Science Pool

Heart failure is a global health challenge that demands innovative therapeutic approaches. Evotec’s pioneering iPSC-derived cardiomyocytes offer a promising solution, addressing the critical challenges of immune rejection, graft-induced arrhythmia, and scalable production.  

The growing global burden of heart failure

Heart failure (HF) remains a leading cause of mortality, affecting millions worldwide​¹​. Current estimations predict that there will be over 8 million people in the United States alone living with HF by 2030^​2​. The condition is exacerbated by the heart’s limited regenerative capacity, as cardiomyocytes — the cells responsible for heart muscle contraction — have minimal ability to regenerate after injury. Consequently, a single myocardial infarction can result in the irreversible loss of billions of cardiomyocytes, leading to heart failure with reduced ejection fraction (HFrEF)​³​. 

Traditional treatments focus on managing symptoms and slowing the progression of HF but fail to address the root cause — the loss of functional cardiac tissue. Over time, patients will reach the end stage of HF, experiencing a vastly diminished quality of life, with ventricular assist devices or rare heart transplantation being final treatment options. As the global population ages and cardiovascular risk factors rise, there is an urgent need for novel therapies that go beyond symptom management, to regenerate lost heart tissue and prevent HF from progressing. 

One promising avenue is regenerative cell therapy using induced pluripotent stem cells (iPSCs). In this article, we explore the current challenges in developing cell therapies for heart failure and Evotec’s innovative approach to overcoming them, with recent findings from their iPSC-derived cardiomyocyte (iCM) Heart Repair program. 

The promise of regenerative cell therapy for HF

Regenerative medicine, especially iPSC-based therapies, holds tremendous potential in addressing the unmet needs of HF patients. iPSCs can be reprogrammed from adult cells, and with unlimited proliferative capacity, they have the unique ability to differentiate into any cell type. 

Thus, iPSCs offer a renewable source of cardiomyocytes. This makes iPSC cell-derived cardiomyocytes (iCM) an ideal candidate for providing novel and curative HF therapies, with the potential to replenish lost heart cells in HF patients and restore cardiac function. However, the path to clinical application is fraught with challenges. 

Challenges in cardiac cell therapy

One of the primary challenges with allogeneic iPSC-based therapies is the prevention of immune rejection. Allogeneic iCM are recognized as foreign by the patient’s immune system. This leads to rejection unless immunosuppressive drugs are used​4​. However, long-term immunosuppression carries significant risks and is not a viable solution for all patients.  

Moreover, there’s the challenge of preventing graft-induced arrhythmias. These can occur when transplanted cardiomyocytes do not integrate properly with the host tissue, leading to irregular heartbeats.  

Another key challenge is successfully scaling up production to meet clinical needs. HF patients require over a billion iCM for effective treatment. However, producing large doses of iCM demands cost-effective, scalable processes. Additionally, ensuring consistent quality and purity of these cells at increased scale is critical for their therapeutic success. 

Evotec’s iCM Heart Repair program, in collaboration with the Medical Center Hamburg-Eppendorf “UKE”, has been developed to address these challenges. This collaborative program focuses on the development of first-class non-immunogenic iCM that can be produced at scale, offering a promising off-the-shelf treatment that can meet the global demand for HF therapies.  

Immune-shielded iCM are protected from T cells and NK cells

As part of Evotec’s iCM Heart Repair program, researchers at Evotec evaluated iCM immune-shielding strategies for tissue replacement in HF patients. In this study, the researchers first developed a robust production process of wildtype (wt) iCM cells, using the fully characterized GMP iPSC line in small-scale GMP-compliant bioreactors. Yield evaluation with flow cytometry demonstrated an exceptionally high purity of >97% cardiomyocyte marker, cardiac troponin T (cTNT).

 To investigate strategies to prevent allogeneic immune rejection, two genetically engineered iCM lines were chosen, human leukocyte antigen (HLA)- encoded class I and II knockout iCM, and iACT iCM​^5​​,6​ (panCELLa Induced Allogenic Cell Tolerance Stealth Cells™). These lines contain genetic modifications to inactivate human leukocyte antigen factors I and II (HLA-I/II KO), or overexpress immune-shielding factors (iACT), which were engineered to the iPSC prior to their differentiation. Thus, both iCM are designed to prevent immune cell activation and cytokine release.  

Using in vitro assays, the two iCM lines were co-cultured with primed T cells or expanded natural killer (NK) cells. The subsequent release of the cytokine IFNγ and killing of the iCM was measured to investigate the immune-suppressing potential of HLA-I/II KO and iACT immune-shielded iCM. 

