An advancement in developmental and reproductive toxicity (DART) risk assessment: evaluation of a bioactivity and exposure-based NAM toolbox

Traditional chemical safety assessment involves identifying the lowest level of a chemical that impacts endpoints measured in standardized animal studies together with appropriate safety factors to establish human exposure limits. In vitro assays have shown promise in providing points of departure that can be protective of human health when combined with exposure predictions into a bioactivity:exposure ratio (BER). Using a combination of broad screening tools and DART-targeted assays, we previously demonstrated high biological coverage of this NAM toolbox against a list of DART-relevant genes and pathways. To fully transition to an animal-free paradigm, it is crucial to establish confidence that these in vitro assays sufficiently represent the DART toxicity mechanisms, ensuring a level of protection that is safe for non-pregnant adults, pregnant women, and fetal populations. In this proof-of-concept study, we have extended the toolbox to include additional in vitro and in silico tools a..., , # An advancement in developmental and reproductive toxicity (DART) risk assessment: evaluation of a bioactivity and exposure-based NAM toolbox

**Contents**

The NAM toolbox described in the manuscript is composed of four new approach methodology (NAM) platforms. These are:

1.      In vitro pharmacological profiling (IPP) to assess activity of chemicals against receptors of pharmacological interest

2.      A cellular stress panel (CSP) to assess potential of chemicals to induce chemical stress

3.      The ReproTracker assay to assess the perturbation of early embryonic development in cardiomyocyte, hepatocyte-like (HLC) and neuronal cell differentiation from human induced pluripotent stem cells (hiPSCs).

4.      The devTox quickPredict assay to predict the concentration which elicts developmental toxicity using hiPSCs.

5.      The H295R steroidogenesis assay with AR/ER CALUX detection described in OECD TG 456.

6.      HTTr metadata of generated data using the TempO-Seq platform (r...,