GCN2 inhibition sensitizes arginine-deprived hepatocellular carcinoma cells to senolytic treatment.
Hepatocellular carcinoma (HCC) is a typically fatal malignancy exhibiting genetic heterogeneity and limited therapy responses. We demonstrate here that HCCs consistently repress urea cycle gene expression and thereby become auxotrophic for exogenous arginine. Surprisingly, arginine import is uniquely dependent on the cationic amino acid transporter SLC7A1, whose inhibition slows HCC cell growth in vitro and in vivo. Moreover, arginine deprivation engages an integrated stress response that promotes HCC cell-cycle arrest and quiescence, dependent on the general control nonderepressible 2 (GCN2) kinase. Inhibiting GCN2 in arginine-deprived HCC cells promotes a senescent phenotype instead, rendering these cells vulnerable to senolytic compounds. Preclinical models confirm that combined dietary arginine deprivation, GCN2 inhibition, and senotherapy promote HCC cell apoptosis and tumor regression. These data suggest novel strategies to treat human liver cancers through targeting SLC7A1 and/or a combination of arginine restriction, inhibition of GCN2, and senolytic agents.
Pubmed ID: 35839757 RIS Download
Research resources used in this publication
- C57BL/6J (RRID:IMSR_JAX:000664) (organism)
- SK-LMS-1 (RRID:CVCL_0628) (cell line)
- MDA-MB-231 (RRID:CVCL_0062) (cell line)
- PC-3 (RRID:CVCL_0035) (cell line)
- c-myc-PT3EF1a (RRID:Addgene_92046) (plasmid)
- Hep-G2 (RRID:CVCL_0027) (cell line)
- Hep 3B2.1-7 (RRID:CVCL_0326) (cell line)
- Huh-7 (RRID:CVCL_0336) (cell line)
- SNU-475 (RRID:CVCL_0497) (cell line)
- SNU-449 (RRID:CVCL_0454) (cell line)
- SNU-423 (RRID:CVCL_0366) (cell line)
- SNU-398 (RRID:CVCL_0077) (cell line)
- SNU-182 (RRID:CVCL_0090) (cell line)
- HEK293T (RRID:CVCL_0063) (cell line)
Additional research tools detected in this publication
Antibodies used in this publication
- Goat Anti-Rabbit IgG Antibody (H+L), Biotinylated (RRID:AB_2313606)
- Arginase-1 (SP156) Rabbit Monoclonal Antibody (RRID:AB_2890926)
- Ki-67 Monoclonal Antibody (SolA15), FITC, eBioscience (RRID:AB_11151330)
- SLC7A1 Polyclonal Antibody (RRID:AB_2805903)
- Rabbit Anti-ASS1 Polyclonal Antibody, Unconjugated, Clone H-231 (RRID:AB_2060474)
- CREB-2 (H-290) (RRID:AB_2058742)
- p27 (F-8) (RRID:AB_628074)
- Anti-Puromycin Antibody, clone 12D10 (RRID:AB_2566826)
- Recombinant Anti-p21 antibody [EPR18021] (RRID:AB_2734729)
- GCN2 (phospho T899) antibody [EPR2320Y] (RRID:AB_1310260)
- Recombinant Anti-GCN2 antibody [EPR5970(2)] (RRID:AB_2890925)
- CPS1 antibody [EPR7493] (RRID:AB_11156290)
- Recombinant Anti-Ornithine Carbamoyltransferase/OTC antibody [EPR19725] (RRID:AB_2876368)
- ASS1 antibody [2B10] (RRID:AB_10975633)
- Anti-mouse IgG, HRP-linked Antibody (RRID:AB_330924)
- Anti-rabbit IgG, HRP-linked Antibody (RRID:AB_2099233)
- Rabbit Anti-GAPDH Monoclonal Antibody, Unconjugated, Clone 14C10 (RRID:AB_561053)
- LC3B Antibody (RRID:AB_915950)
- SQSTM1/p62 Antibody (RRID:AB_10624872)
- Rabbit Anti-AMPK-alpha, phospho (Thr172) Monoclonal Antibody, Unconjugated, Clone 40H9 (RRID:AB_331250)
- Anti-AMPK-alpha Antibody, Unconjugated (RRID:AB_330331)
- Sestrin-2 (D1B6) Rabbit mAb (RRID:AB_11178663)
- eIF2alpha (D7D3) XP Rabbit mAb (RRID:AB_10692650)
- Phospho-eIF2alpha (Ser51) Antibody (RRID:AB_330951)
- Rabbit Anti-S6 Ribosomal Protein Monoclonal Antibody, Unconjugated, Clone 5G10 (RRID:AB_331355)
- Phospho-S6 Ribosomal Protein (Ser235/236) Antibody (RRID:AB_331679)
- p70 S6 Kinase Antibody (RRID:AB_331676)
- Phospho-p70 S6 Kinase (Thr389) Antibody (RRID:AB_330944)
- mTOR (7C10) Rabbit mAb (RRID:AB_2105622)
- Phospho-mTOR (Ser2448) (D9C2) XP Rabbit mAb (RRID:AB_10691552)