Figure 1: Wild type iCM (wt), HLA-I/II KO, and iACT immune-shielded iCM were co-cultured with primed T cells or expanded NK cells of human donors. iCM survival and IFNγ release from immune cells was measured after 6-24 hours.

Results of the in vitro co-culture assays (Figure 1) showed iACT iCM provide efficient protection against NK cells, but only moderate protection against T cells. Contrastingly, HLA-I/II KO iCM were shown to be vulnerable to NK cells due to the “missing-self-response”, but highly efficient against T cells. 

Engineering an improved iCM cell line  

To improve the therapeutic potential of immune-shielded iCM, Evotec has developed its proprietary EvoCloaking iPSC line. The novel cell line is based on HLA-I/II KO, with an additional innovative anti-NK cell strategy to increase its immune-shielding properties. 

In vitro data supports that EvoCloaking iPSC-derived iCM can reduce cytokine release and are protected against both T cells and NK cells. This means the proprietary cell line offers an innovative strategy to ensure the acceptance and persistence of engrafted iCM, improving the safety and long-term function of the therapy, while avoiding the need for immunosuppressants. 

In addition to its immune-shielding properties, EvoCloaking iPSC-derived iCM are genetically engineered to safeguard against tumorgenicity, with a drug inducible kill-switch allowing for the elimination of undesired proliferating cells. Importantly, the proprietary cell line will also contain a genetic modification to avoid graft-induced arrhythmia in derived iCM. This is supported by in vitro data, and is currently tested on arrhythmia-prone pigs.

EvoCloaking iCM are highly pure cells optimized for off-the-shelf product use as a single cell suspension. In heart injury guinea pig models, efficient iCM engraftment has been demonstrated when injected as single cell suspensions. Consequently, catheter-based interventional administration should be compatible with this product format.

Evotec's scalable therapeutics platform

The iCM Heart Repair Program aims to overcome challenges associated with large-scale iCM production. Using Evotec’s GMP-compatible manufacturing infrastructure, scalable and cost-effective bioreactor processes have been established for iPSC expansion and differentiation in 3D. Furthermore, predictive in-process controls and advanced in silico modeling have been implemented into the differentiation process, to optimize cell yield and purity. 

Evotec’s cell therapy pipeline expansion to include immune-shielded iCM is testament to its commitment to allogeneic cell therapeutics. Our unique end-to-end platform and expertise support the development and manufacturing of off-the-shelf iPSC therapies for a range of diseases and conditions, from cancer and autoimmune disease to diabetes and cardiovascular disease. The integrated platform covers development stages from early exploration to GMP compliant scale-up for further research, pre-clinical, and clinical studies. 

Figure 2: An overview of Evotec’s end-to-end platform for iPSC-based therapeutics

The future of heart repair

The development of immune-shielded iPSC-derived cardiomyocytes represents a significant advancement in the field of regenerative medicine. This therapy could revolutionize the treatment of heart failure, offering a solution that addresses the underlying cause of the disease, rather than just managing symptoms. Integrated iCM development and manufacturing processes are also ensuring the scalable, cost-effective production of this therapy, holding great potential for meeting the increasing global demand for curative HF treatment.

Evotec’s commitment to innovation and excellence in the field of regenerative medicine positions it as a leader in the development of next-generation therapies. By addressing the key challenges in iCM therapy, including scalable production, immune rejection, and graft-induced arrhythmias, Evotec is paving the way for a new era in heart failure treatment.

Moreover, the broader impact of this research by Evotec extends beyond heart failure, as the principles of immune-shielding and scalable bioprocessing can be applied to other cell therapies, opening new avenues for treating a range of diseases.

