Report of the First ONTOX Stakeholder Network Meeting: Digging Under the Surface of ONTOX Together With the Stakeholders.

The first Stakeholder Network Meeting of the EU Horizon 2020-funded ONTOX project was held on 13-14 March 2023, in Brussels, Belgium. The discussion centred around identifying specific challenges, barriers and drivers in relation to the implementation of non-animal new approach methodologies (NAMs) and probabilistic risk assessment (PRA), in order to help address the issues and rank them according to their associated level of difficulty. ONTOX aims to advance the assessment of chemical risk to humans, without the use of animal testing, by developing non-animal NAMs and PRA in line with 21st century toxicity testing principles. Stakeholder groups (regulatory authorities, companies, academia, non-governmental organisations) were identified and invited to participate in a meeting and a survey, by which their current position in relation to the implementation of NAMs and PRA was ascertained, as well as specific challenges and drivers highlighted. The survey analysis revealed areas of agreement and disagreement among stakeholders on topics such as capacity building, sustainability, regulatory acceptance, validation of adverse outcome pathways, acceptance of artificial intelligence (AI) in risk assessment, and guaranteeing consumer safety. The stakeholder network meeting resulted in the identification of barriers, drivers and specific challenges that need to be addressed. Breakout groups discussed topics such as hazard versus risk assessment, future reliance on AI and machine learning, regulatory requirements for industry and sustainability of the ONTOX Hub platform. The outputs from these discussions provided insights for overcoming barriers and leveraging drivers for implementing NAMs and PRA. It was concluded that there is a continued need for stakeholder engagement, including the organisation of a 'hackathon' to tackle challenges, to ensure the successful implementation of NAMs and PRA in chemical risk assessment.

OECD workshop consensus report: Ethical considerations with respect to human derived products, specifically human serum, in OECD test guidelines.

The ethical needs and concerns with use and sourcing of human materials, particularly serum, in OECD in vitro test guidelines were explored in a dedicated international workshop held in 2019. The health-related aspects of the donation procedure, including tissue screening, donor health, laboratory work health protection, permission from the donor for commercial use, payment of the donors and the potential for exploitation of low-income populations and data protection of the donors; supply, availability, and competition with clinical needs; traceability of the serum and auditability/GLP needs for the Test Guideline Programme, were examined. Here we provide the recommendations of the workshop with respect to the use of human serum, and potentially other human reagents, specifically with regard to test method development for OECD Test Guideline utility as part of the Mutual Acceptance of Data requirement across all OECD member countries. These include informed donor consent terminology, a checklist of human serum information requirements to be included with the Good Laboratory Practise report, and suitable sources for human serum to ensure waste supplies are used, that can no longer be used for medical purposes, ensuring no competition of supply for essential medical use.

In vitro and integrated in vivo strategies to reduce animal use in genotoxicity testing.

Scientific, financial, and ethical drivers have led to unprecedented interest in implementing human-relevant, mechanistic in vitro and in silico testing approaches. Further, as non-animal approaches are being developed and validated, researchers are interested in strategies that can immediately reduce the use of animals in toxicology testing. Here, we aim to outline a testing strategy for assessing genotoxicity beginning with standard in vitro methods, such as the bacterial reverse mutation test and the in vitro micronucleus test, followed by a second tier of in vitro assays including those using advanced 3D tissue models. Where regulatory agencies require in vivo testing, one demonstrated strategy is to combine genotoxicity studies traditionally conducted separately into a single test or to integrate genotoxicity studies into other toxicity studies. Standard setting organisations and regulatory agencies have encouraged such strategies, and examples of their use can be found in the scientific literature. Employing approaches outlined here will reduce animal use as well as study time and costs.

Limitations and uncertainties of acute fish toxicity assessments can be reduced using alternative methods.

Information about acute fish toxicity is routinely required in many jurisdictions for environmental risk assessment of chem­icals. This information is typically obtained using a 96-hour juvenile fish test for lethality according to OECD test guideline (TG) 203 or equivalent regional guidelines. However, TG 203 has never been validated using the criteria currently required for new test methods including alternative methods. Characterization of the practicality and validity of TG 203 is important to provide a benchmark for alternative methods. This contribution systematically summarizes the available knowledge on limitations and uncertainties of TG 203, based on methodological, statistical, and biological consider­ations. Uncertainties stem from the historic flexibility (e.g., use of a broad range of species) and constraints of the basic test design (e.g., no replication). Other sources of uncertainty arise from environmental safety extrapolation based on TG 203 data. Environmental extrapolation models, combined with data from alternative methods, including mechanistic indicators of toxicity, may provide at least the same level of environmental protection. Yet, most importantly, the 3R advan­tages of alternative methods allow a better standardization, characterization, and an improved basic study design. This can enhance data reliability and thus facilitate the comparison of chemical toxicity, as well as the environmental classifi­cations and prediction of no-effect concentrations of chemicals. Combined with the 3R gains and the potential for higher throughput, a reliable assessment of more chemicals can be achieved, leading to improved environmental protection.

