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A Fit-for-Purpose Method to Measure Circulating Levels of the mRNA Component of a Liposomal-Formulated Individualized Neoantigen-Specific Therapy for Cancer

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Abstract

Autogene cevumeran is an individualized neoantigen-specific therapy (iNeST) under development for the treatment of various solid tumors. It consists of an RNA-Lipoplex (RNA-LPX) in which the encapsulated mRNA molecule encodes up to ten neoepitopes identified from each individual patient. In association with major histocompatibility complex (MHC) class I and MHC class II, these neoantigens can potentially stimulate and expand neoantigen-specific CD4+ and CD8+ T cells, leading to antitumor responses. As part of the pharmacokinetic (PK) property assessment of Autogene cevumeran in patients, both the lipid and mRNA content in circulation are measured. This work focused on our efforts to establish a sensitive and robust method for the measurement of mRNA levels of RNA-LPX in plasma. Due to the chemical characteristics of mRNA, extra precautions are required in order to effectively preserve mRNA integrity in human plasma during sample collection, handling and storage. To this end, a number of sample collection tubes and storage conditions were evaluated in order to inform the most optimal and operationally feasible conditions by which to preserve mRNA integrity during sample collection and upon freeze–thaw. PAXgene Blood ccfDNA tubes successfully prevented mRNA degradation and were subsequently selected for patient sample collection in the clinical trial. A branched DNA (bDNA)-based mRNA PK assay was developed to achieve the desired assay performance. Here, we discuss the evaluation of various sample collection and processing conditions as well as the optimization of the work flow during bDNA PK method development.

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Acknowledgements

We want to thank Chris Petry and Sara Wichner for providing critical reagents. The graphical abstract was designed using BioRender.

Funding

All work was funded by Genentech Inc., a member of the Roche group.

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Authors and Affiliations

  1. Department of BioAnalytical Sciences, Genentech Inc., 1 DNA Way, South San Francisco, California, 94080-4990, USA

    Sebastian Guelman, Ying Zhou, Ann Brady & Kun Peng

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  1. Sebastian Guelman
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  2. Ying Zhou
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  3. Ann Brady
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Contributions

All authors contributed to the experimental design and data analysis. YZ and AB did the experimental work. SG and KP wrote the manuscript.

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Correspondence to Sebastian Guelman.

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All authors are employees of Genentech Inc., a member of the Roche group and Roche shareholders.

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Guelman, S., Zhou, Y., Brady, A. et al. A Fit-for-Purpose Method to Measure Circulating Levels of the mRNA Component of a Liposomal-Formulated Individualized Neoantigen-Specific Therapy for Cancer. AAPS J 24, 64 (2022). https://doi.org/10.1208/s12248-022-00709-x

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