Cell of origin determines tumor phenotype in an oncogenic Ras/p53 knockout transgenic model of high-grade glioma.

Human high-grade gliomas (HGGs) are known for their histologic diversity. To address the role of cell of origin in glioma phenotype, transgenic mice were generated in which oncogenic Ras and p53 deletion were targeted to neural stem/progenitor cells (NSPCs) and mature astrocytes. The hGFAP-Cre/Kras/p53 mice develop multifocal HGGs that vary histopathologically and with respect to the expression of markers associated with NSPCs. One HGG pattern strongly expressed markers of NSPCs and arose near the subventricular zone. Additional nonoverlapping patterns that recapitulate human HGG variants were present simultaneously in the same brain. These neoplastic foci were more often cortical or leptomeningeal based, and the neoplastic cells lacked expression of NSPC markers. To determine whether cell of origin determines tumor phenotype, astrocytes and NSPCs were harvested from neonatal mutant pups. Onorthotopic transplantation, early-passage astrocytes and NSPCs formed tumors that differed in engraftment rates, latency to clinical signs, histopathology, and protein expression. Astrocyte-derivedtumors were more aggressive, had giant-cell histology, and glial fibrillary acidic protein expression. The NSPC-derived tumors retained NSPC markers and showed evidence of differentiation along astrocytic, oligodendroglial, and neuronal lineages. These results indicate that identical tumorigenic stimuli produce markedly different glioma phenotypes, depending on the differentiation status of the transformed cell.

Computational model and microfluidic platform for the investigation of paracrine and autocrine signaling in mouse embryonic stem cells.

Autocrine and paracrine signaling mechanisms are traditionally difficult to study due to the recursive nature of the process and the sub-micromolar concentrations involved. This has proven to be especially limiting in the study of embryonic stem cells that might rely on such signaling for viability, self-renewal, and proliferation. To better characterize possible effects of autocrine and paracrine signaling in the setting of expanding stem cells, we developed a computational model assuming a critical need for cell-secreted survival factors. This model suggested that the precise way in which the removal of putative survival factors could affect stem cell survival in culture. We experimentally tested the predictions in mouse embryonic stem cells by taking advantage of a novel microfluidic device allowing removal of the cell-conditioned medium at defined time intervals. Experimental results in both serum-containing and defined N2B27 media confirmed computational model predictions, suggested existence of unknown survival factors with distinct rates of diffusion, and revealed an adaptive/selective phase in mouse embryonic stem cell response to a lack of paracrine signaling. We suggest that the described computational/experimental platform can be used to identify and study specific factors and pathways involved in a wide variety of paracrine signaling systems.