Mesenchymal Stem Cell Culture – Challenges and solutions for isolation, expansion and maintenance

In this podcast, we talked with Dr. Jennifer Chain, Scientific Director of Research and Development, Oklahoma Blood Institute about the isolation of mesenchymal stem cells form cadaveric bone marrow and the differences between live and cadaveric donors.  We also discussed the expansion and maintenance of mesenchymal stem cells in culture including media design and selection.

CryoVault Freeze and Thaw Platform Provides a Scalable, Robust and Single-Use End-to-End Solution for Bulk Drug Substance

In this podcast, I talked with Max Blomberg, Executive Director of Operations and Andrew Govea, Senior Product Engineer,  Meissner about the challenges of handling bulk drug substance, specifically freeze and thaw and how the need for scalability, flexibility and a robust approach led to the development of CryoVault.  CryoVault offers a unique and intelligently designed end-to-end freeze and thaw process solution

Improved gene therapy analytics permit monitoring of critical quality attributes and increased manufacturing efficiency

Jonathan Royce, Director, Instruments Business Unit, Vironova talks about analytics in gene therapy manufacturing. Specifically, Jonathan details new technologies to monitor critical quality attributes, reduce development time and increase the number of projects that can be run.

Generating actionable data and analysis in complex models using live-cell analysis

In this podcast, we talked with Paul Jantzen, Product Manager for IncuCyte at Sartorius about the benefits of real-time live-cell analysis and image processing workflows. We also discussed how live-cell analysis is enabling the use of neuronal cell models to study cell health, morphology, function and cell dynamics.

New Advancements in Cell and Gene Therapy Implementing Precision Medicine

In this podcast, we talked with Dr. Paul Wotton, Chief Executive Officer, Obsidian Therapeutics about the evolution of cancer therapy and new advancements in the space including precision medicine. Our discussion included Obsidian’s cytoDRiVE™ platform that provides a technology, in which the level and timing of protein activity are fully controlled in a dose-dependent manner by an FDA-approved small molecule.

Using 3D cell-based human liver microtissue models in predicting adverse effects caused by chronic exposure to engineered nanomaterial

In this podcast, we interviewed Shareen Doak is Professor of Genotoxicology and Cancer in Swansea University Medical School and Dr. Wolfgang Moritz, Co-founder and Head of External Collaborations and IP, Insphero about the use of 3D cell-based human liver microtissue models to predict the adverse effects of chronic exposure to nanomaterial . We also discussed the use of liver microtissue models for drug discovery and development.

Live Cell Analysis for Neuroscience Research

In this podcast, Susana Alcantara, Senior R&D Scientist and Neuroscience Program Leader, BioAnalytics, Sartorius talks about the ways real-time live cell analysis is enabling neuroscience research. We also discussed how real-time cell analysis can be used in neurological disease model development and in studying disease pathology to enable drug discovery.

Antibody Drug Conjugate (ADC) Development and Manufacturing Challenges and Solutions

In this podcast, we interviewed Dr. J.J. Luo, Executive Director and Dr. Lily Yin, Head of Biologics Conjugation Development, at WuXi Biologics about the state of the ADC marketplace and the challenges in developing antibody drug conjugates. We then dove into reasons behind why they built a dedicated manufacturing facility for ADCs and other bioconjugates and how a single-source development platform can benefit bioconjugate drug development efforts.

Podcast Notes:

I began the podcast by asking Dr. Luo about WuXi Biologics’ investment in and construction of multiple clinical and commercial scale facilities around the world. I pointed out that nearly all of these facilities are dedicated to the GMP production of Drug Substance and Drug Product of antibody or recombinant protein therapeutics, but their new facility appears to be dedicated for a very specific niche within the biologics industry. JJ explained that the new facility was purpose-built for the manufacture of a wide range of bioconjugates, in which the largest class of these products would be Antibody Drug Conjugates. He went on to say that this new facility is part of WuXi Biologics’ new state-of-the-art integrated Biologics Conjugation Solution Center in Wuxi City, China. The center will consolidate all bioconjugate drug development and commercial manufacturing of bioconjugates in the future.

