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Analysis of Off-target Effects and Risk Assessment Leading from Preclinical to Clinical Trials of Gene-edited Therapeutic Products

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Abstract

Objective

Concerns about off-target effects (OTEs) of genomic DNA cleavages by gene-editing enzymes have been raised, especially for OTEs that go undetected due to technical limitations. Since no explicit guidelines have been in place for risk assessment of OTEs, the regulatory authorities’ concept of an acceptable evaluation scheme for OTEs in the investigational drug application (IND) has not been clear. Here, we clarify the regulatory expectations by examining reports on OTE evaluations of leading gene-editing products that have achieved IND clearance.

Methods

We collected and analyzed the gene-editing products that have entered clinical trials by searching on ClinicalTrials.gov and EU-Clinical-Trial-Registries, and related reports for OTE evaluations from Google Scholar, PubMed, and the developers’ websites.

Results

We found a common two-step verification method used for different products at the preclinical stage. First, numerous potential off-target loci (POLs) are listed with state-of-the-art high-sensitivity detection methods and theoretical screens; Second, these OTEs are checked by amplicon sequencing of the POLs after treatment by enzymes in in vitro models close to clinical use conditions. Only the OTEs that can be detected and verified are addressed in the risk assessment in the translational phase from preclinical to clinical study.

Discussion

Here, we describe a clear scheme for risk assessment of OTEs at the key translational phase, based on the common features in protocols for gene-editing products that were cleared for use in clinical trials. This report will provide a guide for those newly attempting to conduct clinical development in this field.

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Authors and Affiliations

  1. Cooperative Major in Advanced Biomedical Sciences, Joint Graduate School of Tokyo Women’s Medical University and Waseda University, TWIns, 8-1 Kawada-Cho, Shinjuku-Ku, Tokyo, 162-8666, Japan

    Naoki Yamada & Atsushi Aruga

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  1. Naoki Yamada
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NY contributed to the study design, analysis, and interpretation, and he agrees to be accountable for all aspects of this work’s integrity and accuracy. AA contributed to the study design and interpretation, and he agrees to be accountable for all aspects of this work’s integrity and accuracy.

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Correspondence to Atsushi Aruga.

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Conflict of interest

Naoki Yamada is an employee of Fujifilm Pharmaceuticals U.S.A. Inc. However, the views of the author in this paper do not reflect the views of the company. Atsushi Aruga has no conflicts of interest to disclose.

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Yamada, N., Aruga, A. Analysis of Off-target Effects and Risk Assessment Leading from Preclinical to Clinical Trials of Gene-edited Therapeutic Products. Ther Innov Regul Sci 57, 538–551 (2023). https://doi.org/10.1007/s43441-022-00481-2

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