Nanoscale Acoustic Oscillator for Mechanoimmunology: NAOMI
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PROVIDER: PRJNA1294740 | ENA |
REPOSITORIES: ENA
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Project description:Mechanoimmunology provides a powerful framework for understanding how mechanical forces orchestrate immune responses, offering insights into immune cell functions and the mechanisms underlying mechanotransduction. However, a major challenge in this field is the absence of reliable platforms that deliver precise and consistent mechanical stimuli to individual cells while achieving practical and reproducible mechanobiological activation of immune cells. In this work, we introduce a highly sensitive and stable nanoscale acoustic oscillator for mechanoimmunology applications: NAOMI. NAOMI features a micropatterned substrate that promotes uniform cell monolayer formation, seamlessly integrated with an acoustic transducer system capable of generating precise nanoscale oscillations (±1 nm deviation) for up to 72 h of continuous stimulation. Unlike conventional passive mechanobiological platforms that rely on static stiffness or topographical cues, NAOMI enables programmable, uniform 3D oscillations, delivering finely tuned and reproducible mechanical stimulation for diverse experimental applications. Validation studies demonstrate that NAOMI-induced oscillations significantly amplify stress intensity and cell displacement, driving robust M1 pro-inflammatory polarization in macrophages. With its high precision, stability, and tunability, NAOMI offers a powerful and versatile platform for studying mechanoimmunology and holds strong potential to support future research and enable innovative immunotherapy applications.
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Project description:Eukaryotic cells are equipped with multiple mechanosensing systems and perceive wide range of mechanical stimuli from the environment. However, cell-level responses against acoustic waves, which transmits feeble but highly frequent physical perturbation, largely remains uninvestigated, especially with regard to audible range of sound. Here we investigate the effect of acoustic stimulation on gene expression profiles of mammalian cultured cells. A direct sound emission system was set up using a vibrational transducer to directly generate acoustic waves in culture medium. A custom-made vibrating plate made of PEEK (poly ether ether ketone) plastic was used as a diaphragm. A set of sound patterns including single-frequency sound and white noise were generated by NCH Tone Generator software. Sound intensity was directly measured by recording it in water using a hydrophone and the pressure level was calculated. C2C12 myoblasts cultured in a plastic dish with approximately 50% confluency were subjected to acoustic stimulation. 440 Hz and 14k Hz single-frequency sine wave sound were selected as representatives of low and high audible frequencies, and white noise was selected as a random noise pattern. 2 and 24 hours after continuous emission of these sound at 100 Pa, total RNA was extracted and subjected to the gene expression profiling analysis by RNA-sequencing technique. Total 42 early- and 145 late-response genes were identified as sound-sensitive genes in 2 and 24 hours stimulation, respectively. Gene annotation analyses revealed that in addition to the known mechanosensitive activities such as fluid shear stress response, cell migration, cell adhesion and blood vessel development, variety of pathways and processes were identified to be affected by acoustic stimulation.
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Project description:In many organisms, the circadian clock is composed of functionally coupled morning and evening oscillators that regulate the bouts of dawn and dusk activity. In Arabidopsis, oscillator coupling relies on a core loop in which the evening oscillator component TOC1 was proposed to activate a subset of morning-expressed oscillator genes. Our systems-biological approach overturns the current view of the Arabidopsis circadian clock showing that TOC1 does not function as an activator but as a timely-controlled general repressor of morning and evening oscillator components. Repression occurs through rhythmic binding to the promoters of all oscillator genes, suggesting a previously unexpected direct connection between the morning and evening loops. Examination of TOC1 genome-wide binding using TOC1 Minigene (TMG) seedlings expressing the genomic fragment of TOC1 fused to the Yellow Fluorescent Protein in a toc1-2 mutant background (TMG-YFP/toc1-2 seedlings) grown under LD cycles (12h light:12h dark).
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Project description:Transcripts for GH, MHC Class I and II genes, and heavy- and light-chain myosins, as well as many others genes, were differentially regulated in the zebrafish inner ear following overexposure to sound. Following acoustic trauma in the zebrafish inner ear, we used microarray analysis to identify genes involved in inner ear repair following acoustic exposure by comparing the gene expression levels of 2 days and 4 days post-sound exposure (NE-ZF-2d and NE-ZF-4d, respectively) to controls without sound exposure (C-ZF).
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Project description:Proteostasis is vital for cellular health, with disruptions leading to pathologies including aging, neurodegeneration and metabolic disorders. Traditionally, proteotoxic stress responses were studied as acute reactions to various noxious factors; however, recent evidence reveals that many stress-response genes exhibit ~12-hour ultradian rhythms under physiological conditions in mammals. These rhythms, driven by an XBP1s-dependent 12h oscillator, are crucial for managing proteostasis. By exploring the chromatin landscape of the murine 12h hepatic oscillator, we identified RBBP5, a key subunit of the COMPASS complex, as an essential epigenetic regulator of proteostasis. RBBP5 is indispensable for regulating both the hepatic 12h oscillator and transcriptional response to acute proteotoxic stress, acting as a co-activator for master proteostasis transcription factor XBP1s. RBBP5 ablation leads to increased sensitivity to proteotoxic stress, chronic inflammation, and hepatic steatosis in mice, along with impaired autophagy and reduced cell survival in vitro. In humans, lower RBBP5 expression is associated with dampened adaptive stress-response gene expression and hepatic steatosis. Our findings establish RBBP5 as a critical regulator of proteostasis, essential for maintaining mammalian organismal health.
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