Sector	Type of NAMs	Endpoint and/or Use	Region	Author / Year	Comment
Cosmetic	Various in vitro and in silico models	NGRA	EU	[@349394]	Ab initio NGRA case study using NAMs (in vitro assays, PBK models, etc.) combined with other toxicity data (e.g., in silico predictions) to a hypothetical safety assessment of 0.1% coumarin in face cream and body lotion.
Cosmetic	Various in vitro and in silico models	NGRA	EU	[@349515]	This paper presents a toolbox and workflow designed for integrating NAMs in systemic safety assessments. The toolbox incorporates diverse NAMs, such as in vitro assays, PBK models, and in silico predictions. An associated workflow was developed to provide the Bioactivity Exposure Ratio (BER), facilitating the formulation of protective systemic safety decisions without reliance on animal data.
Cosmetic	QSAR Toxtree	NGRA	EU	[@349414]	This paper illustrates the practical application of TTC as a tool for safety assessment, examining case studies involving cosmetic ingredients (Perilla alcohol, Basic Blue 124, Trifolium pratense flowers) with low consumer exposure. It demonstrates how the TTC approach can be effectively applied to ensure the safety of cosmetic ingredients for which toxicity data are scarce, especially when exposure levels are low.
Cosmetic	Various NAMs including RxA and physiologically-based kinetic (PBK) modelling	NGRA	EU	[@349383]	This paper outlines a 10-step framework for implementing RxA in NGRA for cosmetics safety evaluation, particularly applicable when a TTC approach is not feasible. The framework is characterised by an exposure-driven approach and is grounded in Mode of Action (MoA) considerations.
Cosmetic	Various NAMs including RxA and PBK modelling	NGRA	EU	[@349413]	Caffein case study to demonstrates the practical effectiveness of a 10-step RxA framework in practice [@349383]. The authors demonstrate that the NGRA approach for caffeine is sufficiently conservative to protect human health.
Cosmetic	Various NAMs including RxA and physiologically-based kinetic (PBK) modelling	NGRA	EU	(Assaf [b@349585])	Propyl paraben case study to demonstrates the practical effectiveness of a 10-step RxA framework in practice [@349383]. The authors illustrate how NGRA, in this case, offers an improved and more accurate approach for assessing the skin sensitisation potential of substances such as propyl paraben. This method presents a significant improvement over traditional approaches, providing a more refined and reliable risk assessment process.
Cosmetic	Various NAMs including RxA	Skin sensitisation and NGRA	EU	[@349472]	NGRA case study using RxA to predict the skin sensitisation potential of 0.5% vanillin in shower gel and face cream.
Cosmetic	Various in vitro and in silico models	Skin sensitisation and NGRA	EU	[@349560]	NGRA case study using NAMs relevant for the skin sensitisation endpoint combined with novel computational modelling approaches to a hypothetical safety assessment of 0.1% coumarin in face cream and 1% in a non-spray deodorant.
Cosmetic	Read across and metabolomics	Skin sensitisation	EU	[@349487]	This paper discusses the safety evaluation of cosmetics using a RxA approach applied to metabolomics data derived from in vitro skin and liver models (RHE and HepaRG® cells).
Various	Various NAMs including -omics and physiologically-based kinetic (PBK) modelling			[@349574]	NGRA case study using transcriptomics data obtained from human liver spheroids combined with other in silico modelling (PBK and QIVIVE) to a hypothetical safety assessment of Perfluorooctanoic acid (PFOA) The authors also included an estimation of the in vitro free concentrations of PFOA and conducted a pathway enrichment analysis to identify the most sensitive molecular pathway associated with exposure to this chemical.
Various	Read across and metabolomics	Systemic Toxicity	EU	[@349400]	As part of the SEURAT-1 project, this report demonstrates how traditional read-across methods based on structural similarities between source and target substances, can be strengthened by integrating additional evidence from new approach data. This includes insights from "-omics" assays and computational models, ultimately enhancing the likelihood of regulatory acceptance.
Various	Hybrid in vitro/in silico approach	Endocrine disruptors	EU	[@349460]	Case study employing a combination of in vitro and in silico methods to assess the estrogenic potencies of the food mycotoxin ZEN and its metabolites.
Various	RxA workflow integrating NAMs	Various endpoints	EU	[@349462]	As part of the EU-ToxRisk project, this report illustrates a structured RxA workflow that relies on existing experimental and in silico data. It demonstrates how newly generated NAM data can be integrated to substantiate the RxA by testing in a systematic way toxicodynamic and -kinetic properties. The project identified key elements essential for effective RxA and devises strategies to address potential challenges.
Various	In vitro	Endocrine disruptors	EU	[@349516]	This paper illustrates the successful validation, approval and subsequent inclusion in the OECD TG 458 of an in vitro method (AR-CALUX®) for the categorisation of (ant)agonism of the androgen receptor.
Various	In vitro	Endocrine disruptors	US	[@349436]	This study investigated the comparative sensitivity of 16 in vitro assays for estrogen agonist activity using a diverse group of compounds sourced from the USEPA ToxCast dataset. The authors highlight the importance of employing multiple assays with various molecular initiation and signaling events to inform selection, application, and interpretation of in vitro endpoint responses during diagnostic applications.
