In this interview, Marianne Henry, Senior Scientist at the Cell and Gene Therapy Catapult, highlights one of our Challenge Theme research projects involving the identification of a potential marker to help monitor haematopoietic progenitor cell differentiation.
1. Please provide an overview of the work presented in this poster.
The cell and gene therapy industry increasingly needs non-destructive analytical tools for precise monitoring and characterisation of closed processes. The cell secretome, the set of molecules secreted into the extracellular space, includes microRNAs (miRNAs) that are crucial in regulating gene expression during cellular development. These molecules can be easily sampled during a manufacturing process and novel analytical methods applied to enhance process understanding.
To address the need for novel analytical tools to characterise our processes, we employed small RNA sequencing to analyse the cell secretome miRNAs and identified a number of novel markers that could potentially provide insights into the haematopoietic differentiation process.
2. Why is in-process monitoring important in cell therapies research?
To address the issue of scale, new bioreactors are continuously being developed and used by biotechnologists. These help to improve the efficiency of cell therapy manufacturing processes with the aim of delivering these life-changing therapies more rapidly to patients. However, there is still an industry bottleneck due to the lack of analytical tools available to characterise cells in closed bioreactor systems. In-process monitoring can help assist in improving our understanding of the cells that are within the bioreactor. This valuable information can help to reduce the burden on end-product testing and help to ensure product quality is maintained as the process progresses.
3. What are some of the challenges associated with monitoring and controlling the differentiation of stem cells to haematopoietic cells?
One of the main challenges we are facing in the analytical space for haematopoietic progenitor cell differentiation is selecting the correct analytical tools to effectively monitor the cells during the manufacturing process. In cell and gene therapy manufacturing regulations, we need to define the critical quality attributes (CQAs) early in the development stage and identify methods that can measure these. These methods are often lengthy, requiring complex processing, and monitoring of continuously changing cell phenotypes and the relation to final product quality is challenging as these intermediate states are poorly characterised.
4. Improving the scalability and control of allogeneic cell therapies is one of our Challenge Themes - how does this project help us address this challenge?
Sampling the cell secretome for specific biomarkers allows us to monitor the process without disturbing the cells. This will aid in the development of more processes that can utilise closed system bioreactors as it will be possible to more accurately monitor the state of the cells inside. Measurements made from the culture medium are also more amenable to various process analytical technologies (PATs), thereby providing a solution for more automated measurement and data analysis for process monitoring and control.
5. What have been the learnings from this project for CGT Catapult, and how can these be applied to the wider advanced therapies industry?
We have successfully screened our exemplar differentiation process using RNA sequencing on the cell secretome and identified a number of potential markers we can monitor during haematopoietic progenitor cell differentiation. In doing so, we have demonstrated the utility of monitoring miRNA in cell therapy processes, establishing a new tool for use in the research of these products.
6. What are your plans to build on this work to continue increasing the scalability and control of stem cell development processes?
We will continue monitoring the biomarkers we have identified in different bioprocessing scenarios and explore other analytical technologies that can measure the gene expression changes in the cell secretome. We will also employ these learnings in projects with our collaborators to investigate the potential of miRNA’s to monitor their stem cell differentiation processes in scalable bioreactor systems.