Testing the Stability of Drug Resistance on Cryopreserved, Gene-Engineered Human Induced Pluripotent Stem Cells

Abstract

Human induced pluripotent stem cells (hiPSCs) have emerged as a powerful tool for in vitro modelling of diseases with broad application in drug development or toxicology testing. These assays usually require large quantities of hiPSC, which can entail long-term storage via cryopreservation of the same cell charges. However, it is essential that cryopreservation does not oppose durable changes on the cells. In this project, we characterize one parameter of functionality of one that is well established in the field, in a different research context, an applied hiPSC line (iPS11), namely their resistance to a medium size library of chemo interventions (>160 drugs). We demonstrate that cells, before and after cryopreservation, do not change their relative overall drug response phenotypes, as defined by identification of the top 20 interventions causing dose-dependent reduction of cell growth. Importantly, also frozen cells that are exogenously enforced for stable overexpression of oncogenes myelocytomatosis (cMYC) or tumor protein 53 mutation (TP53R175H), respectively, are not changed in their relative top 20 drugs response compared to their non-frozen counterparts. Taken together, our results support iPSCs as a reliable in vitro platform for in vitro pharmacology, further raising hopes that this technology supports biomarker-associated drug development. Given the general debate on ethical and economic problems associated with the reproducibly crisis in biomedicine, our results may be of interest to a wider audience beyond stem cell research.

Progenitor cells derived from gene‐engineered human induced pluripotent stem cells as synthetic cancer cell alternatives for in vitro pharmacology

“…12,19 Moreover, we have recently shown that hiPSC-oncogene models present a functional in vitro platform suitable for long-term storage while ensuring longitudinal reproducibility for in vitro substance testing. 20 This work presents the generation, characterization, and application of isogenic hiPSCs with overexpression of different pan-cancer relevant oncogenes including tumor protein 53 (TP53) R175H, gliomaassociated oncogene 1 (GLI1), c-MYC, and epidermal growth factor receptor variant III (EGFRvIII) and their subsequent differentiation in tissue-specific progenitor cells. Given that malignant brain tumors present a disease class with clinical unmet needs, we chose to differentiate the hiPSC-oncogene models into the neural lineage (referred to as neural progenitorcells/ NPCs) as our example forapplication.…”

“…It is speculated that iPSC models of cancer, derived through synthetic approaches or reprogramming of tumor cells, may be beneficial to homogeneously depict a cell of origin of cancers 12,19 . Moreover, we have recently shown that hiPSC‐oncogene models present a functional in vitro platform suitable for long‐term storage while ensuring longitudinal reproducibility for in vitro substance testing 20 …”