Discover more about Evotec’s innovative iPSC-based therapies

Download the iCM research poster

References

(1)    Savarese, G.; Becher, P. M.; Lund, L. H.; Seferovic, P.; Rosano, G. M. C.; Coats, A. J. S. Global Burden of Heart Failure: A Comprehensive and Updated Review of Epidemiology. Cardiovasc Res 2023, 118 (17), 3272–3287. https://doi.org/10.1093/CVR/CVAC013.
(2)    Golla, M. S. G.; Hajouli, S.; Ludhwani, D. Heart Failure and Ejection Fraction. StatPearls 2024.
(3)    Laflamme, M. A.; Murry, C. E. Regenerating the Heart. Nature Biotechnology 2005 23:7 2005, 23 (7), 845–856. https://doi.org/10.1038/nbt1117.
(4)    Lanza, R.; Russell, D. W.; Nagy, A. Engineering Universal Cells That Evade Immune Detection. Nature Reviews Immunology 2019 19:12 2019, 19 (12), 723–733. https://doi.org/10.1038/s41577-019-0200-1.
(5)    Harding, J.; Vintersten-Nagy, K.; Yang, H.; Tang, J. K.; Shutova, M.; Jong, E. D.; Lee, J. H.; Massumi, M.; Oussenko, T.; Izadifar, Z.; Zhang, P.; Rogers, I. M.; Wheeler, M. B.; Lye, S. J.; Sung, H. K.; Li, C. J.; Izadifar, M.; Nagy, A. Immune-Privileged Tissues Formed from Immunologically Cloaked Mouse Embryonic Stem Cells Survive Long Term in Allogeneic Hosts. Nature Biomedical Engineering 2023 8:4 2023, 8 (4), 427–442. https://doi.org/10.1038/s41551-023-01133-y.
(6)    Harding, J.; Vintersten-Nagy, K.; Shutova, M.; Yang, H.; Tang, J. K.; Massumi, M.; Izaidfar, M.; Izadifar, Z.; Zhang, P.; Li, C.; Nagy, A. Induction of Long-Term Allogeneic Cell Acceptance and Formation of Immune Privileged Tissue in Immunocompetent Hosts. bioRxiv 2019, 716571. https://doi.org/10.1101/716571.

Note: This article was developed for Bio Europe Supplement, EBR Autumn Edition 2024, Samedan Ltd 

Discover how an optimized hit identification strategy, integrating advanced tools – including AI and machine learning – can provide the best possible chemical starting points. Explore how Evotec’s collaborative approach is key to identifying high-quality hits at lightning speed, saving on costs and time, while reducing the risk of attrition. 

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At Evotec, our expert immunologists drive your immuno-oncology projects from target identification to clinical trials. We offer comprehensive support for integrated projects and specialized expertise at critical stages. Our services include bespoke in vitro immunological assays, genetic engineering of primary immune cells, and advanced preclinical models. By integrating cutting-edge technologies and leveraging our extensive clinician network, we ensure the successful translation of innovative cancer immunotherapies from the lab to the clinic. Partner with Evotec to advance your cancer immunotherapy research and achieve breakthrough results.

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Quality Hits At Lightning Speed - Accelerated Hit Finding Through Innovative Screening Approaches.

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In the evolving landscape of cancer treatment, Immuno-Oncology (IO) has been at the forefront for the past 15 years, revolutionizing our approach to battling this formidable disease. Immunotherapy, which harnesses the body's immune system to fight cancer, offers hope where traditional therapies have often fallen short. For instance, in advanced stage IV melanoma, the median survival has improved from 6 months to 6 years over the past decade thanks to immunotherapy.

However, the journey from discovery to clinical application is fraught with challenges. Why do some patients respond to immunotherapy while others do not? What opportunities exist for developing the right combination therapies? How can we better select the right cancer indications for a particular IO target? How can we ensure the success of these treatments across diverse patient populations?

At Evotec, our immunologists are equipped with the expertise and experience to support your IO projects from target identification through to clinical trials. Whether you require a comprehensive and strategic partnership or targeted support at critical stages, our capabilities can synergize with your efforts to make a tangible difference.

Comprehensive Support Across the Immuno-Oncology Spectrum

In-depth Immunology Expertise

Our team’s knowledge spans from innate to adaptive immunity, providing a robust foundation for developing cutting-edge cancer immunotherapies. We offer:

●    Genetic Engineering of Primary Immune Cells: Optimizing target validation for effective therapy development.
●    Bespoke In Vitro Immunological Assays: Custom-designed assays to evaluate immune responses and the potency of immunotherapeutic candidates.
●    Single-Cell Level Immunotherapy Assessment: Utilizing Immunological Synapse technology for precise drug discovery.

Preclinical Animal Models Dedicated to IO

Moving forward with candidates within the drug discovery journey requires robust and tailored in vivo models for IO:
●    Syngeneic Mouse Models: Developed with various tumor-immune phenotypes and responsiveness to checkpoint inhibitors; these models help in assessing therapeutic efficacy, modulation of the immune response (tumor microenvironment and periphery), as well as pharmacodynamics features (PK/PD).
●    Humanized Mouse Models: Presenting a functional human immune system with the possibility of xenografting a human tumor. These models are pivotal for particular therapeutic modalities such as biologic therapeutics (e.g., immune cell engagers) or cell therapies; they could also be interesting for early toxicology studies looking at immune-related adverse events.

Translational Validation Using Relevant Patient Samples

To bridge the gap between in vitro models done with primary human immune cells isolated from healthy donors and in vivo models in the drug discovery process:
●    Broad Clinical Network: Collaborations with hospitals and clinicians to select the relevant samples for the project.
●    Translational Validation of Therapies: Access to cancer patient samples ensures that therapies are effective in a patient context.
●    Enhanced Target Validation: Possibility to develop on-target biomarker assays or validate target expression in a particular indication.

