Cbl and Cbl-b ubiquitin ligases are essential for intestinal epithelial stem cell maintenance

Neha Zutshi; Bhopal C. Mohapatra; Pinaki Mondal; Wei An; Benjamin T. Goetz; Shuo Wang; Sicong Li; Matthew D. Storck; David F. Mercer; Adrian R. Black; Sarah P. Thayer; Jennifer D. Black; Chi Lin; Vimla Band; Hamid Band

iScience (Jun 2024)

Cbl and Cbl-b ubiquitin ligases are essential for intestinal epithelial stem cell maintenance

Neha Zutshi,
Bhopal C. Mohapatra,
Pinaki Mondal,
Wei An,
Benjamin T. Goetz,
Shuo Wang,
Sicong Li,
Matthew D. Storck,
David F. Mercer,
Adrian R. Black,
Sarah P. Thayer,
Jennifer D. Black,
Chi Lin,
Vimla Band,
Hamid Band

Affiliations

Neha Zutshi: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA; Department of Pathology & Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, USA
Bhopal C. Mohapatra: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA; Department of Genetics, Cell Biology & Anatomy, University of Nebraska Medical Center, Omaha, NE 68198, USA
Pinaki Mondal: Department of Surgery, University of Nebraska Medical Center, Omaha, NE 68198, USA
Wei An: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA
Benjamin T. Goetz: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA
Shuo Wang: Department of Radiation Oncology, College of Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA
Sicong Li: Department of Radiation Oncology, College of Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA
Matthew D. Storck: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA
David F. Mercer: Department of Surgery, University of Nebraska Medical Center, Omaha, NE 68198, USA
Adrian R. Black: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA; Fred & Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA
Sarah P. Thayer: Department of Surgery, University of Nebraska Medical Center, Omaha, NE 68198, USA; Fred & Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA
Jennifer D. Black: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA; Fred & Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA
Chi Lin: Department of Radiation Oncology, College of Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA; Fred & Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA
Vimla Band: Department of Genetics, Cell Biology & Anatomy, University of Nebraska Medical Center, Omaha, NE 68198, USA; Fred & Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA; Corresponding author
Hamid Band: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA; Department of Pathology & Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, USA; Department of Genetics, Cell Biology & Anatomy, University of Nebraska Medical Center, Omaha, NE 68198, USA; Fred & Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA; Corresponding author

Journal volume & issue: Vol. 27, no. 6
p. 109912

Abstract

Read online

Summary: Receptor tyrosine kinases (RTKs) control stem cell maintenance vs. differentiation decisions. Casitas B-lineage lymphoma (CBL) family ubiquitin ligases are negative regulators of RTKs, but their stem cell regulatory roles remain unclear. Here, we show that Lgr5+ intestinal stem cell (ISC)-specific inducible Cbl-knockout (KO) on a Cblb null mouse background (iDKO) induced rapid loss of the Lgr5Hi ISCs with transient expansion of the Lgr5Lo transit-amplifying population. LacZ-based lineage tracing revealed increased ISC commitment toward enterocyte and goblet cell fate at the expense of Paneth cells. Functionally, Cbl/Cblb iDKO impaired the recovery from radiation-induced intestinal epithelial injury. In vitro, Cbl/Cblb iDKO led to inability to maintain intestinal organoids. Single-cell RNA sequencing in organoids identified Akt-mTOR (mammalian target of rapamycin) pathway hyperactivation upon iDKO, and pharmacological Akt-mTOR axis inhibition rescued the iDKO defects. Our results demonstrate a requirement for Cbl/Cblb in the maintenance of ISCs by fine-tuning the Akt-mTOR axis to balance stem cell maintenance vs. commitment to differentiation.

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

About the journal

Abstract

Keywords