Tumour-intrinsic PDL1 signals regulate the Chk2 DNA damage response in cancer cells and mediate resistance to Chk1 inhibitors.
Chk2
DDR inhibitors
DNA damage repair
Immune checkpoints
PDL1
Synthetic lethality
Journal
Molecular cancer
ISSN: 1476-4598
Titre abrégé: Mol Cancer
Pays: England
ID NLM: 101147698
Informations de publication
Date de publication:
30 Oct 2024
30 Oct 2024
Historique:
received:
07
02
2024
accepted:
05
10
2024
medline:
31
10
2024
pubmed:
31
10
2024
entrez:
31
10
2024
Statut:
epublish
Résumé
Aside from the canonical role of PDL1 as a tumour surface-expressed immune checkpoint molecule, tumour-intrinsic PDL1 signals regulate non-canonical immunopathological pathways mediating treatment resistance whose significance, mechanisms, and therapeutic targeting remain incompletely understood. Recent reports implicate tumour-intrinsic PDL1 signals in the DNA damage response (DDR), including promoting homologous recombination DNA damage repair and mRNA stability of DDR proteins, but many mechanistic details remain undefined. We genetically depleted PDL1 from transplantable mouse and human cancer cell lines to understand consequences of tumour-intrinsic PDL1 signals in the DNA damage response. We complemented this work with studies of primary human tumours and inducible mouse tumours. We developed novel approaches to show tumour-intrinsic PDL1 signals in specific subcellular locations. We pharmacologically depleted tumour PDL1 in vivo in mouse models with repurposed FDA-approved drugs for proof-of-concept clinical translation studies. We show that tumour-intrinsic PDL1 promotes the checkpoint kinase-2 (Chk2)-mediated DNA damage response. Intracellular but not surface-expressed PDL1 controlled Chk2 protein content post-translationally and independently of PD1 by antagonising PIRH2 E3 ligase-mediated Chk2 polyubiquitination and protein degradation. Genetic tumour PDL1 depletion specifically reduced tumour Chk2 content but not ATM, ATR, or Chk1 DDR proteins, enhanced Chk1 inhibitor (Chk1i) synthetic lethality in vitro in diverse human and murine tumour models, and improved Chk1i efficacy in vivo. Pharmacologic tumour PDL1 depletion with cefepime or ceftazidime replicated genetic tumour PDL1 depletion by reducing tumour Chk2, inducing Chk1i synthetic lethality in a tumour PDL1-dependent manner, and reducing in vivo tumour growth when combined with Chk1i. Our data challenge the prevailing surface PDL1 paradigm, elucidate important and previously unappreciated roles for tumour-intrinsic PDL1 in regulating the ATM/Chk2 DNA damage response axis and E3 ligase-mediated protein degradation, suggest tumour PDL1 as a biomarker for Chk1i efficacy, and support the rapid clinical potential of pharmacologic tumour PDL1 depletion to treat selected cancers.
Sections du résumé
BACKGROUND
BACKGROUND
Aside from the canonical role of PDL1 as a tumour surface-expressed immune checkpoint molecule, tumour-intrinsic PDL1 signals regulate non-canonical immunopathological pathways mediating treatment resistance whose significance, mechanisms, and therapeutic targeting remain incompletely understood. Recent reports implicate tumour-intrinsic PDL1 signals in the DNA damage response (DDR), including promoting homologous recombination DNA damage repair and mRNA stability of DDR proteins, but many mechanistic details remain undefined.
METHODS
METHODS
We genetically depleted PDL1 from transplantable mouse and human cancer cell lines to understand consequences of tumour-intrinsic PDL1 signals in the DNA damage response. We complemented this work with studies of primary human tumours and inducible mouse tumours. We developed novel approaches to show tumour-intrinsic PDL1 signals in specific subcellular locations. We pharmacologically depleted tumour PDL1 in vivo in mouse models with repurposed FDA-approved drugs for proof-of-concept clinical translation studies.