I followed up by asking JJ about the drivers behind building a facility dedicated solely to ADCs and other bioconjugates. He stated that although bioconjugates are a subset of the greater biological therapeutic marketplace, various market research reports have shown a year on year increase in the number of ADCs and other bioconjugates entering clinical trials over the past 5 years by roughly 12% per year worldwide. Some estimate that commercial sales of ADCs will grow 22% annually for the next 5 to 10 years. In addition, based on drug company pipeline data, it appears that this trend will continue into the foreseeable future. JJ made it clear that WuXi Biologics’ mission is to provide open access drug development platforms to ensure that every drug can be made and every disease can be treated. Given WuXi Biologics’ expertise, vast capabilities, and resources they believe that they can provide a single-source product development and supply chain for companies developing these novel therapeutics.

Dr. Yin added that WuXi Biologics has a strong history of providing extensive product development and manufacturing services to clients. They have developed a wide variety of bioconjugates from idea to GMP manufacturing for clinical study. She goes on to share that they have provided full CMC services for 11 IND filings for bioconjugate products in just the last few years and 25% of the total IND filings for ADCs in 2018. With the new bioconjugation facility, WuXi Biologics’ has increased GMP manufacturing capacity for both drug substance and drug product and can meet their client’s needs for clinical supply while still offering additional capacity for new clients.

Next, I asked Dr. Yin about their ability to provide a single-source for all bioconjugate needs. I clarified that ADCs and other bioconjugates are complex molecules with both chemical and biological components. I wanted to know how they would be able to provide all the services required to get from early discovery to clinical supply and if there were any gaps. Lily said that WuXi Biologics is known for their ability to take clients from early discovery and lead generation of antibody therapeutics into development and the clinic. Through WuXi Biologics’ expertise and single-source services they are able to provide this without having to use another service vendor. She went on to say WuXi Biologics is now a separate company, but it is still a member of the WuXi AppTec Group of companies. They still work very closely with the Group and utilize its vast resources and capabilities in discovery chemistry, development and GMP manufacture of small molecule payloads and linkers. In addition, WuXi Biologics, along with WuXi AppTec, can provide the essential preclinical services such as DMPK, oncology research, early stage IND-enabling toxicology amongst other critical services. Couple these single-source services with the fact all of these services and the entire supply chain are within minutes to a couple hours from each other and WuXi Biologics can provide a significant advantage in time and also risk reduction for any company wanting to develop these type of drugs. Lily encouraged listeners to go to their website to learn more about their comprehensive capabilities.

Then we discussed how some of the new bioconjugate modalities often contain highly potent or highly toxic payloads thus requiring development and manufacture of these intermediates and the subsequent conjugation to be done in special facilities equipped to handle these compounds. They often also require additional oversight from regulatory authorities. I asked JJ how WuXi Biologics is addressing these types of intermediates and compounds. JJ described how their new facility has been purpose-built to handle highly potent and toxic materials. The design and construction were set to meet U.S. FDA, EMA and China National Medical Products Administration (NMPA) standards for those compounds.

We then focused on how ADCs have historically had many challenges in development and also challenges in demonstrating efficacy in humans. I asked Lily what is the industry doing to help mitigate these development challenges, reduce risks, and help ensure success to move more of these treatments into the clinic and hopefully to commercialization. Lily acknowledged that there have been set-backs and challenges in the field, but stated that bioconjugates are still a relatively young field with a lot left to learn on how we can more effectively develop successful drugs. She went on to say that the good news is that there are 5 ADCs on the market with approximately another 10 in late stage clinical trials with over half of those having obtained “Fast-track” status. WuXi Biologics believes that this demonstrates that the industry is beginning to turn the corner with better understanding of how to develop these molecules for clinical success. She shared that there is a better understanding of how to engineer antibodies to find suitable targets, how to select an active toxin, and how to use better linker chemistry to create better targeted binding characteristics, improved in vivo half-life, more consistently controlled drug-to-antibody ratio, residual free drug levels and better payload release to control drug levels in vivo.

