distinct responses. Specifically, as evidenced by the slight upregulation of the Arg1 marker, large pore sizes promoted an anti-inflammatory phenotype. Conversely, iNOS expression measurements indicated that small pores, which impose spatial constraints on macrophages, favoured a massive pro-inflammatory state. Our results demonstrate that 3D microstructures are versatile tools for multiple applications. Their precisely tunable architecture enables fine control over macrophage behaviour and immunomodulation, opening new avenues both for tissue engineering, by preventing fibrosis and promoting anti-inflammatory and pro-regenerative responses in vivo, and for the development of in vitro platforms to model inflamed tissues for screening anti-inflammatory drugs.

bioengineering↗

Exploiting an Epigenetic Resistance Mechanism to PI3 Kinase Inhibition in Leukemic Stem Cells

Acquired non-genetic resistance mechanisms to existing therapies contribute to poor outcomes for acute myeloid leukemia (AML) patients, and inability to target leukemic stem cells (LSCs) can lead to relapse. To overcome these challenges, we tested whether LSCs have dependencies on PI3 kinase (PI3K). We found that LSCs are susceptible to isoform-selective targeting of PI3K and are particularly dependent on the P110 alpha isoform of PI3K. We discovered that PI3K inactivation leads to dynamic changes in EZH2/PRC2 function in leukemic cells, and we uncovered downregulation of EZH2 protein levels as a resistance mechanism in response to PI3K inhibition. We found that PI3K inhibition in AML cells can lead to compensatory upregulation of EZH1, and that EZH1 knockdown can sensitize AML cells to PI3K inhibition. We leveraged this resistance mechanism by combining a PI3K inhibitor with an EZH1/2 dual inhibitor, which successfully overcomes the acquired resistance and leads to sustained targeting of AML cells ex vivo and in murine AML and PDX models in vivo. This study identifies a promising novel therapeutic regimen for targeting LSCs in AML.

cancer biology↗

Evaluation of natural killer cell tumor homing and effector function in response to CDK4/6 and AURKA inhibition in a melanoma tumor-on-a-chip platform

Natural killer (NK) cells have emerged as an important clinical tool cellular immunotherapy. Whereas immune checkpoint blockade (ICB) or chimeric antigen receptor (CAR) T-cell therapy (CAR-T) therapy have been adopted as a first line treatments in different malignancies, such as melanoma, these approaches do not work for all patients. T cells require proper antigen presentation on tumor cells for recognition and to carry out their corresponding cytotoxic functions. Deficiency of tumor antigens, or high variability in those present, make T cell-based CAR-T and ICB ineffective. By contrast, NK cells are not limited by antigen presentation deficiencies, offering a potential alternative approach, yet their efficacy can suffer from immunosuppressive signals. Herein, we sought to develop in vitro and on-chip platforms to identify strategies for enhance, rather than suppress, NK cell homing to tumor cells. We explored the use of inhibition of kinases such as CK4/6 and AURKA to induce tumor cell production of chemokines that NK cells migrate towards in aggressive melanoma models. We evaluated chemokine-aided NK cell migration-homing capabilities and their therapeutic efficacy and found that treatment of both melanoma cell line and patient-tumor constructs (PTCs) with CDK4/6 and AURKA generally resulted in improved NK cell homing to tumor cells and accompanying tumor cell killing. Interestingly, this chemokine-guided NK cell migration did not generate as effective outcomes in models using a mildly aggressive melanoma cell line. For our studies, we used 3D tumor constructs in both static Transwell models and then in a bioengineered NK cell-functionalized tumor-on-a-chip (NK-TOC) platform. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=83 SRC="FIGDIR/small/662476v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@17bf445org.highwire.dtl.DTLVardef@e1e1d9org.highwire.dtl.DTLVardef@1b29c2corg.highwire.dtl.DTLVardef@12b1f03_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

Lead optimization against drug-resistant Leishmania donovani infection: Semi-synthetic derivatives of ethyl linoleate isolated from Indian edible mushroom Meripilus giganteus.

Visceral leishmaniasis, the most severe form, affects thousands of people annually. Current drugs in practice fail to provide an absolute cure. Globally, treatment relies on a single dose of liposomal amphotericin B, which requires a costly setup and cold chain to maintain. Natural molecules remain the vital sources of therapeutic ingredients in modern medicine. Meripilus giganteus is traditionally consumed by the people of the North-eastern regions of the Himalayas, India. In this study, we planned for the stepwise bioactivity-guided isolation of natural molecules from M. giganteus and to synthesize more potential therapeutic derivatives against the drug-resistant Leishmania infection. HPLC chromatogram of ethyl acetate extract obtained from M. giganteus revealed the presence of ethyl linoleate as the most active anti-leishmanial molecule. Several derivatives of ethyl linoleate were synthesized and evaluated against Leishmania promastigotes. Among them, a single epoxygenated variant of ethyl linoleate, ethyl (Z)-8-(3-(oct-2-en-1-yl) oxiran-2-yl) octanoate (EL3), showed promising anti-leishmanial activity against both drug-sensitive and drug-resistant Leishmania donovani. The lead derivative disrupts the biosynthesis of trypanothione from glutathione and spermidine, making the parasite vulnerable to the oxidative burst inside the host. Interestingly, the lead derivative was found to be more efficient against the drug-resistant L. donovani, in vivo. Besides this, it has almost no toxicity towards host cells. Considering the urgency for new drug development, we propose this novel semi-synthetic derivative as a potent therapeutic lead.