RESULTS
RESULTS
We show that tumour-intrinsic PDL1 promotes the checkpoint kinase-2 (Chk2)-mediated DNA damage response. Intracellular but not surface-expressed PDL1 controlled Chk2 protein content post-translationally and independently of PD1 by antagonising PIRH2 E3 ligase-mediated Chk2 polyubiquitination and protein degradation. Genetic tumour PDL1 depletion specifically reduced tumour Chk2 content but not ATM, ATR, or Chk1 DDR proteins, enhanced Chk1 inhibitor (Chk1i) synthetic lethality in vitro in diverse human and murine tumour models, and improved Chk1i efficacy in vivo. Pharmacologic tumour PDL1 depletion with cefepime or ceftazidime replicated genetic tumour PDL1 depletion by reducing tumour Chk2, inducing Chk1i synthetic lethality in a tumour PDL1-dependent manner, and reducing in vivo tumour growth when combined with Chk1i.
CONCLUSIONS
CONCLUSIONS
Our data challenge the prevailing surface PDL1 paradigm, elucidate important and previously unappreciated roles for tumour-intrinsic PDL1 in regulating the ATM/Chk2 DNA damage response axis and E3 ligase-mediated protein degradation, suggest tumour PDL1 as a biomarker for Chk1i efficacy, and support the rapid clinical potential of pharmacologic tumour PDL1 depletion to treat selected cancers.
Identifiants
pubmed: 39478560
doi: 10.1186/s12943-024-02147-z
pii: 10.1186/s12943-024-02147-z
doi:
Substances chimiques
Checkpoint Kinase 1
EC 2.7.11.1
Checkpoint Kinase 2
EC 2.7.1.11
B7-H1 Antigen
0
CHEK1 protein, human
EC 2.7.11.1
Protein Kinase Inhibitors
0
CHEK2 protein, human
EC 2.7.11.1
CD274 protein, human
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
242Subventions
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Organisme : NIH HHS
ID : CA239390, GM113896, CA241801, CA054174, CA268641, CA023108
Pays : United States
Informations de copyright
© 2024. The Author(s).
Références
Topalian SL, Taube JM, Anders RA, Pardoll DM. Mechanism-driven biomarkers to guide immune checkpoint blockade in cancer therapy. Nat Rev Cancer. 2016;16:275–87. https://doi.org/10.1038/nrc.2016.36 .
doi: 10.1038/nrc.2016.36
pubmed: 27079802
pmcid: 5381938
Taube JM, et al. Colocalisation of inflammatory response with B7–h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape. Sci Transl Med. 2012;4:127ra137. https://doi.org/10.1126/scitranslmed.3003689 https://doi.org:4/127/127ra37 .
doi: 10.1126/scitranslmed.3003689
Topalian, S. L., Drake, C. G. & Pardoll, D. M. J. C. o. i. i. Targeting the PD-1/B7-H1 (PD-L1) pathway to activate anti-tumour immunity. 24, 207–212 (2012).
Paterson AM, et al. The programmed death-1 ligand 1:b7–1 pathway restrains diabetogenic effector T cells in vivo. J Immunol. 2011;187:1097–105. https://doi.org/10.4049/jimmunol.1003496 https://doi.org:jimmunol.1003496 .
doi: 10.4049/jimmunol.1003496
pubmed: 21697456
Dong H, et al. tumour-associated B7–H1 promotes T-cell apoptosis: A potential mechanism of immune evasion. Nat Med. 2002;8:793–800.
doi: 10.1038/nm730
pubmed: 12091876
Brahmer JR, et al. Safety and activity of anti-PD-L1 antibody in patients with advanced cancer. N Engl J Med. 2012;366:2455–65. https://doi.org/10.1056/NEJMoa1200694 .
