ABSTRACT
Several investigators have shown the utility of systemically delivered optical imaging probes to image tumors in small animal models of cancer. Here we demonstrate an innovative method for imaging tumors and tumor margins during surgery. Specifically, we show that optical imaging probes topically applied to tumors and surrounding normal tissue rapidly differentiate between tissues. In contrast to systemic delivery of optical imaging probes which label tumors uniformly over time, topical probe application results in rapid and robust probe activation that is detectable as early as 5 minutes following application. Importantly, labeling is primarily associated with peri-tumor spaces. This methodology provides a means for rapid visualization of tumor and potentially infiltrating tumor cells and has potential applications for directed surgical excision of tumor tissues. Furthermore, this technology could find use in surgical resections for any tumors having differential regulation of cysteine cathepsin activity.
Subject(s)
Brain Neoplasms/diagnosis , Diagnostic Imaging , Fluorescent Dyes , Molecular Imaging/methods , Monitoring, Intraoperative/methods , Administration, Topical , Brain Neoplasms/pathology , Brain Neoplasms/surgery , Cathepsins/metabolism , Cell Line, Tumor , Cysteine/metabolism , Humans , Molecular Imaging/instrumentation , Monitoring, Intraoperative/instrumentationABSTRACT
We recently found that normal human brain and low-grade astrocytomas express the receptor protein tyrosine phosphatase mu (PTPmu) and that the more invasive astrocytomas, glioblastoma multiforme (GBM), downregulate full-length PTPmu expression. Loss of PTPmu expression in GBMs is due to proteolytic cleavage that generates an intracellular and potentially a cleaved and released extracellular fragment of PTPmicro. Here, we identify that a cleaved extracellular fragment containing the domains required for PTPmicro-mediated adhesion remains associated with GBM tumor tissue. We hypothesized that detection of this fragment would make an excellent diagnostic tool for the localization of tumor tissue within the brain. To this end, we generated a series of fluorescently tagged peptide probes that bind the PTPmu fragment. The peptide probes specifically recognize GBM cells in tissue sections of surgically resected human tumors. To test whether the peptide probes are able to detect GBM tumors in vivo, the PTPmu peptide probes were tested in both mouse flank and intracranial xenograft human glioblastoma tumor model systems. The glial tumors were molecularly labeled with the PTPmu peptide probes within minutes of tail vein injection using the Maestro FLEX In Vivo Imaging System. The label was stable for at least 3 hours. Together, these results indicate that peptide recognition of the PTPmu extracellular fragment provides a novel molecular diagnostic tool for detection of human glioblastomas. Such a tool has clear translational applications and may lead to improved surgical resections and prognosis for patients with this devastating disease.
Subject(s)
Brain Neoplasms/diagnosis , Glioblastoma/diagnosis , Molecular Diagnostic Techniques/methods , Peptide Fragments/analysis , Protein Tyrosine Phosphatases/analysis , Amino Acid Sequence , Animals , Biomarkers, Tumor/analysis , Biomarkers, Tumor/metabolism , Brain Neoplasms/metabolism , Extracellular Space/chemistry , Extracellular Space/metabolism , Female , Fluorescence , Glioblastoma/metabolism , Humans , Isoenzymes/analysis , Isoenzymes/chemistry , Isoenzymes/metabolism , Mice , Mice, Nude , Models, Molecular , Molecular Sequence Data , Neoplasm Transplantation , Peptide Fragments/isolation & purification , Peptide Fragments/metabolism , Protein Tyrosine Phosphatases/chemistry , Protein Tyrosine Phosphatases/metabolism , Transplantation, Heterologous , Tumor Cells, CulturedABSTRACT
Both pro- and antioncogenic properties have been attributed to EphA2 kinase. We report that a possible cause for this apparent paradox is diametrically opposite roles of EphA2 in regulating cell migration and invasion. While activation of EphA2 with its ligand ephrin-A1 inhibited chemotactic migration of glioma and prostate cancer cells, EphA2 overexpression promoted migration in a ligand-independent manner. Surprisingly, the latter effects required phosphorylation of EphA2 on serine 897 by Akt, and S897A mutation abolished ligand-independent promotion of cell motility. Ephrin-A1 stimulation of EphA2 negated Akt activation by growth factors and caused EphA2 dephosphorylation on S897. In human astrocytoma, S897 phosphorylation was correlated with tumor grades and Akt activation, suggesting that the Akt-EphA2 crosstalk may contribute to brain tumor progression.
Subject(s)
Brain Neoplasms/pathology , Cell Movement/physiology , Neoplasm Invasiveness , Proto-Oncogene Proteins c-akt/metabolism , Receptor, EphA2/metabolism , Brain Neoplasms/metabolism , Cell Movement/genetics , Disease Progression , Enzyme Activation , Humans , Ligands , PTEN Phosphohydrolase/metabolism , Phosphorylation , Phosphoserine/metabolism , Polymorphism, Single Nucleotide , Receptor, EphA1/metabolism , Receptor, EphA2/geneticsABSTRACT
Therapies targeting glioma cells that diffusely infiltrate normal brain are highly sought after. Our aim was to identify novel approaches to this problem using glioma spheroid migration assays. Lithium, a currently approved drug for the treatment of bipolar illnesses, has not been previously examined in the context of glioma migration. We found that lithium treatment potently blocked glioma cell migration in spheroid, wound-healing, and brain slice assays. The effects observed were dose dependent and reversible, and worked using every glioma cell line tested. In addition, there was little effect on cell viability at lithium concentrations that inhibit migration, showing that this is a specific effect. Lithium treatment was associated with a marked change in cell morphology, with cells retracting the long extensions at their leading edge. Examination of known targets of lithium showed that inositol monophosphatase inhibition had no effect on glioma migration, whereas inhibition of glycogen synthase kinase-3 (GSK-3) did. This suggested that the effects of lithium on glioma cell migration could possibly be mediated through GSK-3. Specific pharmacologic GSK-3 inhibitors and siRNA knockdown of GSK-3alpha or GSK-3beta isoforms both reduced cell motility. These data outline previously unidentified pathways and inhibitors that may be useful for the development of novel anti-invasive therapeutics for the treatment of brain tumors.
Subject(s)
Antineoplastic Agents/pharmacology , Brain Neoplasms/enzymology , Cell Movement/drug effects , Glioma/enzymology , Glycogen Synthase Kinase 3/metabolism , Lithium Chloride/pharmacology , Animals , Blotting, Western , Brain Neoplasms/pathology , Cell Cycle/drug effects , Cell Line, Tumor , Enzyme Inhibitors/pharmacology , Glioma/pathology , Glycogen Synthase Kinase 3/drug effects , Humans , Mice , Neoplasm Invasiveness/physiopathology , Neurons/drug effects , Organ Culture TechniquesABSTRACT
Primary glioblastoma multiforme is an aggressive brain tumor that has no cure. Current treatments include gross resection of the tumor, radiation and chemotherapy. Despite valiant efforts, prognosis remains dismal. A promising new technique involves the use of oncolytic viruses that can specifically replicate and lyse in cancers, without spreading to normal tissues. Currently, these are being tested in relevant preclinical models and clinical trials as a therapeutic modality for many types of cancer. Results from recent clinical trials with oncolytic viruses have revealed the safety of this approach, although evidence for efficacy remains elusive. Oncolytic viral strategies are summarized in this review, with a focus on therapies used in brain tumors.
