Tuberous Sclerosis Complex Axis Controls Renal Extracellular Vesicle Production and Protein Content

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2020 Mar 7

PMID 32138326

Citations 12

Authors

Fahad Zadjali

Prashant Kumar

Ying Yao

Daniel Johnson

Aristotelis Astrinidis

Peter Vogel

Kenneth W Gross

John J Bissler

Affiliations

Soon will be listed here.

Abstract

The tuberous sclerosis complex (Tsc) proteins regulate the conserved mTORC1 growth regulation pathway. We identified that loss of the gene in mouse inner medullary collecting duct (mIMCD) cells induced a greater than two-fold increase in extracellular vesicle (EV) production compared to the same cells having an intact axis. We optimized EV isolation using a well-established size exclusion chromatography method to produce high purity EVs. Electron microscopy confirmed the purity and spherical shape of EVs. Both tunable resistive pulse sensing (TRPS) and dynamic light scattering (DLS) demonstrated that the isolated EVs possessed a heterogenous size distribution. Approximately 90% of the EVs were in the 100-250 nm size range, while approximately 10% had a size greater than 250 nm. Western blot analysis using proteins isolated from the EVs revealed the cellular proteins Alix and TSG101, the transmembrane proteins CD63, CD81, and CD9, and the primary cilia Hedgehog signaling-related protein Arl13b. Proteomic analysis of EVs identified a significant difference between the -intact and -deleted cell that correlated well with the increased production. The EVs may be involved in tissue homeostasis and cause disease by overproduction and altered protein content. The EVs released by renal cyst epithelia in TSC complex may serve as a tool to discover the mechanism of TSC cystogenesis and in developing potential therapeutic strategies.

Citing Articles

Tuberous Sclerosis Complex and the kidneys: what nephrologists need to know.

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Physiologically Based Pharmacokinetic Modeling of Extracellular Vesicles.

Kumar P, Mehta D, Bissler J Biology (Basel). 2023; 12(9).

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Tuberous Sclerosis Complex cell-derived EVs have an altered protein cargo capable of regulating their microenvironment and have potential as disease biomarkers.

Bhaoighill M, Falcon-Perez J, Royo F, Tee A, Webber J, Dunlop E J Extracell Vesicles. 2023; 12(6):e12336.

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Arginine depletion attenuates renal cystogenesis in tuberous sclerosis complex model.

Amleh A, Pri Chen H, Watad L, Abramovich I, Agranovich B, Gottlieb E Cell Rep Med. 2023; 4(6):101073.

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Specific Features of Focal Cortical Dysplasia in Tuberous Sclerosis Complex.

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