Advanced Technological Integration

Our approach integrates cutting-edge technologies to de-risk your drug development strategy:
●    High-Throughput Imaging and Analysis: Tools like the ImageStream X and Operetta provide high-speed, high-resolution insights into immune cell interactions.
●    Complex Flow-Cytometry and Functional Assays: These analyses on fresh human tumor samples facilitate target engagement validation and biomarker identification.
●    Omics Technologies: Including scRNAseq, TCR sequencing, and proteomics, to uncover novel biomarkers and therapeutic targets.

Precision Medicine and Biomarker Strategies

By incorporating biomarker strategies early in the discovery process, we enhance the understanding of disease mechanisms and improve patient stratification for clinical trials. Our efforts in precision medicine are aimed at identifying the right therapeutic interventions for the right patients, thereby increasing the likelihood of clinical success.

The Evotec Advantage

Evotec’s experienced IO team supports your projects from the lab bench to the patient bedside. Our comprehensive services include:
●    Functional In Vitro Immunological Assays: Supporting programs for small molecules, biologics, and cell therapies.
●    In Vivo Rodent Models: Essential for preclinical testing of therapeutic efficacy and safety.
●    Translational Research and Patient Sample Access: Enhancing the relevance and translatability of preclinical findings.

Driving Success in Immuno-Oncology

Our integrated approach, combining deep immunological expertise with advanced technologies and translational capabilities, ensures that we address the complex challenges of IO drug discovery. By partnering with Evotec, you gain access to a wealth of knowledge and resources designed to propel your project forward, from the initial stages of discovery to the clinical validation of novel therapies.
For more information on how Evotec can support your Immuno-Oncology projects, visit our website or contact us at info@evotec.com.

Conclusion

The revolution in cancer immunotherapy is just beginning, and the journey to effective treatments requires collaboration, innovation, and expertise. At Evotec, we are committed to advancing the field of Immuno-Oncology, providing the tools and support necessary to transform groundbreaking discoveries into life-saving therapies. Join us in this mission and let’s make a difference in the fight against cancer.
Contact us today to speak with one of our immunologists and explore how we can progress your IO projects together.

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Targeting RNA represents a paradigm shift for drug discovery. The ability to seek out and destroy, or modify, a faulty RNA template, before the toxic protein has even been made, has only recently begun to be harnessed for the benefit of patients. 
At the time of writing, 21 Oligonucleotide drugs have been approved for human use, with an exponential increase in clinical trials and development projects involving this new modality. 
There exist several different mechanisms of action for Oligonucleotide drugs, all of which are transient and reversible and do not lead to alteration of patient DNA, unlike Gene therapy. 

Antisense Oligonucleotides harness endogenous systems already existing within a cell to achieve their purpose, with the only limitation being accessibility of the target tissue.
Once bound with great specificity to its RNA target, a short synthetic Oligonucleotide can trigger degradation, upregulation of the translated protein, or alteration of a splicing event leading to a correctly folded protein. Longer Oligonucleotides can fold into 3 dimensional shapes called Aptamers with similar target affinities and applications as antibodies, and shorter Oligonucleotides can act as MicroRNA   mimetics or antagonists to alter multiple targets or pathways concurrently with subtle but broader effect. 

The precision of an Oligonucleotide and its ability to correct a faulty RNA produced by an error in the genetic code, lends itself to applications in the fields of rare disease therapeutics and toxic gain of function mutations. The field of Oligonucleotide therapeutics is developing to address this as a whole and to pioneer a new preclinical and regulatory path that could be adapted for these unique disease biologies to make this type of therapeutic innovation more accessible. 

Evotec is a leader in integrated end-to-end Research and Development and has built substantial drug discovery expertise and technical capabilities that can drive new innovative and diverse modalities into the clinic. In addition, Evotec has developed a deep internal knowledge base in key therapeutic areas including neuroscience, pain, immunology, respiratory, women’s health, aging, fibrosis, inflammation, oncology, metabolic and infectious diseases. Leveraging these skills and expertise, Evotec successfully delivers on superior science-driven discovery and development alliances with pharmaceutical and biotechnology companies.

The global interest in this new modality area has led to high demand in Oligonucleotide synthesis and related chemistry applications, from modified Oligonucleotides to conjugates and complex formulations. 
Evotec offers Oligonucleotide research and development capabilities as well as ligand and linker chemistry expertise to support projects from discovery through to development.