doi: 10.1056/NEJMoa1200694
pubmed: 22658128
pmcid: 3563263
Zou W, Wolchok JD, Chen L. PD-L1 (B7–H1) and PD-1 pathway blockade for cancer therapy: Mechanisms, response biomarkers, and combinations. Sci Transl Med. 2016;8:328rv324. https://doi.org/10.1126/scitranslmed.aad7118 .
doi: 10.1126/scitranslmed.aad7118
Sharma P, Hu-Lieskovan S, Wargo JA, Ribas A. Primary, Adaptive, and Acquired Resistance to Cancer Immunotherapy. Cell. 2017;168:707–23. https://doi.org/10.1016/j.cell.2017.01.017 .
doi: 10.1016/j.cell.2017.01.017
pubmed: 28187290
pmcid: 5391692
Kornepati AVR, Vadlamudi RK, Curiel TJ. Programmed death ligand 1 signals in cancer cells. Nat Rev Cancer. 2022;22:174–89. https://doi.org/10.1038/s41568-021-00431-4 .
doi: 10.1038/s41568-021-00431-4
pubmed: 35031777
pmcid: 9989967
Clark CA, et al. tumour-Intrinsic PD-L1 Signals Regulate Cell Growth, Pathogenesis, and Autophagy in Ovarian Cancer and Melanoma. Cancer Res. 2016;76:6964–74. https://doi.org/10.1158/0008-5472.CAN-16-0258 .
doi: 10.1158/0008-5472.CAN-16-0258
pubmed: 27671674
pmcid: 5228566
Gupta HB, et al. tumour cell-intrinsic PD-L1 promotes tumour-initiating cell generation and functions in melanoma and ovarian cancer. Signal Transduct Target Ther. 2016;1:1–9. https://doi.org/10.1038/sigtrans.2016.30 .
doi: 10.1038/sigtrans.2016.30
Zhu H, et al. BET Bromodomain Inhibition Promotes Anti-tumour Immunity by Suppressing PD-L1 Expression. Cell reports. 2016;16:2829–37. https://doi.org/10.1016/j.celrep.2016.08.032 .
doi: 10.1016/j.celrep.2016.08.032
pubmed: 27626654
Wu B, et al. Adipose PD-L1 Modulates PD-1/PD-L1 Checkpoint Blockade Immunotherapy Efficacy in Breast Cancer. Oncoimmunology. 2018;7:e1500107. https://doi.org/10.1080/2162402X.2018.1500107 .
doi: 10.1080/2162402X.2018.1500107
pubmed: 30393583
pmcid: 6209395
Liang J, et al. Verteporfin Inhibits PD-L1 through Autophagy and the STAT1-IRF1-TRIM28 Signaling Axis. Exerting Antitumour Efficacy Cancer immunology research. 2020;8:952–65. https://doi.org/10.1158/2326-6066.CIR-19-0159 .
doi: 10.1158/2326-6066.CIR-19-0159
Murray, C. et al. Pharmacologic tumour PDL1 Depletion with Cefepime or Ceftazidime Promotes DNA Damage and Sensitivity to DNA-Damaging Agents. Int J Mol Sci 23 (2022). https://doi.org/10.3390/ijms23095129
Kornepati AVR, et al. tumour-intrinsic PD-L1 promotes DNA repair in distinct cancers and suppresses PARP inhibitor-induced synthetic lethality. Cancer Res. 2022;82:2156–70. https://doi.org/10.1158/0008-5472.CAN-21-2076 .
doi: 10.1158/0008-5472.CAN-21-2076
pubmed: 35247877
pmcid: 9987177
Tu X, et al. PD-L1 (B7–H1) Competes with the RNA Exosome to Regulate the DNA Damage Response and Can Be Targeted to Sensitize to Radiation or Chemotherapy. Mol Cell. 2019;74:1215–26. https://doi.org/10.1016/j.molcel.2019.04.005 . e1214.
