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Bacterial extracellular vesicles as recyclable nutrient reservoirs

ABSTRACT: Bacterial extracellular vesicles (EVs) are known to mediate intercellular communication, virulence, and immune modulation. Here we show that bacteria can utilise EVs also as recyclable nutrient reservoirs. Using Bacillus cereus as a model organism, we demonstrate that EVs exhibit distinct dynamics depending on growth conditions: EVs produced in complex nutrient-rich media undergo time-dependent degradation, while those produced in defined nutrient-limited conditions remain stable and accumulate. We observe similar EV degradation patterns in Staphylococcus aureus. Time-resolved multi-omics profiling reveals that EVs containing the lipid sphingomyelin undergo progressive degradation. Using pharmacological inhibition, knockout mutants, and enzymatic complementation, we show that this process is driven by secreted sphingomyelinase (SMase). This enzyme contributes to degradation of sphingomyelin-containing EVs, thereby releasing their biomolecular cargo which can be used as a nutrient source. Growth assays confirm that SMase-mediated EV degradation provides a growth advantage when nutrients become depleted, thus establishing EVs as dynamic nutrient reservoirs.

INSTRUMENT(S):

ORGANISM(S): Bacillus Cereus Ah187

SUBMITTER: Christina Ludwig

LAB HEAD: Christina Ludwig

PROVIDER: PXD065751 | Pride | 2026-02-17

REPOSITORIES: Pride

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			Action	DRS
	BBM_737_P257_04_LUB_001.raw	Raw
	BBM_737_P257_04_LUB_002.raw	Raw
	BBM_737_P257_04_LUB_003.raw	Raw
	BBM_737_P257_04_LUB_007.raw	Raw
	BBM_737_P257_04_LUB_009.raw	Raw

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Project description:Mesenchymal stromal cells (MSCs) are multipotent stromal cells with regenerative, immunomodulatory, and anti-inflammatory properties, making them a promising tool in cell-based therapies. Increasing evidence suggests the therapeutic effects of MSCs are largely mediated by secreted paracrine factors and extracellular vesicles (EVs), which facilitate extracellular communication by transferring bioactive molecules such as proteins, lipids, and nucleic acids to target cells. The lipid composition of EVs is critical for their stability, uptake, and functional activity. Sphingomyelinase (SMase), an enzyme that hydrolyzes sphingomyelin into ceramide, plays a key role in EV biogenesis by influencing membrane curvature allowing for invagination and budding of multivesicular bodies (MVBs). However, the impact of SMase treatment on the characteristics and therapeutic potential of MSC-derived EVs remains largely unexplored. In this study, human MSCs were cultured with SMase for 24 hours and EVs were isolated via ultracentrifugation. EV size distribution was analyzed using nanoparticle tracking analysis (NTA), microRNA (miRNA) profile was determined using RNA-sequencing, while lipidomic and proteomic profiling were assessed using mass spectrometry. Functional assays were conducted to assess changes in bioactivity. NTA revealed a shift in EV production, with a higher vesicle concentration in SMase induced MSCs compared to untreated cells. SMase treatment significantly altered MSC-EV composition, leading to increased ceramide and glycosphingolipids. In addition, the modifications in protein and miRNA EV cargo were found to be associated with pathways involving TNF-α signaling, vesicle trafficking, wound healing, and angiogenesis. Functional assays indicated greater suppression of TNF-α in activated macrophages and greater tubular network formation in HUVECS. These findings provide new insights into distinct biophysical and biochemical changes in MSC-derived EVs following SMase stimulation, potentially enhancing their therapeutic properties and may inform strategies for optimizing future EV-based therapies.

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Bacterial extracellular vesicles as recyclable nutrient reservoirs

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