The Kuopio Center for Gene and Cell Therapy is home to the research and development of new innovative medicines.
The Kuopio Center for Gene and Cell Therapy (KCT) is a significant, international, newly established research organisation in Kuopio, Finland, focused on developing novel gene and cell therapy products in the class known as advanced therapy medicinal products (ATMPs).
The centre’s niche is in the interface between academic, basic research and the commercial biotechnology industry. KCT provides an exciting environment for applied research of ATMPs, discovering, establishing and developing them through Phase I clinical trials and utilising some of the most advanced analytical and mechanistic proof-of-principle approaches in the sector.
About the Kuopio Center for Gene and Cell Therapy
The driving forces and founders of the Kuopio Center for Gene and Cell Therapy, Professor Seppo Ylä-Herttuala and Dr Nigel Parker, are both acknowledged experts and pioneers in their respective areas of gene and cell therapy. Their teams have been responsible for many notable ‘firsts’ in the ATMP sector, and they have founded well-established companies such as the leading ATMP manufacturer FinVector Oy and gene therapy developer FKD Therapies Oy, located in the same research campus in Kuopio.
The Kuopio Center for Gene and Cell Therapy’s laboratories provide modern facilities for state-of-the-art molecular biology, gene and cell therapy as well as for the extensive range of advanced analytics now needed in molecular medicine. Current programmes are mostly at the discovery stage, are basic science or are moving through early translational stages with a view to entering clinical development. Translational-stage work encompasses manufacturing process development, analytical development, mechanistic proof of principle and preclinical animal studies. Currently, KCT employs 25 people forming experienced gene and cell therapy scientific teams. In the next 2-3 years, KCT expects to have 40-50 employees and has already started to expand its operations. KCT has a unique location in the Kuopio Campus, which is strongly focused on health science. It has established working links with the A.I. Virtanen Institute for Molecular Sciences, the University of Eastern Finland (UEF), the National Laboratory Animal Center, and Kuopio University Hospital’s (KUH) Gene Therapy Unit. These are all situated on the same campus, the capability of which to undertake ATMP research is made complete by the presence of the office of the Finnish Medicines Agency (Fimea).
Close collaborations with academic groups in Finland and abroad are highly valued in the Kuopio Center for Gene and Cell Therapy, accelerating the progress of science and extending the repertoire of knowledge and partners. KCT offers a unique research environment to foster new ideas and scientific breakthroughs and move them into clinical development. Educational practices include opportunities for applied research experience for master’s thesis students and staff involvement in higher education teaching.
Successful development of gene and cell therapy is uniquely complex, and each product needs its own strategy from discovery to market. Roadblocks such as manufacturing and regulatory requirements are well-known major hurdles to progress with ATMPs, and the KCT team is one of the longest established in the sector, with extensive experience of all stages of ATMP development. Its exclusive access to FinVector’s GMP (good manufacturing practice) manufacturing and regulatory capability can rapidly move novel science through these difficult stages and into human trials. In ATMPs the ‘manufacturing is the product’.
FinVector has four GMP production suites authorised by the European Medicines Agency (EMA) for the production of gene therapy products from experimental through all stages of clinical trials and for commercial supply. It is crucial to properly record all such research work from the beginning to ensure the smooth transfer of the project to the GMP and clinical trials world. FinVector’s quality teams support KCT in the required documentation.
Current status in the field of ATMPs
During the last couple of years, gene and cell therapy has achieved many breakthroughs. The Food and Drug Administration (FDA) has given marketing authorisation for two T-cell products. These products are made from the patient’s own white blood cells, which are modified in a laboratory. Kymriah is a treatment for B-cell precursor acute lymphoblastic leukaemia where the patient’s T-cells are transduced with lentivirus. For non-Hodgkin lymphoma (Yescarta), patients’ T-cells are transduced with γ-retrovirus. On the other hand, Luxturna is an adeno-associated viral-based treatment for RPE65 mutation-associated retinal dystrophy, which is an inherited disease of the eye.
