Vol. 17 No. 2 (2024), EXPERIMENTAL STUDIES
Vol. 17 No. 2 (2024)
EXPERIMENTAL STUDIES

Xenotransplantation of Human Hematopoietic Stem Cells into NBSGW Mice: A Basic Model for Preclinical Development of Gene Therapy Approaches

Published 01.04.2024
Alena Igorevna Shakirova
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
I.N. Gaponenko
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
Ya.V. Komarova
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
O.S. Epifanovskaya
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
D.A. Senichkina
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
V.S. Sergeev
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
A.R. Muslimov
VA Almazov National Medical Research Center, 2 Akkuratova ul., Saint Petersburg, Russian Federation, 197341
A.V. Onopchenko
VA Almazov National Medical Research Center, 2 Akkuratova ul., Saint Petersburg, Russian Federation, 197341
E.V. Shchelina
VA Almazov National Medical Research Center, 2 Akkuratova ul., Saint Petersburg, Russian Federation, 197341
S.A. Osipova
VA Almazov National Medical Research Center, 2 Akkuratova ul., Saint Petersburg, Russian Federation, 197341
O.G. Bredneva
VA Almazov National Medical Research Center, 2 Akkuratova ul., Saint Petersburg, Russian Federation, 197341
M.L. Vasyutina
VA Almazov National Medical Research Center, 2 Akkuratova ul., Saint Petersburg, Russian Federation, 197341
Ya.G. Toropova
VA Almazov National Medical Research Center, 2 Akkuratova ul., Saint Petersburg, Russian Federation, 197341
K.V. Lepik
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
M.O. Popova
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
I.S. Moiseev
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
A.D. Kulagin
RM Gorbacheva Scientific Research Institute of Pediatric Oncology, Hematology and Transplantation; IP Pavlov First Saint Petersburg State Medical University, 6/8 L’va Tolstogo ul., Saint Petersburg, Russian Federation, 197022
PDF_2024-17-2_82-93 (Russian)

Keywords

xenograft model
immunodeficient NBSGW mice
transplantation
hematopoietic stem cells
preclinical development of gene and cell therapy products

How to Cite

1.
Shakirova A.I., Gaponenko I.N., Komarova Y.V., et al. Xenotransplantation of Human Hematopoietic Stem Cells into NBSGW Mice: A Basic Model for Preclinical Development of Gene Therapy Approaches. Клиническая онкогематология. 2024;17(2):82-93. doi:10.21320/2500-2139-2024-17-2-82-93
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Abstract

Background. The gene therapy based on hematopoietic cell xenotransplantation is becoming a powerful and universally applied therapeutic strategy in an ever-expanding range of human diseases. One of the current issues in implementing the techniques of genome modification in hematopoietic stem cells (HSCs) into clinical practice is to assure the quality and safety of gene and cell therapy products for human use. This is achieved by animal model testing at the stage of preclinical studies. With this purpose in view, NBSGW mice seem to be a unique and promising model for human HSC engraftment without pre-conditioning.

Aim. To test the NBSGW mouse model for human HSC engraftment, to optimize the methods of assessing the state of the animals and monitoring the chimerism level for translational preclinical development of HSC-based products for gene and cell therapy.

Materials & Methods. The xenograft models of NBSGW mice were generated using the samples of the selected peripheral blood CD34+ HSCs from a healthy donor. Serial transplantation was performed by intravenous injection of bone marrow cells from primary recipients with a high chimerism level. Engraftment efficiency was evaluated by flow cytofluorometry (FCF) and droplet digital PCR (ddPCR). Subpopulation pattern of human cell engraftment was assessed by FCF.

Results. The tested HSC transplantation regimen is characterized by favorable toxicity profile. In the entire study sample of mice, the FCF analysis showed a long-term engraftment of human cells with a high chimerism level (23.5–93.6 %) in the bone marrow of the animals, also after serial transplantation, which was confirmed by ddPCR. The B-lineage differentiation cells predominated in all tested samples (of peripheral blood, bone marrow, and spleen) from mice after primary and serial transplantation. The ddPCR assay can be used as an additional tool for validating the level of human cell engraftment determined by FCF.

Conclusion. NBSGW mice present a promising reference model for preclinical development of gene and cell therapy products based on human primary HSCs with a modified genome.

PDF_2024-17-2_82-93 (Russian)

