Large scale lentiviral vector production

Lead Research Organisation: King's College London
Department Name: Cancer Studies

Abstract

Lentiviral vectors (LV) are remarkable in their ability to insert their genetic payload into a target cell's genome. This affects a
permanent genetic change in the target which is propagated through to its progeny. While there are several means of
genetic modification, few are capable of permanent modification of target cells. Amongst all vectors for gene delivery, LV
are unique in unparalleled efficiency, safety, lack of toxicity and ability to modify non-dividing target cells. They have
therefore come to be recognised as a key reagent required for the efficient development of the burgeoning cell therapy, as
well as gene therapy industries. However, although LV represent a well understood and robust technology, there is no
manufacturing methodology for very large-scale LV production. This is now an acknowledged bottle-neck both for clinical
trials and for commercial exploitation of many cell and gene therapy products in current development. We propose to
address this unmet need

Technical Summary

* Lentiviral vectors (LV) are used to genetically modify target cells. The main commercial application of LVs is in vitro
production of cellular therapeutics, although they can be used as therapeutic agents in their own right and for generation of
protein producer cells. While small-scale production is well established, a process for very large-scale production has not
been developed. Recently, commercial demand for LV has increased greatly. Large-scale production is a major unmet
need and its lack a bottleneck esp. in the marketing of cellular therapies. The challenge of large-scale LV production is
multifaceted, requiring a deep understanding of lentiviral biology, as well as skilful genetic and cellular engineering,
appreciation of regulatory / GMP considerations, industrial/pharmaceutical scale production issues as well as an
understanding of the down-stream clinical applications.

Planned Impact

As described in proposal submitted to IUK
 
Description The single most important contribution of this project has been effective process development and manufacture of reagents (cells and vectors) needed for the manufacture of gamma-retrovirus and lentivirus vectors that are used for various clinical applications of cell and gene therapy.
Exploitation Route The findings of two previous BBSRC grants (BB/E005896/1 and BB/D014301/1) and the current project (BB/N003853/1) and to a smaller extent BB/K013785/1, have underpinned the development of a major manufacturing activity (GMP Production of viral vectors for clinical use) at King's College London. We are now in active collaborations with a number of pharmaceutical (Cellectis, as well as Pfizer and Servier in active discussions for further collaborations) and biotech companies (Autolus and LiFT), as well as the Cell Therapy Catapult, for the further development and commercial exploitation of these manufacturing facilities and experties, including IP.
Sectors Education,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description This research, in combination with BB/K013785/1 and BB/E005896/1 has underpinned a great deal of innovation in the manufacture and production of Lentivirus and retrovirus vectors for clinical trials. This IP, combined with a GLP/GMP facility supported by grants from NIHR and CRUK have enabled the attraction of a number of major research grants including £2.5M from Roche Pharmaceuticals for the evaluation of glycoengineered antibodies and the evaluation of immunological markers in patients receiving these antibodies in phase-i/II clinicaal trials. The IP generated, the facilities and the experience of working on the Roche supported projects have in turn resulted in the attraction of manufacturing contracts from a US Biotech company (Northwest Bio - 1.2M), from a French company focused on the generation of allogeneic T cells expressing Chimeric antigen Receptors (Cellectis, over £11M between 2016 and 2019). The contract with this compnay, Cellectis, is worth in total about £15M over the next 3 years (2016-2019). We are also engaged in a fruther collaborations with a new biotech company in UK (Autolous) for the production of retroviral vectors that are used in the generation of other CAR-T cells for clinical studies. Finally, a recently initiated collaboration with Cell and Gene Therapy Catapult is developing procedures and products for the industrialisation of cell and gene therapy products, including the development of a scalable strategy for suspension cell based manufacture of retrovirus vectors encoding specific T cell receptors.
First Year Of Impact 2016
Sector Education,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
Impact Types Economic

 
Description Next generation CAR19 studies - Collaboration with Martin Pule et al at UCL
Amount £900,000 (GBP)
Funding ID II-C3-0714-20005 
Organisation National Institute for Health Research 
Department NIHR i4i Invention for Innovation (i4i) Programme
Sector Public
Country United Kingdom
Start 05/2016 
End 04/2019
 
