A High-Throughput Screen Identifies That CDK7 Activates Glucose Consumption in Lung Cancer Cells

Nat Commun. 2019 Nov 29;10(1):5444. doi: 10.1038/s41467-019-13334-8.

Abstract

Elevated glucose consumption is fundamental to cancer, but selectively targeting this pathway is challenging. We develop a high-throughput assay for measuring glucose consumption and use it to screen non-small-cell lung cancer cell lines against bioactive small molecules. We identify Milciclib that blocks glucose consumption in H460 and H1975, but not in HCC827 or A549 cells, by decreasing SLC2A1 (GLUT1) mRNA and protein levels and by inhibiting glucose transport. Milciclib blocks glucose consumption by targeting cyclin-dependent kinase 7 (CDK7) similar to other CDK7 inhibitors including THZ1 and LDC4297. Enhanced PIK3CA signaling leads to CDK7 phosphorylation, which promotes RNA Polymerase II phosphorylation and transcription. Milciclib, THZ1, and LDC4297 lead to a reduction in RNA Polymerase II phosphorylation on the SLC2A1 promoter. These data indicate that our high-throughput assay can identify compounds that regulate glucose consumption and that CDK7 is a key regulator of glucose consumption in cells with an activated PI3K pathway.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Carcinoma, Non-Small-Cell Lung / metabolism*
  • Cell Line, Tumor
  • Class I Phosphatidylinositol 3-Kinases / drug effects
  • Class I Phosphatidylinositol 3-Kinases / metabolism
  • Cyclin-Dependent Kinases / antagonists & inhibitors*
  • Cyclin-Dependent Kinases / metabolism
  • Glucose / metabolism*
  • Glucose Transporter Type 1 / drug effects*
  • Glucose Transporter Type 1 / metabolism
  • High-Throughput Screening Assays
  • Humans
  • Lung Neoplasms / metabolism*
  • Phenylenediamines / pharmacology
  • Phosphorylation
  • Protein Kinase Inhibitors / pharmacology*
  • Pyrazoles / pharmacology
  • Pyrimidines / pharmacology
  • Quinazolines / pharmacology
  • RNA Polymerase II / drug effects
  • RNA Polymerase II / genetics
  • RNA Polymerase II / metabolism
  • RNA, Messenger / metabolism
  • Signal Transduction
  • Triazines / pharmacology

Substances

  • Glucose Transporter Type 1
  • LDC4297
  • N,1,4,4-tetramethyl-8-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4,5-dihydro-1H-pyrazolo(4,3-h)quinazoline-3-carboxamide
  • Phenylenediamines
  • Protein Kinase Inhibitors
  • Pyrazoles
  • Pyrimidines
  • Quinazolines
  • RNA, Messenger
  • THZ1 compound
  • Triazines
  • Class I Phosphatidylinositol 3-Kinases
  • PIK3CA protein, human
  • Cyclin-Dependent Kinases
  • cyclin-dependent kinase-activating kinase
  • RNA Polymerase II
  • Glucose