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Epigenetic repression of gonadotropin gene expression via a GnRH-mediated DNA delivery system

Gene Therapy volume 29, pages 294–303 (2022)Cite this article

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Abstract

The reproductive axis is activated by gonadotropin-releasing hormone (GnRH), which stimulates the pituitary gonadotropes to secrete hormones that drive gonadal function and steroidogenesis. Thus repression of this axis, which is conserved across mammals and sexes, can reduce steroid levels and/or prevent reproduction. Steroid-dependent pathologies, including various cancers, are commonly treated with GnRH super-analogs which have long-term side-effects, while humane solutions for controlling reproduction in domestic and wild animal populations are lacking. GnRH-conjugated toxins are undergoing clinical trials for GnRHR-expressing cancer cells, and have been examined for gonadotrope ablation in animals, but showed low and/or transient effects and administration of toxins has many potential complications. Here we exploit GnRH targeting to gonadotropes to deliver DNA encoding an effector that induces gonadotropin gene repressive epigenetic modifications which are perpetuated over time. Several layers of specificity are endowed through targeting to GnRHR-expressing cells and due to local cleavage of the peptide packaging the DNA; the DNA-encoded effector is expressed and directed to the target genes by the DNA binding domain of a highly specific transcription factor. This design has multiple advantages over existing methods of shutting down the reproductive axis, and its modular design should allow adaptation for broad applications.

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Fig. 1: Overview of the approach.
Fig. 2: A GnRH-targeted carrier protein delivers DNA to gonadotropes specifically.
Fig. 3: Repression of gonadotropin gene expression by KRAB-Sf1 and KRAB-Egr1 chimeric proteins.
Fig. 4: Binding of the chimeric effector proteins to the Cga, Lhb and Gnrhr gene promoters.
Fig. 5: KRAB-Sf1 induces chromatin compaction at Cga and Lhb.
Fig. 6: KRAB-Sf1 induces repressive epigenetic modifications at Cga and Lhb.
Fig. 7: The delivery protein loaded with DNA encoding KRAB-Sf1 represses Cga expression in mixed cultures.

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Funding

This research was supported by the Office of the Chief Scientist, Ministry of Agriculture (14-16-0004), POLAK Fund for Applied Research and Hittman Family Foundation Biomedical Innovation Fund.

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  1. Faculty of Biology, Technion-Israel Institute of Technology, Haifa, 32000, Israel

    Lilach Pnueli & Philippa Melamed

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  1. Lilach Pnueli
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LP and PM conceived the project, analyzed the results and wrote the manuscript. LP performed the experiments. PM acquired the funding.

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Correspondence to Philippa Melamed.

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LP and PM are inventors on US Provisional Patent Application No. 63/221,056.

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Pnueli, L., Melamed, P. Epigenetic repression of gonadotropin gene expression via a GnRH-mediated DNA delivery system. Gene Ther 29, 294–303 (2022). https://doi.org/10.1038/s41434-022-00325-6

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