- Cleaved Caspase-3 (Asp175) Antibody (RRID:AB_2341188)
- anti-Cyclin E1 (HE12) Mouse monoclonal Antibody (RRID:AB_2071200)
- Cyclin D1 (92G2) Rabbit mAb (RRID:AB_2259616)
- Cyclin B1 Antibody (RRID:AB_2072132)
- p16 INK4A (D7C1M) Rabbit mAb (RRID:AB_2799960)
- p21 Waf1/Cip1 (12D1) Rabbit monoclonal antibody (RRID:AB_823586)
Associated grants
- Agency: NCI NIH HHS, United States
Id: P01 CA104838 - Agency: NCI NIH HHS, United States
Id: R35 CA220483 - Agency: NCI NIH HHS, United States
Id: T32 CA009140
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This is a list of tools and resources that we have found mentioned in this publication.
FlowJo (tool)
RRID:SCR_008520
Software for single-cell flow cytometry analysis. Its functions include management, display, manipulation, analysis and publication of the data stream produced by flow and mass cytometers.
View all literature mentionsGoat Anti-Rabbit IgG Antibody (H+L), Biotinylated (antibody)
RRID:AB_2313606
This polyclonal targets IgG
View all literature mentionsArginase-1 (SP156) Rabbit Monoclonal Antibody (antibody)
RRID:AB_2890926
This monoclonal targets Arginase-1
View all literature mentionsKi-67 Monoclonal Antibody (SolA15), FITC, eBioscience (antibody)
RRID:AB_11151330
This monoclonal targets Ki-67
View all literature mentionsSLC7A1 Polyclonal Antibody (antibody)
RRID:AB_2805903
This unknown targets SLC7A1
View all literature mentionsRabbit Anti-ASS1 Polyclonal Antibody, Unconjugated, Clone H-231 (antibody)
RRID:AB_2060474
This polyclonal targets ASS1
View all literature mentionsp27 (F-8) (antibody)
RRID:AB_628074
This monoclonal targets Epitope corresponds to AA 1-197 representing full length p27 of mouse origin
View all literature mentionsAnti-Puromycin Antibody, clone 12D10 (antibody)
RRID:AB_2566826
This monoclonal targets Puromycin from Streptomyces alboniger
View all literature mentionsRecombinant Anti-p21 antibody [EPR18021] (antibody)
RRID:AB_2734729
This recombinant targets p21
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RRID:AB_1310260
This monoclonal targets Human GCN2 (phospho T899)
View all literature mentionsRecombinant Anti-GCN2 antibody [EPR5970(2)] (antibody)
RRID:AB_2890925
This recombinant monoclonal targets GCN2
View all literature mentionsCPS1 antibody [EPR7493] (antibody)
RRID:AB_11156290
This monoclonal targets CPS1 antibody [EPR7493]
View all literature mentionsRecombinant Anti-Ornithine Carbamoyltransferase/OTC antibody [EPR19725] (antibody)
RRID:AB_2876368
This recombinant monoclonal targets Ornithine Carbamoyltransferase/OTC
View all literature mentionsASS1 antibody [2B10] (antibody)
RRID:AB_10975633
This monoclonal targets ASS1 antibody [2B10]
View all literature mentionsAnti-mouse IgG, HRP-linked Antibody (antibody)
RRID:AB_330924
This polyclonal targets IgG
View all literature mentionsAnti-rabbit IgG, HRP-linked Antibody (antibody)
RRID:AB_2099233
This polyclonal secondary targets IgG
View all literature mentionsRabbit Anti-GAPDH Monoclonal Antibody, Unconjugated, Clone 14C10 (antibody)
RRID:AB_561053
This recombinant monoclonal targets GAPDH
View all literature mentionsSQSTM1/p62 Antibody (antibody)
RRID:AB_10624872
This polyclonal targets SQSTM1/p62
View all literature mentionsRabbit Anti-AMPK-alpha, phospho (Thr172) Monoclonal Antibody, Unconjugated, Clone 40H9 (antibody)
RRID:AB_331250
This monoclonal targets AMPK-alpha, phospho (Thr172)
View all literature mentionsAnti-AMPK-alpha Antibody, Unconjugated (antibody)
RRID:AB_330331
This polyclonal targets AMPK-alpha
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RRID:AB_11178663
This monoclonal targets Sestrin-2 (D1B6) Rabbit mAb
