Endothelial Cells Stably Infected with Recombinant Kaposi’s Sarcoma-Associated Herpesvirus Display Distinct Viscoelastic and Morphological Properties

TL;DR

This study shows that Kaposi’s sarcoma-associated herpesvirus changes the shape and stiffness of infected endothelial cells, which could help diagnose or treat the disease.

Contribution

The study identifies specific morphological and viscoelastic changes in endothelial cells infected with KSHV, suggesting potential diagnostic and therapeutic applications.

Findings

rKSHV-infected cells showed increased deformability with higher mitochondrial MSD compared to uninfected cells.

Infected cells exhibited a spindloid shape with reduced roundness and circularity.

Cytosolic viscosity increased in rKSHV-infected cells, indicated by decreased MSD power law exponents.

Abstract

Kaposi’s sarcoma-associated herpesvirus (KSHV) is a γ-herpesvirus that has a tropism for endothelial cells and leads to the development of Kaposi’s sarcoma, especially in people living with HIV. The present study aimed to quantify morphological and mechanical changes in endothelial cells after infection with KSHV to assess their potential as diagnostic and therapeutic markers.

Vascular (HuARLT2) and lymphatic endothelial cells (LEC) were infected with recombinant KSHV (rKSHV) by spinoculation, establishing stable infections (HuARLT2-rKSHV and LEC-rKSHV). Cellular changes were assessed using mitochondria-tracking microrheology and morphometric analysis.

rKSHV infection increased cellular deformability, indicated by higher mitochondrial mean squared displacement (MSD) for short lag times. Specifically, MSD at τ = 0.19 s was 49.4% and 42.2% higher in HuARLT2-rKSHV and LEC-rKSHV, respectively, compared to uninfected controls. There were 23.9% and 36.7% decreases in the MSD power law exponents for HuARLT2-rKSHV and LEC-rKSHV, respectively, indicating increased cytosolic viscosity associated with rKSHV infection. Infected cells displayed a marked spindloid phenotype with an increase in aspect ratio (29.7%) and decreases in roundness (26.1%) and circularity (25.7%) in HuARLT2-rKSHV, with similar changes observed in LEC-rKSHV.

The quantification of distinct KSHV-induced morpho-mechanical changes in endothelial cells demonstrates the potential of these changes as diagnostic markers and therapeutic targets.

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