Summary of Progress (18 month)

A strong evidence base for the effective and safe use of iPSC-based strategies for regenerative treatment of IDD, restoring the IVD to a functional unit.

Standard operating procedures (SOPs) were developed to isolate disc progenitors, where after, by the emerging non-integrative RNA-based reprogramming strategies, iPSC were obtained and iPS-notochordal- like cells (iPS-NLCs) were generated. Much effort was placed by the Partners on consensus methodology optimisation and standardisation of culture, tools and methods for cell, tissues, and biomaterial characterisation. Importantly, culture conditions and induction methods of disc degeneration ex vivo were established to mimic the environment of the healthy and degenerated disc. These will be used in the battery of in vitro and ex vivo models with increasing complexity mimicking the in vivo environment employed in iPSpine. Collaborative efforts merged the expertise of partners to develop and characterise (semi)synthetic, natural, and hybrid biomaterials that meet the demands of the degenerated disc environment.

Enabling technologies for the development, characterisation, and validation of the iPSCbased strategy.

Different complementary methods and tools are developed and optimised including flow cytometry, immunostainings, DigiWest proteomics technology and transcriptomic analyses, iPS reporter cell lines to assess iPS-NLC differentiation efficiency, phenotypic identity, and purity. Furthermore, RNA-based non-integrating reprogramming for the generation of GMP-compliant iPSC lines was developed. By making the intracellular delivery (iTOP) technology compatible with the adapted CRISPR-Cas9 systems, a game changer will be introduced for (epi)genetic reprogramming of (pluripotent) stem cells enabling lineage commitment. Furthermore, novel equipment was designed and construction of the portable spine biomechanics tester and the dynamic loading bioreactor for the human disc tissue has begun. A deep machine learning algorithm for staging disc degeneration/regeneration is being developed for objective evaluation of the iPSpine therapy at the tissue level. With the iPSpine therapy as point of convergence, partners worked on identifying the key aspects and parameters that must be addressed for the safety of ATMPs that are employing iPS-NLCs, as an example.  Lastly, Engineering requirements for the open access knowledge-sharing platform have been established.

A recommendation on ethical and regulatory affairs to enhance and accelerate further development and implementation of iPSC-based therapeutic strategies.

Hard and soft impacts were mapped to widen the understanding of the ethical implications of stem cell research at different levels (scientists, regulators, and patients). Furthermore, two focus groups with stem cell iPSpine researchers were conducted and a study started on the evolving policy frameworks governing stem cell research. The Partners received training by regulatory experts and in the iPSpine research activities, regulatory advice was embedded to guide the consortium through the regulatory field of ATMP during development up to non-clinical proof of concept (PoC). Hereby, iPSpine envisions to set an example on long term, fast forwarding, preclinical research that is closer to the patient in need. This is enhanced by the open digital management platform that guides the design of preclinical studies on a high-level that are conducted in a battery of increasing complexity of in vitro, ex vivo, and in vivo studies for advanced therapies based on regulatory requirements. In preparation for advice by the authorities, a template Candidate ATMP Report was drafted that will also assist in identifying any gaps in the development of the iPSpine therapy. Since the project start, its structure was established including project management (i.e. scientific, financial, risk, ethical, IP aspects) and the executive steering committee. The Partners have been developing channels for dissemination and means for effective communication and engagement of stakeholders. We have developed the iPSpine website and built a social media presence (Twitter and LinkedIn). The Patient Advisory Board (PAB) was established as one of its first tasks, the PAB, together with the Consortium, developed an animation to tell the iPSpine story . These dissemination activities and channels, along with utilization of the stakeholder relationships, have set the stage for effective, engaged dissemination and communication for the remainder of the project (and stages beyond).

Progress beyond the state of the art, expected results until the end of the project and potential impacts.

Upon successful implementation of the iPSpine therapy, we envision improved outcome and quality of life for patients with disc-related LBP, through effective regeneration of the degenerated tissue with restored biomechanical function of the disc, and long-lasting reduction of LBP. Altogether, this will result in reduced LBP-related premature retirement and improved socio-economic contribution.  The novel and extended knowledge, tools and technologies that will be the outcome of the iPSpine project, will provide unique platforms of knowledge, testing, and exploitation with broad applicability. These platforms will contribute to acceleration, innovation, and implementation of novel technologies and ATMPs for treatment of large patient groups. Hereby, iPSpine realizes its ambition to make a significant contribution by reducing translational bottlenecks through open innovation and taking European leadership in the development of ATMPs.