The Society of Biomolecular Imaging and Informatics (SBI2; www.sbi2.org) is an international community for promoting high-content imaging at all stages of experimental development, technological advancement, and discovery. SBI2 is focused on providing education to interrogate biological models at the cellular level. The third annual meeting of SBI2 was held during September 12–14 at the Joseph B. Martin Conference Center at Harvard Medical School in Boston, MA. This 3-day meeting brought together scientists from the academic and industrial arenas, along with commercial vendors providing tools and reagents. One of the most important differentiating features was the full day of short courses focused on high-content screening and analysis, and the very popular and intense open forum colloquium, focusing on 3D imaging and analysis. Details of these events and the podium sessions may be found in the editorial and conference program published in the September 2016 SBI2 Special Issue of ASSAY and Drug Development Technologies (http://online.liebertpub.com/toc/adt/14/7).
This second Special Issue for 2016 includes podium and poster abstracts from the September SBI2 conference not included in the earlier issue, as well as a series of research articles on high content screening (HCS) and high content analysis (HCA) to add to those published in September. A report on the 3D imaging and analysis colloquium at the September conference, along with new articles from the SBI2 membership and wider HCS community, will follow in a Special Focus Section in the January 2017 issue of ASSAY and Drug Development Technologies. A summary of the articles in this October issue is as follows.
High-content screening generates a large amount of image and quantitative data that can be a challenge to navigate for the novice and experienced user alike. Mining of biological data can present a major roadblock in the correct interpretation of large data sets and software can be of considerable expense and require having access to an experienced bioinformatics expert. Omta
et al.
present a publication on their web-based tool for high-content data analysis, StratoMineR. They demonstrate its utility in the analysis of RNAi and compound screens in performing quality control, data reduction and exploration, as well as hit picking and clustering based on multiparameter high-content end points.
Approximately one in seven men in Western countries will develop prostate cancer, and of these, 20% will develop castration-resistant prostate cancer (CRPT) after treatment, resulting in a high-mortality rate. It is therefore important to find new treatments for these tumors. Fancher
et al.
demonstrate the use of their previously developed androgen receptor (AR)/transcriptional intermediary factor 2 (TIF2) protein–protein interaction (PPI) assay to rescreen active compounds in a disease-relevant prostate cancer cell line model. In CRPT, TIF2 levels are significantly elevated, compensating for antiandrogen treatment and making TIF2 an attractive drug target. The AR/TIF2 PPI assay used AR-RFP and TIF2-GFP, expressed using a recombinant adenovirus. PPI was measured as colocalization of AR-RFP with TIF2-GFP in the nucleolus, which is the main site for TIF2 in the cell.
Loss of apical basal polarity in epithelial cells is a key hallmark in tumor progression and is associated with processes such as cell migration, metastasis, and endothelialmesenchymal transition. The mislocalization of key proteins is a key driver for these cellular processes. Smith
et al.
have developed a high-content assay to measure the membrane localization of scribble, which is normally localized to the basolateral membrane in confluent MCF10A mammary epithelial cells. Loss of scribble membrane localization can be seen as an indicator of tumor progression, and membrane localization has been shown to block oncogenic Ras transformation. The authors describe the application of this assay to measure the colocalization of scribble with a membrane marker in a screen for a custom siRNA library. The screen had an excellent Z′ and the authors highlighted the genes IGFR1 and MYH9 as hits.
Pagliero
et al.
took a two-tiered screening approach to identify compounds regulating lysosomal cell death, an alternative cell death pathway that represents a potential therapeutic strategy. This high-throughput screen was designed to identify lysosomal cell death by measuring lysosomal membrane permeabilization, using Galectin-3 as a reporter. Targets that caused apoptotic cell death as measured by a propidium iodide cytotoxicity assay were excluded from further analysis, and compounds were then stratified through live imaging and quantitation of Gal-3 puncta. This assay was highly specific and validated by detection of cathepsin release in the cytosol, identifying known lysosomal cell death regulators in addition to novel compounds.
Together, the September and October SBI2 Special Issues comprise an exciting collection of groundbreaking research that will significantly advance progress in high-content imaging and informatics.