In particular, since 2015 the discovery chemistry group is equipped with MerMade 48X synthesizers (Biosearch Technologies, see Fig. 1) with associated purification, post processing and QC platforms (at the Evotec sites of Toulouse and Verona, see Fig 2), while the development team, since 2022, is equipped with a Cytiva ÄKTA oligosyntTM synthesizer associated to the ÄKTA pureTM 150 and ÄKTA flux 6 purification platforms and with a Virtis Lyophilizer at Evotec’s state-of-the-art Verona site (see Fig. 3 to Fig. 6).
	Figure 1: Biosearch MerMade 48X	Figure 2: Agilent Preparative HPLC for purification
	Figure 3: Cytiva ÄKTA oligosyntTM	Figure 4: Cytiva ÄKTA pureTM 150
	Figure 5: Cytiva ÄKTA flux 6TM	Figure 6: SP Virtis Advantage Pro Lyophilizer
	Figure 7: UPLC with a Thermo ScientificTM Orbitrap ExplorisTM 120 mass spectrometer for oligonucleotides characterization

This fully integrated suite of capabilities allows for the synthesis, purification, isolation, and quality control of complex modified Oligonucleotides (ASOs, siRNAs, etc) on a scale from milligrams up to 25 g (up to 12 millimoles). The objective is to support Oligonucleotide drug discovery and development projects from the earliest phases of discovery, such as the generation of screening libraries, up to the selection of a preclinical development candidate followed by manufacture and release of material to support initial preclinical development studies. 

All these activities are supported by an experienced Oligonucleotide chemistry team operating across two sites and at different scales, to ensure flexible support for projects with highly efficient information and process transfer.
Evotec capabilities also include expert analysts for Analytical Development and QC, capable of developing and validating the analytical procedures needed for a full characterization and routine testing of Oligonucleotide drug substances up to and including IND enabling studies. In addition, Evotec’s support can encompass the release of preclinical batches according to regulatory requirements, including stability and formulation studies.

The journey to commercialization can be challenging. Scaling up production while maintaining process consistency, product quality, and regulatory compliance, requires expert process development capabilities, and the adoption of innovative science and risk management methodologies. A common pitfall for the Sponsor of an innovative therapy is to under-estimate the complexity and intricacy of this enterprise, which involves the coordinated optimization of strategies for process control, risk management, data management, and supply chain management.

With ever-evolving regulatory requirements and the increasing urge to shorten drug development timelines, getting your drug to market can seem like a daunting undertaking. That’s why taking some of the pressure off your organization by outsourcing your drug development and manufacturing activities to an expert partner can be the smartest decision. This will ensure your drug is commercialized in the fastest and most cost-efficient way possible, utilizing expertise, facilities, equipment, and processes to anticipate and overcome any challenges thrown at your program with ease.

Evotec offers an integrated end-to-end solution for innovative drug R&D, with the capabilities to support all phases of your drug development program. Your projects are in safe hands with our team of expert scientists who are pioneers in QbD, process design, scale-up, and validation, operating to full cGMP within FDA, MHRA, AIFA and BfArM approved facilities.

Our experts are just a click away! 

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Autoimmune diseases (ADs) represent a significant challenge in healthcare, affecting millions worldwide. These conditions arise when the body's immune system mistakenly attacks healthy tissues, leading to chronic inflammation and tissue damage. While current treatments for ADs focus on immunosuppression, they often come with significant side effects and provide only symptomatic relief. In recent years, there has been growing interest in cell-based therapies, particularly CAR (chimeric antigen receptor) cell therapy, as a potential solution to address the underlying causes of ADs.  

Autoimmune diseases encompass a wide range of conditions, including rheumatoid arthritis, lupus, multiple sclerosis and inflammatory bowel disease, among others. Despite their diverse manifestations, these diseases share a common underlying pathology of immune dysregulation. In healthy individuals, the immune system is finely tuned to distinguish between self and non-self antigens, but in autoimmune disorders, this distinction becomes blurred, leading to an attack on the body's own tissues. 

CAR cell therapy offers a promising approach to treating ADs by depleting disease-driving immune cells and rebalancing immune homeostasis. CAR T cells, which are engineered to express synthetic receptors targeting antigens expressed on the surface of pathogenic cells, have shown remarkable success in treating certain cancers. Now, researchers are exploring the potential of CAR T cells and their counterparts, CAR iNK (natural killer) cells, in targeting the aberrant autoreactive cells implicated in autoimmune diseases to reset the immune sytsem¹. We believe that cell therapy approaches could provide long-lasting drug-free remission and potentially a curative treatment for AD patients.

Understanding CAR Therapeutics

CAR T cell therapy first emerged as a groundbreaking treatment modality for hematological malignancies, such as leukemia and lymphoma. Building on this clinical success, researchers have recently turned their attention to applying CAR cell therapy to treat autoimmune diseases. 

