Whitepapers

Effective drug discovery and development rely heavily on the availability of predictive preclinical models. For decades, target-based drug discovery has focused on immortalized cells to identify and optimize inhibitory or activating molecules. Testing on more complex biological systems takes place only during late stages in the drug development pipeline. Bringing the most relevant biology into the pipeline earlier would help to mitigate late-stage failures due to safety or efficacy concerns. However, complex biological systems are rarely available in the scale required for high throughput screening. Recent developments in human induced pluripotent stem cell (hiPSC) technologies hold great promise to overcome these limitations.

hiPSCs retain patient-specific genetic background information, differentiate into functional cell types, and closely mimic human pathophysiology. Ncardia has developed a system for scaled expansion and differentiation of hiPSCs, generating large batches of cells that can be cryopreserved until use. This enables the same batch of hiPSC-derived cells to be used for both hit identification and lead optimization.

Successful drug efficacy screening and validation studies require not only a physiologically relevant cell model, but also validated high throughput screening (HTS) protocols to test the effects of drug candidates. To achieve this, Ncardia has automated its cell culture processes in a 384-well microplate format, as well as its assay readouts and data handling.

This article outlines how the Fluent workstation was used to automate cell culture for Ncardia’s beating, hiPSC-derived cardiomyocytes. It also describes the use of these cultured cells to screen >3,500 small molecules in a chemically induced hypertrophy disease model, using a validated phenotypic assay.

Effective drug discovery and development relies in large part on the availability of predictive preclinical model systems. Application of human cellular models from tissues which are difficult to access, such as cardiomyocytes and neurons, is still challenging. Technologies based on human induced pluripotent stem cells (hiPSC) hold great promise to overcome this challenge. Their routine use in industrial drug research requires a constant supply of stem cell-derived cells of consistent high quality.

Researchers from Ncardia developed a bioprocess for the large-scale manufacturing of cardiomyocytes derived from a variety of healthy and diseased hiPSC lines for implementation into their integrated Drug Discovery platform. They expanded hiPSCs as cell aggregates, in a DASbox® Mini Bioreactor System. Aggregate size, hiPSCmarker expression during the expansion phase, and differentiation to the desired cell type were reproducible in three batches.

In a proof of concept study, the researchers scaled-up the process using a BioFlo® 320 bioprocess control system. The cells retained key iPSC markers during the expansion phase, providing confidence that stirred-tank bioreactors are suitable to scale-up the production of hiPSC-derived cellular models. The production of several billion cardiomyocytes per batch will enable the screening of large libraries of compounds against phenotypic cellular models representing human biology.

Chronic neuroinflammation, a feature common to neurodegenerative diseases such as Alzheimer’s Disease, Parkinson’s Disease, and Amyotrophic Lateral Sclerosis, may be responsive to therapeutic intervention. Recent research has identified glial cells as mediators of neurodegeneration, driving disease onset and progression.

The aim of neuroscience research in this area is to acquire an in-depth understanding of the chronological events that lead to neuroinflammation, in order to promote the neuroprotective pathways.

To overcome the limitations of primary or animal-derived cellular models regarding translatability, reproducibility and availability, Ncardia has developed fully functional hiPSC-derived (disease) models and phenotypic assays for drug discovery. An example is their neuroinflammation platform which is based on combining Ncyte Astrocytes (hiPSC-derived astrocytes) and HTRF® technology.

This collaborative study demonstrates that Ncyte Astrocytes combined with HTRF cellular assays represent a straightforward and physiologically relevant solution for the study of neuroinflammation pathways that are altered in various neurodegenerative disorders.