GENT-08. HUMAN GLIOBLASTOMA ASTROCYTES RESEMBLE FETAL ASTROCYTE PRECURSOR CELLS AS REVEALED BY IMMUNOPANNING AND RNAseq
Year: 2016
Type: conference-abstract
Abstract: Genes and pathways active in glioblastoma astrocytes yet quiescent in mature astrocytes hold vast therapeutic potential. We hypothesized that glioblastoma astrocytes have reverted to a ‘precursor’ state, as defined by the expression of fetal astrocyte genes. We have developed an innovative method leveraging primary human tissue to study in culture the individual cell subtypes comprising normal brain and glioblastoma. Prior studies in the field have relied on bulk glioblastoma tissue in which more numerous non-tumor or bystander cells drown out the glioblastoma astrocyte gene signature. We applied novel immunopanning separation methods (HepaCAM) to isolate human astrocytes from fetal, adult, and glioblastoma specimens. Previous cell sorting techniques (e.g. FACS) kill the primary brain cells quickly, lead to reactive states, are contaminated with other cell types, or can only be done on fetal rodent cells. Human fetal, adult, and glioblastoma astrocytes then underwent RNA-seq and culture. We compared transcriptional similarities between glioblastoma and normal mature and fetal astrocytes, all using fresh, human tissue. Once isolated, we found our astrocytes were minimally reactive, highly pure, and could be maintained in primary cell culture with serum-free media. Unsupervised hierarchical clustering of gene expression profiles showed glioblastoma astrocytes grouped with human astrocyte precursor cells (fetal) compared to mature astrocytes. Genes that distinguished astrocyte precursor cells from mature human astrocytes were shared by glioblastoma astrocytes. The molecular profiles of astrocyte precursor cells were overwhelmingly immature and proliferative, in sharp distinction to the human mature resting astrocyte profiles. Human glioblastoma astrocytes have a similar gene expression profile to fetal astrocyte precursor cells. Application of the immunopanning technique will allow us to study in vitro the transcriptional profiles and phenotypes of brain and glioblastoma cellular subtypes, reliably recapitulating the human disease and suggesting therapeutic targets.
Source: Neuro-Oncology
Institutions: University of California, Los Angeles, Stanford University
Language: English
Subfield: Genetics
Field: Medicine
Domain: Health Sciences
Open Access status: bronze
OpenAlex ID: W2587949966