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Dominant selection of hematopoietic progenitor cells with retroviral MDR1 co-expression vectors

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Dominant selection of hematopoietic progenitor cells with retroviral MDR1 co-expression vectors. / Hildinger, M; Fehse, B; Hegewisch-Becker, S; John, J; Rafferty, J R; Ostertag, W; Baum, Christopher.

In: HUM GENE THER, Vol. 9, No. 1, 01.01.1998, p. 33-42.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Hildinger, M, Fehse, B, Hegewisch-Becker, S, John, J, Rafferty, JR, Ostertag, W & Baum, C 1998, 'Dominant selection of hematopoietic progenitor cells with retroviral MDR1 co-expression vectors', HUM GENE THER, vol. 9, no. 1, pp. 33-42. https://doi.org/10.1089/hum.1998.9.1-33

APA

Hildinger, M., Fehse, B., Hegewisch-Becker, S., John, J., Rafferty, J. R., Ostertag, W., & Baum, C. (1998). Dominant selection of hematopoietic progenitor cells with retroviral MDR1 co-expression vectors. HUM GENE THER, 9(1), 33-42. https://doi.org/10.1089/hum.1998.9.1-33

Vancouver

Hildinger M, Fehse B, Hegewisch-Becker S, John J, Rafferty JR, Ostertag W et al. Dominant selection of hematopoietic progenitor cells with retroviral MDR1 co-expression vectors. HUM GENE THER. 1998 Jan 1;9(1):33-42. https://doi.org/10.1089/hum.1998.9.1-33

Bibtex

@article{c4ff64a1a9fc44c2a8bda947bdef85d3,
title = "Dominant selection of hematopoietic progenitor cells with retroviral MDR1 co-expression vectors",
abstract = "When transferring the human multidrug resistance 1 (MDR1) cDNA, FMEV retroviral vectors mediate high-dose multidrug resistance and, thus, background-free selection in primary human hematopoietic progenitor cells. Here, we analyzed strategies for co-expression of a second gene from an FMEV:MDR1 vector. When linking the cDNAs with the internal ribosomal entry site (IRES) of poliovirus or retroviral splice signals, almost all multidrug-resistant hematopoietic colonies simultaneously coexpressed the 3' positioned second gene, neomycin-phosphotransferase (neoR). The IRES strategy allowed functional co-transfer of a 4.2-kb lacZ-neoR fusion gene, resulting in a total proviral genome size of 11 kb, corresponding to the packaging limit of retroviral vectors. Preselection based on multidrug resistance elevated the expression of the second gene in IRES constructs, but not in splice vectors. Moreover, three intriguing observations were made. First, up to 30% of cells preselected for functional transfer of the 3' positioned cDNA (neoR) showed infunctional MDR1; this occurred irrespective of the linking principle and was associated with instability of the MDR1 transcription unit. Second, the levels of multidrug resistance achieved with the co-expression vectors were moderately lower (15-30% reduced) than those mediated by the monocistronic counterpart. Third, transduction with FMEV:MDR1 co-expression vectors still resulted in high-dose cancer drug resistance and background-free selection of hematopoietic progenitor cells (including primary human CD34+ colony-forming units). Thus, for the first time, we describe MDR1 co-expression vectors that maintain their desired function in early and primary human hematopoietic cells. However, careful interpretation of the data reveals that further vector improvements are required to obtain clinically useful MDR1 co-expression vectors.",
keywords = "Antigens, CD34, Cell Culture Techniques, Cell Separation, Gene Expression, Gene Transfer Techniques, Genes, MDR, Genetic Vectors, Hematopoietic Stem Cells, Humans, Retroviridae, Selection, Genetic, Transcription, Genetic",
author = "M Hildinger and B Fehse and S Hegewisch-Becker and J John and Rafferty, {J R} and W Ostertag and Christopher Baum",
year = "1998",
month = jan,
day = "1",
doi = "10.1089/hum.1998.9.1-33",
language = "English",
volume = "9",
pages = "33--42",
journal = "HUM GENE THER",
issn = "1043-0342",
publisher = "Mary Ann Liebert Inc.",
number = "1",

}

RIS

TY - JOUR

T1 - Dominant selection of hematopoietic progenitor cells with retroviral MDR1 co-expression vectors