A multi-faceted approach to achieving the global acceptance of animal-free research methods.

In 2015, the PETA International Science Consortium Ltd. was awarded the Lush Training Prize for its broad approach to education and training on the effective use of human-relevant, non-animal research techniques. The prize was awarded for work that included hosting workshops and webinars, initiating in-person training sessions and developing educational resources. The Consortium works closely with industry and regulatory agencies to identify and overcome barriers to the validation and use of alternatives to animal testing, by using an approach that identifies, promotes and verifies the implementation of these methods. The Consortium's recent activities toward replacing tests on animals for nanomaterials, pesticides and medical devices, are described, as examples of projects with broad applicability aimed at large-scale regulatory change.

A campaign to end animal testing: introducing the PETA International Science Consortium Ltd.

The successful development and validation of non-animal techniques, or the analysis of existing data to satisfy regulatory requirements, provide no guarantee that this information will be used in place of animal experiments. In order to advocate for the replacement of animal-based testing requirements, the PETA International Science Consortium Ltd (PISC) liaises with industry, regulatory and research agencies to establish and promote clear paths to validation and regulatory use of non-animal techniques. PISC and its members use an approach that identifies, promotes and verifies the implementation of good scientific practices in place of testing on animals. Examples of how PISC and its members have applied this approach to minimise the use of animals for the Registration, Evaluation, Authorisation and Restriction of Chemicals regulation in the EU and testing of cosmetics on animals in India, are described.

Using routinely gathered data to evaluate locally led service improvements.

Background Between 2009 and 2010 NHS Ealing tested the feasibility of a) combining data from more than one data-domain at the same time to quantify patient movement across the primary care/acute hospital boundary, and b) establishing online analyses so they can be constantly updated with near real-time data to compare different subsets of patients. The reports allowed us to see: changes in hospital admissions before and after referral to community matrons of patients with complex conditions from one practice-based commissioning (PBC) groupchanges in hospital bed-days of all patients from one practice or PBC group during a complex intervention designed to assist inter-disciplinary collaboration. Results The teams leading the projects found that the reports gave them confidence in the projects and helped to influence local policy. Discussion GP consortia need to evaluate complex service improvements in order to contain costs and improve quality. They will find such reports helpful to give ongoing feedback and this may help to keep people engaged. Present plans for data warehousing in London do not have this ability - they do not combine data from across the whole health economy, and are focused either on claims validation or risk stratification.

Inter-organisational communication for patients being case managed by community matrons.

Background Poor communication between community matrons (CMs), in-hours and out-of-hours (OoH) general practitioners (GPs) causes uncertainty and inefficiencies. Setting A practice-based commissioning group in West London and the associated CMs who case manage high users of hospital services. Question What helps good communication between CMs, GPs and OoH services to ensure that the right patients are case managed and hospital admissions are avoided? Methods Whole system participatory action research, with four stages: 1) identify communication problems as perceived by a wide range of stakeholders; 2) draw a diagram of the existing communication system, and with stakeholders redraw this to overcome its weaknesses; 3) pilot the changes proposed; 4) gain consensus among stakeholders about policy. Results Stakeholders agreed that standards should be adopted to improve communication for the care of patients who are case managed by CMs. Routine passage of information between GP, CMs and the OoH services would achieve this, and is feasible. Specifically: routine information (termed Special Patient Notes) should be sent to the OoH service about vulnerable patients, including those who are case managed by CMsclear information about CM attachment to general practices and how to refer to them should be easily accessibleGPs and CMs should meet quarterly for mutual learning and to discuss patientsthe OoH service electronically should cascade information to GPs, CMs and others named in the Special Patient Notescommissioners should routinely gather data to compare clusters of general practices for i) referrals to CMs, ii) posting Special Patient Notes, iii) unscheduled consultations and hospital admissions of all patients including those being case managed. Discussion This project revealed system-wide communication problems for the care of patients being case managed by CMs, and ways to overcome these. Commissioners could insist that these are adopted locally, and gather data to prompt compliance and evaluate the consequential cost savings.