Biology subjects

Storm, J. A.

Publications and source records attributed to Storm, J. A..

4 recordsLinked to original sources

bioRxiv2025

Milk-derived extracellular vesicles mitigate NF-κB pathway and NLRP3 inflammasome formation in Long Evans neonates.

Exposure to a maternal high fat diet (HFD) during perinatal (prenatal and postnatal combined) life increases offsprings risk of developing metabolic diseases (obesity, type II diabetes and hypertension), impairs immunity, behaviour, and neurodevelopment. Exclusive breast/chest milk feeding is a potential solution to reduce the negative developmental effects of HFD, mainly chronic systemic pro-inflammation. This study focuses on analyzing anti-inflammatory effects of a group of biological nanovesicles found in human milk, entitled milk-derived extracellular vesicles (MEVs). Specifically, we characterized the modulation of the nuclear factor {kappa}B (NF-{kappa}B) signaling pathway and NLR family pyrin domain containing 3 (NLRP3) inflammasome formation by MEVs in male and female neonatal rats with perinatal HFD exposure in the liver and hypothalamus. Female Long Evans dams were placed on a HFD or a control diet (CHD), with matching sucrose levels, 4 weeks before breeding and remained on the diets through gestation and lactation. HFD and CHD offspring received human MEVs through oral gavage twice a day from postnatal day (PND) 4 to 11. Transcript and protein abundance of candidate targets in the NF-{kappa}B signaling pathway and NLRP3 inflammasome were measured by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and western immunoblotting, respectively. Our results indicate that MEV treatment attenuates the activation of NF-{kappa}B pathway and NLRP3 inflammasome formation at critical checkpoints, in males and females with perinatal HFD exposure in liver and the hypothalamus. Taken together, our data suggests that MEVs may elicit anti-inflammatory benefits postnatally that mitigates gestational HFD exposure.

developmental biology↗

bioRxiv2025

Human milk-derived extracellular vesicle treatment promotes the heat shock response in neonates with perinatal high fat diet exposure.

Maternal consumption of a high-fat diet (mHFD) during perinatal life (the collective prenatal and postnatal periods) influences neonatal development, initiates hypothalamic-pituitary-adrenal (HPA) axis activation, and impacts the long-term physiological and metabolic health of offspring. Milk-derived extracellular vesicles (MEVs) are lipid-coated nanovesicles found in mammalian milk that survive intestinal degradation and cross complex biological barriers, including the blood-brain barrier. MEVs have known cytoprotective activity in peripheral organs; however, their pro-survival functions in response to chronic pro-inflammation stemming from early life nutrient stress remain unknown in the neonatal brain. Further, sex differences resulting from MEV treatment require investigation, as male and female neonates illicit variable responses to early life nutrient stress. We investigated whether MEVs promote the heat shock response (HSR), a principal pro-survival mechanism responsible for refolding or degrading misfolded protein aggregates through the action of heat shock protein (HSP) chaperones. We investigated the interaction between MEVs and the HSR in the liver, hypothalamus, and prefrontal cortex in male and female neonatal rats exposed to perinatal mHFD within the stress hyporesponsive period at postnatal day 11. MEV treatment robustly modulated the HSR in female neonates with the largest response recorded in the prefrontal cortex. Specifically, in the prefrontal cortex, MEV treatment led to an upregulation of the main transcription factor (HSF1), while downregulating the negative regulators of HSF1 (Hsp70 and Hsp90). These results suggest that MEVs may influence pro-survival outcomes in the prefrontal cortex by activating HSF1-mediated pro-survival in a sex specific manner in response to mHFD.

molecular biology↗

bioRxiv2025

Human milk-derived extracellular vesicles promote the heat shock response in polarized microglia.

Proteotoxic stress induces microglia polarization and attenuates cytoprotective, pro-survival cellular cascades. Milk-derived extracellular vesicles (MEVs) are lipid-coated nanovesicles that combat pro-inflammation in peripheral cells and tissues; however, the cytoprotective potential of MEVs remains unknown in brain macrophages. We investigated whether MEVs reduce neuroinflammation in human microglia by activating the heat shock response (HSR). The HSR triggers the upregulation of molecular chaperones (heat shock proteins; HSPs) to restore proteostasis by refolding or degrading misfolded aggregates. MEVs were isolated from unpasteurized human donor milk. Human microglia clone 3 (HMC3) cells were primed with 10 ng/mL IFN-{gamma} to induce polarization, and a subset of cells were supplemented with 200 {micro}g of MEVs. The abundance of HSF1 and candidate HSPs (Hsp70, Hsp90, Hsp40, Hsp27) were analyzed via RT-qPCR and western immunoblotting at 6h, 12h, and 24h post-MEV supplementation. We found that MEV supplementation promoted the HSR in polarized microglia, compared to homeostatic cells. Furthermore, MEVs increased the duration of the HSR in response to pro-inflammatory stress, exerting robust and continued pro-survival benefits.

molecular biology↗

bioRxiv2024

The glia club: Validation of polarization biomarkers for human microglia (HMC3) using quantitative real time RT-qPCR.

Microglia are the primary immune cells of the brain and play critical roles in neurodevelopment, neuroprotection, the maintenance of homeostasis, and neurotoxicity. Classification of microglia polarization, however, remains contentious. Identifying suitable biomarkers for gene expression analysis of microglia polarization is crucial for characterizing the biological significance of microglia in health and disease. In this study, we use human microglia clone 3 (HMC3) cells to validate and test suitable internal controls (GAPDH, PKM, 18S, ACTB, PGK1, TKT1, TPI1), homeostatic (CD68, TGF-{beta}, IBA1, BIN1, RGS10), proinflammatory (IL-6, CXCL10, CCL5, SAA, IL-1{beta}) and anti-inflammatory (CCL2, SOCS3, IL-10, CD200R1, ARG1) markers along with their transcriptional profiles upon interferon-gamma (IFN-{gamma}) stimulation to characterize the microglia polarization spectrum. Our study is the first to present a comprehensive list of biomarkers with detailed methodology on gene selection, primer design, RT-qPCR parameters, and transcript abundance in baseline and polarized HMC3 cells. Our study will increase the rigor of gene expression analysis and target selection in a widely used brain macrophage.

cell biology↗