PluriCon 2024

Postscript

We are thrilled to share a wrap up of the iPSC-targeted discussions and insights from PluriCon 2024, Pluristyx’s inaugural iPSC Innovation Symposium held in Seattle on September 10, 2024.

Translating Today’s Discoveries into Tomorrow’s Therapies: Observations and experiences in the complexities of moving academic iPSC research into the clinic.

Designing a Better Cell Therapy – Revealing cutting edge approaches to iPSC differentiation and scalability,

Testing for Success – Identifying strategies to identify iPSC genetic instability. 

Culture Consistency – Understanding variability in iPSC cell culture to ensure developmental success.

Regulatory Challenges and Opportunities – Navigating the global regulatory environment of highly complex iPSC-based therapies.

The Last Mile – Identifying the critical role cryopreservation plays in iPSC genetic stability and commercialization. 

The Last Mile: Cryopreservation Challenges for iPSC Products

Dr. Brian Hawkins
Chief Technology Officer, Pluristyx

Cryopreservation is an often overlooked, but absolutely critical, component of development that can make or break the success of a promising iPSC-derived therapy, from research through large-scale clinical manufacturing.

Key insights from Dr. Hawkins’ talk included:

  • Learn from the past – Cryopreservation, especially when performed improperly, clearly impacts iPSC genetic stability. Dr. Hawkins utilized real-world examples along with ideas on how to leverage yesterday’s experiences in CAR T-cell therapies for Tomorrow’s iPSC breakthroughs.
  • Cryopreservation is more than just freezing – The science behind cell cryopreservation is complex and a continuum.  What happens both BEFORE and AFTER the freezing event are the key to preventing cell damage and improving post-thaw function.
  • Customization is key – Groups should not rely simply on historical protocols, but rather identify all the critical process parameters, collect the relevant data, and then implement proper controls to enable consistent cryopreservation outcomes.  This optimization should occur early in development to ensure iPSCs remain genetically stable and can be transformed into breakthrough living medicines.  
  • Always consider the clinic – The goal of any cell therapy research should be the patient.  Proper consideration should be given to cryopreservation that can easily transition to the clinic with minimal impact on cryopreservation efficacy.
  • Go Big – Proper consideration should be given to cryopreservation processes that can follow iPSC development from laboratory to industrial scale.  Procrastinating scale up until the last minute is a recipe for disaster for complex and expensive iPSC therapies.

 

Dr. Hawkins' presentation was a timely reminder that cryopreservation is essential for iPSC therapies and should be treated as a continuous process, ensuring scalability and effectiveness of Tomorrow’s Cell Therapies, Today®.

   

"Cryopreservation is a stress point. And any time you're trying to work on a process, if you make a process without an idea towards commercialization or large scale manufacture, in my mind, it's a bit of a wasted effort. You need to think about how you're moving it forward to product."
Brian Hawkins

Safeguarding Genomic Stability ​ in iPSC Line Development

Dr. Raluca Marcu
Director of Cell Line Development, Pluristyx

Dr. Marcu, Director of Cell Line Development at Pluristyx, discussed the challenges and solutions involved in maintaining genomic stability during the development of unmodified and engineered pluripotent stem cell lines. Her talk focused on identifying sources of genomic instability, improving testing methods, and optimizing workflows to create stable, genetically robust iPSC lines suitable for clinical applications.

Key insights from Dr. Marcu’s talk included:

