A NOVEL EXPANSION OF PRIMARY LEUKEMIA STEM CELLS -
DESCRIPTION (provided by applicant): Arteriocyte's long-term objective is to develop a platform for ex vivo expansion of leukemic stem cell (LSC) based on cytokine-enriched medium and the company's proprietary nanofiber culture scaffold (NANEX). Cancer stem cell research is of significant importance because, this approach has the potential to create an effective therapy that overcomes challenges of metastasis and relapse for leukemia and other cancers. However, current capabilities for research of leukemia stem cell (LSCs) expansion are hampered by lack of an effective in vitro culture technology. Because leukemia stem cells lose clonogenic phenotype in ex vivo culture, current method for propagation of LSC relies on NOD/SCID mice transplantation model. This severely limits researcher's ability on LSCs. In this project, we will test the feasibility of an ex vivo expansion of acute myeloid leukemia (AML) LSC. This study will be tested by the following specific aims: 1. To establish ex vivo expansion of AML LSC in a functionalized nanofiber based NANEX culture system. We will conduct a series of primary culture of human CD34+ AML MNCs with NANEX scaffold and cytokines to determine feasibility and effectiveness of clonogenic LSC expansion in a controlled, duplicated study. Immunophenotyping, CFU assay, and cytogenic analysis will be performed to measure culture clonogenic potential and phenotype/karyotype drift. 2. To establish in vivo study for biological validation of ex vivo expanded AML LSC in NOD/SCID mice model. We will verify propagation of LSC sub-population in NANEX-expanded AML MNCs using NOD/SCID mice transplantation model. Preservation of SCID-leukemia initiating cells (SL-ICs) in NANEX-cultured AML cells will be verified. Immunophenotyping, CFU assay, cytogenic and DNA analyses will be used to measure phenotype/karyotype profile, clonogenic potential, and graft reconstitution in animal. If successful, the NANEX platform can be developed and commercialized as in vitro model for leukemic cell research, which will greatly extend scientist's ability to manipulate and monitor LSCs for research on cell signaling pathways, drug screening, and genetic modifications.
PUBLIC HEALTH RELEVANCE: This project aims to develop a nanofiber-based ex vivo culture to propagate leukemic stem cells. A cost-effective, well defined, accessible in vitro model of LSC will enhance scientists' ability to manipulate and observe LSCs for leukemia research and drug development.