I then asked Lily what specifically WuXi Biologics is doing to help further the ADC field. She said that they have now worked with so many different antibody or other biological molecules, linker and payload chemistries and combinations thereof, that they are uniquely qualified to advise clients on development strategies that will best suit their needs and ensure success. They have strong manufacturability and developability strategies when evaluating the various technologies and bioconjugation options to provide sound data driven decisions for lead selection and throughout development in order to generate a robust GMP process. Looking at many of the challenges and pitfalls of certain linker and bioconjugation schemes WuXi Biologics has continually developed their own new technology platforms for linkers and in some cases proprietary modifications to enhance certain payloads to increase their effectiveness for first in human trials.

I followed up by asking Lily to elaborate on the proprietary technologies she mentioned and also to discuss the benefits of WuXi Biologics’ IP. Lily said that they have developed a novel linker for Lysine-based conjugation that demonstrates higher reactivity, better solubility and a more flexible range of conjugation temperatures and the ability to conjugate this linker to other molecules beyond IgGs. Their unique payload chemistry involves more homogenous drug loading for cysteine conjugation. She went on to say that they would be happy to discuss more under a CDA with any company who is interested.

Next I asked JJ if these technologies were scale-able. He clarified that WuXi Biologics designed all of these technologies with an eye for ease of formulation and to be able to scale up for clinical trial scale and beyond.

I followed up with JJ and asked what manufacturing scales could be achieved at the new facility. He said that the new facility could produce up to 50 batches per year with up to 2 kilograms per batch for drug substance. He said that they can perform both liquid and lyophilization for Drug Product fills with a maximum of 100,000 2R vials per batch for liquid fills and 25,000 2R vials per batch for lyophilized fills. In addition, they are able to provide GMP lyophilization of ADCs in various vial sizes including 2R, 6R, 10R, 20R and 50R. He also mentioned that the facility could perform filling of other non-bioconjugate products such as small molecule or peptide therapeutics as long as they require liquid or lyo type fills. He added that since this is a multi-product and multi-use facility, they are utilizing single-use systems including single-use compounding vessels, tubing, bags, and filling needles for the entire bioconjugation and drug product compounding, filling and finishing process. They do this to greatly reduce risk during the GMP manufacturing runs and provide the highest level of quality assurance for their clients.

I asked that since they are working with highly potent and toxic compounds in this facility were there any special equipment requirements. JJ shared that isolator-based systems are widely utilized in the industry for working with highly potent or highly toxic materials, so they utilize isolator-based systems for payload and linker dispensing and dissolution, filling and lyophilization. In addition, fully automated equipment is used for vial washing and depyrogenation, filling, and for automated loading and unloading the drug product containers into and out of the lyophilizer. Filling is operated by an automated peristaltic pump dosing system and weight check is performed using on-line weighing system for automatic fill weight adjustment. If required, the filling line is also equipped with Nitrogen blanketing capabilities and light protected material handling is also available from start of fill processing to final package steps.

Lastly, I asked when they will start GMP ADC manufacturing activities. JJ said that they have just started operating the facility and their first GMP production will be in the third quarter of this year. There are nearly 20 lots that are scheduled including 10 GMP lots by the end of this year. So, although he said that they will be busy, there is still capacity for this year and next year as well.

I closed by thanking JJ and Lily for their time. The interview was very informative and provided good insight into the current state of the ADC/bioconjugate marketplace and how the industry and WuXi Biologics is working to overcome the historical challenges. It looks like a bright future for bioconjugated therapeutics is ahead of us.

Mustang Bio’s gene therapy has tremendous potential to cure XSCID or Bubble Boy Disease based on study results from St. Jude

In this podcast, we interviewed Dr. Manny Litchman, President and CEO, Mustang Bio and Dr. Knut Niss, Chief Technology Officer, Mustang Bio about the exciting study results for their gene therapy candidate to treat X-SCID, why this disease is a good fit for gene therapy and next steps.

Show Notes

To begin the podcast, I asked Dr. Litchman about the study results in treating X-SCID or “bubble boy disease”, including current standard of care and why gene therapy is a good fit for this disease. He explained that X-SCID is an X linked rare genetic disease with mutation in gene responsible for insufficient immune system generation at birth. As a result, severe, recurrent and opportunistic infections occur and usually results in death by age one if left untreated. Current standard of care is Immune reconstitution by allogeneic stem cell transplant, however the transplant must be from a matched donor and the best donors are siblings. There are only about 15% matched sibling donor transplants at this time. Gene therapy would eliminate the need for a matched donor and would improve survival and quality of life for the patient.

