Post Learning Implementation Plan (PLIP)

Pre-course

• Access to the Educo Training Portal
• Submit your needs analysis and objectives for training by completing a simple online form

Action Plan

• At the end of the training you will create an action plan to help you implement and develop your new skills and knowledge

Reinforcement Session

• Delivered 3 to 4 weeks after the training
• It is a 1 hour virtual session enabling the group to reinforce the key learnings from the training and cover and subsequent questions and challenges

Introductory Online Module

FREE to Attend | Live & Recorded

24 September 2024 | 1:00 PM (UK)

Delivered Online

Overview of viruses used in cell and gene therapies

• Review various types of viruses used in CGT
• Learn how virus selection is dependent on application
  • Learn why and some of the challenges
  • Integration with host cells
  • Understanding that a virus must be integrative but non-replicative
• Examine examples of viruses used in CGT applications
  • Lentivirus
  • Adenovirus
  • Review gene therapy viruses
  • Examine cell therapy viruses
• Examine the differences between viral vectors used in gene therapies and cell therapies

Session 1

180 mins | 12:00 PM UK

Understanding underlying virology and cell biology

• Explain the wide variety of virus types and how this can be utilised to choose the best virus for your application
• Review the native targeting ability for some virus types
• Describe the development of the different GT viruses that have been and are used today
• What virus is good for some and not for other applications with details on the genetics and infectious cycle of example virus types to illustrate the reasons. The following will be considered:

• • The ability of viruses to integrate with the host genome and insert viral DNA
  • Engineering of viruses so they become integrative but non-replicative in order to ensure patient safety
  • What are the viruses we use for CGT applications: some strands of the HIV virus have been modified over time, lentivirus, adenovirus etc.
  • CAR-T cell therapy

Viral Vector manufacturing and therapeutic cell manufacturing principles

• Elimination of bacterial DNA sequences from plasmids and how different DNA sequences can cause immune reactions and cell death plus other considerations of plasmid design.
• Considerations of scaling up from the lab to the clinical scale plus media composition and cell line considerations due to potential contaminations causing immune reactions when injected into the patient
• Design of production facilities with positive pressure and strict protocols for manufacturing etc.
• Plasmid design
• Growth in HEK293 Cells and expansion to reach desired titration, 50l, 200l
• Industrialisation of cell therapies
• Understanding the different technologies available and when to use what
• Facility requirements – operating in closed systems saves time and money as you can operate in a grade C/D facility

Group Exercise
This will be the start of a rolling case study. Delegates in groups will be presented with a scenario where they develop a CGT application which needs viral vectors for delivery or
engineering. Delegates will consolidate the previous two sessions and consider key steps for engineering a viral vector for their application.

Session 2

180 mins | 12:00 PM UK

Choosing the most appropriate vector for your application

• Understand the importance of having a deep knowledge of the disease to be able to design the best therapy ie choose the right vector system and genetic control systems for the therapeutic genetic elements
• Explain the difference in ex vivo and in vivo GT and the design of the different approaches
• Discuss the use of naked nucleic acid therapies v/s vector-based therapies
• Examine the difference in how VV (Viral Vectors) are used in gene-modified cell therapy and gene therapy – GMCT – vector is the raw material whereas in gene therapy the vector is the final product

Choosing the most appropriate viral vector for your application

• Size of the gene which needs to be transduced
• Transduction efficiency
• Serotype efficiency
• Examine the issues with large genes and also how the distant enhancers can cause issues in vector design
• Consider the target cells having a low level of available receptors for the infection and how this can affect the design of the therapy and overall design
• Brainstorm common factors which influence VV selection

Group exercise and feedback
Groups will consider important factors and design a viral vector for their cell and
gene therapy application and feedback their thoughts to the group.

Session 3

180 mins | 12:00 PM UK

Purification requirements

• Host cell protein requirements
• Residual empty capsid
• Residual host cell DNA
• How these purification requirements differ between gene therapy and gene-modified cell therapy
• Understand the importance of using optimal culture media and certified producer cells
• How it can be good and bad to “push” a producer cell line to high titters as it can be causing an issue with empty v/s full capsids
• Discuss how bacterial and producer cell DNA can cause negative effects as well as other impurities from manufacturing

Purification techniques

• ELISA
• q PCR
• HPLC
• TCID50
• Replication competent lentivirus detection
• QC of cell therapeutic products
• Discuss what techniques to use at what stage of production and why they should be used
• Examine what can be seen and what cannot be seen and how this can affect the overall design of a therapy
• Discuss the different ways to titre viruses and how this can be related to the specific characteristics of the virus itself
• Examine the inducible as well as SIV approach and the issues related to each methodology

Group exercise 
In groups, delegates will consolidate the previous talks and brainstorm the purification
requirements and techniques they need to use for their particular viral vector or gene therapy
methodology rolling on from the previous group sessions.

Dr.Lars Brandén has long experience from the fields of viral and non-viral gene delivery, high throughput screening and cell based assays. Dr. Lars Brandén has a career that encompasses start-up of gene therapy companies, building and operating several high throughput cell biology centres and has worked within the life science sector as an international consultant. During his tenure in academia Dr.Brandén was mentored by professor James E. Rothman (Nobel prize winner, 2013) and recruited to several director positions with Columbia University and Yale university. Dr.Brandén has also spent a year in art school and has had international art exhibitions. Dr.Brandén is now working as Science Advisor for a global provider of automated laboratory instruments and solutions in Switzerland