In the context of ADs, CAR T cells have been investigated for their ability to target autoreactive T cells or B cells. B cells, in particular, play a central role in many autoimmune disorders by producing autoantibodies and driving inflammation. One promising target for CAR cell therapy in ADs is the cell surface protein CD19. High-level expression of CD19 is maintained throughout the majority of B-cell differentiation stages. By targeting CD19-positive B cells, CAR T cells or CAR NK cells can selectively eliminate the autoreactive B cell populations responsible for driving autoimmune responses². 

A recent early clinical study showed promising results for using CD19 CAR T cell therapy to treat ADs. Patients with systemic sclerosis, severe Systemic Lupus Erythematosus (SLE), and idiopathic inflammatory myositis showed remission following therapy³. However, CAR NK cell therapy may be beneficial over CAR T cell therapy, particularly in ADs that have dysfunctional T cells. CAR T cell therapy may also result in side-effects such as GvHD (Graft-versus-host disease, worsening the AD symptoms), neurotoxicity, and cytokine release syndrome; CAR NK cell therapy may offer a safer alternative⁴. 

Despite the potential of CAR cell therapy in ADs, several challenges remain, including scalability, persistence, and off-target effects. Evotec's innovative approach to addressing these challenges involves the use of induced pluripotent stem cells (iPSCs) to generate allogeneic off-the-shelf CAR iNK cells with enhanced scalability and precision. iPSCs, reprogrammed from adult somatic cells, offer a potentially inexhaustible source of immune cells that can be genetically engineered and differentiated into various cell types to tackle a range of diseases.

iPSC-Derived CD19 CAR iNK Cells for Targeted B Cell Depletion

A study by researchers at Evotec investigated the performance of iPSC-derived CD19 CAR iNK cells as a novel therapeutic approach for ADs. Using Evotec’s validated GMP iPSC line, researchers produced genetically modified cells with a knock-in of CD19-CAR. The modified cells successfully differentiated into CD19 CAR iNK cells using a feeder-free 3D differentiation process, which could be confirmed by flow cytometry. The established protocol can ensure high purity and functionality of the resulting cells. Results showed that iNKs generated from the GMP iPSC line were homogenous and phenotypically comparable to blood-derived (BD) NK cells form healthy donors.  

Figure 1: Cytotoxicity against SLE patient B cells. NK killing assays of effector cells - iNK without CAR (WT), CD19 CAR iNK or healthy donor BD NK cells (BD-NK) co-cultured 1:1 (E:T) with SLE patient B cells, + 10μg/ml anti-CD20 antibody (Obinutuzumab (Obi)) or isotype control (Iso) (n=2). 

In vitro experiments demonstrated the cytotoxic effector function of CD19 CAR iNK cells in selectively targeting and eliminating CD19-positive B cells. Co-culture assays using patient-derived primary B cells from patients with SLE autoimmune disease, showed robust cytotoxicity of CD19 CAR iNK cells. The CD19 CAR iNK cells were more efficient than iNK cells without a CAR or BD NK cells in depleting the patient-derived primary B cells. These findings highlight the therapeutic potential of CD19 CAR iNK cells in treating ADs, offering a targeted and scalable alternative to conventional immunosuppressive therapies.

Evotec's Scalable Therapeutics Approach

Evotec's commitment to allogeneic cell therapy innovation is exemplified by its scalable therapeutics approach, which leverages cutting-edge technologies and infrastructure to develop next-generation therapies for ADs. Central to this approach is the use of iPSCs as a platform for generating CAR iNK cells with enhanced scalability and precision. By introducing CARs targeting CD19 into iPSCs and differentiating them into iNK cells, Evotec aims to create scalable and precise off-the-shelf therapies for ADs. 

Figure 2: Evotec’s pipeline to co-create iPSC-based cell therapeutics with partners in Inflammation & Immunology.

Evotec's end-to-end process for iPSC-based therapeutics encompasses differentiation, gene editing, preclinical and clinical development, ensuring the efficient generation and characterization of CAR iPSC-derived cells. By utilizing validated GMP iPSC lines and GMP-compatible differentiation protocols, Evotec ensures the safety and quality of its allogeneic cell therapy products, paving the way for clinical translation.

Figure 3: Evotec’s end-to-end process for iPSC-based therapeutics. 

The scalability of Evotec's approach enables the production of large quantities of CAR iNK cells, making them suitable for widespread use in treating ADs. Additionally, the precision of iPSC-derived CAR iNK cells allows for targeted and personalized therapies tailored to individual patients' needs, reducing the risk of off-target effects and enhancing treatment efficacy.