Also, a number of cord blood products have been approved for use in unrelated donor hematopoietic progenitor cell transplantation in patients with disorders affecting the hematopoietic system. Cell therapy products for cartilage defects of the knee can be treated with a scaffold product containing cultured chondrocytes. Provenge is a prostate cancer treatment based on prostate antigen and growth factor-induced peripheral blood mononuclear cells. This product has been withdrawn from the European market but is still available in the USA. In Europe, Strimvelis reached markets for treating inherited immune deficiency ADA-SCID with lentiviral vector modified autologous CD34+ cells. The first allogenic stem cell treatment to achieve marketing approval in Europe was Alofisel to treat perianal fistulas in Crohn’s disease. Also, a number of positive results from clinical trials have received significant attention in devastating disorders such as epidermolysis bullosa, haemophilia and spinal muscular atrophy (SMA).
Despite these success stories and increasing numbers of clinical trials, there is still much work to be done before ATMPs become routine medicines. The biological complexity of these products, manufacturing scale up and regulatory issues remain examples of the challenges faced for their development. Today, every product in the market is the result of decades of pioneering research, development and investment.
KCT’s charter is to discover and develop ATMPs into Phase 1 clinical trials. The combination of high-level science, the latest technologies and the specialist capabilities of experienced staff is the key to KCT and, as such, KCT looks forward to an exciting future in the ATMP sector.
Kuopio Center for Gene and Cell Therapy carries out top-level basic and translational research. Professional teams and laboratories with cutting-edge equipment provide scientific knowhow and modern technologies to develop advanced therapies. Science is undertaken in collaboration with academic groups around the world. The link with FinVector’s GMP manufacturing, quality and regulatory teams ensures a fast product path from research to patients.
KCT Gene Therapy Unit
The Kuopio Center for Gene and Cell Therapy is functionally divided into a Gene Therapy Unit (GTU), a Cell Therapy Unit (CTU) and the Analytics Core Unit (ACU). Today, the GTU’s research is focusing on the most commonly used gene therapy vectors: adenoviral, AAV and lentiviral vectors. Over the years, adenoviral vectors have remained the leading vectors in the number of clinical trials conducted. Adenoviruses are efficient in transducing dividing and non-dividing cells, capable of being produced at large scale, with short-term transgene expression, and the virus is immunogenic. Adenoviruses have proven to be excellent in diseases where short-term therapeutic protein expression is required, e.g. for cancer treatment and to induce angiogenesis. Lentiviral vectors and AAVs are promising tools for long-lasting gene therapies. Lentiviral vectors have been extensively used in ex vivo approaches. Non-pathogenic AAV has shown its efficacy especially in the eye, brain and spinal cord. Non-integrating lentiviral vectors are an option for short-term therapy with lower immune response compared to adenovirus. KCT’s vector development programmes focus on improving the current vectors to be safer, regulated and more efficient, which ultimately will relate to the quality of the final product.
The manufacturing of viral vectors is key when heading towards clinical trials. The preclinical and toxicological studies should be conducted using the same vector that will be used in the later clinical trials. Thus, there is intense pressure for manufacturing process development in the early stage of the product lifecycle. The Kuopio Center for Gene and Cell Therapy founds its translational research on supporting the early manufacturing process development. Two main platforms developed are an adherent production system utilising customised iCELLis technology from Pall and a serum-free suspension platform in disposable stirred tank bioreactors. In upstream process development, the aim is to determine the main process parameters for production, for example where cell growth characteristics, feeding strategies, mixing, infection/transduction conditions or harvest are optimised. The downstream process usually incorporates filtration and chromatographic technologies for clarification, capture, polishing and buffer exchange. During the downstream process development, the product-specific conditions such as filters, columns, membranes, buffers, flow rate and pressures are established. The focus is to define a process which is scalable, disposable and transferable to GMP manufacturing. The last step is to find a final formulation buffer to support the product’s stability during the storage and which is suitable for final administration to the patient and commercial supply.