View all literature mentionseIF2alpha (D7D3) XP Rabbit mAb (antibody)
RRID:AB_10692650
This monoclonal targets eIF2alpha
View all literature mentionsPhospho-eIF2alpha (Ser51) Antibody (antibody)
RRID:AB_330951
This polyclonal targets Phospho-eIF2alpha (Ser51)
View all literature mentionsRabbit Anti-S6 Ribosomal Protein Monoclonal Antibody, Unconjugated, Clone 5G10 (antibody)
RRID:AB_331355
This monoclonal targets S6 Ribosomal Protein
View all literature mentionsPhospho-S6 Ribosomal Protein (Ser235/236) Antibody (antibody)
RRID:AB_331679
This polyclonal targets Phospho-S6 Ribosomal Protein (Ser235/236)
View all literature mentionsp70 S6 Kinase Antibody (antibody)
RRID:AB_331676
This polyclonal targets p70 S6 Kinase
View all literature mentionsPhospho-p70 S6 Kinase (Thr389) Antibody (antibody)
RRID:AB_330944
This polyclonal targets Phospho-p70 S6 Kinase (Thr389)
View all literature mentionsmTOR (7C10) Rabbit mAb (antibody)
RRID:AB_2105622
This monoclonal targets mTOR
View all literature mentionsPhospho-mTOR (Ser2448) (D9C2) XP Rabbit mAb (antibody)
RRID:AB_10691552
This monoclonal targets Phospho-mTOR (Ser2448)
View all literature mentionsCleaved Caspase-3 (Asp175) Antibody (antibody)
RRID:AB_2341188
This polyclonal targets Cleaved Caspase-3 (Asp175)
View all literature mentionsanti-Cyclin E1 (HE12) Mouse monoclonal Antibody (antibody)
RRID:AB_2071200
This monoclonal targets Cyclin E1
View all literature mentionsCyclin D1 (92G2) Rabbit mAb (antibody)
RRID:AB_2259616
This monoclonal targets Cyclin D1
View all literature mentionsCyclin B1 Antibody (antibody)
RRID:AB_2072132
This unknown targets Cyclin B1
View all literature mentionsp16 INK4A (D7C1M) Rabbit mAb (antibody)
RRID:AB_2799960
This monoclonal targets p16-INK4A
View all literature mentionsp21 Waf1/Cip1 (12D1) Rabbit monoclonal antibody (antibody)
RRID:AB_823586
This monoclonal targets p21 Waf1/Cip1
View all literature mentionsC57BL/6J (organism)
RRID:IMSR_JAX:000664
Mus musculus with name C57BL/6J from IMSR.
View all literature mentionsSK-LMS-1 (cell line)
RRID:CVCL_0628
Cell line SK-LMS-1 is a Cancer cell line with a species of origin Homo sapiens (Human)
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Cell line MDA-MB-231 is a Cancer cell line with a species of origin Homo sapiens (Human)
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RRID:CVCL_0035
Cell line PC-3 is a Cancer cell line with a species of origin Homo sapiens (Human)
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RRID:CVCL_0027
Cell line Hep-G2 is a Cancer cell line with a species of origin Homo sapiens (Human)
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RRID:CVCL_0326
Cell line Hep 3B2.1-7 is a Cancer cell line with a species of origin Homo sapiens (Human)
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RRID:CVCL_0336
Cell line Huh-7 is a Cancer cell line with a species of origin Homo sapiens (Human)
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RRID:CVCL_0497
Cell line SNU-475 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsSNU-449 (cell line)
RRID:CVCL_0454
Cell line SNU-449 is a Cancer cell line with a species of origin Homo sapiens (Human)
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RRID:CVCL_0366
Cell line SNU-423 is a Cancer cell line with a species of origin Homo sapiens (Human)
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RRID:CVCL_0077
Cell line SNU-398 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsSNU-182 (cell line)
RRID:CVCL_0090
Cell line SNU-182 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsHEK293T (cell line)
RRID:CVCL_0063
Cell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)
View all literature mentions
