The purpose of the present protocol is to develop an in vitro means by which human bone marrow can be cleared of metastatic foci of carcinoma. Preliminary in vitro experiments performed with CELLMAXtm hollow fiber cell culture systems demonstrated that 1) normal human marrow can be grown to produce pluripotent cells yielding all hematopoietic cell types and 2) human T lymphocytes can be expanded to massive numbers for clinical therapy. Activated T cells destroy neoplastic cells both in vivo and in vitro. We will culture aliquots of bone marrow from ten patients, each containing metastatic neuroblastoma, breast or prostate carcinoma. These marrows will bcmaintained within continuously perfused hollow fiber bioreactors for 3 weeks during which time, the endogenous T cells will be activated and expanded by addition of rIL-2. Cells will then be harvested from the extra-fiber space and the numbers of both malignant cells and colony-forming bone marrow stem cells determined. Phenotypic and in vitro cytotoxicity profiles will also be determined. Successful depletion of malignant cells with concomitant maintenance of the colony-forming cells within harvested marrow could enable patients having such metastic malignancies to be treated with marrow-destroying doses of chemotherapy and radiation therapy and then be rescued by re-infusion and cngraftment of their own marrow.Awardee's statement of the potential commercial applications of the research:The ability to remove neoplastic cells from bone marrow while simultaneously maintaining its viability in vitro during the period that the patient is receiving high dose chemotherapy or radiation therapy will provide the option of autologous bone marrow transplantation to more than 100,000 patients annually in the United States. Each treatment will utilize one hollow fiber bioreactor and disposable tubing thus resulting in the development and manufacture of a new medical device and the development of a new paramedical support staff at university and community hospitals.National Cancer Institute (NCI)
