Recently, Axiogenesis hosted an American user symposium titled, “iPSC Derived Cells & Assays Come of Age.” Held in Boston, Massachusetts (June 13, 2016), the symposium explored how experts in the pharmaceutical industry and academia use Axiogenesis’ cells and assays within drug discovery, safety/toxicology screening, and bioengineering applications.
It also presented current approaches to establish and define standards for these cellular models. Widely attended by representatives from the pharmaceutical sector, symposium attendees had the valuable opportunity to view case studies of iPSC-derived cells utilized in drug development decision-making and biomedical research.
To capture highlights from the event and facilitate thought-provoking conversation, below is a list of the top 10 talks from the recent Axiogenesis’ iPSC user symposium.
Top 10 Talks from Axiogenesis’ American iPSC User Symposium
KIT PARKER, Harvard School of Engineering and Applied Science / Wyss Institute – “Standards in hiPSC / Human Cells for Organs on a Chip”
Kit Parker’s enthusiastic talk featured comparisons of engineered myocardium from embryonic stem cell-derived versus rat neonatal cardiomyocytes. His lab data demonstrated that the unequal forces generated by stem cell-derived and native rat cardiomyocytes are sufficient to induce the formation of cellular adhesions that can dissipate force into the cells’ surroundings.
Using their muscle on-a-chip technology featuring Axiogenesis and primary cardiomyocytes on long contractile cantilevers, the Parker lab is trying to uncover mechanisms for improving the mechanical coupling of stem cell-derived cardiomyocytes to ultimately improve efficacy of stem cell transplantation for clinical purposes.
In concert with the theme of this meeting, Kit also presented his recently published work that calls for the establishment of iPS-cardiomyocyte (iPS-CM) standards and performance metrics, not only for commercial purposes, but also for clinical and regulatory applications.
Importance of Findings: Working towards a functional organ-on-a-chip system utilizing iPSC-derived cardiomyocytes; defining a standard for iPSC-derived cardiomyocytes.
GARY GINTANT, Abbvie – “Stem-Cell Derived Cardiomyocytes and Preclinical Safety: Comprehensive In Vitro Proarrhythmia Assay (CiPA) Update”
Gary Gintant provided an in-depth overview of the current CiPA initiative, including the role of cardiac ion channel patch clamp data, human clinical pharmacology, in silico cardiac action potential simulations, and importantly, the use of human iPS-cardiomyocytes in preclinical cardiac safety assessment.
Axiogenesis has been an important contributor in the development of protocols and finalizing test compound concentrations, as well as providing the Cor.4U cardiomyocytes as part of the CiPA myocyte subteam. Gary presented retrospective data from the first CiPA pilot study conducted last year.
Gary also summarized the basic structure of the core groups, compounds to be run, and instrumentation to be used in the 2016 CiPA validation studies to be performed later this year.
Importance of Findings: Successful round 1 of CiPA experiments in a multi-site study showing high predictivity of iPSC-derived cardiomyocytes.
GREG LUERMAN, Axiogenesis – “Axiogenesis Fresh Cor.4U Cells: A User Experience”
Greg Luerman presented a case study of a large pharma and collaborator of Axiogenesis who have implemented fresh ready to use shipments of Cor.4U cardiomyocytes pre-plated on Axion Maestro 96 well MEA plates. He gave an overview of how the cells are plated, packaged, and shipped to U.S. customers, making them ready for immediate use by the end-user.
Greg was able to show customer data quantifying plate-to-plate and shipment-to-shipment variability of measureable MEA parameters. This parameter variability was demonstrated to be very low.
Pre-plated Cor.4U-MEA shipments, in conjunction with the help of robotics, are helping to reduce variability as well as eliminating many of the tedious steps associated with these handling-intensive assays, thus freeing up a user’s time for other critical activities.
Importance of Findings: Successful implementation of 37⁰C shipment of ready to use Cor.4U on MEA plates to increase efficiency of drug development process.
ELENA KFOURY, Axiogenesis – “A Six Sigma Case: Reduction of Variance in Fresh Cell Shipments”
As part of Abbvie’s continued collaboration with Axiogenesis, Elena presented internal data around the optimization of the pre-plated Axion MEA shipments. The steps to reduce process variability and risk/costs while increasing process capabilities were presented within the framework of Six Sigma.
This methodology has broad applicability, especially as it relates to high-throughput screening and the use of stem cell derived products for regulatory enabling studies (i.e. CiPA). Axiogenesis plans to use this methodology to assess compound (IC50) drift across multiple cell lots, plates, days, etc.
Again, these well-controlled optimization steps are necessary precursors to establishing global standards for cardiomyocyte diagnostic assays.
Importance of Findings: Establishment of stringent quality control and optimization of shipment of 37⁰C shipment of ready to use MEA plates.