doi: 10.1016/j.molcel.2019.04.005
pubmed: 31053471
pmcid: 6737939
Marechal, A. & Zou, L. DNA damage sensing by the ATM and ATR kinases. Cold Spring Harbor perspectives in biology 5 (2013). https://doi.org/10.1101/cshperspect.a012716
Cleary JM, Aguirre AJ, Shapiro GI, D’Andrea AD. Biomarker-Guided Development of DNA Repair Inhibitors. Mol Cell. 2020;78:1070–85. https://doi.org/10.1016/j.molcel.2020.04.035 .
doi: 10.1016/j.molcel.2020.04.035
pubmed: 32459988
pmcid: 7316088
Smith, J., Tho, L. M., Xu, N. & Gillespie, D. A. The ATM-Chk2 and ATR-Chk1 pathways in DNA damage signaling and cancer. Adv Cancer Res 108, 73–112 (2010). https://doi.org/10.1016/B978-0-12-380888-2.00003-0
Gudmundsdottir, K. & Ashworth, A. The roles of BRCA1 and BRCA2 and associated proteins in the maintenance of genomic stability. Oncogene 25, 5864–5874 (2006). https://doi.org/10.1038/sj.onc.1209874
Zhang, D. et al. Bladder cancer cell-intrinsic PD-L1 signals promote mTOR and autophagy activation that can be inhibited to improve cytotoxic chemotherapy. Cancer medicine 10, 2137–2152 (2021). https://doi.org: https://doi.org/10.1002/cam4.3739
Rodier, F. et al. Persistent DNA damage signaling triggers senescence-associated inflammatory cytokine secretion. Nat Cell Biol 11, 973–979 (2009). https://doi.org/10.1038/ncb1909
Xu G, et al. REV7 counteracts DNA double-strand break resection and affects PARP inhibition. Nature. 2015;521:541–4. https://doi.org/10.1038/nature14328 .
doi: 10.1038/nature14328
pubmed: 25799992
pmcid: 4671316
duplicate with ref 22
Bartek J, Lukas J. Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer Cell. 2003;3:421–9.
doi: 10.1016/S1535-6108(03)00110-7
pubmed: 12781359
Zaugg, K. et al. Cross-talk between Chk1 and Chk2 in double-mutant thymocytes. 104, 3805–3810 (2007).
Gay CM, et al. Patterns of transcription factor programs and immune pathway activation define four major subtypes of SCLC with distinct therapeutic vulnerabilities. Cancer cell. 2021;39:346–60. https://doi.org/10.1016/j.ccell.2020.12.014 . e347.
doi: 10.1016/j.ccell.2020.12.014
pubmed: 33482121
pmcid: 8143037
Hennessey RC, et al. Ultraviolet radiation accelerates NRas-mutant melanomagenesis: A cooperative effect blocked by sunscreen. Pigment Cell Melanoma Res. 2017;30:477–87. https://doi.org/10.1111/pcmr.12601 .
doi: 10.1111/pcmr.12601
pubmed: 28544727
Gato-Cañas, M. et al. PDL1 signals through conserved sequence motifs to overcome interferon-mediated cytotoxicity. 20, 1818–1829 (2017).
Burr ML, et al. CMTM6 maintains the expression of PD-L1 and regulates anti-tumour immunity. Nature. 2017;549:101–5. https://doi.org/10.1038/nature23643 .
doi: 10.1038/nature23643
pubmed: 28813417
pmcid: 5706633
Mezzadra R, et al. Identification of CMTM6 and CMTM4 as PD-L1 protein regulators. Nature. 2017;549:106–10. https://doi.org/10.1038/nature2366 .
doi: 10.1038/nature2366
pubmed: 28813410
pmcid: 6333292
Gao Y, et al. Acetylation-dependent regulation of PD-L1 nuclear translocation dictates the efficacy of anti-PD-1 immunotherapy. Nat Cell Biol. 2020;22:1064–75. https://doi.org/10.1038/s41556-020-0562-4 .
