Barcoding of Macaque Hematopoietic Stem and Progenitor Cells: A Robust Platform to Assess Vector Genotoxicity

Overview

Journal Mol Ther Methods Clin Dev

Publisher Cell Press

Date 2018 Dec 15

PMID 30547048

Citations 7

Authors

Idalia M Yabe

Lauren L Truitt

Diego A Espinoza

Chuanfeng Wu

Samson Koelle

Sandhya Panch

Marcus A F Corat

Thomas Winkler

Kyung-Rok Yu

So Gun Hong

Aylin Bonifacino

Allen Krouse

Mark Metzger

Robert E Donahue

Cynthia E Dunbar

Affiliations

Soon will be listed here.

Abstract

Gene therapies using integrating retrovirus vectors to modify hematopoietic stem and progenitor cells have shown great promise for the treatment of immune system and hematologic diseases. However, activation of proto-oncogenes via insertional mutagenesis has resulted in the development of leukemia. We have utilized cellular bar coding to investigate the impact of different vector designs on the clonal behavior of hematopoietic stem and progenitor cells (HSPCs) during expansion, as a quantitative surrogate assay for genotoxicity in a non-human primate model with high relevance for human biology. We transplanted two rhesus macaques with autologous CD34+ HSPCs transduced with three lentiviral vectors containing different promoters and/or enhancers of a predicted range of genotoxicities, each containing a high-diversity barcode library that uniquely tags each individual transduced HSPC. Analysis of clonal output from thousands of individual HSPCs transduced with these barcoded vectors revealed sustained clonal diversity, with no progressive dominance of clones containing any of the three vectors for up to almost 3 years post-transplantation. Our data support a low genotoxic risk for lentivirus vectors in HSPCs, even those containing strong promoters and/or enhancers. Additionally, this flexible system can be used for the testing of future vector designs.

Citing Articles

Long-term tracking of haematopoietic clonal dynamics and mutations in non-human primate undergoing transplantation of lentivirally barcoded haematopoietic stem and progenitor cells.

Hosuru R, Yang J, Zhou Y, Gin A, Hayal T, Hong S Br J Haematol. 2024; 205(6):2487-2497.

PMID: 39523608 PMC: 11637732. DOI: 10.1111/bjh.19889.

Impact of CRISPR/HDR editing versus lentiviral transduction on long-term engraftment and clonal dynamics of HSPCs in rhesus macaques.

Lee B, Gin A, Wu C, Singh K, Grice M, Mortlock R Cell Stem Cell. 2024; 31(4):455-466.e4.

PMID: 38508195 PMC: 10997443. DOI: 10.1016/j.stem.2024.02.010.

Clonal reconstruction from co-occurrence of vector integration sites accurately quantifies expanding clones in vivo.

Wagner S, Baldow C, Calabria A, Rudilosso L, Gallina P, Montini E Nat Commun. 2022; 13(1):3712.

PMID: 35764632 PMC: 9240075. DOI: 10.1038/s41467-022-31292-6.

Comparative engraftment and clonality of macaque HSPCs expanded on human umbilical vein endothelial cells versus non-expanded cells.

Srivastava S, Truitt L, Wu C, Glaser A, Nolan D, Ginsberg M Mol Ther Methods Clin Dev. 2021; 20:703-715.

PMID: 33738325 PMC: 7937567. DOI: 10.1016/j.omtm.2021.02.009.

The TRACE-Seq method tracks recombination alleles and identifies clonal reconstitution dynamics of gene targeted human hematopoietic stem cells.

Sharma R, Dever D, Lee C, Azizi A, Pan Y, Camarena J Nat Commun. 2021; 12(1):472.

PMID: 33473139 PMC: 7817666. DOI: 10.1038/s41467-020-20792-y.

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