PAYAL RANA, Pfizer – “Using Peripheral Neurons in Neurotox Applications”
Payal provided a case study of how Pfizer Drug Safety R&D is using Axiogenesis’ Peri.4U human iPS-derived peripheral neurons for assessment of neurotox liability for new chemical matter.
Initially, Payal compared Peri.4Us versus Pfizer’s established neurite outgrowth screening assay that featured NG108 cells (immortalized rat neuroblastoma/glioma cell line). Peri.4U cells demonstrated significantly enhanced sensitivity to compounds associated with chemotherapeutic induced peripheral neuropathy (CIPN).
This is a very hot subject matter, because the oncology market is booming and novel, sensitive assays to address this painful problem well prior to clinical trials is essential. Payal reported that the Peri.4Us provided a large assay window to examine neurite-specific toxicity versus overt cytotoxicity (which previous models were not able to provide).
Additionally, Payal shared that Pfizer’s collaborative work with Axiogenesis will yield a publication that provides a mechanistic framework to identify drug induced peripheral neuron toxicity.
Importance of Findings: High predictivity and sensitivity of iPSC neurons in chemotherapeutic/oncology compounds for neurotoxicity/ peripheral neuropathy.
ALINA BERDICHEVSKY, Novartis Institute for Biomedical Research – “Target Validation Using an iPS-Cardiomyocytes-Based System”
Alina is a laboratory head representing a large project group at Novartis in Cardiovascular Metabolism. Alina’s lab is utilizing iPS-cardiomyocytes to investigate the impairment of contractility due to several disease related mechanisms, including hypertrophy, atrophy, electrical remodeling, metabolism effects, fibrosis, and necrosis.
Different methods of modulation or inducement, like siRNA transfection, small molecule application, and peptides, were used to elicit the phenotype.
Readouts of Calcium flux, MEA, contractility, and metabolic analysis assays were used, and the expected in vivo effect was shown in vitro.
Importance of Findings: Identified phenotypic relevance of iPSC-derived cardiomyocytes in disease models.
HERIBERT BOHLEN, CEO of Axiogenesis – “Closing the iPSC Cardiac Circle”
Heribert Bohlen, CEO of Axiogenesis, gave the keynote speech of the Axiogenesis User Symposium. He provided a detailed development of stem cell derived cells and cardiac tissue and how the quality and utility has continually been improved at Axiogenesis since 2001 – from the initial differentiation of mouse cardiomyocytes, to Yamanaka paving the way for human iPSC-CMs, isogenic cardiac fibroblasts, selection of ventricular, atrial, or nodal cells, and beyond.
Heribert also detailed new work in the “tools” space, such as Axiogenesis cell lines (neurons, cardiomyocytes) with integrated calcium or voltage sensors and channel rhodopsins.
Finally, he presented work on a skeletal muscle development project – a large unmet need in the iPSC drug discovery world.
Importance of Findings: Axiogenesis has a powerful lineup of new products, including assays and tools designed in collaboration with industry.
JORDAN LANCASTER, Avery Therapeutics / University of Arizona – “Pre-Clinical Studies of iPSC Cell Therapies for Heart Failure”
Jordan Lancaster gave a fascinating presentation, as his program is facilitating direct clinical translation of iPS cell technologies. His group is aiming to address chronic heart failure with surgically implantable heart patches loaded with iPS derived cardiac and angiogenic tissue.
More specifically, his group is using a bio-absorbable (or absorptive) mesh as a scaffold to attach iPS-derived fibroblasts (FibroCor.4U) and ventricular cardiomyocytes (vCor.4U). Jordan showed off their initial POC work, including IHC and a mesh loaded with cardiomyocytes that was “jumping out of the dish” due to contractile forces.
Additionally, they have begun to implant these patches into their rat infarct model and have seen functional improvements.
Importance of Findings: Application of iPSC cardiomyocytes in a cardiac patch destined for clinical applications showing high contractile forces and physiological tissue structures.
FRANCESCO PASQUALINI, Wyss Translational Center – “Engineering the Next-Generation of Cardiac Cell Therapies On-A-Chip”
At the Wyss Translational Center and the Institute for Regenerative Medicine (University of Zurich), Francesco is developing in-vitro and in-silico strategies to predict the quality of regenerative medicine interventions.
He pointed out that there are high-quality control standards in raw materials (cellular medium, chemical matter, etc). However, as we move towards regenerative medicine, we do not have models to predict the quality of iPS cells for therapeutic purposes.
Francesco’s work has established quality metrics for stem cell cardiomyocytes using a multi-parametric assessment of various measureable endpoints (i.e. genetic, physiologic, structural/morphological, and similar), including a contractile architecture (sarcomere characteristics) as a marker of myocyte maturity.
