PLXY321 PROTECTS AGAINST ARA-C OR TAXOL-INDUCED APOPTOSIS AND LOSS OF CLONOGENIC SURVIVAL OF NORMAL HUMAN BONE-MARROW PROGENITOR CELLS

By suppressing apoptosis, hemopoietic growth factors (HGFs) promote the survival of CD34(+), HLA-DR(+) marrow cells that are enriched for hemopoietic progenitor cells (HPC). In the present studies, we have examined the effects of pIXY321, a genetically engineered fusion protein of GM-CSF and IL-3 (GM-CSF/IL-3), on high-dose Ara-C (HIDAC) and taxol-induced apoptosis and survival of a multilineage HPC, the CFU-GEMM. Exposure to 1.0 mu mol/l taxol for 24 h or HIDAC greater than or equal to 10 mu mol/l for 4 h induced internucleosomal DNA fragmentation and the morphologic features of apoptosis in CD34(+), HLA-DR(+) cells. These treatments were associated with greater than or equal to 50% inhibition of the assayable CFU-GEMM colony numbers. Incubation in serum-free medium (SFM) alone for 24 h also induced apoptosis of CD34(+), HLA-DR(+) cells, which was associated with reduced intracellular levels of the bcl-2 gene product p26BCL-2. Go-treatment with pIXY321 (10 ng/ml) inhibited apoptosis of CD34(+), HLA-DR(+) cells incubated in SFM, without significantly increasing the intracellular p26BCL-2 levels. Furthermore, co-treatment with pIXY321 significantly reduced taxol- and Ara-C-induced apoptosis and promoted the survival of CFU-GERMM (P < 0.05). Taxol and Ara-C-mediated apoptosis of CD34(+), HLA-DR(+) cells, and its inhibition by pIXY321, was not accompanied by any significant alteration in the intracellular p26BCL-2 levels. By demonstrating that cc-treatment with pIXY321 confers significant protection against apoptosis of CD34(+), HLA-DR(+) cells as well as promotes survival of normal HPC exposed to clinically relevant schedules and concentrations of taxol or Ara-C, these results support the design of chemotherapy regimens incorporating pIXY321 plus taxol and/or high-dose Ara-C for solid tumors and/or acute leukemias. It is hoped that the use of such a cytokine might maintain normal HPC numbers following chemotherapy, therefore avoiding prolonged suppression.