Abundance of Flt3 and its ligand in astrocytic tumors

Abstract

BackgroundMolecular targeted therapies for astrocytic tumors are the subject of growing research interest, due to the limited response of these tumors, especially glioblastoma multiforme, to conventional chemotherapeutic regimens. Several of these approaches exploit the inhibition of receptor tyrosine kinases. To date, it has not been elucidated if fms-like tyrosine kinase-3 (Flt3) and its natural ligand (Flt3L) are expressed in astrocytic tumors, although some of the clinically intended small-molecule receptor tyrosine kinase inhibitors affect Flt3, while others do not. More importantly, the recent proof of principle for successful stimulation of the immune system against gliomas in preclinical models via local Flt3L application requires elucidation of this receptor tyrosine kinase pathway in these tumors in more detail. This therapy is based on recruitment of Flt3-positive dendritic cells, but may be corroborated by activity of this signaling pathway in glioma cells.MethodsReceptor and ligand expression was analyzed by real-time polymerase chain reaction in 31 astrocytic tumors (six diffuse and 11 anaplastic astrocytomas, 14 glioblastomas) derived from patients of both genders and in glioblastoma cell lines. The two most common activating mutations of the Flt3 gene, ie, internal tandem duplication and D835 point mutation, were assessed by specific polymerase chain reaction.ResultsA relatively high abundance of Flt3L mRNA (4%–6% of the reference, b2 microglobulin) could be demonstrated in all tumor samples. Flt3 expression could generally be demonstrated by 40 specific polymerase chain reaction cycles and gel electrophoresis in 87% of the tumors, including all grades, although the small quantities of the receptor did not allow reliable quantification. Expression of both mRNAs was verified in the cell lines, excluding a derivation solely from contaminating lymphocytes or macrophages. No activating mutations were found.ConclusionOur results warrant further analysis of endogenous Flt3 signaling in these tumors prior to application of immunotherapy in human patients.

Receptor Tyrosine Kinase Signaling and Targeting in Glioblastoma Multiforme

“…Fms-like tyrosine kinase 3 (FLT3) is normally expressed by hematopoietic stem or progenitor cells and plays an important role in the early stages of both myeloid and lymphoid lineage development [ 123 ]. In GBM, mRNA expression of FLT3 and its natural ligand FLT3L is detected in astrocytic tumors as well as GBM cell lines [ 124 ]; however, the exact mechanism by which FLT3 contributes to GBM occurrence/progression remains unknown. In the TCGA-GBM dataset, FLT3 has copy number loss in all subtypes (29.2–36.8%), including in the G-CIMP (57.1%) tumor subset, and is mutated in the CL (2.6%) and ME (2.1%) subtypes.…”