Future Potential with Evotec

Evotec's iPSC-derived CD19 CAR iNK cells represent a promising new approach to treating autoimmune diseases. By harnessing the power of iPSC and CAR technology, allogeneic cell therapy can help revolutionize the treatment landscape for ADs, offering patients a targeted, scalable and potentially curative alternative to conventional therapies. 

As research in this field continues to advance, Evotec remains at the forefront of allogeneic cell therapy innovation, driving the development of next-generation treatments for ADs (Figure 2). Evotec’s GMP-compliant production pipelines provides an efficient, reproducible, and scalable way to produce CAR iNK cells derived from iPSC for clinical development. With its commitment to precision medicine and scalable therapeutics, Evotec is well-positioned to meet the growing demand for effective and accessible off-the-shelf therapies for autoimmune diseases.

See more iPSC-based Cell Therapies for I&I Diseases

To discover more about this research, download our scientific poster

References: 

(1) Blache, U.; Tretbar, S.; Koehl, U.; Mougiakakos, D.; Fricke, S. CAR T Cells for Treating Autoimmune Diseases. RMD Open 2023, 9 (4), e002907. https://doi.org/10.1136/rmdopen-2022-002907.

(2) Jin, X.; Xu, Q.; Pu, C.; Zhu, K.; Lu, C.; Jiang, Y.; Xiao, L.; Han, Y.; Lu, L. Therapeutic Efficacy of Anti-CD19 CAR-T Cells in a Mouse Model of Systemic Lupus Erythematosus. Cell. Mol. Immunol. 2021, 18 (8), 1896–1903. https://doi.org/10.1038/s41423-020-0472-1.

(3) Müller Fabian; Taubmann Jule; Bucci Laura; Wilhelm Artur; Bergmann Christina; Völkl Simon; Aigner Michael; Rothe Tobias; Minopoulou Ioanna; Tur Carlo; Knitza Johannes; Kharboutli Soraya; Kretschmann Sascha; Vasova Ingrid; Spoerl Silvia; Reimann Hannah; Munoz Luis; Gerlach Roman G.; Schäfer Simon; Grieshaber-Bouyer Ricardo; Korganow Anne-Sophie; Farge-Bancel Dominique; Mougiakakos Dimitrios; Bozec Aline; Winkler Thomas; Krönke Gerhard; Mackensen Andreas; Schett Georg. CD19 CAR T-Cell Therapy in Autoimmune Disease — A Case Series with Follow-Up. N. Engl. J. Med. 2024, 390 (8), 687–700. https://doi.org/10.1056/NEJMoa2308917.

(4) Műzes, G.; Sipos, F. CAR-Based Therapy for Autoimmune Diseases: A Novel Powerful Option. Cells 2023, 12 (11), 1534. https://doi.org/10.3390/cells12111534.

Inflammatory Bowel Disease (IBD) is the umbrella term for a group of diseases characterized by chronic, idiopathic and remitting inflammation of the gastrointestinal tract. Common symptoms include diarrhea, abdominal pain, and fatigue. However, extraintestinal manifestations such as inflammatory arthralgias/arthritides, primary sclerosing cholangitis (PSC), ocular or cutaneous involvement are often present and add to the complexity of the clinical picture.

The two most common forms of IBD are Crohn's disease and ulcerative colitis. Crohn's disease can cause inflammation anywhere in the gastrointestinal tract from the mouth to the anus, while ulcerative colitis is usually confined to the colon, where it can cause inflammation and ulceration. In about one in ten people, the two diseases cannot be clearly distinguished, and the condition is called indeterminate colitis. With approximately 6-8 million IBD patients worldwide (2 million in Europe and 1.5 million in North America), there is a huge unmet medical need.

IBD is an immune-mediated disease, but the exact causes are unknown. Crohn's disease, for example, involves a complex interplay of genetic predisposition, immune dysregulation, environmental influences, and microbial factors.

This complexity poses many challenges for drug development, as exemplified by the recent failure of a drug approved for ulcerative colitis that failed in late-stage clinical trials for Crohn's disease. The important lesson here is the need for proper patient stratification.

With its multimodal approach and patient stratification tools, Evotec is well positioned to develop innovative medicines. The Company is focused on development of IBD medicines through in-house research and through collaborations. Evotec’s strategy is modality agnostic, utilizing Evotec’s entire scope of technology platforms - from small molecules to biologics as well as iPSC-based cell therapies.

The Evotec approach

Drug discovery at Evotec always starts with patient data. Evotec's drug discovery efforts are based on its proprietary panOmics approach and a proprietary portfolio of molecular patient databases (E.MPD). panOmics combines both data generation and analysis platforms to industrialize OMICs data generation and AI/ML-based omics data analysis. Based on proprietary molecular patient data, panOmics fundamentally improves the understanding of disease processes, in vitro and in vivo disease modeling, identification of novel high value targets, biomarker discovery and patient selection.

