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. 

Cancer is the second most common cause of death in Germany. For years now, the number of new cancer diagnoses has seen a steady rise. There continues to be an enormous need for new therapeutic options to treat all kinds of cancer. In 2017 and 2018 respectively, the US Food and Drug Administration (FDA) and the European Commission approved the first ever form of cell and gene therapy for the treatment of cancer – CAR-T cell therapy. CAR stands for chimeric antigen receptor, which is expressed through a genetic modification on the T cells. The receptor identifies and binds specific antigens on cancer cells, subsequently activating a targeted immune response. 

This revolutionary type of therapy also lies at the heart of the ROR-1 CAR-T research project. ROR-1 is a tyrosine-protein kinase transmembrane receptor, which is strongly expressed during embryonic development, but rarely on healthy adult cells. A high level of ROR-1 expression has been detected on mantle cell lymphoma tumor cells as well as in the case of breast cancer. CAR-T cells designed to target surface molecule ROR-1 are to be used to address hematological tumors such as mantle cell lymphoma but also solid tumors such as breast and lung cancer.

In order to manufacture this kind of therapeutic agent, immune cells are taken from the patient’s body by means of leukapheresis. T helper cells and cytotoxic T cells are then selected by magnetic cell separation. The genetic material for the CAR is introduced into the genome of the T cells via a non-viral gene transfer using the “Sleeping Beauty” transposon system (jumping gene). This reprograms the T cells in such a way that they perceive ROR-1-positive cancer cells as “foreign” and eliminate them by releasing cytotoxic messengers. The reprogrammed cells are multiplied and administered to the patient intravenously.

The project is being funded as a pilot project under the proof-of-concept initiative launched by the Fraunhofer-Gesellschaft, the Helmholtz Association and Deutsche Hochschulmedizin in order to promote the translation of innovative research projects. With the help of this funding, preclinical trials investigating the safety and efficacy of the ROR-1 CAR-T cells have been conducted at Fraunhofer IZI and the pharmaceutical manufacture of the therapeutic agent has been established, with the aim of achieving clinical translation in a phase I / II study (first in man).

Test batches were produced in the project to begin with; these batches were used to optimize the process in line with the stringent production requirements under GMP conditions and to qualify the necessary equipment. After successfully establishing the process and determining the required specifications, three successful validation batches were produced in the clean room and the analytical methods were established. 

Using the cell products generated in these validation batches, the analytical methods relevant to microbiological safety (mycoplasmas, sterility, bacterial endotoxins) and the verification of genomic safety (determining the vector copy number) were also validated. Finally, a product stability test was performed, which culminated in the successful realization of an additional batch. Once the validations were complete, an application was made to and granted by the competent authority to include the investigational medicinal product in the existing manufacturing authorization pursuant to Section 13 of the German Medicinal Products Act (AMG).

CAR-T cell therapy is a type of cancer immunotherapy. It uses the patient’s own T cells to fight certain types of cancer. In order to do this, the cells are collected in the clinic by leukapheresis before being genetically reprogrammed in vitro in such a way that they can use a chimeric antigen receptor to recognize cancer cells that have a special antigen on the cell surface. Following lymphodepleting chemotherapy, the reprogrammed cells are administered to the patient by way of infusion, where they are able to proliferate and trigger the immune response. 

Fraunhofer IZI has been assisting and supporting the company Novartis since as early as 2015 with the manufacture and further development of the CAR-T cell therapy Kymriah® (tisagenlecleucel). 

After successfully establishing routine manufacture at Novartis’ manufacturing facilities, the continued collaboration will now focus on manufacturing novel next-generation CAR-T investigational drugs developed by Novartis. A key feature of the T-Charge™ platform is the preservation and / or improvement of T cell stemness, i.e. the T cells’ ability to self-renew and mature. This leads to a product with a greater proliferative potential and fewer exhausted T cells, which is likely closely connected to its therapeutic potential.

With T-Charge™, CAR-T cell expansion occurs primarily in the patient’s body (in vivo), eliminating the need for an extended culture time outside of the body (ex vivo). In addition to the altered cell properties, the T-Charge™ products will therefore be more readily available to patients compared with the conventional CAR-T technology due to their simplified processes and optimized quality control. Not only patients, but also the health-care system as a whole would benefit from this.

Press releases and further information

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.