ABSTRACT. The ability to transfer genes into a hematopoietic stem cell and to achieve regulation of their expression in lymphoid or myeloid lineages should open many new therapeutic opportunities. Besides gene transfer mediated by virus vectors like retrovirus or adenovirus, non viral systems have the theoretical advantage of being safe and easy to manage. We developed a new family of cationic lipids called phosphonolipids, synthesized 24 new molecules, and then in a first step we tested their potential to transfer genes in human hematopoietic cell lines (K562 and TF1). A LacZ plasmid under the control of a strong viral promoter was used as a reporter gene and a FACS-Gal assay and a quantitative test CPRG assay evaluated the beta gal expression. The targeted cells were analyzed 48 hours after transfection. The present work shows that seven novel molecules display a high transfer efficiency. One of them is nine-fold more efficient than the commercially available cationic lipids. The results obtained ex vivo on CD34 cells with the FACS-Gal assay show that at day 10 after transfection, 45 percent of cells are expressing gal.
Keywords: Non viral vectors, gene transfer, hematopoietic cells, cationic phosphonolipids, gene therapy.
Reprint requests to: Dr. Claude Férec, Centre de Biogénétique, ETSBO, BP 454, 29275 Brest Cedex, France, phone: (33) 98 44 50 64, fax: (33) 98 43 05 55, email: claude.ferec@univ-brest.fr.
ABSTRACT. Several studies carried out between 1965 and 1985 showed that G-6-PD deficiency in Mexico is heterogeneous at the biochemical level and that the G-6-PD A- phenotype is relatively common. We have now investigated the molecular basis of G-6-PD deficiency in Mexico. Up-to-date 60 chromosomes with G6PD mutations have been studied, 16 in previous studies and 44 in the present work. Molecular analysis of DNA from G-6-PD deficient Mexican mestizos and their relatives show that G-6-PD A- genotypes are relatively common but also that in Mexico G-6-PD deficiency is heterogeneous at the DNA level. Thus, five different genotypes have been observed: G-6-PD A- 202A/376G (41 chromosomes), G-6-PD A- 376G/968C (14 chromosomes), G-6-PD Seattle 844C (3 chromosomes), G-6-PD "Mexico City" 680A (1 chromosome) and G-6-PD Guadalajara 1159T (1 chromosome). The G-6-PD A- 202A/376G, G-6-PD A- 376G/968C and G-6-PD Seattle 844C mutations in Mexico are on the same Pvu II/ Pst I/ 1311 / Nla III haplotypes as found in individuals from Africa, Spain and the Canary Islands. Consequently,these mutations were probably imported to Mexico through African slaves and/or the Spanish immigrants during and after the colonization.
Keywords: G6PD, erythrocyte, population genetics.
Reprint requests to: Dr. Gerardo Vaca, Division de Genetica, Centro de Investigacion Biomedica de Occidente, Instituto Mexicano del Seguro Social, Apartado postal 1-3838, Guadalajara, Jalisco, Mexico, phone: (913) 618 17 56, fax: (913) 618 17 56.