The E3 ubiquitin ligase Cul5 regulates hematopoietic stem cell function for steady-state hematopoiesis in mice

Siera A. Tomishima; Dale D. Kim; Nadia Porter; Ipsita Guha; Asif A. Dar; Yohaniz Ortega-Burgos; Jennifer Roof; Hossein Fazelinia; Lynn A. Spruce; Christopher S. Thom; Robert L. Bowman; Paula M. Oliver

doi:10.1172/JCI180913

The E3 ubiquitin ligase Cul5 regulates hematopoietic stem cell function for steady-state hematopoiesis in mice

Siera A. Tomishima,¹ Dale D. Kim,¹ Nadia Porter,² Ipsita Guha,² Asif A. Dar,² Yohaniz Ortega-Burgos,¹ Jennifer Roof,³ Hossein Fazelinia,³ Lynn A. Spruce,³ Christopher S. Thom,⁴ Robert L. Bowman,⁵ and Paula M. Oliver¹

Published June 26, 2025 - More info

View PDF

Abstract

The balance of hematopoietic stem cell (HSC) self-renewal versus differentiation is essential to ensure long-term repopulation capacity while allowing response to events that require increased hematopoietic output. Proliferation and differentiation of HSCs and their progeny is controlled by the JAK/STAT pathway downstream of cytokine signaling. E3 ubiquitin ligases, like Cullin 5 (Cul5), can regulate JAK/STAT signaling by degrading signaling intermediates. Here we report that mice lacking Cul5 in hematopoietic cells (Cul5^Vav-Cre) have increased numbers of HSPCs, splenomegaly, and extramedullary hematopoiesis. Differentiation in Cul5^Vav-Cre mice is myeloid- and megakaryocyte-biased, resulting in leukocytosis, anemia and thrombocytosis. Cul5^Vav-Cre mice increased HSC proliferation and circulation, associated with a decrease in CXCR4 surface expression. In bone marrow cells, we identified LRRC41 co-immunoprecipitated with CUL5, and vice versa, supporting that CRL5 forms a complex with LRRC41. We identified an accumulation of LRRC41 and STAT5 in Cul5^Vav-Cre HSCs during IL-3 stimulation, supporting their regulation by Cul5. Whole cell proteome (WCP) analysis of HSPCs from Cul5^Vav-Cre bone marrow identified upregulation of many STAT5 target genes and associated pathways. Finally, JAK1/2 inhibition with ruxolitinib normalized hematopoiesis in Cul5^Vav-Cre mice. These studies demonstrate the function of Cul5 in HSC function, stem cell fate decisions, and regulation of IL-3 signaling.

The E3 ubiquitin ligase Cul5 regulates hematopoietic stem cell function for steady-state hematopoiesis in mice

Siera A. Tomishima,¹ Dale D. Kim,¹ Nadia Porter,² Ipsita Guha,² Asif A. Dar,² Yohaniz Ortega-Burgos,¹ Jennifer Roof,³ Hossein Fazelinia,³ Lynn A. Spruce,³ Christopher S. Thom,⁴ Robert L. Bowman,⁵ and Paula M. Oliver¹

Article tools

Metrics

Go to

The E3 ubiquitin ligase Cul5 regulates hematopoietic stem cell function for steady-state hematopoiesis in mice

Siera A. Tomishima,1 Dale D. Kim,1 Nadia Porter,2 Ipsita Guha,2 Asif A. Dar,2 Yohaniz Ortega-Burgos,1 Jennifer Roof,3 Hossein Fazelinia,3 Lynn A. Spruce,3 Christopher S. Thom,4 Robert L. Bowman,5 and Paula M. Oliver1

Article tools

Metrics

Go to

Sign up for email alerts

Siera A. Tomishima,¹ Dale D. Kim,¹ Nadia Porter,² Ipsita Guha,² Asif A. Dar,² Yohaniz Ortega-Burgos,¹ Jennifer Roof,³ Hossein Fazelinia,³ Lynn A. Spruce,³ Christopher S. Thom,⁴ Robert L. Bowman,⁵ and Paula M. Oliver¹