HLA-homozygous iPSC-cardiomyocytE Aggregate manufacturing technoLogies for allogenic cell therapy to the heart

Project Information

HEAL

Grant agreement ID: 101056712

DOI

10.3030/101056712

EC signature date 23 May 2022

Start date 1 September 2022

End date 28 February 2027

Funded under

• Health

Total cost

€ 5 824 074,75

EU contribution

€ 5 824 074,00

Coordinated by

MEDIZINISCHE HOCHSCHULE HANNOVER
Germany

Periodic Reporting for period 2 - HEAL (HLA-homozygous iPSC-cardiomyocytE Aggregate manufacturing technoLogies for allogenic cell therapy to the heart)

Reporting period: 2024-03-01 to 2025-08-31

Summary of the context and overall objectives of the project

Background
The project “HLA-homozygous iPSC-cardiomyocyte Aggregate manufacturing technologies for allogenic cell therapy to the heart” (HEAL) is focused on overcoming numerus hurdles to human induced pluripotent stem cell (iPSC)-based therapies to heart failure, which remains a major global cause of morbidity and mortality.

Aims of the project
The overarching goal is to establish new tools, technologies and expertise for accelerating the development of advanced iPSC-based therapies. The project also aims at overcoming scientific, regulatory and in particular safety hurdles, necessary to initiate a first in-man (FIM) clinical trial for the therapeutic administration of allogeneic, human leukocyte antigen (HLA)-homozygous iPSC-CMAs, for improving the function of damaged hearts in heart failure patient for whom very few alternative therapies currently exist. Project outcome will facilitate decision-making by authorities and clinicians in a secure and ethical manner, for respecting individual integrity and underpinned with public acceptance and trust.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

WP 1:
• Created cGMP-compliant culture and differentiation media with 12-18 month shelf-life
• Achieved scalable iPSC expansion and cardiomyocyte production from 20mL to 500mL cultures
• Established robust residual iPSC assay, developed potency assay
• Created GMP-compliant SOPs, developed 24-hour cold transport protocols
• Successfully achieved cryopreservation

WP2:
• established standardized quality control protocols
• Adapted co-culture protocols to assess T cell alloresponses
• enabled standardized immune profiling protocols across multiple research sites, and created reference datasets for AI-based immune prediction algorithms
• Provided essential immune data for developing predictive models and advancing clinical translation of HLA-homozygous iPSC-CM therapies with robust immune characterization and standardized protocols

WP3:
• developed comprehensive mutation categorization system for therapeutic safety
• Successfully created HLA-homozygous iPSC suicide lines
• Developed scalable GMP-compliant gene, and validated production protocols for iPSC-derived cardiomyocyte aggregates
• Completed tumorigenicity studies in NSG mice and pig models, demonstrated no life-threatening arrhythmias in animal studies, and established comprehensive quality control for clinical-grade cell products

WP4:
• Established Target Product Profile (TPP), and created structured documentation system for IMPD development
• Completed preliminary documentation for PEI Scientific Advice request
• Developed SOPs for toxicity and carcinogenicity studies
• Established regulatory-ready documentation for clinical trial application

WP5:
• Conducted comprehensive FTO analysis, and established ongoing IP monitoring and strategic guidance
• Optimized GMP-compliant iPSC banking process to produce banks of up to 360 cryovials
• Created cost-effectiveness analysis framework for therapeutic development

WP6:
• Organized General Assembly meetings and quarterly consortium meetings, and maintained transparent communication with EU authorities
• Managed project website with multimedia content, organized HEAL symposium at GSCN 2024 conference, produced 24 scientific publications and multiple conference presentations, and maintained active social media presence across LinkedIn, X, and YouTube
• Implemented comprehensive risk monitoring and mitigation framework
• Maintained comprehensive project documentation on SharePoint, and ensured timely regulatory submissions and progress reports

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

- PoC for the newly designed ACF CM differentiation formulation demonstrated for R26_6 cells
- Substantial process upscaling for iPS-CMA production in STBRs achieved.
- SOPs development resulted into publication of “Standardized production of hPSC-derived cardiomyocyte aggregates in stirred spinner flasks”; Kriedemann et al. Nat. Prot. (in print 2024).
- Central tumorigenicity assay established and validated.
- Novel 60- and 354-marker single-cell level deep immunophenotyping prototype established.
- Defining high rate of cancer mutations in progenies of hPSC published in Nat. Biotech. doi: 10.1038/s41587-023-02090-2.
- Unique tool demonstrating the epigenetic status of hPSC published in Nat. Prot. doi: 10.1038/s41596-023-00898-5).

llustration of the preclinical platform for IPS-CMA developments and the assays and tools generated

Safety Measures for Heart Repair using HLA-homozygous iPSC-derivde cardiac aggregates

Bioprocess development for iPS-CMA production applying calculated process upscaling in Stirred Tank