doi: 10.1038/s41556-020-0562-4
pubmed: 32839551
pmcid: 7484128
Hou J, et al. PD-L1-mediated gasdermin C expression switches apoptosis to pyroptosis in cancer cells and facilitates tumour necrosis. Nat Cell Biol. 2020;22:1264–75. https://doi.org/10.1038/s41556-020-0575-z .
doi: 10.1038/s41556-020-0575-z
pubmed: 32929201
pmcid: 7653546
Zannini L, Delia D, Buscemi G. CHK2 kinase in the DNA damage response and beyond. Journal of molecular cell biology. 2014;6:442–57. https://doi.org/10.1093/jmcb/mju045 .
doi: 10.1093/jmcb/mju045
pubmed: 25404613
pmcid: 4296918
Zannini, L., Delia, D. & Buscemi, G. J. J. o. m. c. b. CHK2 kinase in the DNA damage response and beyond. 6, 442–457 (2014).
Satelli, A. et al. Potential role of nuclear PD-L1 expression in cell-surface vimentin positive circulating tumour cells as a prognostic marker in cancer patients. Scientific reports 6, 28910 (2016). https://doi.org/10.1038/srep28910
Wei Y, et al. The local immune landscape determines tumour PD-L1 heterogeneity and sensitivity to therapy. J Clin Invest. 2019;129:3347–60. https://doi.org/10.1172/JCI127726 .
doi: 10.1172/JCI127726
pubmed: 31112529
pmcid: 6668685
Bohgaki, M. et al. The E3 ligase PIRH2 polyubiquitylates CHK2 and regulates its turnover. Cell Death Differ 20, 812–822 (2013). https://doi.org/10.1038/cdd.2013.7
Labay, E. et al. Repurposing cephalosporin antibiotics as pro-senescent radiosensitizers. Oncotarget 7, 33919–33933 (2016). https://doi.org/10.18632/oncotarget.8984
Sen T, et al. Targeting DNA Damage Response Promotes Antitumour Immunity through STING-Mediated T-cell Activation in Small Cell Lung Cancer. Cancer Discov. 2019;9:646–61. https://doi.org/10.1158/2159-8290.CD-18-1020 .
doi: 10.1158/2159-8290.CD-18-1020
pubmed: 30777870
pmcid: 6563834
Farkkila, A. et al. Immunogenomic profiling determines responses to combined PARP and PD-1 inhibition in ovarian cancer. Nature communications 11, 1459 (2020). https://doi.org/10.1038/s41467-020-15315-8
Drerup JM, et al. CD122-Selective IL2 Complexes Reduce Immunosuppression, Promote Treg Fragility, and Sensitize tumour Response to PD-L1 Blockade. Cancer Res. 2020;80:5063–75. https://doi.org/10.1158/0008-5472.CAN-20-0002 .
doi: 10.1158/0008-5472.CAN-20-0002
pubmed: 32948605
pmcid: 7669742
Pilie PG, Tang C, Mills GB, Yap TA. State-of-the-art strategies for targeting the DNA damage response in cancer. Nat Rev Clin Oncol. 2019;16:81–104. https://doi.org/10.1038/s41571-018-0114-z .
doi: 10.1038/s41571-018-0114-z
pubmed: 30356138
pmcid: 8327299
dup. of ref. 45
Li, Q. et al. A new wave of innovations within the DNA damage response. Signal Transduct Target Ther 8, 338 (2023). https://doi.org/10.1038/s41392-023-01548-8
Ditano JP, Eastman A. Comparative Activity and Off-Target Effects in Cells of the CHK1 Inhibitors MK-8776, SRA737, and LY2606368. ACS Pharmacol Transl Sci. 2021;4:730–43. https://doi.org/10.1021/acsptsci.0c00201 .
