Enhancing prime editing in hematopoietic stem and progenitor cells by modulating nucleotide metabolism

Sébastien Levesque; Andrea Cosentino; Archana Verma; Pietro Genovese; Daniel E Bauer

doi:10.1038/s41587-024-02266-4

Enhancing prime editing in hematopoietic stem and progenitor cells by modulating nucleotide metabolism

Nat Biotechnol. 2024 May 28. doi: 10.1038/s41587-024-02266-4. Online ahead of print.

Authors

Sébastien Levesque^{1

2

3

4

5}, Andrea Cosentino^{1

2

5

6}, Archana Verma^{1

2

3

4

5}, Pietro Genovese^{1

2

3

4

5}, Daniel E Bauer^{7

8

9

10

11}

Affiliations

¹ Division of Hematology/Oncology, Boston Children's Hospital, Boston, MA, USA.
² Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.
³ Harvard Stem Cell Institute, Cambridge, MA, USA.
⁴ Broad Institute, Cambridge, MA, USA.
⁵ Department of Pediatrics, Harvard Medical School, Boston, MA, USA.
⁶ Milano-Bicocca University, Milan, Italy.
⁷ Division of Hematology/Oncology, Boston Children's Hospital, Boston, MA, USA. [email protected].
⁸ Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA. [email protected].
⁹ Harvard Stem Cell Institute, Cambridge, MA, USA. [email protected].
¹⁰ Broad Institute, Cambridge, MA, USA. [email protected].
¹¹ Department of Pediatrics, Harvard Medical School, Boston, MA, USA. [email protected].

PMID: 38806736
DOI: 10.1038/s41587-024-02266-4

Abstract

Therapeutic prime editing of hematopoietic stem and progenitor cells (HSPCs) holds great potential to remedy blood disorders. Quiescent cells have low nucleotide levels and resist retroviral infection, and it is possible that nucleotide metabolism could limit reverse transcription-mediated prime editing in HSPCs. We demonstrate that deoxynucleoside supplementation and Vpx-mediated degradation of SAMHD1 improve prime editing efficiency in HSPCs, especially when coupled with editing approaches that evade mismatch repair.

Grants and funding

MPI RO1 - 1R01HL170629/U.S. Department of Health & Human Services | National Institutes of Health (NIH)