Development of a gene therapy based bone marrow purging system for leukemias.
Although viable gene therapy based methods have been reported for the selective removal or purging of contaminating epithelial derived cancer cells from stem cell grafts, similar strategies for the purging of leukemia cells have been significantly less efficient. Hematopoietic cells are difficult targets for transduction with currently available vectors. Polylysine based molecular conjugate vectors (MCV) were previously found to effectively transduce both normal and malignant hematopoietic cells. A panel of human leukemia cell lines as well as CD34+ selected primary human hematopoietic progenitor cells (HPC) were tested for differential gene expression utilizing different promoters. Reporter gene expression under the control of RSV and SV40 promoters showed a 6-log fold increase in leukemia cells when compared to primary HPC. Using a polylysine based recombinant molecular conglomerate vector (recMCV) encoding the HSV-tk suicide gene under control of RSV, we demonstrated effective and specific cell killing in all leukemia cell lines as well as in primary human leukemia cells derived from chemotherapy refractory patients, while HPC survived under the same conditions at approximately 20% viability. These proof of principle experiments demonstrate that gene therapy technology could be utilized to successfully purge leukemia cells from HPC.
Huang
Weitao
W
Gene Therapy Program, Louisiana State University Health Sciences Center, New Orleans, Louisiana 70122, USA.
Tan
Weihong
W
Zhong
Qiu
Q
Schwarzenberger
Paul
P
D018952:Antigens, CD34; D016465:Bone Marrow Purging / Q000379:methods*; D002470:Cell Survival; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D041722:Genes, Transgenic, Suicide; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells; D006801:Humans; D007938:Leukemia / Q000235:genetics* / Q000628:therapy*; D009360:Neoplastic Cells, Circulating; D011107:Polylysine; D014162:Transfection; D014407:Tumor Cells, Cultured
D018952:Antigens, CD34; D011107:Polylysine