TY - JOUR
T1 - Histone deacetylase-10 liberates spermidine to support polyamine homeostasis and tumor cell growth
AU - Stewart, Tracy Murray
AU - Foley, Jackson R.
AU - Holbert, Cassandra E.
AU - Klinke, Glynis
AU - Poschet, Gernot
AU - Steimbach, Raphael R.
AU - Miller, Aubry K.
AU - Casero, Robert A.
N1 - Funding Information:
DFMO was kindly provided by Dr Patrick Woster at the Medical University of South Carolina, and Trimer44NMe was a gift from Dr Otto Phanstiel, IV at the University of Central Florida. We thank Dr David Christianson at the University of Pennsylvania for initially spawning our interest in HDAC10-mediated polyamine deacetylation. T. M. S. and A. K. M. conceptualization; T. M. S. J. R. F. G. K. G. P. and R. A. C. methodology; T. M. S. validation; T. M. S. and R. R. S. formal analysis; T. M. S. J. R. F. G. K. G. P. and R. R. S. investigation; T. M. S. writing – original draft; T. M. S. and R. R. S. visualization; C. E. H. G. K. G. P. R. R. S. A. K. M. and R. A. C. writing – review and editing; A. K. M. and R. A. C. resources; A. K. M. and R. A. C. supervision; R. A. C. funding acquisition. Work in the Casero and Stewart laboratory is supported by grants from the US National Institutes of Health, United States (CA204345 and CA235863), the Samuel Waxman Cancer Research Foundation, United States, the University of Pennsylvania Orphan Disease Center, United States Million Dollar Bike Ride (MDBR-20–135-SRS), the Chan Zuckerberg Initiative, and a research contract from Panbela Therapeutics, Inc. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
Work in the Casero and Stewart laboratory is supported by grants from the US National Institutes of Health, United States ( CA204345 and CA235863 ), the Samuel Waxman Cancer Research Foundation, United States , the University of Pennsylvania Orphan Disease Center, United States Million Dollar Bike Ride (MDBR-20–135-SRS), the Chan Zuckerberg Initiative, and a research contract from Panbela Therapeutics, Inc. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2022 The Authors
PY - 2022/10
Y1 - 2022/10
N2 - Cytosolic histone deacetylase-10 (HDAC10) specifically deacetylates the modified polyamine N8-acetylspermidine (N8-AcSpd). Although intracellular concentrations of N8-AcSpd are low, extracellular sources can be abundant, particularly in the colonic lumen. Extracellular polyamines, including those from the diet and microbiota, can support tumor growth both locally and at distant sites. However, the contribution of N8-AcSpd in this context is unknown. We hypothesized that HDAC10, by converting N8- AcSpd to spermidine, may provide a source of this growth-supporting polyamine in circumstances of reduced polyamine biosynthesis, such as in polyamine-targeting anticancer therapies. Inhibitors of polyamine biosynthesis, including α-difluoromethylornithine (DFMO), inhibit tumor growth, but compensatory uptake of extracellular polyamines has limited their clinical success. Combining DFMO with inhibitors of polyamine uptake have improved the antitumor response. However, acetylated polyamines may use different transport machinery than the parent molecules. Here, we use CRISPR/Cas9-mediated HDAC10-knockout cell lines and HDAC10-specific inhibitors to investigate the contribution of HDAC10 in maintaining tumor cell proliferation. We demonstrate inhibition of cell growth by DFMO-associated polyamine depletion is successfully rescued by exogenous N8-AcSpd (at physiological concentrations), which is converted to spermidine and spermine, only in cell lines with HDAC10 activity. Furthermore, we show loss of HDAC10 prevents both restoration of polyamine levels and growth rescue, implicating HDAC10 in supporting polyamine-associated tumor growth. These data suggest the utility of HDAC10-specific inhibitors as an antitumor strategy that may have value in improving the response to polyamine-blocking therapies. Additionally, the cell-based assay developed in this study provides an inexpensive, high-throughput method of screening potentially selective HDAC10 inhibitors.
AB - Cytosolic histone deacetylase-10 (HDAC10) specifically deacetylates the modified polyamine N8-acetylspermidine (N8-AcSpd). Although intracellular concentrations of N8-AcSpd are low, extracellular sources can be abundant, particularly in the colonic lumen. Extracellular polyamines, including those from the diet and microbiota, can support tumor growth both locally and at distant sites. However, the contribution of N8-AcSpd in this context is unknown. We hypothesized that HDAC10, by converting N8- AcSpd to spermidine, may provide a source of this growth-supporting polyamine in circumstances of reduced polyamine biosynthesis, such as in polyamine-targeting anticancer therapies. Inhibitors of polyamine biosynthesis, including α-difluoromethylornithine (DFMO), inhibit tumor growth, but compensatory uptake of extracellular polyamines has limited their clinical success. Combining DFMO with inhibitors of polyamine uptake have improved the antitumor response. However, acetylated polyamines may use different transport machinery than the parent molecules. Here, we use CRISPR/Cas9-mediated HDAC10-knockout cell lines and HDAC10-specific inhibitors to investigate the contribution of HDAC10 in maintaining tumor cell proliferation. We demonstrate inhibition of cell growth by DFMO-associated polyamine depletion is successfully rescued by exogenous N8-AcSpd (at physiological concentrations), which is converted to spermidine and spermine, only in cell lines with HDAC10 activity. Furthermore, we show loss of HDAC10 prevents both restoration of polyamine levels and growth rescue, implicating HDAC10 in supporting polyamine-associated tumor growth. These data suggest the utility of HDAC10-specific inhibitors as an antitumor strategy that may have value in improving the response to polyamine-blocking therapies. Additionally, the cell-based assay developed in this study provides an inexpensive, high-throughput method of screening potentially selective HDAC10 inhibitors.
KW - N8-acetylspermidine
KW - colorectal cancer
KW - difluoromethylornithine (DFMO)
KW - histone deacetylase-10 (HDAC10)
KW - metabolism
KW - microbiome
KW - polyamine
KW - polyamine-blocking therapy
KW - spermidine
KW - tumor microenvironment (TME)
UR - http://www.scopus.com/inward/record.url?scp=85138048439&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85138048439&partnerID=8YFLogxK
U2 - 10.1016/j.jbc.2022.102407
DO - 10.1016/j.jbc.2022.102407
M3 - Article
C2 - 35988653
AN - SCOPUS:85138048439
SN - 0021-9258
VL - 298
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 10
M1 - 102407
ER -