Comparative engraftment and clonality of macaque HSPCs expanded on human umbilical vein endothelial cells versus non-expanded cells
| Autor: | Samson J. Koelle, Idalia M. Yabe, Michael Ginsberg, Diego A. Espinoza, Chuanfeng Wu, Kyung-Rok Yu, Adam Glaser, So Gun Hong, Lauren L. Truitt, Mark E. Metzger, Allen E. Krouse, Sandhya R. Panch, Cynthia E. Dunbar, Stephanie Sellers, Pradeep K. Dagur, Sandeep Kumar Srivastava, Robert E. Donahue, Aylin C. Bonifacino, Daniel J. Nolan |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
lcsh:QH426-470 Genetic enhancement CD34 clonal expansion Macaque 03 medical and health sciences 0302 clinical medicine ex-vivo expansion biology.animal Genetics Autologous transplantation lcsh:QH573-671 Progenitor cell Molecular Biology HUVECs biology lcsh:Cytology hematopoiesis Cell biology genetic barcoding Endothelial stem cell lcsh:Genetics Haematopoiesis 030104 developmental biology 030220 oncology & carcinogenesis Cord blood Molecular Medicine Original Article rhesus macaque |
| Zdroj: | Molecular Therapy. Methods & Clinical Development Molecular Therapy: Methods & Clinical Development, Vol 20, Iss, Pp 703-715 (2021) |
| ISSN: | 2329-0501 |
| DOI: | 10.1016/j.omtm.2021.02.009 |
| Popis: | Ex vivo hematopoietic stem and progenitor cell (HSPC) expansion platforms are under active development, designed to increase HSPC numbers and thus engraftment ability of allogeneic cord blood grafts or autologous HSPCs for gene therapies. Murine and in vitro models have not correlated well with clinical outcomes of HSPC expansion, emphasizing the need for relevant pre-clinical models. Our rhesus macaque HSPC competitive autologous transplantation model utilizing genetically barcoded HSPC allows direct analysis of the relative short and long-term engraftment ability of lentivirally transduced HSPCs, along with additional critical characteristics such as HSPC clonal diversity and lineage bias. We investigated the impact of ex vivo expansion of macaque HSPCs on the engineered endothelial cell line (E-HUVECs) platform regarding safety, engraftment of transduced and E-HUVEC-expanded HSPC over time compared to non-expanded HSPC for up to 51 months post-transplantation, and both clonal diversity and lineage distribution of output from each engrafted cell source. Short and long-term engraftment were comparable for E-HUVEC expanded and the non-expanded HSPCs in both animals, despite extensive proliferation of CD34+ cells during 8 days of ex vivo culture for the E-HUVEC HSPCs, and optimization of harvesting and infusion of HSPCs co-cultured on E-HUVEC in the second animal. Long-term hematopoietic output from both E-HUVEC expanded and unexpanded HSPCs was highly polyclonal and multilineage. Overall, the comparable HSPC kinetics of macaques to humans, the ability to study post-transplant clonal patterns, and simultaneous multi-arm comparisons of grafts without the complication of interpreting allogeneic effects makes our model ideal to test ex vivo HSPC expansion platforms, particularly for gene therapy applications. Graphical Abstract The rhesus macaque barcoded autologous competitive repopulation preclinical transplantation model was utilized to study the engraftment and safety of hematopoietic stem and progenitor cells expanded in co-culture with E4ORF-1 gene transduced human umbilical vein endothelial cell as compared to non-expanded cells, documenting polyclonal long-term multilineage hematopoiesis. |
| Databáze: | OpenAIRE |
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