Periodic Reporting for period 2 - AutoCRAT (Automated Cellular Robot-Assisted Technologies for translation of discovery-led research in Osteoarthritis)
Periodo di rendicontazione: 2021-07-01 al 2022-12-31
Osteoarthritis (OA), a degenerative disease of joints, is one of the most significant causes of disability worldwide and is associated with severe pain and loss of function. OA can have a profound impact on quality of life for those with the condition. Recent data indicates that over 651 million people worldwide are suffering from knee OA alone (A. Cui, et al 2020, doi: 10.1016/j.eclinm.2020.100587). Joint replacement surgery does not always provide a complete solution in the long term and surgery carries associated risks and expenses. Non-surgical options may involve the long-term use of non-steroidal anti-inflammatory drugs and/or the use of opioids in pain management. These treatments aim to provide pain relief and control inflammation, but they do not cure or treat the causes of the condition. Novel stem cell based therapeutics may deliver better and more efficient treatment options than the palliative options currently available.
The use of Mesenchymal stromal cells (MSCs) as a treatment for OA, has been studied for many years with early phase clinical trials showing promising results. The production of MSCs from tissues such as bone marrow, adipose tissue or umbilical cord obtained from healthy donors or patients means there is a limited, heterogeneous supply, and in the case of bone marrow and adipose tissue acquisition, an invasive procedure is required. Current manual processes to produce MSCs are labour intensive and expensive and produce inconsistent results. The promise of MSC therapies dictates the need to produce cells at scale and in a consistent and affordable manner to ensure a sustainable supply of high quality, therapeutic cells.
AutoCRAT aims to develop novel sustainable cell and cell-based therapies for OA. The project is using human induced pluripotent stem cells (hiPSC) to generate articular chondrocytes for cartilage repair (hiCHO) and human induced MSCs (hiMSC) for the prevention and treatment of established OA. In addition, the project will investigate the potential of the MSC cell secretome as a next-generation therapy, specifically exploring the therapeutic effect of extracellular vesicles (or EVs) secreted by cells. We aim to produce the therapeutics identified in the project using cost-effective, automated processes with incorporated product testing in a novel manufacturing system to expedite translation to patients.
The use of Mesenchymal stromal cells (MSCs) as a treatment for OA, has been studied for many years with early phase clinical trials showing promising results. The production of MSCs from tissues such as bone marrow, adipose tissue or umbilical cord obtained from healthy donors or patients means there is a limited, heterogeneous supply, and in the case of bone marrow and adipose tissue acquisition, an invasive procedure is required. Current manual processes to produce MSCs are labour intensive and expensive and produce inconsistent results. The promise of MSC therapies dictates the need to produce cells at scale and in a consistent and affordable manner to ensure a sustainable supply of high quality, therapeutic cells.
AutoCRAT aims to develop novel sustainable cell and cell-based therapies for OA. The project is using human induced pluripotent stem cells (hiPSC) to generate articular chondrocytes for cartilage repair (hiCHO) and human induced MSCs (hiMSC) for the prevention and treatment of established OA. In addition, the project will investigate the potential of the MSC cell secretome as a next-generation therapy, specifically exploring the therapeutic effect of extracellular vesicles (or EVs) secreted by cells. We aim to produce the therapeutics identified in the project using cost-effective, automated processes with incorporated product testing in a novel manufacturing system to expedite translation to patients.
The project has now been running for 36 months. Although we have experienced some delays due to the COVID-19 pandemic, we have made significant progress towards achieving our aims.
We have generated sustainable, reproducible, cell sources for hiMSCs and have characterized these cells, confirming similarity to primary hMSCs. Our hiMSCs have been studied (e.g. in a human cartilage explant model), demonstrating beneficial effects. In addition, results to date from studies with hiCHOs show promising therapeutic potential. EVs harvested from hiMSC from early to late passages thus far appear biologically effective in vitro.
We have successfully tested a thermosensitive injectable hydrogel for intra-articular injection in a pre-clinical OA model, demonstrating its potential use as a carrier for cells and EVs in the treatment of OA. The hydrogel has the ability to provide sustained release of therapeutic product at the cartilage site. Results from studies to date show an improvement in the regeneration of the OA cartilage, with better therapeutic effects and long-term retention of cells and EVs at the site by using a combination of hydrogel + cells or EVs, though further studies are needed.
Scalable, automatable and adaptive AutoCRAT production processes have been devised for the cultivation of hiPSCs, the differentiation of hiPSCs to hiMSCs, the culture of hiMSCs at scale and the differentiation of hiPSCs to hiCHOs. Work is ongoing to optimise protocols for the isolation of hiMSC derived hiEVs.
Our planned technology for inline assessment of therapeutic products during manufacture is well advanced. A prototype of ValitaCell’s ChemStress® Fingerprinting MSC characterisation tool is currently in production. We also continue to work on identifying a candidate probe for our planned fluorescence polarization (FP) EV quantification assay.
The build of the automated AutoCRAT platforms continues to progress well (despite COVID-19 delays) and the automated platforms will soon be ready for process implementation in the next phase of the project.
A dedicated team within AutoCRAT ensures that due regard is taken of GMP and GAMP requirements in the design of the AutoCRAT manufacturing platforms and production processes, developing appropriate GMP documentation/protocols. Work is also ongoing on the model for the economic evaluation of GMP manufacturing and in line testing using the AutoCRAT system.
