A concerted mechanism involving ACAT and SREBPs by which oxysterols deplete accessible cholesterol to restrict microbial infection.

Most of the cholesterol in the plasma membranes (PMs) of animal cells is sequestered through interactions with phospholipids and transmembrane domains of proteins. However, as cholesterol concentration rises above the PM's sequestration capacity, a new pool of cholesterol, called accessible cholesterol, emerges. The transport of accessible cholesterol between the PM and the endoplasmic reticulum (ER) is critical to maintain cholesterol homeostasis. This pathway has also been implicated in the suppression of both bacterial and viral pathogens by immunomodulatory oxysterols. Here, we describe a mechanism of depletion of accessible cholesterol from PMs by the oxysterol 25-hydroxycholesterol (25HC). We show that 25HC-mediated activation of acyl coenzyme A: cholesterol acyltransferase (ACAT) in the ER creates an imbalance in the equilibrium distribution of accessible cholesterol between the ER and PM. This imbalance triggers the rapid internalization of accessible cholesterol from the PM, and this depletion is sustained for long periods of time through 25HC-mediated suppression of SREBPs and continued activation of ACAT. In support of a physiological role for this mechanism, 25HC failed to suppress Zika virus and human coronavirus infection in ACAT-deficient cells, and Listeria monocytogenes infection in ACAT-deficient cells and mice. We propose that selective depletion of accessible PM cholesterol triggered by ACAT activation and sustained through SREBP suppression underpins the immunological activities of 25HC and a functionally related class of oxysterols.

A mechanistic relative biological effectiveness model-based biological dose optimization for charged particle radiobiology studies.

In charged particle therapy, the objective is to exploit both the physical and radiobiological advantages of charged particles to improve the therapeutic index. Use of the beam scanning technique provides the flexibility to implement biological dose optimized intensity-modulated ion therapy (IMIT). An easy-to-implement algorithm was developed in the current study to rapidly generate a uniform biological dose distribution, namely the product of physical dose and the relative biological effectiveness (RBE), within the target volume using scanned ion beams for charged particle radiobiological studies. Protons, helium ions and carbon ions were selected to demonstrate the feasibility and flexibility of our method. The general-purpose Monte Carlo simulation toolkit Geant4 was used for particle tracking and generation of physical and radiobiological data needed for later dose optimizations. The dose optimization algorithm was developed using the Python (version 3) programming language. A constant RBE-weighted dose (RWD) spread-out Bragg peak (SOBP) in a water phantom was selected as the desired target dose distribution to demonstrate the applicability of the optimization algorithm. The mechanistic repair-misrepair-fixation (RMF) model was incorporated into the Monte Carlo particle tracking to generate radiobiological parameters and was used to predict the RBE of cell survival in the iterative process of the biological dose optimization for the three selected ions. The post-optimization generated beam delivery strategy can be used in radiation biology experiments to obtain radiobiological data to further validate and improve the accuracy of the RBE model. This biological dose optimization algorithm developed for radiobiology studies could potentially be extended to implement biologically optimized IMIT plans for patients.