Description Cellectis: Production of viral vectors (primarily lentivirus), and gene modified cells, for clinical applications of cell and gene therapy 
Organisation Cellectis
Country France 
Sector Private 
PI Contribution Development and production of multiple lentivirus and retrovirus vectors for a range of clinical studies in collaboration with both academic and industry partners, the largest of which is the collaboration with Cellectis culminating in over £1.6 million of funding todate, plus a new contract for £10.1 million over the next 3 years.
Collaborator Contribution Provision of funding and know-how in specific areas (e.g. site directed endonuclease mediated inhibition of endogenous T cell receptors, in order to allow the generation of allogeneic (off-the-shelf) Chimeric Antigen Receptor (CAR) T cells for the treatment of malignant disease. This project is directly supported by BB/N003853/1 and assisted by the outputs from our previous BBSRC grants: BB/E005896/1, BB/D014301/1 and BB/K013785/1.
Impact The development of allogeneic CAR-T cells (referred to as UniCAR-T) for the treatment of malignant disease. There has also been substantial inward investment (over £11,000,000 between 2016 and 2019 from Cellectis alone) underpining further developments that we expect to culminate in substantially larger collaborations with other pharmaceuticaal companies (active discussions in progress with Cell Therapy Catapult, Pfizer and Servier). This collaboration has also resulted in a separate collaboration with a UK based start-up company - Autolus (reported as a separate collaboration).
Start Year 2015
 
Description Collaboration with the University fo Lausanne for the development and production of lentivirus lectors 
Organisation University of Lausanne
Country Switzerland 
Sector Academic/University 
PI Contribution We have set up a new collaboration with the University of Lausanne for the development of lentivirus vectors and their GMP manufacture over the next 3 years (2018 to 2020). University of Lausanne has provided a contract of £2.6M of which the first instalment of 20% has already been paid.
Collaborator Contribution The development of vectors that they have produced for a number of gene therapy based clinical studies, and the use of the vectors made under GMP at King's College London, in these clinical trials.
Impact Contracts of Collaboration Signed
Start Year 2018
 
Description Vector Industrialisation Project 
Organisation Cell Therapy Catapult
Country United Kingdom 
Sector Academic/University 
PI Contribution The aim of this recently initiated project is the development of GMP compatible procedures for the industrialisation of gene therapy products. The main focus of this specific project is the development of cell line/s with characteristics needed for large scale manufacture of a retrovirus vector encoding a specific T cell receptor (TCR).
Collaborator Contribution Funding of the initial studies, providing the vector manufacturing cell line and expertise in project management, accurate calculation of costs of goods. Cell and Gene Therapy Catapult is also providing expertise in the development of strategies aimed at reducing the cost of goods, risk-reduction for manufacturing campaigns and strategies for efficient large scale manufacture of clinical grade vectors. This project is underpinned by two previous BBSRC grants and directly affected by our current BBSRC supported project.
Impact This project is contributing to the development of new therapies and therapeutic strategies, particularly with respect of the industrial scale manufacture of cell and gene therapy vectors, thus contributing both to better health care and to creation of wealth, including inward investment from outside the UK.
Start Year 2016
 
Title Development of GMP compliant manufacturing strategies for the production of clinical grade viral vectors 
Description The production of viral vectors, in particular lentivirus and gamma-retrovirus in sufficient quantities and able to meet the regulatory standards of quality is particularly challenging. Using the technologies that were developed as part of our BBSRC supported projects, we have established a range of manufacturing, purification and concentration strategies that have enabled us to manufacture the largest number (academia or industry) of retroviral and lentivirus vectors for regulatory approved clinical trials in Europe. This extensive research and development programme has now culminated in over £15 million pound of income (2012 to 2019) for King's College London from overseas based companies. 
IP Reference  
Protection Protection not required
Year Protection Granted 2016
Licensed Yes
Impact The background manufacturing IP and know-how is licensed (non-exclusive) to Cellectis and to Cell Therapy Catapult. Discussions are in progress with other organisations in taking similar non-exclusive licenses.
 
Title The processes developed in the course of this study have directly contributed to the success of subsequent contracts with the Industry, including Autolus and Cellectis (biotech and pharmaceutical companies. 
Description We have developed procedures for the fast manufacture of retrovirus and lentivirus vectors in compliance with the regulatory requirements for clinical use (GMP compliant procedures). These highly optimised procedures have enabled the production of high titre vectors (about 50,000 million infectious units of vector) from relatively small scale cultures (circa 10 litres), with greater than 50% recovery (frequently in excess of 70%) and minimal quantities of contaminating proteins and nucleic acids. This knowhow has recently been licensed on non-exclusive deals to the industry (Cellectis) in contracts producing in excess of £15 million pounds of income over the next 3 years. 
IP Reference  
Protection Protection not required
Year Protection Granted 2016
Licensed Yes
Impact We have produced, for regulatory approved clinical trials, the largest number of lenti- and retroviral vectors in Europe. Each of the 4 BBSRC supported projects have contributed to this outcome. We are now extending this expertise with a view to similarly innovative manufacture of Adeno Associated Virus (AAV) manufacture for clinical use.
 
Title CAR T cells 
Description The GMP vectors produced with the support of this BBSRC grant are now being used in a range of clinical trials based at Uuniversity College Hospital, Great Ormod Street Hospital and King's College Hospital 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Early clinical assessment
Year Development Stage Completed 2016
Development Status Under active development/distribution
Clinical Trial? Yes
Impact The vectors produced with the support of this project are now being used in multiple clinical trials - all Phase-1, all first-in-man