# Mutational profiling in human cells as an in vitro alternative to in vivo mutagenicity assessments

> **NIH NIH R43** · INTEGRATED LABORATORY SYSTEMS, LLC · 2021 · $249,903

## Abstract

Project Summary
A significant shift in the safety assessment of new drug candidates and environmental chemicals that can
revolutionize the practice of regulatory toxicology is ongoing. This shift includes a reduction, or in some cases
elimination, of traditional toxicity testing in animals with the implementation of higher-throughput testing schemes
using human cell systems. Knowledge regarding the potential of new chemicals, food additives, and
pharmaceuticals to damage the human genome and cause mutations remains critical to public health. Mutations
are heritable changes in the cellular genome and are key events in the induction of cancer, birth defects, and
neurological diseases. Screening chemicals for their potential to cause mutation in a human relevant assay offers
an effective strategy for improving public health. The lack of a mutagenicity bioassay in human cells is a major
data gap in the genetic toxicology test battery used by regulatory agencies for hazard identification and
quantitative risk assessments. Our novel methods, initially developed to identify genetic subclones within
cancers, permits analytical assessment of mutagenicity and precise quantification of these very rare events (1
in 100,000 - 1,000,000). These methods can be integrated with human cells as a genetic toxicology assay that
can replace 50 yr. old clonal selection techniques to assess mutation. Although duplex sequencing is highly
innovative and a “game changer”, without integration into a well-defined human cell-based system and careful
validation studies, its application in regulatory Genetic Toxicology will be limited. The focus of this SBIR Phase
I application is to develop a New Approach Methodologies (NAM) combining human cells and duplex sequencing
as an in vitro alternative to in vivo mutation assays. We will accomplish this by conducting “proof of principle”
experiments using a well-established human cell line, as outlined in two specific aims. In Specific Aim 1, we will
determine the time course for the induction of mutations at multiple loci using duplex sequencing for a prototypic
mutagenic compound. In Specific Aim 2, we will determine the dose response for the induction of mutations at
multiple loci using duplex sequencing for two prototypic mutagenic compounds. Completion of this Phase I SBIR
will lead to development of a human cell-based mutation assay that can be used as a follow up to bacterial
mutation assays and as a NAM to reduce reliance on current in vivo gene mutation assays in rodents. This data
will also support a Phase II application to validate this assay and adapt these methods to quantify mutation in
human CD34+ cells and HepaRG™ cells.

## Key facts

- **NIH application ID:** 10155923
- **Project number:** 1R43ES032751-01
- **Recipient organization:** INTEGRATED LABORATORY SYSTEMS, LLC
- **Principal Investigator:** James Todd Auman
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $249,903
- **Award type:** 1
- **Project period:** 2021-02-17 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10155923

## Citation

> US National Institutes of Health, RePORTER application 10155923, Mutational profiling in human cells as an in vitro alternative to in vivo mutagenicity assessments (1R43ES032751-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10155923. Licensed CC0.

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