RT Journal Article
SR Electronic
T1 An Approach to Measuring Protein Turnover in Human Induced Pluripotent Stem Cell Organoids by Mass Spectrometry
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.09.30.462679
DO 10.1101/2021.09.30.462679
A1 Anthony Duchesne
A1 Jing Dong
A1 Andrew N. Bayne
A1 Nguyen-Vi Mohamed
A1 Wei Yi
A1 Meghna Mathur
A1 Edward A. Fon
A1 Thomas M. Durcan
A1 Jean-François Trempe
YR 2021
UL http://biorxiv.org/content/early/2021/10/01/2021.09.30.462679.abstract
AB Patient-derived organoids from induced pluripotent stem cells have emerged as a model for studying human diseases beyond conventional two-dimensional (2D) cell culture. Briefly, these three-dimensional organoids are highly complex, capable of self-organizing, recapitulate cellular architecture, and have the potential to model diseases in complex organs, such as the brain. For example, the hallmark of Parkinson’s disease - proteostatic dysfunction leading to the selective death of neurons in the substantia nigra - present a subtle distinction in cell type specificity that is simply lost in 2D cell culture models. As such, the development of robust methods to study global proteostasis and protein turnover in organoids will remain a critical need as organoid models evolve. To solve this problem, we have designed a workflow to extract proteins from organoids and measure global protein turnover using mass spectrometry and stable isotope labeling using amino acids in cell culture (SILAC). This allowed us to measure the turnover rates of 844 proteins and compare protein turnover to previously reported data in primary cell cultures and in vivo models. Taken together, this method will facilitate the study of proteostasis in organoid models of human disease and will provide an analytical and statistical framework to measure protein turnover in organoids of all cell types.Competing Interest StatementJ.-F.T. is a member of the scientific advisory board of Mitokinin Inc.