Francesco stressed that you need to understand your necessity for a certain phenotype – rather than finding an assay system that is fit for purpose. This led to one of his last point that in the race for cardiomyocyte maturity, you may actually “price yourself out” of a viable assay system due to the increased costs associated with the production of highly matured myocytes.
Importance of Findings: Working towards a definition of which iPSC-derived cells and assays to use for which application.
HUAN (SHARON) WANG, Harvard Medical School – “Molecular Network Modeling of Drug-Induced Cardiotoxicity in the Space of Dose and Time”
Sharon presented multi-disciplinary work that aims to determine how oncology drugs may result in cardiotoxicity and how we can develop therapeutic alternatives by better understanding the mechanisms of their toxicity.
Sharon and colleagues in Peter Sorger’s lab are working toward identifying biomarkers of cardiotoxic stress (associated with oncology on- and off-target effects) using genomic and proteomic profiling of Cor.4U cardiomyocytes treated with various tyrosine kinase and broad spectrum kinase inhibitors.
Additionally, her group has looked at the cardiotoxic effects of these various compounds in cell death and calcium transient assays to help guide some of their ‘omics work. By plugging in this multiparametric analysis data into advanced computer models, the Sorger lab may one day be able to predict how a drug would work, or not work, for individual patients and tailor a therapeutic strategy specific to the patient.
Importance of Findings: Demonstrated utility of iPSC-derived cells for cardiotoxicity analysis of oncology/chemotherapeutic compounds.
Bonus: Panel Discussion – “Establishing Global Standards for Commercial hiPSC”
In addition to the ten powerful talks above, Axiogenesis’ symposium “iPSC Derived Cells & Assays Come of Age” included a panel discussion in which industry thought leaders gave their opinions on important concerns to the iPS community, including questions about cell maturity, establishment of stem cell quality metrics, price points, and the value add for commercially-derived iPS cell types.
Gary Gintant (Abbvie) provided an insightful statement about iPS-cardiomyocytes being “fit for purpose”. He suggested that even though there is a major push for cells with greater maturity, a perfect reconstruction of the adult human ventricular myocardium may not be necessary for the vast majority of iPS-CM users.
Morrie Ruffin (SCB, ARM) provided insight into the Standards Coordinating Body (SCB) and how/why it was recently organized. Multiple working groups have now been developed within the SCB – with funding from FDA, industry, and academia – in order to identify gaps and establish standards around the characterization of stem cells, including how to derive potency, ensure identity and purity, count cells, and more. This is critical as it relates to stem cells in drug discovery.
Kit Parker (Harvard, Wyss Institute) identified the reach of the SCB and its relation to NIST (National Institute of Standards & Technology). Interestingly, Kit suggested that there is a need to incorporate more framework around the development and training of quality engineers for manufacturing in science.
Laszlo Urban (Novartis) provided some commentary on the need for large compound test sets to truly validate stem cell assays, such as what is occurring currently in the CiPA initiative. Additionally, he had some concerns about the translation of human stem cell assays to preexisting animal model data, and suggested that you must fully understand your test system before adopting it.
Elena Kfoury (Axiogenesis) indicated that there is conservatism within the market as users wait for “burden of proof.” Therefore, early adopters may be the ones supporting the development of iPSC products and establishing the standards within the market, while others wait for this process to play out.
Importance of Panel Discussion: This discussion was a rare opportunity for pharmaceutical stakeholders to discuss the standards required for iPSC-derived cardiomyocytes to be suitable for use within the industry.
Summary of Axiogenesis’ American iPSC User Symposium
This user meeting brought together a diverse range of stakeholders for effective exchange of ideas and opinions, and provided a rare opportunity for industry participants to view case studies of iPS cells utilized within a range of drug development applications. Such case studies demonstrate the predictive value of iPSC-derived cells, which ultimately improves drug candidate selection, lowers costs, increases drug safety and thus accelerates overall drug development.
The symposium also underpinned Axiogenesis’ commitment in establishing global standards for iPSC-derived cell products – a topic that is critical to a wide range of stem cell industry stakeholders, including representatives from across the pharmaceutical, biotechnology, and chemical sectors. According to Axiogenesis, the goal of developing standards is to efficiently support sector regulatory acceptance for submission to improve the predictivity, as well as cost and time resources for product development and approval. Therefore, creating standards will support a more uniform compliance environment and will address and assist in future efforts for harmonization internationally of the regulatory framework for submission across the globe.
Ultimately, global standards for iPS cells will support, or may even be a prerequisite, for widespread adoption of these cell types in pharma preclinical drug development.
To learn more about Axiogenesis or to participate in future user symposiums, visit www.axiogenesis.com.