Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening.

Human brain cells generated by in vitro cell programming provide exciting prospects for disease modeling, drug discovery and cell therapy. These applications frequently require efficient and clinically compliant tools for genetic modification of the cells. Recombinant adeno-associated viruses (AAVs) fulfill these prerequisites for a number of reasons, including the availability of a myriad of AAV capsid variants with distinct cell type specificity (also called tropism). Here, we harnessed a customizable parallel screening approach to assess a panel of natural or synthetic AAV capsid variants for their efficacy in lineage-related human neural cell types. We identified common lead candidates suited for the transduction of directly converted, early-stage induced neural stem cells (iNSCs), induced pluripotent stem cell (iPSC)-derived later-stage, radial glia-like neural progenitors, as well as differentiated astrocytic and mixed neuroglial cultures. We then selected a subset of these candidates for functional validation in iNSCs and iPSC-derived astrocytes, using shRNA-induced downregulation of the citrate transporter SLC25A1 and overexpression of the transcription factor NGN2 for proofs-of-concept. Our study provides a comparative overview of the susceptibility of different human cell programming-derived brain cell types to AAV transduction and a critical discussion of the assets and limitations of this specific AAV capsid screening approach.

Where contrast agent concentration really matters - a comparison of CT and MRI.

OBJECTIVES: First pass contrast-enhanced magnetic resonance imaging (MRI) and computed tomography (CT) are influenced by parameters that characterize the injected bolus. The aim of this study was to assess the role of contrast agent concentration and the differences between MRI and CT. MATERIAL AND METHODS: We systematically evaluated the published literature to define the differences between MRI and CT with regards to the influence of contrast agent concentration and flow rate on signal enhancement and image quality. Subsequently, we used a simulation model to simulate bolus dispersion in the human body for contrast agents with different concentration. We performed this simulation for different injection times (3-25 seconds) as well as for single and double contrast agent dose, and calculated the effect of contrast agent concentration and dose on the increase of local contrast agent concentration. RESULTS: Although CT studies have shown that even a moderate increase in contrast agent concentration leads to higher peak concentration in the tissue or artery of interest, MRI studies have failed to show a marked benefit of higher concentration. The simulation demonstrated that the use of high concentrated contrast agent leads to an increase in local contrast agent concentration within the tissue or artery of interest, only if injection time is long (in CT commonly >10 seconds) compared with the time constant of bolus dispersion (about 5 seconds in humans). If the injection time is shorter (in MRI commonly 1-4 seconds), the local contrast agent concentration is mainly affected by the injected dose. CONCLUSION: Contrast agent concentration is a key parameter for the optimization of dynamic imaging techniques such as angiography or perfusion in CT, whereas in dynamic MRI, contrast agent dose and relaxivities are the leading parameters.