GMP Cell and Gene Therapy

The aim of the project is to validate a manufacturing process, including safety-relevant quality controls, for a novel cell therapeutic to be used in the treatment of focal cartilage defects in the knee.

Developed by BioPlanta GmbH, the product is based on mesenchymal stem cells derived from the umbilical cord. This kind of cell boasts an exceptionally high level of immune tolerance, making it suitable for allogeneic therapy concepts. The immunomodulating properties of mesenchymal stem cells have an anti-inflammatory effect, activate regenerative processes, and help restore hyaline cartilage. The project's therapeutic goals are therefore to relieve pain, improve mobility and reduce symptoms of arthrosis in the knee.

The collaboration project will also establish the scientific and technical requirements for the pharmaceutical manufacture of the investigational medicinal product and for dispensing it to patients as part of a clinical trial. The product is to be classified as an advanced therapy medicinal product (ATMP) and falls under the “tissue engineered product” category.

In line with both German and European regulations, validation of the manufacturing process and the respective safety-relevant analytical methods (quality controls) are central to verifying the safe, robust and reproducible manufacture of the ATMP. This includes testing for sterility and bacterial endotoxins. A secondary aim of the project is to update the assortment list covered by Fraunhofer IZI’s existing manufacturing authorization pursuant to Section 13 of the German Medicinal Products Act (AMG).

The product has already undergone extensive testing under GLP conditions in the GLP test facility at Fraunhofer IZI with regard to potential undesirable biodistribution and tumorigenicity. 

In 2017 and 2018 respectively, the US FDA and the European Commission approved the first CAR cell therapy. This was the first time a gene therapy was approved for cancer treatment. This revolutionary treatment form is also the focus of the ROR1 CAR T research project. The chimeric antigen receptor (CAR) developed at Würzburg University Hospital recognises the ROR1 molecule, which is, e.g., expressed by cancer cells in leukaemia as well as breast and lung cancer. 

The patient’s own cells are collected using leukapheresis to produce the cell product. Afterwards, T helper cells and cytotoxic T cells are selected by magnetic cell separation. The genetic material for the CAR is transferred into the T cell genome by non-viral gene transfer with help of the “Sleeping Beauty” transposon system (jumping gene). This re-programmes the T cells so that they recognise ROR1-positive cancer cells as being “foreign” and kill these by releasing cytotoxic messenger substances. The reprogrammed cells are in vitro expanded and intravenously administered to the patient. 

This project is sponsored as a pilot project by the Proof-of-Concept Initiative, which was established by Fraunhofer Society, Helmholtz Association and the Association of German Medical Faculties to promote the translation of innovative research projects. This funding was used to prepare pre-clinical studies on the safety and effectiveness of ROR1 CAR T cells; moreover, the clinical translation into a phase I/II study (first-in-man study) is to be prepared.

In this project, test batches were initially produced. These were then used to optimise the process with regard to the demanding production under GMP conditions and qualify the required equipment. After the successful establishment of the process and the required specifications, three successful validation batches were produced under cleanroom conditions and the analytical methods were established. Moreover, the cell products generated with these validation batches were used to validate the analytical methods relevant for microbiological safety (mycoplasma, sterility, bacterial endotoxins) and to commence the validation of the proof of genomic safety (determination of vector copy number). Following the conclusion of these validations, an application for the inclusion of the investigational medicinal product in the existing production authorization according to section 13 of the German Drug Act is to be submitted to the competent authority. 

CAR-T cell therapy is a cancer immuno­therapy. It uses the patient‘s own T cells to fight certain types of cancer. To do this, the cells are harvested in the clinic by leukapheresis and genetically reprogrammed in vitro to recognize cancer cells that carry a specific antigen on the cell surface by means of a chimeric antigen receptor. After lymphodepleting chemo­therapy, the reprogrammed cells are infused into the patient, where they can proliferate and start the immune response. In August 2017, Kymriah® (CTL019 / tisagenlecleucel), the first CAR (chimeric antigen receptor) -T cell therapy, became available in the US. Kymriah® received FDA approval for children and young adults up to 25 years of age with B-cell acute lymphoblastic leukemia (ALL) who have not responded or have already relapsed to usual therapies, and in May 2018 for adult patients with diffuse large B-cell lymphoma (DLBCL) who have relapsed or not responded to therapies after two or more lines of systemic therapy. In August 2018, Novartis announced the approval of the EU Commission for these two indications following a corresponding recommendation by the European Medicines Agency (EMA). Fraunhofer IZI has been a manufacturing site for this CAR-T cell therapy in Europe for several years and in 2021 handed over the long-standing very successful collaboration with Novartis regarding the manufacture of CTL019/Kymriah® in accordance with the contract. Personnel experienced in the manufacturing and quality control of CTL019/Kymriah® are now handling the process transfer and manufacturing of novel ”next generation” CAR-T investigational products developed by Novartis. This T-Charge™ program was first presented by Novartis in December 2021 at the 63rd American Society of Hematology Annual Meeting (ASH). The T-Charge™ platform preserves T-cell stemness (the ability of T cells to self-renew and mature), an important property of T cells that is closely linked to their therapeutic potential. This results in a product with greater proliferation potential and fewer exhausted T cells. With T-Charge™, CAR T cell expansion occurs primarily in the patient‘s body (in vivo), eliminating the need for extended culture time outside the body (ex vivo). Beside those changes in the cells’ biology, the T-Charge platform will be faster and more reliable compared to traditional CAR-T technology by simplifying processes and optimizing quality control. 

Press releases and further information

The US American Food and Drug Administration FDA approved for the first time in 2017 a gene therapy with so-called CAR-T cell therapy. Clinical studies of cancer diseases show that impressive success has already been achieved in the treatment of cancer using this form of therapy. This revolutionary therapy is also the focus of a research project that the PoC initiative will be supporting with about 2.8 million euros. The chimeric antigen receptors (CAR) developed at the University Hospital of Würzburg detect a certain molecule (ROR1), which is barely present in healthy cells, but which occurs all the more on cancerous cells from leukemia, breast or lung cancer. In the case of the research project now being funded by the PoC initiative, the non-viral gene transfer takes place by means of the so-called "Sleeping Beauty" transposon system (SB100X). The funding is to be used to complete pre-clinical studies on the safety and efficacy of the ROR1 CAR-T cells and to achieve the clinical translation into a Phase I study (First-in-Man).

Thy project is funded by the Proof-of-Concept initiative. The initiative was instituted by the Helmholtz Association, the Fraunhofer-Gesellschaft and the German university medical departments, to promote translation of innovative, promising research projects into clinic.

Partners

The Department GMP Cell and Gene Therapy of Fraunhofer is collaborating with Iovance Biotherapeutics Inc. (San Carlos, CA, USA) on the technology transfer and subsequent continuous manufacturing of allogeneic feeder cells used in Iovances’ Tumor Infiltrating Lymphocyte manufacturing process. This collaboration supports Iovance’s upcoming European clinical trials for the treatment of solid tumors in metastatic melanoma and cervical carcinoma.

Iovance Biotherapeutics, Inc. is a clinical-stage biotechnology company focused on the development of cancer immunotherapy products for the treatment of various cancers. The Company's lead product candidate is an adoptive cell therapy using tumor-infiltrating lymphocyte (TIL) technology being investigated for the treatment of patients with metastatic melanoma, recurrent and/or metastatic squamous cell carcinoma of the head and neck and recurrent and metastatic or persistent cervical cancer. For more information, please visit www.iovance.com.