KCT Cell Therapy Unit
Treatment of diseases with living cells is called cell therapy (or cellular therapy). Following EU regulation, there is a division based on the level of manipulations of the cells, e.g. minimally manipulated transplants and transfusions have different legislation than substantially manipulated cells, which are regulated as medicines. Most of the current cell therapies are experimental, using patients’ own cells (autologous) or donor cells (allogenic) as starting material to treat a number of diseases including cancer, neurological and cardiovascular indications.
A specific challenge for cell therapy medicines is their aseptic manufacturing since the end-product cannot be sterilised. Therefore, the cell therapy field is now developing towards closed, disposable and automated manufacturing processes, these being KCT’s focuses in the process development area. Advances in closed and disposable technologies will allow production in clean room areas, which are more economic than needed for processes with open steps. Additionally, logistics become easily complicated as the starting material is often from the patients or from a healthy donor, and has to be transported to facilities for production and the manufactured product taken back to the patient for administration.
Often cell therapy products cannot be cryopreserved, implying the injection to the patient within 48 hours. In the future, the traditional centralised manufacturing of cell therapy products could be amended with production proximal to the patients. Such an option needs early translational development to meet the requirements of this decentralised model. In contrast to conventional medicines or even to other ATMPs not including living cells, i.e. gene therapy vectors, any change in the manufacturing process of a cell therapy product may result in a significantly different phenotype and characteristics. Therefore, controlled manufacturing and phenotypic characterisation linked to cell functionality are investigated at KCT at the very early stage of developing any cell therapy product.
Currently, there are three active projects on cell therapy at the Kuopio Center for Gene and Cell Therapy. The product in translational phase is differentiated from the donor’s mononuclear cells using a designed combination of cytokines and growth factors. The manufacturing process is developed from an operator-dependent manual process to a semi-automated process utilising disposable elements with minimal manipulations and manufacturing time. Furthermore, the product is characterised using the latest high-throughput technologies alongside traditional methods for characterisation and functionality. KCT is developing specialised viral vectors providing a unique environment for ex vivo cell editing. Currently, in two projects, integrating and non-integrating viral vectors are used to carry desired genes into cells to improve their cancer targeting and anti-inflammatory potential. Additionally, key research is undertaken to define sub-populations of cells likely to be optimal for ex vivo gene transfer inherently having anti-cancer and anti-inflammatory potential.
Kuopio Center for Gene and Cell Therapy:
• Modern research laboratory facilities for gene and cell therapy;
• Excellent location in Kuopio University campus focused on health sciences;
• Experienced and professional personnel;
• Cutting-edge technologies;
• Possibility to advance clinical studies;
• Track record on regulatory knowhow in the EU and USA; and
• Access to the University of Eastern Finland’s extensive in vivo facilities with up to porcine capability.
KCT Analytical Core Unit
Only relevant and precise analytics can show the critical quality attributes of the products and ensure refinement of the exact conditions during the development. Analytics finally confirm that the manufacturing process is robust, and it must also be cost-effective. Gene and cell therapy products for clinical trials are tested for safety, identity, quality and purity using an extensive battery of qualified or validated assays. In the research phase, typically remarkable broader ranges of analytics are performed to deepen the understanding of the product. The function of KCT’s Analytical Core Unit is to support daily routine-level analytics but also to develop increasingly precise new assay approaches.
For viral vectors, functional viral particles have traditionally been analysed/titered with a cell-based assay analysing transduced cells by flow cytometer or qPCR. Empty/non-functional particles of viral vectors have been analysed by techniques like enzyme-linked immunosorbent assays (ELISA), OD260, Western blotting, qPCR or high-performance liquid chromatography (HPLC). Alongside these traditional techniques, KCT employs novel innovative and high-throughput technologies such as Droplet Digital PCR (ddPCR), enabling the precise quantification of the template based on a water-oil emulsion droplet without the need for additional reference standards. The Kuopio Center for Gene and Cell Therapy is developing many new viral assays via ddPCR. In addition to viral tittering, in-process analytics are needed to follow the clearance of unwanted components in the product preparation or product-related issues such as virus integrity, empty particles, viral aggregation or morphology.
