Neuronal calcium sensor 1 (NCS1) promotes motility and metastatic spread of breast cancer cells in vitro and in vivo

Abstract

Increased levels of the calcium‐binding protein neuronal calcium sensor 1 (NCS1) predict an unfavorable patient outcome in several aggressive cancers, including breast and liver tumors. Previous studies suggest that NCS1 overexpression facilitates metastatic spread of these cancers. To investigate this hypothesis, we explored the effects of NCS1 overexpression on cell proliferation, survival, and migration patterns in vitro in 2‐ and 3‐dimensional (2/3‐D). Furthermore, we translated our results into an in vivo mouse xenograft model. Cell‐based proliferation assays were used to demonstrate the effects of overexpression of NCS1 on growth rates. In vitro colony formation and wound healing experiments were performed and 3‐D migration dynamics were studied using collagen gels. Nude mice were injected with breast cancer cells to monitor NCS1‐dependent metastasis formation over time. We observed that increased NCS1 levels do not change cellular growth rates, but do significantly increase 2‐ and 3‐D migration dynamics in vitro. Likewise, NCS1‐overexpressing cells have an increased capacity to form distant metastases and demonstrate better survival and less necrosis in vivo. We found that NCS1 preferentially localizes to the leading edge of cells and overexpression increases the motility of cancer cells. Furthermore, this phenotype is correlated with an increased number of metastases in a xenograft model. These results lay the foundation for exploring the relevance of an NCS1‐mediated pathway as a metastatic biomarker and as a target for pharmacologic interventions.—Apasu, J. E., Schuette, D., LaRanger, R., Steinle, J. A., Nguyen, L. D., Grosshans, H. K., Zhang, M., Cai, W. L., Yan, Q., Robert, M. E., Mak, M., Ehrlich, B. E. Neuronal calcium sensor 1 (NCS1) promotes motility and metastatic spread of breast cancer cells in vitro and in vivo. FASEB J. 33, 4802–4813 (2019). http://www.fasebj.org

Neuronal Calcium Sensor 1 is up‐regulated in response to stress to promote cell survival and motility in cancer cells

“…We found that this mechanism of NCS1 up‐regulation is conserved in the pathophysiology of human breast cancer, and we showed that NCS1 has essential functions for cell survival and migration through altering intracellular Ca 2+ signaling and Akt signaling. The novel finding that extracellular stressors lead to NCS1 up‐regulation helps us understand how high NCS1 expression leads to more aggressive behavior of tumor cells and worse patient outcome (Apasu et al , 2019; Moore et al , 2017; Schuette et al , 2018). In conclusion, we describe a novel role for NCS1 as stress response protein linking the tumor microenvironment and cancer progression.…”

“…Here, we focus on the Ca 2+ ‐binding protein neuronal calcium sensor 1 (NCS1), which shares multiple characteristics with the S100 protein family. Like S100 proteins, NCS1 belongs to the EF‐hand superfamily of Ca 2+ ‐binding proteins (Burgoyne, 2007; Donato et al , 2013), is increased in cancerous tissues, and contributes to a more aggressive phenotype of these tumors (Apasu et al , 2019; Moore et al , 2017; Moore et al , 2018; Schuette et al , 2018). It binds Ca 2+ with high affinity and contains four EF‐hand domains, three of which bind Ca 2+ , and one of which, in the N‐terminal domain, is unable to bind Ca 2+ (Boeckel and Ehrlich, 2018; Bourne et al , 2001).…”

Neuronal calcium sensor 1 (NCS1) dependent modulation of neuronal morphology and development

“…55,56 A role for NCS1 in regulating neuronal morphology is supported by previous studies describing that NCS1 enhances neurite outgrowth [16][17][18] and synaptic transmission, specifically neurotransmitter release, LTP, and LTD. [12][13][14][15] Similarly, NCS1 regulates exocytosis in adrenal chromaffin cells, 57 morphology of cardiomyocytes, 52 and motility of cancer cells. 50,58 The gross morphology of the brain has been reported to be unaltered in Ncs1 −/− mice. 21 Reduced dendritic morphology in Ncs1 −/− mice may be due to several, not mutually exclusive mechanisms.…”

Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling

“…Abnormalities in the expression and function of NCS-1, as well as hereditary mutations in its gene, are associated with neurodegenerative and neuropsychiatric diseases, including schizophrenia, autism, Alzheimer's disease, and Parkinson's disease [23]. In addition, NCS-1 is aberrantly expressed in certain cancers, and the level of its expression correlates with tumor aggressiveness and patients' survival rates [24,25].…”