Ethics requirements are a key consideration across all project activities.
Finally, we continue to disseminate our results and innovations so that our message reaches our target audiences (e.g. see https://www.autocrat.eu/ and https://twitter.com/AutoCRAT2020).
We have generated sustainable, reproducible, cell sources for hiMSCs and have characterized these cells, confirming similarity to primary hMSCs. Our hiMSCs have been studied (e.g. in a human cartilage explant model), demonstrating beneficial effects. In addition, results to date from studies with hiCHOs show promising therapeutic potential. EVs harvested from hiMSC from early to late passages thus far appear biologically effective in vitro.
We have successfully tested a thermosensitive injectable hydrogel for intra-articular injection in a pre-clinical OA model, demonstrating its potential use as a carrier for cells and EVs in the treatment of OA. The hydrogel has the ability to provide sustained release of therapeutic product at the cartilage site. Results from studies to date show an improvement in the regeneration of the OA cartilage, with better therapeutic effects and long-term retention of cells and EVs at the site by using a combination of hydrogel + cells or EVs, though further studies are needed.
Scalable, automatable and adaptive AutoCRAT production processes have been devised for the cultivation of hiPSCs, the differentiation of hiPSCs to hiMSCs, the culture of hiMSCs at scale and the differentiation of hiPSCs to hiCHOs. Work is ongoing to optimise protocols for the isolation of hiMSC derived hiEVs.
Our planned technology for inline assessment of therapeutic products during manufacture is well advanced. A prototype of ValitaCell’s ChemStress® Fingerprinting MSC characterisation tool is currently in production. We also continue to work on identifying a candidate probe for our planned fluorescence polarization (FP) EV quantification assay.
The build of the automated AutoCRAT platforms continues to progress well (despite COVID-19 delays) and the automated platforms will soon be ready for process implementation in the next phase of the project.
A dedicated team within AutoCRAT ensures that due regard is taken of GMP and GAMP requirements in the design of the AutoCRAT manufacturing platforms and production processes, developing appropriate GMP documentation/protocols. Work is also ongoing on the model for the economic evaluation of GMP manufacturing and in line testing using the AutoCRAT system.
Ethics requirements are a key consideration across all project activities.
Finally, we continue to disseminate our results and innovations so that our message reaches our target audiences (e.g. see https://www.autocrat.eu/ and https://twitter.com/AutoCRAT2020).
Delivery of new regenerative therapies to benefit patients with OA is the over-arching aim of AutoCRAT.
The expected innovations we intend to achieve include:
1. Scalable, automated production of hiMSC as a treatment for moderate to severe OA and to prevent development after joint injury.
2. Scalable, automated production of hiCHO for application as a treatment for cartilage repair.
3. Scalable, automated production of EV derived from hiMSC for treatment of OA and joint injury to prevent development of OA.
4. Assessment, development and production of cell-derived therapies.
5. A novel stem cell characterisation platform based on ChemStress® fingerprinting technology.
6. Novel methods for EV analysis.
7. The Automated Regenerative Medicine Factory (ARM-F).
In addition to the above, we are also exploring innovations through the use of a novel thermosensitive injectable hydrogel for intra-articular injection of therapeutic product and the use of a xeno-free cell culture medium (PurStem 2).
We believe that AutoCRAT innovations can ultimately lead to:
• Benefits for patients, clinicians and healthcare systems - initially for people with OA, clinicians and the communities around them, but ultimately we also hope to benefit people with other conditions, healthcare providers and healthcare systems.
• Innovation in the cell therapy industry - improving consistency and efficiency and reducing costs.
• Economic benefits - through reduced costs of cell therapy manufacturing and a reduction in the burden of OA disease (and potentially other conditions).
• Technological impacts - through optimisation of existing technologies and the development of new innovations.
• Impacts on research, knowledge and science – advancing the state of the art and researcher capacity.
We believe that our work to date in AutoCRAT has laid the necessary foundations for the ultimate achievement of the impacts outlined above.
The expected innovations we intend to achieve include:
1. Scalable, automated production of hiMSC as a treatment for moderate to severe OA and to prevent development after joint injury.
2. Scalable, automated production of hiCHO for application as a treatment for cartilage repair.
3. Scalable, automated production of EV derived from hiMSC for treatment of OA and joint injury to prevent development of OA.
4. Assessment, development and production of cell-derived therapies.
5. A novel stem cell characterisation platform based on ChemStress® fingerprinting technology.
6. Novel methods for EV analysis.
7. The Automated Regenerative Medicine Factory (ARM-F).
In addition to the above, we are also exploring innovations through the use of a novel thermosensitive injectable hydrogel for intra-articular injection of therapeutic product and the use of a xeno-free cell culture medium (PurStem 2).
We believe that AutoCRAT innovations can ultimately lead to:
• Benefits for patients, clinicians and healthcare systems - initially for people with OA, clinicians and the communities around them, but ultimately we also hope to benefit people with other conditions, healthcare providers and healthcare systems.
• Innovation in the cell therapy industry - improving consistency and efficiency and reducing costs.
• Economic benefits - through reduced costs of cell therapy manufacturing and a reduction in the burden of OA disease (and potentially other conditions).
• Technological impacts - through optimisation of existing technologies and the development of new innovations.
• Impacts on research, knowledge and science – advancing the state of the art and researcher capacity.
We believe that our work to date in AutoCRAT has laid the necessary foundations for the ultimate achievement of the impacts outlined above.