AU - Hildinger, M

AU - Fehse, B

AU - Hegewisch-Becker, S

AU - John, J

AU - Rafferty, J R

AU - Ostertag, W

AU - Baum, Christopher

PY - 1998/1/1

Y1 - 1998/1/1

N2 - When transferring the human multidrug resistance 1 (MDR1) cDNA, FMEV retroviral vectors mediate high-dose multidrug resistance and, thus, background-free selection in primary human hematopoietic progenitor cells. Here, we analyzed strategies for co-expression of a second gene from an FMEV:MDR1 vector. When linking the cDNAs with the internal ribosomal entry site (IRES) of poliovirus or retroviral splice signals, almost all multidrug-resistant hematopoietic colonies simultaneously coexpressed the 3' positioned second gene, neomycin-phosphotransferase (neoR). The IRES strategy allowed functional co-transfer of a 4.2-kb lacZ-neoR fusion gene, resulting in a total proviral genome size of 11 kb, corresponding to the packaging limit of retroviral vectors. Preselection based on multidrug resistance elevated the expression of the second gene in IRES constructs, but not in splice vectors. Moreover, three intriguing observations were made. First, up to 30% of cells preselected for functional transfer of the 3' positioned cDNA (neoR) showed infunctional MDR1; this occurred irrespective of the linking principle and was associated with instability of the MDR1 transcription unit. Second, the levels of multidrug resistance achieved with the co-expression vectors were moderately lower (15-30% reduced) than those mediated by the monocistronic counterpart. Third, transduction with FMEV:MDR1 co-expression vectors still resulted in high-dose cancer drug resistance and background-free selection of hematopoietic progenitor cells (including primary human CD34+ colony-forming units). Thus, for the first time, we describe MDR1 co-expression vectors that maintain their desired function in early and primary human hematopoietic cells. However, careful interpretation of the data reveals that further vector improvements are required to obtain clinically useful MDR1 co-expression vectors.

AB - When transferring the human multidrug resistance 1 (MDR1) cDNA, FMEV retroviral vectors mediate high-dose multidrug resistance and, thus, background-free selection in primary human hematopoietic progenitor cells. Here, we analyzed strategies for co-expression of a second gene from an FMEV:MDR1 vector. When linking the cDNAs with the internal ribosomal entry site (IRES) of poliovirus or retroviral splice signals, almost all multidrug-resistant hematopoietic colonies simultaneously coexpressed the 3' positioned second gene, neomycin-phosphotransferase (neoR). The IRES strategy allowed functional co-transfer of a 4.2-kb lacZ-neoR fusion gene, resulting in a total proviral genome size of 11 kb, corresponding to the packaging limit of retroviral vectors. Preselection based on multidrug resistance elevated the expression of the second gene in IRES constructs, but not in splice vectors. Moreover, three intriguing observations were made. First, up to 30% of cells preselected for functional transfer of the 3' positioned cDNA (neoR) showed infunctional MDR1; this occurred irrespective of the linking principle and was associated with instability of the MDR1 transcription unit. Second, the levels of multidrug resistance achieved with the co-expression vectors were moderately lower (15-30% reduced) than those mediated by the monocistronic counterpart. Third, transduction with FMEV:MDR1 co-expression vectors still resulted in high-dose cancer drug resistance and background-free selection of hematopoietic progenitor cells (including primary human CD34+ colony-forming units). Thus, for the first time, we describe MDR1 co-expression vectors that maintain their desired function in early and primary human hematopoietic cells. However, careful interpretation of the data reveals that further vector improvements are required to obtain clinically useful MDR1 co-expression vectors.

KW - Antigens, CD34

KW - Cell Culture Techniques

KW - Cell Separation

KW - Gene Expression

KW - Gene Transfer Techniques

KW - Genes, MDR

KW - Genetic Vectors

KW - Hematopoietic Stem Cells

KW - Humans

KW - Retroviridae

KW - Selection, Genetic

KW - Transcription, Genetic

U2 - 10.1089/hum.1998.9.1-33

DO - 10.1089/hum.1998.9.1-33

M3 - SCORING: Journal article

C2 - 9458240

VL - 9

SP - 33

EP - 42

JO - HUM GENE THER

JF - HUM GENE THER

SN - 1043-0342

IS - 1

ER -