Another integral aspect of drug development at Evotec is precision medicine. For this approach, Evotec has developed a comprehensive patient stratification toolbox via its panOmics-driven diagnostic approach – EVOgnostic.  

Therefore, IBD patients are an integral part of a pilot study in which Evotec is performing plasma and metabolomic analyses on samples from autoimmune disease patients to combine these clinical data with experimental data and data science approaches to identify potentially novel biomarker signatures.

Evotec aims to develop medicines that allow an early intervention and or, ideally, a cure for patients suffering from IBD. Evotec is engaged in several drug discovery programs tackling various aspects of IBD such as restoration of epithelial barrier function, modulation of inflammation, or resolution of intestinal fibrosis. Depending on the different aspects of the disease. Evotec’s experts are engaged in the IBD community and you can see our recent poster summarizing our IBD activities here. 

Selected collaborations

Evotec also is constantly seeking to enhance its capabilities through strategic investments and collaborations. For example, in 2022 it invested in IMIDomics Inc., a company focused on immune-mediated inflammatory diseases (IMIDs). IBD constitutes a large part of IMIDs. The aim is to jointly develop and use IMIDomics' Precision Discovery™ Engine. This technology enables a deep understanding of how inflammatory diseases work in patients. It applies a combination of clinical and computational expertise to clinical data and biological samples from more than 17,000 patients in a biobank, generating proprietary biomolecular signatures. 

With the Crohn's & Colitis Foundation, Evotec is advancing drug discovery for two novel IBD drug targets originating from academic research. The targets address fibrosis, the excessive accumulation of scar tissue in the intestinal wall, and the restoration of intestinal barrier function in IBD to reduce the increased intestinal permeability and chronic intestinal inflammation often seen in IBD patients.

Efficacy models in IBD

Another challenge in IBD is the lack of adequate animal models. While more than two dozen mouse and rat models of colitis have been developed and implemented, the multifactorial etiology and highly heterogeneous manifestations of the disease have prevented the development of a model that fully represents the pathophysiology of human IBD and related complications. Each mouse model has its strengths in elucidating the pathogenesis of colonic inflammation, fibrosis, or CAC, but each has a self-limiting nature and shows marked variability in drug development. While these IBD models cannot fully recapitulate the disease features commonly seen in humans (genetic and environmental influences, gut microbiota interactions, etc.), they have led to the identification of three key elements for disease etiology: T lymphocytes (T cells) mediate chronic intestinal inflammation; intestinal inflammation is initiated and maintained by certain commensal intestinal bacteria; the onset and severity of the disease is largely dependent on the genetic background.

Therefore, Evotec has carefully selected several preclinical models that recapitulate key aspects of IBD: inflammation, leukocyte trafficking, breakdown of epithelial barrier integrity, T cell-mediated injury. These models are routinely used and complemented by the current gold standard: the T cell transfer model of chronic colitis. This mouse model best reflects the clinical pathology observed in IBD and dissects the initiation, induction, and regulation of T cell-mediated immunopathology. 

In summary, Evotec is advancing breakthrough solutions for IBD using the most advanced technologies and platforms available. The Company's primary focus is on precision medicine and leading-edge approaches with the goal of providing tailored, effective, and minimally invasive treatments by taking into account the unique characteristics of each patient.

Download our poster from the IBD Innovate Conference

IBD is a collective term for a range of clinical phenotypes caused by chronic, idiopathic and remitting inflammation of gastrointestinal tract. Crohn’s disease and ulcerative colitis are the two most common forms. Despite many advancements in the treatment of IBD, there remains a high unmet medical need to provide patients with an early intervention of highly effective therapy, preferably with curative potential. 

Evotec is currently engaged in several drug discovery programs tackling various aspects of the disease such as restoration of epithelial barrier function, modulation of inflammation and intestinal fibrosis.

Given the heterogeneity of the disease, we actively invest into efforts leading to increased disease understanding and stratification of patients based on the disease endotypes.

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Evotec specializes in antifungal drug discovery with a focus on WHO and CDC priority human pathogens such as Aspergillus fumigatus, Candida spp, and Cryptococcus neoformans. Their track record in late-stage drug discovery is proven, aided by a range of established in vivo models of fungal infection that facilitate seamless progression from in vitro to in vivo studies.

Their expertise encompasses various areas, including in vitro compound characterization, invasive and localized in vivo fungal infection models, PK/PD mathematical modeling, and custom assay development tailored to individual project needs. They conduct comprehensive antifungal susceptibility testing to industry standards and offer specialized modalities such as combination testing to determine synergy or antagonism.

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