References

  1. Koniali L, Lederer CW, Kleanthous M. Therapy development by genome editing of hematopoietic stem cells. Cells. 2021;10(6):1492. doi: 10.3390/cells10061492.
  2. Cring MR, Sheffield VC. Gene therapy and gene correction: targets, progress, and challenges for treating human diseases. Gene Ther. 2022;29(1–2):3–12. doi: 10.1038/s41434-020-00197-8.
  3. Sagoo P, Gaspar HB. The transformative potential of HSC gene therapy as a genetic medicine. Gene Ther. 2023;30(3–4):197–215. doi: 10.1038/s41434-021-00261-x.
  4. Morgan RA, Gray D, Lomova A, Kohn DB. Hematopoietic stem cell gene therapy: progress and lessons learned. Cell Stem Cell. 2017;21(5):574–90. doi: 10.1016/j.stem.2017.10.010.
  5. Radtke S, Humbert O, Kiem HP. Mouse models in hematopoietic stem cell gene therapy and genome editing. Biochem Pharmacol. 2020;174:113692. doi: 10.1016/j.bcp.2019.113692.
  6. Mian SA, Anjos-Afonso F, Bonnet D. Advances in Human Immune System Mouse Models for Studying Human Hematopoiesis and Cancer Immunotherapy. Front Immunol. 2021;11:619236. doi: 10.3389/fimmu.2020.619236.
  7. McDermott SP, Eppert K, Lechman ER, et al. Comparison of human cord blood engraftment between immunocompromised mouse strains. Blood. 2010;116(2):193–200. doi: 10.1182/blood-2010-02-271841.
  8. McIntosh BE, Brown ME, Duffin BM, et al. Nonirradiated NOD,B6.SCID Il2rγ-/- Kit(W41/W41) (NBSGW) mice support multilineage engraftment of human hematopoietic cells. Stem Cell Reports. 2015;4(2):171–80. doi: 10.1016/j.stemcr.2014.12.005.
  9. Sun G, Liang X, Qin K, et al. Functional analysis of KIT gene structural mutations causing the porcine dominant white phenotype using genome edited mouse models. Front Genet. 2020;11:138. doi: 10.3389/fgene.2020.00138.
  10. Rojas-Sutterlin S, Lecuyer E, Hoang T. Kit and Scl Regulation of Hematopoietic Stem Cells. Curr Opin Hematol. 2014;21(4):256–64. doi: 10.1097/moh.0000000000000052.
  11. Cosgun KN, Rahmig S, Mende N, et al. Kit regulates HSC engraftment across the human-mouse species barrier. Cell Stem Cell. 2014;15(2):227–38. doi: 10.1016/j.stem.2014.06.001.
  12. Theocharides AP, Rongvaux A, Fritsch K, et al. Humanized hemato-lymphoid system mice. Haematologica. 2016;101(1):5–19. doi: 10.3324/haematol.2014.115212.
  13. Ito M, Hiramatsu H, Kobayashi K, et al. NOD/SCID/gamma(c)(null) mouse: an excellent recipient mouse model for engraftment of human cells. Blood. 2002;100(9):3175–82. doi: 10.1182/blood-2001-12-0207.
  14. Pattabhi S, Lotti SN, Berger MP, et al. In vivo outcome of homology-directed repair at the HBB gene in HSC using alternative donor template delivery methods. Mol Ther Nucleic Acids. 2019;17:277–88. doi: 10.1016/j.omtn.2019.05.025.
  15. Wu Y, Zeng J, Roscoe BP, et al. Highly efficient therapeutic gene editing of human hematopoietic stem cells. Nat Med. 2019;25(5):776–83. doi: 10.1038/s41591-019-0401-y.
  16. Metais JY, Doerfler PA, Mayuranathan T, et al. Genome editing of HBG1 and HBG2 to induce fetal hemoglobin. Blood Adv. 2019;3(21):3379–92. doi: 10.1182/bloodadvances.2019000820.
  17. Everette KA, Newby GA, Levine RM, et al. Ex vivo prime editing of patient haematopoietic stem cells rescues sickle-cell disease phenotypes after engraftment in mice. Nat Biomed Eng. 2023;7(5):616–28. doi: 10.1038/s41551-023-01026-0.
  18. Leonard A, Yapundich M, Nassehi T, et al. Low-dose busulfan reduces human CD34(+) cell doses required for engraftment in c-kit mutant immunodeficient mice. Mol Ther Methods Clin Dev. 2019;15:430–7. doi: 10.1016/j.omtm.2019.10.017.
  19. Hess NJ, Lindner PN, Vazquez J, et al. Different human immune lineage compositions are generated in non-conditioned NBSGW mice depending on HSPC source. Front Immunol. 2020;11:573406. doi: 10.3389/fimmu.2020.573406.
  20. Adigbli G, Hua P, Uchiyama M, et al. Development of LT-HSC- reconstituted nonirradiated NBSGW mice for the study of human hematopoiesis in vivo. Front Immunol. 2021;12:642198. doi: 10.3389/fimmu.2021.642198.
  21. Amoah A, Keller A, Emini R, et al. Aging of human hematopoietic stem cells is linked to changes in Cdc42 activity. Haematologica 2022;107(2):393–402. doi: 10.3324/haematol.2020.269670.
  22. Karuppusamy KV, Demosthenes JP, Venkatesan V, et al. The CCR5 gene edited CD34+CD90+ hematopoietic stem cell population serves as an optimal graft source for HIV gene therapy. Front Immunol. 2022;13:792684. doi: 10.3389/fimmu.2022.792684.
  23. Magis W, DeWitt MA, Wyman SK, et al. High-level correction of the sickle mutation is amplified in vivo during erythroid differentiation. iScience 2022;25(6):104374. doi: 10.1016/j.isci.2022.104374.
  24. Suchy FP, Nishimura T, Seki S, et al. Streamlined and quantitative detection of chimerism using digital PCR. Sci Rep. 2022;12(1):10223. doi: 10.1038/s41598-022-14467-5.
  25. Maganti HB, Bailey AJM, Kirkham AM, et al. Persistence of CRISPR/Cas9 gene edited hematopoietic stem cells following transplantation: A systematic review and meta-analysis of preclinical studies. Stem Cells Transl Med. 2021;10(7):996–1007. doi: 10.1002/sctm.20-0520.
  26. Fiorini C, Abdulhay NJ, McFarland SK, et al. Developmentally-faithful and effective human erythropoiesis in immunodeficient and Kit mutant mice. Am J Hematol. 2017;92(9):Е513–E519. doi: 10.1002/ajh.24805.
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