entrectinib and Neuroblastoma

RTK plays important roles in many cellular signaling processes involved in cancer growth and development. ALK, TRKA, TRKB, TRKC, and ROS1 are RTKs involved in several canonical pathways related to oncogenesis. These proteins can be genetically altered in malignancies, leading to receptor activation and constitutive signaling through their respective downstream pathways. Neuroblastoma (NB) is the most common extracranial solid tumor in childhood, and despite intensive therapy, there is a high mortality rate in cases with a high-risk disease. Alterations of ALK and differential expression of TRK proteins are reported in a proportion of NB. Several inhibitors of ALK or TRKA/B/C have been evaluated both preclinically and clinically in the treatment of NB. These agents have had variable success and are not routinely used in the treatment of NB. Entrectinib (RXDX-101) is a pan-ALK, TRKA, TRKB, TRKC, and ROS1 inhibitor with activity against tumors with

Topics: Anaplastic Lymphoma Kinase; Animals; Benzamides; Humans; Indazoles; Neuroblastoma; Protein Kinase Inhibitors; Receptor Protein-Tyrosine Kinases

Neuroblastoma (NB) is the second leading extracranial solid tumor of early childhood with about two-thirds of cases presenting before the age of 5, and accounts for roughly 15 percent of all pediatric cancer fatalities in the United States. Treatments against NB are lacking, resulting in a low survival rate in high-risk patients. A repurposing approach using already approved or clinical stage compounds can be used for diseases for which the patient population is small, and the commercial market limited. We have used Bayesian machine learning, in vitro cell assays, and combination analysis to identify molecules with potential use for NB. We demonstrated that pyronaridine (SH-SY5Y IC

Topics: Bayes Theorem; Cell Line, Tumor; Child; Child, Preschool; Crizotinib; Drug Repositioning; Etoposide; Fingolimod Hydrochloride; Humans; Neuroblastoma; Niclosamide

Topics: Anaplastic Lymphoma Kinase; Benzamides; Chromosomes, Human, Pair 2; Cytokines; Genetic Variation; Germ Cells; Humans; Indazoles; Infant; Male; Neuroblastoma; Protein Kinase Inhibitors; Receptor, trkA

Topics: Alternative Splicing; Benzamides; Calmodulin; Cell Line, Tumor; Crizotinib; Doxorubicin; Humans; Indazoles; Neuroblastoma; Oxygen; Proto-Oncogene Proteins c-akt; Receptor, trkA; Ryanodine Receptor Calcium Release Channel

Topics: Animals; Apoptosis; Benzamides; Biomarkers, Tumor; Cell Proliferation; Drug Resistance, Neoplasm; Gene Expression Regulation, Neoplastic; Humans; Indazoles; Mice; Mice, Nude; Neuroblastoma; Protein Kinase Inhibitors; Proteome; RNA-Seq; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Neuroblastoma (NB) is a threatening childhood malignancy. Its prognosis is affected by several morphological, and biological characteristics, including the constitutive expression of ALK tyrosine kinase. In this study we examined the therapeutic potential of a novel ALK inhibitor, entrectinib, in obliterating NB tumor cells. Entrectinib showed the growth-inhibitory effects on NB cells with a 50% inhibitory concentration range of 0.03-5 μM. In the ALK-dependent cells, entrectinib mediated G1-arrest, which was associated with modified expression of multiple cell-cycle regulators. Down-regulation of Ki-67, and attenuated phosphorylation of ERK1/2, and STAT3, correlated with observed antiproliferative capacity of entrectinib. Initial cytostatic activity of entrectinib was followed by concentration-dependent apoptotic cell death, and Caspase-3 activation. However, we delineated a reduced sensitivity of ALK mutated NB cells to entrectinib, and demonstrated strong activation of autophagy in SH-SY5YF1174L NB cell line. Abrogation of autophagy by chloroquine increased significantly the toxicity of entrectinib, as confirmed by enhanced death rate, and PARP protein cleavage in SH-SY5YF1174L cells. In aggregate, our data show that entrectinib inhibits proliferation, and induces G1-arrest, and apoptosis in NB cells. We propose entrectinib for further consideration in treatment of NB, and recommend pharmacological inhibition of autophagy to be explored for a combined therapeutic approach in NB patients that might develop resistance to entrectinib.

Topics: Anaplastic Lymphoma Kinase; Apoptosis; Autophagy; Benzamides; Blotting, Western; Cell Cycle; Cell Movement; Cell Proliferation; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; Indazoles; Neuroblastoma; Phosphorylation; Real-Time Polymerase Chain Reaction; Receptor Protein-Tyrosine Kinases; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Tumor Cells, Cultured; Wound Healing

Neuroblastoma (NB) is one of the most common and deadly childhood solid tumors. These tumors are characterized by clinical heterogeneity, from spontaneous regression to relentless progression, and the Trk family of neurotrophin receptors plays an important role in this heterogeneous behavior. We wanted to determine if entrectinib (RXDX-101, Ignyta, Inc.), an oral Pan-Trk, Alk and Ros1 inhibitor, was effective in our NB model. In vitro effects of entrectinib, either as a single agent or in combination with the chemotherapeutic agents Irinotecan (Irino) and Temozolomide (TMZ), were studied on an SH-SY5Y cell line stably transfected with TrkB. In vivo growth inhibition activity was studied in NB xenografts, again as a single agent or in combination with Irino-TMZ. Entrectinib significantly inhibited the growth of TrkB-expressing NB cells in vitro, and it significantly enhanced the growth inhibition of Irino-TMZ when used in combination. Single agent therapy resulted in significant tumor growth inhibition in animals treated with entrectinib compared to control animals [p < 0.0001 for event-free survival (EFS)]. Addition of entrectinib to Irino-TMZ also significantly improved the EFS of animals compared to vehicle or Irino-TMZ treated animals [p < 0.0001 for combination vs. control, p = 0.0012 for combination vs. Irino-TMZ]. We show that entrectinib inhibits growth of TrkB expressing NB cells in vitro and in vivo, and that it enhances the efficacy of conventional chemotherapy in in vivo models. Our data suggest that entrectinib is a potent Trk inhibitor and should be tested in clinical trials for NBs and other Trk-expressing tumors.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Benzamides; Camptothecin; Cell Line, Tumor; Cell Proliferation; Dacarbazine; Dose-Response Relationship, Drug; Humans; Indazoles; Irinotecan; Membrane Glycoproteins; Mice, Nude; Neuroblastoma; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Receptor, trkB; Signal Transduction; Temozolomide; Time Factors; Transfection; Tumor Burden; Xenograft Model Antitumor Assays