An essential role of CBL and CBL-B ubiquitin ligases in mammary stem cell maintenance

Abstract

CBL and CBL-B ubiquitin ligases are negative regulators of tyrosine kinase signaling with established roles in the immune system. However, their physiological roles in epithelial tissues are unknown. Here we used the MMTV-Cre-mediated Cbl gene deletion on a Cbl-b-null background as well as a tamoxifen-inducible mammary stem cell (MaSC)-specific Cbl/Cbl-b double knockout (DKO), using Lgr5-GFP-CreERT, to demonstrate a mammary epithelial cell-autonomous requirement of CBL and CBL-B in the maintenance of MaSCs. Using a newly engineered tamoxifen (TAM)-inducible Cbl/Cbl-b deletion model with a dual fluorescent reporter (Cblflox/flox; Cbl-bflox/flox; Rosa26-CreERT; mT/mG), we show that Cbl/Cbl-b DKO in mammary organoids leads to hyper-activation of AKT-mTOR signaling with depletion of MaSCs. Chemical inhibition of AKT or mTOR rescued MaSCs from Cbl/Cbl-b DKO induced depletion. Our studies reveal a novel, cell-autonomous, requirement of CBL and CBL-B in epithelial stem cell maintenance during organ development and remodeling through modulation of mTOR signaling.

Document Details

Document Type
Pub Defense Publication
Publication Date
Jan 01, 2017
Source ID
10.1242/dev.138164

Entities

People

  • Benjamin Goetz
  • Bhopal Mohapatra
  • Hamid Band
  • Insha Mustaq
  • Jane L. Meza
  • Matthew D Storck
  • Neha Zutshi
  • Priyanka Arya
  • Timothy A. Bielecki
  • Wei An

Organizations

  • Office of Extramural Research
  • United States Department of Defense
  • University of Nebraska Medical Center
  • University of Nebraska–Lincoln

Tags

Fields of Study

  • Biology
  • Chemistry

Readers

  • Breast cancer cell signaling and growth regulation.
  • Molecular Biology and Genetics

Technology Areas

  • Biotechnology