Various	In vitro	Biokinetics predictions	EU	[@349555]	Workshop report that analyses the available NAMs for human-relevant biokinetics and the developments that are needed to increase regulatory use and acceptance of NAMs for biokinetics in risk evaluations
Various	Physiologically based kinetic (PBK) modeling	Biokinetics predictions	EU	[@349384]	This paper illustrates three case studies (ibuprofen, amiodarone, and chlorpyrifos) to assess the extent drug-induced side effects or chemical-induced adverse effects could be quantitatively predicted using in vitro data. The study showed the applicability of QIVIVE and its reliability when compared against in vivo data. The authors identified three key points: Careful consideration of in vitro conditions compared to in vivo situations, particularly regarding protein binding. Accounting for the inhibition of metabolising enzymes by formed metabolites in vitro. Extrapolating from the measured intracellular concentration in vitro, rather than the nominal concentration, to estimate tissue/organ concentration for relevant QIVIVE regarding adverse effects.
Various	Battery of in vitro tests	DNT / screening/prioritisation	EU	[@349393]	Workshop report that analyses the current landscape of in vitro DNT testing, validation and readiness of NAM-based approaches and the development of an IATA designed for DNT screening and prioritisation purposes. The conclusion offers recommendations for test readiness criteria, highlighting the challenge of covering potential adverse outcomes, some of which are not fully characterised in terms of mechanism of action or adverse outcome pathways (AOP).
Various	Battery of in vitro tests	repeated dose toxicity (RDT) to key organs and developmental toxicity (DART)	EU	[@349503]	Communication from the EU-ToxRisk project, presenting a case study which evaluated 23 in-vitro test methods for assessing repeated dose toxicity to key organs (RDT) and developmental toxicity (DART) using 19 compounds. The authors identified important issues that requiring further development: Using readiness criteria of test methods as a basis for fit-for-purpose evaluations. enhancing transparency in (meta)data handling and processing better defining and documenting procedures for test compound management, and clearly defining study procedures' objectives before initiation in a traceable manner (e.g., pre-registration).
Food	Whole genome sequencing (WGS)	characterisation of foodborne pathogens	US	[@349385]	Review by FDA presenting evidence of the varied and impactful roles that genomic sciences play in advancing food safety. The paper explores how genomic techniques are applied to enhance food safety by understanding the genetic makeup of foodborne pathogens. The goal is to improve detection, prevention, and control strategies for foodborne illnesses.
Food	(Q)SAR models	genotoxicity and carcinogenicity / screening	EU	[@349417]	This paper evaluates the effectiveness of non-commercial (Q)SAR models for predicting genotoxicity and carcinogenicity within EFSA's databases. The study finds that (Q)SAR models exhibit strong predictive capabilities for genotoxicity, particularly in terms of the bacterial reverse mutation test, achieving an accuracy rate of nearly 90%. However, the predictive accuracy for in vivo micronucleus tests falls short, hovering around 50%. Regarding carcinogenicity assessment, the best models demonstrate an accuracy rate close to 70% in prediction. Additionally, the paper offers an example illustrating the potential application of in silico models for conducting preliminary screenings of the genotoxicity properties of botanicals intended for use as food supplements.
Pesticides	PBK modeling and QIVIVE	NGRA	EU	[@349422]	Case study on Fenitrothion (FNT) to predict dose levels that would not pose a risk of acute neurotoxicity in humans due to erythrocyte AChE inhibition resulting from oral exposure to FNT. This study employed a PBK modelling for humans and rats, coupled with QIVIVE. The research aimed to support Next Generation Risk Assessment, showing promise in deriving PODs for the risk and safety evaluation of organophosphate pesticides in line with the 3R principles. Additionally, it could potentially facilitate the development of a generic PBK model capable of predicting acute toxicity for numerous organophosphate pesticides.
Pesticide	Battery of in vitro tests and in silico analysis combined with transcriptomics	NGRA	EU	(Sprenger et al., 2022)	Case study on the pesticidal active substances (imazalil, thiacloprid, and clothianidin) to predict genotoxicity using a battery of in vitro tests and omics techniques. This study illustrated possible challenges resulting from the use of batteries of new approach methods and omics techniques. Transcriptome analysis from animals treated with the three pesticidal compounds indicated genotoxicity in rat liver for clothianidin (CTD). In contrast, CTD was positive in two out of three in vitro clastogenicity assays and negative in a number of follow-up in vivo tests. Computational approaches supported the assumption of a clastogenic potential for CTD. However, transcript signature of CTD in human HepaRG liver cells and Comet assay did not show a clear-cut classification of CTD as genotoxic or non-genotoxic. The authors employed a WoE decision-making approach, considering regulatory studies alongside findings in human cells and the exceptionally high doses of CTD administered in the animal study used for the transcriptomic analysis. They concluded that CTD most likely does not pose a genotoxic risk to consumers exposed to residues of CTD via the diet.