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Characterization of small intracellular vesicles uncovers their potential as drug carriers and therapeutic agents

ABSTRACT:

Small extracellular vesicles (sEVs) have been extensively investigated as a source of clinically relevant cargo. Alongside the secretion of EVs, such as exosomes, cells house a multitude of internal vesicles involved in various physiologic processes and metabolic activities. Herein, we developed protocols to isolate small intracellular vesicles (sIVs) from different cell types and investigated their biological characteristics and potential therapeutic applications. SIVs are smaller in size, have higher yield, and display a markedly greater uptake in vitro and in vivo than sEVs. The composition of proteins, RNAs, and lipids of sIVs differed from that of sEVs. Additionally, sIVs derived from mesenchymal stem cells outperformed sEVs in ameliorating retinal degeneration by mitigating endoplasmic reticulum stress and providing neuroprotective factors in an in vivo model. This study identified a distinct functional intracellular nanoparticle as a suitable alternative to sEVs in clinical translation

INSTRUMENT(S): Liquid Chromatography MS - negative - reverse phase, Liquid Chromatography MS - positive - reverse phase

PROVIDER: MTBLS12923 | MetaboLights | 2025-09-02

REPOSITORIES: MetaboLights

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			Action	DRS
	a_MTBLS12923_LC-MS_negative_reverse-phase_metabolite_profiling.txt	Txt
	a_MTBLS12923_LC-MS_positive_reverse-phase_metabolite_profiling.txt	Txt
	i_Investigation.txt	Txt
	m_MTBLS12923_LC-MS_negative_reverse-phase_metabolite_profiling_v2_maf.tsv	Tabular
	m_MTBLS12923_LC-MS_positive_reverse-phase_metabolite_profiling_v2_maf.tsv	Tabular

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Project description:Amyotrophic lateral sclerosis (ALS) is a terminal neurodegenerative disease. Clinical and molecular observations suggest that ALS pathology originates at a single site and spreads in an organized and prion-like manner, possibly driven by extracellular vesicles. Extracellular vesicles transfer cargo molecules associated with ALS pathogenesis, such as misfolded and aggregated proteins and dysregulated microRNAs (miRNAs). However, it is poorly understood whether altered levels of circulating extracellular vesicles or their cargo components reflect pathological signatures of the disease. In this study, we used immuno-affinity-based microfluidic technology, electron microscopy, and NanoString miRNA profiling to isolate and characterize extracellular vesicles and their miRNA cargo from frontal cortex, spinal cord, and serum of sporadic ALS (n=15) and healthy control (n=16) participants. We found larger extracellular vesicles in ALS spinal cord versus controls and smaller sized vesicles in ALS serum. However, there were no changes in the number of extracellular vesicles between cases and controls across any tissues. Characterization of extracellular vesicle-derived miRNA cargo in ALS compared to controls identified significantly altered miRNA levels in all tissues; miRNAs were reduced in ALS frontal cortex and spinal cord and increased in serum. Two miRNAs were dysregulated in all three tissues: miR-342-3p was increased in ALS, and miR-1254 was reduced in ALS. Additional miRNAs overlapping across two tissues included miR-587, miR-298, miR-4443, and miR-450a-2-3p. Predicted targets and pathways associated with the dysregulated miRNAs across the ALS tissues were associated with common biological pathways altered in neurodegeneration, including axon guidance and long-term potentiation. A predicted target of one identified miRNA (N-deacetylase and N-sulfotransferase 4; NDST4) was likewise dysregulated in an in vitro model of ALS, verifying potential biological relevance. Together, these findings demonstrate that circulating extracellular vesicle miRNA cargo mirror those of the central nervous system disease state in ALS, and thereby offer insight into possible pathogenic factors and diagnostic opportunities.

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Characterization of small intracellular vesicles uncovers their potential as drug carriers and therapeutic agents

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