doi: 10.1021/acsptsci.0c00201
pubmed: 33860197
pmcid: 8033610
Toledo LI, et al. A cell-based screen identifies ATR inhibitors with synthetic lethal properties for cancer-associated mutations. Nat Struct Mol Biol. 2011;18:721–7. https://doi.org/10.1038/nsmb.2076 .
doi: 10.1038/nsmb.2076
pubmed: 21552262
pmcid: 4869831
Buisson R, Boisvert JL, Benes CH, Zou L. Distinct but Concerted Roles of ATR, DNA-PK, and Chk1 in Countering Replication Stress during S Phase. Mol Cell. 2015;59:1011–24. https://doi.org/10.1016/j.molcel.2015.07.029 .
doi: 10.1016/j.molcel.2015.07.029
pubmed: 26365377
pmcid: 4575890
Saldivar JC, Cortez D, Cimprich KA. The essential kinase ATR: ensuring faithful duplication of a challenging genome. Nat Rev Mol Cell Biol. 2017;18:622–36. https://doi.org/10.1038/nrm.2017.67 .
doi: 10.1038/nrm.2017.67
pubmed: 28811666
pmcid: 5796526
Kornepati AVR, Rogers CM, Sung P, Curiel TJ. The complementarity of DDR, nucleic acids and anti-tumour immunity. Nature. 2023;619:475–86. https://doi.org/10.1038/s41586-023-06069-6 .
doi: 10.1038/s41586-023-06069-6
pubmed: 37468584
Chakravarty D, et al. Extranuclear functions of ER impact invasive migration and metastasis by breast cancer cells. Cancer Res. 2010;70:4092–101. https://doi.org/10.1158/0008-5472.CAN-09-3834 .
doi: 10.1158/0008-5472.CAN-09-3834
pubmed: 20460518
pmcid: 2889925
Hasty P, et al. eRapa Restores a Normal Life Span in a FAP Mouse Model. Cancer Prev Res (Phila). 2014;7:169–78. https://doi.org/10.1158/1940-6207.CAPR-13-0299 .
doi: 10.1158/1940-6207.CAPR-13-0299
pubmed: 24282255
Clark CA, Gupta HB, Curiel TJ. tumour cell-intrinsic CD274/PD-L1: A novel metabolic balancing act with clinical potential. Autophagy. 2017;13:987–8. https://doi.org/10.1080/15548627.2017.1280223 .
doi: 10.1080/15548627.2017.1280223
pubmed: 28368722
pmcid: 5446070
Lin, C. et al. Ceftazidime is a potential drug to inhibit SARS-CoV-2 infection in vitro by blocking spike protein-ACE2 interaction. Signal Transduct Target Ther 6, 198 (2021). https://doi.org/10.1038/s41392-021-00619-y
Ramon-Garcia, S. et al. Repurposing clinically approved cephalosporins for tuberculosis therapy. Scientific reports 6, 34293 (2016). https://doi.org/10.1038/srep34293
de la Pena Avalos, B. & Dray, E. Visualization of DNA Repair Proteins Interaction by Immunofluorescence. Journal of visualized experiments : JoVE (2020). https://doi.org/10.3791/61447
Cardnell RJ, et al. Proteomic markers of DNA repair and PI3K pathway activation predict response to the PARP inhibitor BMN 673 in small cell lung cancer. Clin Cancer Res. 2013;19:6322–8. https://doi.org/10.1158/1078-0432.CCR-13-1975 .
doi: 10.1158/1078-0432.CCR-13-1975
pubmed: 24077350
Gradia SD, et al. MacroBac: New Technologies for Robust and Efficient Large-Scale Production of Recombinant Multiprotein Complexes. Methods Enzymol. 2017;592:1–26. https://doi.org/10.1016/bs.mie.2017.03.008 .
doi: 10.1016/bs.mie.2017.03.008
pubmed: 28668116
pmcid: 6028233
Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15, 550 (2014). https://doi.org/10.1186/s13059-014-0550-8