immunology↗

Phagosome Maturation in Macrophages is Enhanced by p38α MAPK Signaling

In macrophages, it is generally assumed that without active escape mechanisms, all phagocytosed cargo eventually reaches lysosomes. Yet, the influence of specific ligands, like lipopolysaccharide (LPS), on phagosome maturation is unclear. Here, using sterile, non-immunogenic particles as model cargo phagocytosed by macrophages, we show that in the absence of an external signal, less than half the phagosomes fuse with lysosomes. By quantifying phagosome maturation in terms of lysosomal localization and mapping lumenal pH dynamics, we find that early events triggered by LPS-induced signaling enhance both cargo delivery to lysosomes and phagosome acidification rates. We demonstrate that stress-activated p38 alpha mitogen-activated protein kinase (p38 MAPK) is important for this signaling event. Other signals known to activate p38 MAPK such as flagellin, IgG and albumin also enhance lysosomal delivery of phagocytosed cargo. Our work indicates that the maturation of phagosomes in macrophages is enhanced by the presence of specific ligands on the cargo, which activate cell surface receptors that signal via p38 MAPK.

cell biology↗

The microcephaly protein Abnormal Spindle has an essential role in symmetrically dividing neural precursors to promote brain growth and development

Neurogenesis must be coordinated in time and space to ensure proper neural development. Defects in this process can lead to a variety of brain malformations that affect tissue size and architecture. Here we examine the spatiotemporal requirements of the Abnormal Spindle (Asp) protein and its human ortholog Abnormal Spindle-Like, Microcephaly Associated (ASPM) in a fly model of human microcephaly (MCPH). By utilizing the fly optic lobe, whose neurogenic program parallels that of mammals, and the genetic tools available in this organism, we uncover the progenitor cell types and critical neurogenesis window necessary for proper brain size and architecture. Asp has an essential function in symmetrically dividing neuroepithelial precursors, yet its expression in asymmetrically dividing neuroblasts alone is not sufficient for optic lobe development. Asp is highly expressed during both embryonic and larval neurogenesis stages, but embryonic expression is dispensable for making a properly sized adult brain. We also show that Asp/ASPMs interphase nuclear localization is not required for its ability to promote brain development. Expression of the human ASPM N-terminus can also significantly rescue fly brain defects, suggesting conserved mechanisms of function in brain growth control. These results support a model where Asp expression in symmetrically dividing precursors is required to generate a sufficient pool of progenitors prior to the cell fate switch to asymmetrically dividing neuroblasts in order to generate enough neuronal cells to make a properly sized brain.

developmental biology↗

Fibroblasts drive differential response in immunotherapy via IL-6 in bioengineered 3D cancer constructs

Despite various therapeutic advancements in cancer, heterogeneity in the tumor microenvironment is a major hindrance to the overall efficacy of treatment. In this study, we focus on the most abundantly found cellular stromal components - fibroblasts - which can be tumor-promoting or tumor-suppressing based on the extent of disease progression and numerous additional factors. In the studies described herein, we evaluated their role in influencing chemotherapy as well as in immunotherapy using colorectal cancer (CRC) cell line-derived 3D tumor constructs. Our initial efforts centered on common chemotherapeutic agents utilized in CRC. However, while the presence of fibroblasts often did influence chemotherapy efficacy, a clear and consistent increase or decrease in efficacy was not obvious. We then focused subsequent studies on natural killer (NK) cell-based immunotherapy efficacy. There are numerous components and pathways involved in tumorigenesis and tumor-immune cell interactions, and the crosstalk between fibroblasts and immune cells like NK cells is still not well-defined. Fibroblasts have been documented to be a major player with respect to immune modulation and understanding this in greater detail can elucidate more options for anti-tumor immunity. Specifically, we used an extracellular matrix-based hydrogel platform to generate 3D CRC constructs to evaluate how fibroblasts affect tumor cell viability in presence and absence of NK cells. In general, fibroblasts aid the tumor cells in our models. We then identify a cytokine-based pathway (interleukin 6 [IL-6]) via which activated fibroblasts engage with the tumor cells and show how NK cell effector function can be restored when the IL-6 pathway is blocked in a melanoma-based tumor-on-a-chip platform. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=118 SRC="FIGDIR/small/660473v1_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@53797forg.highwire.dtl.DTLVardef@4dbc83org.highwire.dtl.DTLVardef@19346deorg.highwire.dtl.DTLVardef@11d834_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

Parental exposure to wet and dry conditions shapes the viability and thermotolerance of eggs in Aedes aegypti

Aedes aegypti, a primary vector of dengue, Zika, and chikungunya, displays remarkable adaptability across ecological gradients. Central to this resilience is the egg stage, which must withstand fluctuating moisture and temperature conditions. Environmental transitions, particularly changes in moisture availability, significantly influence egg hatching success in mosquitoes. This study investigates how parental exposure to variable hydration conditions shapes key reproductive traits in Ae. aegypti. Using four environmental regimes, continuous wet, continuous dry, wet-to- dry, and dry-to-wet, we assessed egg output, hatching success, thermotolerance, and egg nutrient composition across three Ae. aegypti populations. Our results show that oviposition timing and egg production are significantly affected by the hydration environment experienced by the parental generation. While the wet, dry, and dry-to-wet groups exhibited a consistent oviposition peak beginning four days post-blood feeding, the wet-to-dry group showed delayed reproductive investment, with peak egg production occurring later. Egg output was highest under continuous wet conditions and significantly reduced in the dry and wet-to-dry treatments across all populations. Interestingly, the wet-to-dry group showed significantly higher egg-thermotolerance than any other group, and this pattern was consistent across all three populations under high- temperature stress conditions (41{degrees}C and 45{degrees}C). Nutritional composition showed an increased glycogen level in eggs when parents were exposed to wet conditions before blood feeding. By integrating physiological and ecological metrics such as hatching rates and thermal stress resilience, we demonstrate how parental environments shape subsequent egg performance, highlighting adaptive responses that enable Ae. aegypti persistence under increasing climate variability.

physiology↗