Miki Lab/Division of Cellular and Molecular Biology  Department of Cellular Regulation

Most cancers originate from epithelial cells. Normal epithelial cells form a sheet-like tissue structure in which cells are tightly attached to each other and to the basement membrane. Through malignant progression, cells proliferate and expand by invading surrounding tissues. Furthermore, cells metastasize to distant organs via blood vessels, forming often incurable tumors. Our aim is to elucidate the mechanism underlying this mysterious process of cancer development.

Role of PRL in malignant progression of cancers

PRL is highly expressed in malignant tumors and promotes cancer metastasis. We discovered that PRL associates with CNNM4, a Mg2+ transporter, and inhibits its Mg2+ transporting activity. Moreover, we also found that intestinal polyps became malignant and invaded the surrounding muscle tissue when CNNM4 was disrupted. At present, we are investigating the functional relationship between cancer malignancy and Mg2+ dyshomeostasis caused by CNNM4 inhibition.
In normal epithelial tissues, cells are attached to each other and collectively maintain their structure; these characteristics are disrupted in cancer tissues. Forced expression of PRL in epithelial cells cultured on matrix gels induced a marked change in their morphology; some cells invaded into the matrix only when PRL-expressing cells were surrounded by non-expressing cells. These results suggest that physical interaction between PRL-expressing cells and non-expressing cells stimulates invasive behavior during malignant progression. We are trying to clarify the molecular mechanism underlying this function of PRL.

Functional analyses based on organoid culture of intestinal epithelia

A method of culturing intestinal epithelia in 3D matrix gels that mimic in vivo situations was recently developed; the system is called organoid culture. In this system, intestinal epithelial cells differentiate and form a structure comprising a monolayer sheet of cells. We are using this organoid culture system to investigate the role of PRL/CNNM in cell proliferation, differentiation, and cancerous transformation.
Many oncogenes and anti-oncogenes involved in regulating cell proliferation and survival have been identified. By contrast, characteristics involved in transformation of epithelial cells in a 3D space, which accompany architectural changes (such as invasion and metastasis) in tissues, remain unclear. For example, how do cancer cells exit the epithelial tissue in which they are “born” and expand their territory by invading surrounding tissues? We are tackling these problems and trying to identify the mechanisms underlying cancer development.

  • Fig. 1. Macroscopic images of the intestine (left) and histological images of polyps (right) in the indicated genetically engineered mice. CNNM4-deficient mice develop adenocarcinomas that invade the muscle layer (arrows).

  • Fig. 2. Organoid culture of intestinal epithelia from the indicated genetically engineered mice. CNNM4 deficiency causes morphological abnormalities.

Staff

  • Prof. : Hiroaki Miki
  • Assoc. Prof. : Daisuke Yamazaki
  • Asst. Prof. : Yosuke Funato
  • Postdoc. : Osamu Hashizume

Website

Publications

  • (1) Phosphocysteine in the PRL-CNNM pathway mediates magnesium homeostasis. Gulerez et al., EMBO Rep. (2016) 17(12):1890-1900
    (2) Mg2+ Extrusion from Intestinal Epithelia by CNNM Proteins Is Essential for Gonadogenesis via AMPK-TORC1 Signaling in Caenorhabditis elegans. Ishii T., et al., PLoS Genet. (2016) 12(8):e1006276
    (3) Membrane protein CNNM4-dependent Mg2+ efflux suppresses tumor progression. Funato Y., et al., J Clin Invest. (2014) 124(12):5398-5410
    (4) Basolateral Mg2+ extrusion via CNNM4 mediates transcellular Mg2+ transport across epithelia: a mouse model. Yamazaki D., et al., PLoS Genet. (2013) 9(12):e1003983
    (5) Thioredoxin mediates oxidation-dependent phosphorylation of CRMP2 and growth cone collapse. Morinaka A., et al., Sci Signal. (2011) 4(170):ra26
    (6) Nucleoredoxin sustains Wnt/β-catenin signaling by retaining a pool of inactive dishevelled protein. Funato Y., et al., Curr Biol. (2010) 20(21):1945-1952