title: Cinacalcet inhibits neuroblastoma tumor growth and upregulates cancer-testis antigens.
abstract: The calcium-sensing receptor is a G protein-coupled receptor that exerts cell-type specific functions in numerous tissues and some cancers. We have previously reported that this receptor exhibits tumor suppressor properties in neuroblastoma. We have now assessed cinacalcet, an allosteric activator of the CaSR approved for clinical use, as targeted therapy for this developmental tumor using neuroblastoma cell lines and patient-derived xenografts (PDX) with different MYCN and TP53 status. In vitro, acute exposure to cinacalcet induced endoplasmic reticulum stress coupled to apoptosis via ATF4-CHOP-TRB3 in CaSR-positive, MYCN-amplified cells. Both phenotypes were partially abrogated by phospholipase C inhibitor U73122. Prolonged in vitro treatment also promoted dose- and time-dependent apoptosis in CaSR-positive, MYCN-amplified cells and, irrespective of MYCN status, differentiation in surviving cells. Cinacalcet significantly inhibited tumor growth in MYCN-amplified xenografts and reduced that of MYCN-non amplified PDX. Morphology assessment showed fibrosis in MYCN-amplified xenografts exposed to the drug. Microarrays analyses revealed up-regulation of cancer-testis antigens (CTAs) in cinacalcet-treated MYCN-amplified tumors. These were predominantly CTAs encoded by genes mapping on chromosome X, which are the most immunogenic. Other modulated genes upon prolonged exposure to cinacalcet were involved in differentiation, cell cycle exit, microenvironment remodeling and calcium signaling pathways. CTAs were up-regulated in PDX and in vitro models as well. Moreover, progressive increase of CaSR expression upon cinacalcet treatment was seen both in vitro and in vivo. In summary, cinacalcet reduces neuroblastoma tumor growth and up-regulates CTAs. This effect represents a therapeutic opportunity and provides surrogate circulating markers of neuroblastoma response to this treatment.
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Carlos J
authors.initials: CJ
authors.lastname: Rodríguez-Hernández
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Silvia
authors.initials: S
authors.lastname: Mateo-Lozano
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: García
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Carla
authors.initials: C
authors.lastname: Casalà
authors.affiliation: Statistics and Bioinformatics Unit, Vall d'Hebron Research Institute, Barcelona, Spain.
authors.forename: Ferran
authors.initials: F
authors.lastname: Briansó
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Nerea
authors.initials: N
authors.lastname: Castrejón
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Rodríguez
authors.affiliation: Department of Pathology, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Mariona
authors.initials: M
authors.lastname: Suñol
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Angel M
authors.initials: AM
authors.lastname: Carcaboso
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.|Department of Oncology, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Lavarino
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.|Department of Oncology, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Jaume
authors.initials: J
authors.lastname: Mora
authors.affiliation: Developmental Tumor Biology Laboratory, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.|Department of Oncology, Institut de Recerca Pediàtrica - Hospital Sant Joan de Déu, Esplugues de Llobregat, Barcelona, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: de Torres
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000187:drug effects; D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D000069449:Cinacalcet / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D000071447:N-Myc Proto-Oncogene Protein / Q000235:genetics; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology*; D044169:Receptors, Calcium-Sensing / Q000819:agonists / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D015854:Up-Regulation; D023041:Xenograft Model Antitumor Assays
chemical_list: D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; C095550:CASR protein, human; C459801:MYCN protein, human; D000071447:N-Myc Proto-Oncogene Protein; D044169:Receptors, Calcium-Sensing; D016159:Tumor Suppressor Protein p53; D000069449:Cinacalcet
keywords: ER stress; calcium-sensing receptor; cancer-testis antigens; cinacalcet; neuroblastoma