Transcriptional profiling of single fiber cells in a transgenic paradigm of an inherited childhood cataract reveals absence of molecular heterogeneity

• Published in: The Journal of biological chemistry 2019-09, Vol.294 (37), p.13530-13544
• Main Authors: Bhat, Suraj P., Gangalum, Rajendra K., Kim, Dongjae, Mangul, Serghei, Kashyap, Raj K., Zhou, Xinkai, Elashoff, David
• Format: Article
• Language: English
• Subjects: Animals; Animals, Genetically Modified; cataract; Cataract - genetics; Cataract - metabolism; Cell Differentiation - genetics; crystallin; crystallin gene expression; Crystallins - genetics; Crystallins - metabolism; Eye Proteins - genetics; Eye Proteins - metabolism; Female; gene expression; Gene Expression Profiling - methods; genetic cataract; Humans; lens; Lens, Crystalline - metabolism; Male; Mice; Mice, Inbred C57BL; Molecular Bases of Disease; Retina - embryology; RNA, Messenger - genetics; single cells; Single-Cell Analysis - methods; spatial transcriptomics; transcription; Transcriptome - genetics; vision
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.RA119.008853

Our recent single-cell transcriptomic analysis has demonstrated that heterogeneous transcriptional activity attends molecular transition from the nascent to terminally differentiated fiber cells in the developing mouse lens. To understand the role of transcriptional heterogeneity in terminal differentiation and the functional phenotype (transparency) of this tissue, here we present a single-cell analysis of the developing lens, in a transgenic paradigm of an inherited pathology, known as the lamellar cataract. Cataracts hinder transmission of light into the eye. Lamellar cataract is the most prevalent bilateral childhood cataract. In this disease of early infancy, initially, the opacities remain confined to a few fiber cells, thus presenting an opportunity to investigate early molecular events that lead to cataractogenesis. We used a previously established paradigm that faithfully recapitulates this disease in transgenic mice. About 500 single fiber cells, manually isolated from a 2-day-old transgenic lens were interrogated individually for the expression of all known 17 crystallins and 78 other relevant genes using a Biomark HD (Fluidigm). We find that fiber cells from spatially and developmentally discrete regions of the transgenic (cataract) lens show remarkable absence of the heterogeneity of gene expression. Importantly, the molecular variability of cortical fiber cells, the hallmark of the WT lens, is absent in the transgenic cataract, suggesting absence of specific cell-type(s). Interestingly, we find a repetitive pattern of gene activity in progressive states of differentiation in the transgenic lens. This molecular dysfunction portends pathology much before the physical manifestations of the disease.