Traditional transmission electron microscopy (TEM) provides high-resolution images of the viruses but requires time-consuming operations, special laboratory facilities and time-consuming interpretation of the pictures. Vironova has recently brought onto the market an advanced MiniTEM system. It has been designed to quickly detect small particles, like viruses, with high-resolution low-voltage electron optics and with automated image acquisition and particle detection to provide high-resolution images with reliable quantitative data. In collaboration with FinVector, KCT has been developing AAV and adenovirus assays for viral particle quantification, virus aggregation, empty viral particle analysis and removal of impurities.
Cell characterisation by expression of cell-specific markers is routinely analysed using qPCR, ddPCR and FACS flow cytometry. KCT’s FACS core offers the ability to sort single cells for further expansion and analysis. Automated cell culture and analysis systems as well as fluorescent microscopy are essential in cell kinetics and imaging studies, and high-throughput assays, including multiplex detection of growth factors and cytokines, are informative when studying the activation status of the cells. These are all illustrative of what the Kuopio Center for Gene and Cell Therapy maintains in its ACU. Today, the so-called ‘next-generation’ high-throughput sequencing is a cutting-edge technique revolutionising the study of genomes. This technology is powerful for genomics, epigenomics and transcriptomics, as is viral genome sequencing to study genetic diversity of viruses, integration sites and defining adventitious viruses from the vector preparation.
KCT utilises next-generation sequencing combined with proteomics in the early product characterisation and bases on determining the possible mode of action of gene and cell therapy products. Bioinformatics in KCT is essential in analysing and integrating the large datasets from several sources, and active interaction with the analysing scientists is essential to understand the biological importance of the results.
As part of establishing that early proof of principle, mechanistic methodologies and relevant, predictive disease models are essential. KCT works closely with UEF and its comprehensive Lab Animal Centre to undertake in vivo testing. The proximity of such infrastructure provides an extensive expert capability for testing the products in development. UEF has capabilities in everything from mice, rats and rabbits to large pigs, as well as an extensive range of disease models in many therapeutic areas. This animal work is supported by high-technology imaging systems such as angiography, PET or hyperpolarised magnetic resonance imaging (MRI) in a 9.4T magnet.
Kuopio Center for Gene and Cell Therapy facilitates long-term, top-level research at the interface of the academic and commercial biotechnology sectors and aims to attract high level scientific projects and leading scientists with their teams from around the world to work on projects that can lead to significant discoveries and introduction of novel cell or gene therapy drugs into the markets.
Searching for new projects
The Kuopio Center for Gene and Cell Therapy is currently searching for novel scientific projects on cell and gene therapy. We aim to attract the best international scientists with an innovative basic or translational project requiring further development. The applicants are supported during the project proposal preparation to align with KCT’s overall strategic goals. Accepted projects are granted adequate funding, modern facilities with cutting-edge technology and an experienced and professional scientific team, and opportunities for collaboration with the University of Eastern Finland, manufacturing via FinVector, and clinical trials at Kuopio University Hospital. Within the same campus in Kuopio, a unique cluster of expertise comes together, allowing programmes to be taken from discovery into clinical trials.
• Is located in northern Europe;
• Is home to a population of 5.5 million;
• Has been deemed the happiest country in the world (UN Report);
• Boasts some of the best English-speaking skills in the world;
• Is home to thousands of lakes and beautiful green Nature;
• Is headed by the capital Helsinki, which is the best mid-sized European region for international future investments (Financial Times fDi Intelligence); and
• Is home to Kuopio, the ninth biggest city in Finland. Kuopio has been named the best place to live in Finland according to the EPSI Rating and is a forerunner in wellbeing, environmental and healthcare issues.
Tuija Kekarainen and Hanna Lesch
KCT Kuopio Center for Gene and Cell Therapy
This is a commercial article that will appear in Health Europa Quarterly issue 6, which will be published in August, 2018.