Analysis of a mechanism of Girdin-mediated regulation of cancer cell metabolism

• Project/Area Number: 17K17791
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Experimental pathology; Pathological medical chemistry
• Research Institution: Nagoya University
• Principal Investigator: 翁 良 名古屋大学, 医学系研究科, 特任助教 (20729280)
• Project Period (FY): 2017-04-01 – 2018-03-31
• Project Status: Discontinued (Fiscal Year 2017)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: cancer metabolism / Girdin / actin cytoskeleton / PKM2 / glycolysis

Outline of Annual Research Achievements

The previous work from our laboratory characterized the actin-binding protein Girdin as a critical regulator of migration of endothelial cells, cancer cells, and neuroblasts, which depends on extracellular cues including growth factor stimulation. We also previously found that Girdin also interacts with amino acids transporters to regulate amino acids-induced mTORC1 activation. To further investigate the role of Girdin in cancer metabolism, we employed affinity column chromatography to isolate Girdin-interacting proteins, and identified Pyruvate Kinase M2 (PKM2), a key enzyme that regulates the Warburg effect that is necessary for tumor growth, as a candidate. The interaction of PKM2 with Girdin was confirmed by co-immunoprecipitation (co-IP), western blot analysis and immunofluorescence. Next, to check on the role of Girdin in cancer metabolism, we evaluated the effect of Girdin depletion on ATP and lactate production, and glucose consumption by commercially available biochemical kits. We also investigated the role of Girdin in PKM2 Y105 phosphorylation, which is highly related to PKM2 enzyme activity. We also analyzed the role of Girdin in lung cancer progression through IHC, where we found that the expression of Girdin is correlated with the expression of PKM2 in lung cancer tissue. In summary, the data we got from the study revealed the role of Girdin in cancer metabolism and cancer progression, and showed the possibility that Girdin is involved in the modulation of chemotherapy effect through affecting aerobic glycolysis.

Research Products

• [Journal Article] Negative regulation of amino acid signaling by MAPK-regulated 4F2hc/Girdin complex (2018)
  • Author(s): Liang Weng , Yi-Peng Han, Atsushi Enomoto, Yasuyuki Kitaura, Shushi Nagamori, Yoshikatsu Kanai, Naoya Asai, Jian An, Maki Takagishi, Masato Asai, Shinji Mii, Takashi Masuko, Yoshiharu Shimomura, Masahide Takahashi.
  • Journal Title: PLoS Biology Volume: 16 Issue: 3 Pages: e2005090-e2005090
  • DOI: 10.1371/journal.pbio.2005090
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Daple Coordinates Planar Polarized Microtubule Dynamics in Ependymal Cells and Contributes to Hydrocephalus (2017)
  • Author(s): Maki Takagishi, Masato Sawada, Shinya Ohata, Naoya Asai, Atsushi Enomoto, Kunihiko Takahashi, Liang Weng, Kaori Ushida, Hosne Ara, Shigeyuki Matsui, Kozo Kaibuchi,3Kazunobu Sawamoto, and Masahide Takahashi.
  • Journal Title: Cell Reports Volume: 20 Issue: 4 Pages: 960-972
  • DOI: 10.1016/j.celrep.2017.06.089
  • Peer Reviewed / Open Access
• [Presentation] Girdin regulates the collective migration of cancer cells by interacting with beta-catenin/alpha-catenin (2017)
  • Author(s): Xiaoze Wang、Atsushi Enomoto、Liang Weng、Naoya Asai、Masahide Takahashi
  • Organizer: 第76回日本癌学会学術総会、パシフィコ横浜（神奈川県横浜市）