High-content image-based drug screen identifies a clinical compound against cell transmission of adenovirus

Abstract

14 15Human adenoviruses (HAdVs) are fatal to immune-suppressed people, but no effective 16 anti-HAdV therapy is available. Here, we present a novel image-based high-throughput 17 screening (HTS) platform, which scores the full viral replication cycle from virus entry to 18 dissemination of progeny. We analysed 1,280 small molecular compounds of the Prestwick 19Chemical Library (PCL) for interference with HAdV-C2 infection in a quadruplicate blinded 20 format, followed by robust image analyses, and hit identification. We present the entire set 21 of image-based screening data including all the images, and the image analysis and data 22 processing pipelines, as deposited at the Image Data repository (IDR) 1 , accession number 23 idr0081. We identified Nelfinavir mesylate as an inhibitor of HAdV plaque formation, in 24 agreement with the previous notion that Nelfinavir is ineffective in single round HAdV 25 infection assays. Nelfinavir has been FDA-approved for anti-retroviral therapy in humans. 26Our results underscore the power of image-based multi-round infection assays in 27 identifying viral inhibitors with clinical potential. 28 29

A high-content image-based drug screen of clinical compounds against cell transmission of adenovirus

“…The problem is exacerbated by the shortage of a suitable small animal model for HAdV disease, although Syrian Hamsters are susceptible to HAdV-C infection and give rise to Blinding. The PCL compound arrangement as dispensed by EPFL in four subset plates A -D comprising each screening set replicate 1-4 was blinded and replaced by UZH with internal identifier (Raw Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH and Processed Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH 29 , compoundIdentifier 1 to 1280). The identity of the compounds was only disclosed after the screening process and hit filtering (Raw Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH and Processed Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH 29 and Table 1, PCL_ID Prestw-1 to Prestw-1804 and compoundName).…”

“…Fig. 2 Project data structure available at IDR, accession number idr0081 33 . (a) In the data provided for download, there are three sub-folders for 1-prePlates, 2-ZPlates and 3-Screen.…”

“…The latter two were imaged on two different microscopes. We provide the parameters used for Plaque2.0 image analysis, and the code for the subsequent hit filtering in R. The data available for download at the IDR, accession number idr0081 33 , are structured as outlined in Fig. 2a.…”

“…2b. Additionally, an annotated list of the PCL compounds as well as raw and scored screening data are available on figshare 29 as.txt files.…”

“…1. Raw and scored infection phenotypes are shown for UZH and EPFL analyses (Raw Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH, Processed Plaque-2.0 infection scores of the PCL screen, imaged and analysed at UZH and Raw Plaque-2.0 infection scores of the PCL screen, imaged and analysed at EPFL, Processed Plaque-2.0 infection scores of the PCL screen, imaged and analysed at EPFL, respectively 29 ). Both dry-lab pipelines confirmed the high assay quality (Tables 2 and 3).…”