  • Sources of Genomic Instability in iPSCs – Genomic instability can arise from mutations that pre-exist in the donor material or are acquired during iPSC derivation and expansion, particularly at population bottlenecks that introduce stress on the genome (reprogramming, gene editing, cloning). Mutations presented in the form of DNA sequence changes and/or genome structural changes can result in altered gene expression and impact biological functions of iPSC.
  • The Importance of Comprehensive Testing – Standard genomic stability tests like G-band karyotyping have limitations in specificity, resolution, and sensitivity. Dr. Marcu presented a case where G-band initially missed genomic instability in iPSC Seed Banks, leading to failures in later-stage Master Cell Banks. Recent ISSCR and FDA guidelines stress the need for detailed genomic characterization to detect mutations and mosaicism in iPSCs. Testing methods should be chosen based on detection capability, resolution, sensitivity, and timing of characterization. Advanced methods like digital PCR and targeted NGS now allow earlier issue detection, preventing costly downstream failures.
  • Pluristyx’ Optimized Workflow – Pluristyx has developed an optimized iPSC derivation workflow to reduce culture-induced stress and enhance genomic stability. This is achieved by minimizing passage numbers, eliminating bottlenecks, standardizing culture protocols and integrating enhanced in-process and bank testing. Utilizing efficient mRNA-based reprogramming and bulk iPSC passaging, this workflow includes gene editing of polyclonal pools to maintain genetic and epigenetic diversity, with cloning deferred until later stages. A stress test on iPSC Seed Banks is conducted to evaluate the long-term genomic stability of iPSCs under anticipated usage conditions.

Dr. Marcu concluded by emphasizing the need for rigorous, multi-step testing and process optimization to generate genetically stable iPSC lines, which are critical for clinical success in cell therapy development.

  

"Our recommendation is to try to minimize the risk of a downstream process failure by paying particular attention to two things. First, try to standardize the expansion protocols in order to minimize those culture-acquired mutations... [Secondly,] increase characterization of the starting material and the seed banks. If there are problems that are there, you want to know early."
Raluca Marcu

Regulatory Landscape for Cell Therapies

Dr. Richard McFarland
Chief Regulatory Officer, ARMI

Dr. McFarland’s key advice was for companies to work closely with regulators and to quantify risks as early as possible to create actionable data.   

While his talk underscored the complexities of navigating the regulatory landscape for cell therapies, there were actionable takeaways to communicate clearly with regulatory bodies, build a robust manufacturing process, and perform rigorous risk assessments when bringing iPSC-based therapies to market. 

  • FDA’s Focus is on Safety and Risk-Benefit - The FDA prioritizes the patient above all else when reviewing cell therapies and all decisions at the FDA are made through a risk/benefit to the patient lens, with regulators continuously assessing whether the benefits of a therapy justify the risks. Reviewers are trained to carefully weigh the risks of potential adverse reactions against the potential benefits of therapies, especially in life-threatening conditions like cancer. This is particularly true for high-risk therapies like gene and cell therapies such as CAR-T therapies, where the FDA allows for some level of risk if the alternative is death or serious harm. 
  • Regulatory Pathways and Uncertainty - While the regulatory path for cell therapies is clearly defined as an IND to a BLA, the specific expectations for individual products remain uncertain due to the variability in scientific data and product processes. This is compounded by the FDA's need to base policies on reviewed data, which takes time to accumulate.  Regulatory hurdles are often a result of unmet product development milestones or miscommunication between sponsors and regulators; therefore, it is advisable to communicate early with regulators to avoid delays.
  • Genetic Drift and Patient Safety - Genetic drift during cell culture is a major concern in iPSC therapies. As even minor growth advantages in some cells can lead to significant product variability over multiple generations, it is important to monitor for genetic mutations throughout the manufacturing process. Similarly, preclinical testing is essential for identifying and mitigating these risks before products reach patients. 
  • The Role of Standards - Industry standards can bridge gaps between the FDA’s vague guidance documents and the specific needs of emerging cell therapies. For example, the ISO cell counting standard helps ensure consistent product quality and reduces the need for each company to independently develop testing protocols.  Companies are encouraged to participate in the development of industry standards to ensure their processes align with regulatory expectations.

   

"This is the FDA IND reviewer standpoint. Statutory authority dictates and bounds our mission. Keep unsafe products off the market and allow potentially beneficial on the market. They see safety and risk everywhere... You've got to know what your risk is. You've got to quantify it and be able to explain it, even if you don't expect that it's going to be there."
Richard McFarland

Contact us for copies of slides from these talks

(where available)


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