Next I asked Dr. Niss about previous attempts at treating this disease with gene therapy and how their therapy is different. He described that in the early days of gene therapy gamma retro virus was sometimes used as the vehicle for delivery.  For the current gene therapy a lentiviral construct is used, which has safety features that the gamma retrovirus didn’t provide. In the previous attempt to use gene therapy for X-SCID, the patients were cured of X-SCID, but developed leukemia due to the use of the gamma retrovirus. Today gene therapies build many safety features into the construct, many of which are incorporated into the vector used in the current study.

I then asked Dr. Litchman to elaborate on the St. Jude study and the very positive results. He explained that the results were published in the New England Journal of Medicine, “Lentiviral Gene Therapy Combined with Low-Dose Busulfan in Infants with SCID-X1,” by St. Jude. The study was a single arm study in newborn patients who were diagnosed with X-SCID. The results were striking as all patients had multi-lineage immune reconstitution. In addition, all patients that had infections prior to gene therapy cleared completely, with levels of IgM that are produced in B cells normalized in 7 patients. Four patients were able to stop IV immunoglobulin (pooled antibodies that these patients receive to prevent infections) and 3 patients had a normal response to childhood vaccinations. The safety profile was excellent with no leukemia and no transfusions required.

Next I asked Dr. Litchman if he could tell us about the two populations receiving the gene therapy currently and other patients who may benefit. Manny said that the therapy is currently being given to newborns with the disease. The incidence of the disease is very low with 1 in 225,000 live births globally. This results in about 20 patients in US. There is a larger population of about 400 who have received hematopoetic stem cell therapy and are eligible now or in the future for gene therapy because their condition has deteriorated due to infections. The National Institutes of Health (NIH) has a current trial running for these patients. Five patients have been treated and they are enrolling more patients and currently have a wait list.

I followed up by asking Manny what are the next steps for this therapy. He said that they will complete the technology transfer of the cell processing from St. Jude to Mustang’s cell processing facility in Massachusetts. It is a state of the art processing center run by Dr. Kniss. They will also continue the ongoing trial and will file for approval. At the same time, they will expand the number of sites sites for the NIH study to a multi center clinical trial.

I then switched gears by asking about their biomanufacturing approach. I began by asking Dr. Niss why they selected a lentiviral approach. Knut clarified that St. Jude selected lentivirus. He explained that it is well recognized that gamma retrovirus not the best approach for stem cell based therapies. He said we understand the integration of the lentivirus into the genome at a much higher level than the gamma retrovirus and gamma retrovirus integrates in a true random pattern where lentivirus has a defined integration pattern. For a therapy like this where you want durable outcomes, it is important to have stable integration into the genome so that the gene is expressed long term.

I then asked what were the biggest successes they found in manufacturing and what their plans are for scaling up in the future. Knut said that they don’t anticipate a problem because it is a fairly simple process. He explained that they isolate CD34 cells from the patient add the lentivirus and prepare the therapy for infusion. Capacity can be built up very quickly. He said that tehy are constantly scouting new technologies looking for the next technology breakthrough. In general, he said that cell and gene therapy products are now not the ultra rare niche technology. Technologies specifically developed for the needs of cell and gene therapy manufacturing have been developed and that has improved cell processing greatly

I asked what is next for Mustang. Manny said that they are looking for other in-licesning opportunities from the academic world of late pre-clinical or early clinical rare diseases that could take advantage of our cell processing facility. In the near term, they  have a rich portfolio in CAR T for oncology with 3 therapies for hematological malignancies and 3 for solid tumors. They will be  filing 2 INDs this year for two of those programs. One for AML and one for a multiple myeloma target.

I closed the interview by asking if there was anything else that they would like to add for our listeners. Manny said that they were very happy to have completed their capital raise and have at least two years of cash to execute on all their clinical programs. They are also grateful to their collaborators, St. Jude, City of Hope, Fred Hutchinson Cancer Center, and Nationwide Children’s Hospital.