Genome wide CRISPR/Cas9 screen identifies the coagulation factor IX (F9) as a regulator of senescence.


Journal

Cell death & disease
ISSN: 2041-4889
Titre abrégé: Cell Death Dis
Pays: England
ID NLM: 101524092

Informations de publication

Date de publication:
19 02 2022
Historique:
received: 08 04 2021
accepted: 17 01 2022
revised: 22 12 2021
entrez: 20 2 2022
pubmed: 21 2 2022
medline: 9 4 2022
Statut: epublish

Résumé

During this last decade, the development of prosenescence therapies has become an attractive strategy as cellular senescence acts as a barrier against tumour progression. In this context, CDK4/6 inhibitors induce senescence and reduce tumour growth in breast cancer patients. However, even though cancer cells are arrested after CDK4/6 inhibitor treatment, genes regulating senescence in this context are still unknown limiting their antitumour activity. Here, using a functional genome-wide CRISPR/Cas9 genetic screen we found several genes that participate in the proliferation arrest induced by CDK4/6 inhibitors. We find that downregulation of the coagulation factor IX (F9) using sgRNA and shRNA prevents the cell cycle arrest and senescent-like phenotype induced in MCF7 breast tumour cells upon Palbociclib treatment. These results were confirmed using another breast cancer cell line, T47D, and with an alternative CDK4/6 inhibitor, Abemaciclib, and further tested in a panel of 22 cancer cells. While F9 knockout prevents the induction of senescence, treatment with a recombinant F9 protein was sufficient to induce a cell cycle arrest and senescence-like state in MCF7 tumour cells. Besides, endogenous F9 is upregulated in different human primary cells cultures undergoing senescence. Importantly, bioinformatics analysis of cancer datasets suggest a role for F9 in human tumours. Altogether, these data collectively propose key genes involved in CDK4/6 inhibitor response that will be useful to design new therapeutic strategies in personalised medicine in order to increase their efficiency, stratify patients and avoid drug resistance.

Identifiants

pubmed: 35184131
doi: 10.1038/s41419-022-04569-3
pii: 10.1038/s41419-022-04569-3
pmc: PMC8858321
doi:

Substances chimiques

Factor IX 9001-28-9
Cyclin-Dependent Kinase 4 EC 2.7.11.22
Cyclin-Dependent Kinase 6 EC 2.7.11.22

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

163

Subventions

Organisme : RCUK | Biotechnology and Biological Sciences Research Council (BBSRC)
ID : BB/P000223/1
Organisme : Office of the Royal Society
ID : RG170399
Organisme : Medical Research Council
ID : MR/K501372/1
Pays : United Kingdom

Informations de copyright

© 2022. The Author(s).

Références

Wagner V, Gil J. Senescence as a therapeutically relevant response to CDK4/6 inhibitors. Oncogene. 2020;39:5165–76.
pubmed: 32541838 pmcid: 7610384 doi: 10.1038/s41388-020-1354-9
Dorr JR, Yu Y, Milanovic M, Beuster G, Zasada C, Dabritz JH, et al. Synthetic lethal metabolic targeting of cellular senescence in cancer therapy. Nature. 2013;501:421–5.
pubmed: 23945590 doi: 10.1038/nature12437
Spring LM, Wander SA, Andre F, Moy B, Turner NC, Bardia A. Cyclin-dependent kinase 4 and 6 inhibitors for hormone receptor-positive breast cancer: past, present, and future. Lancet. 2020;395:817–27.
pubmed: 32145796 doi: 10.1016/S0140-6736(20)30165-3
Salvador-Barbero B, Alvarez-Fernandez M, Zapatero-Solana E, El Bakkali A, Menendez MDC, Lopez-Casas PP, et al. CDK4/6 Inhibitors Impair Recovery from Cytotoxic Chemotherapy in Pancreatic Adenocarcinoma. Cancer Cell. 2020;38:584.
pubmed: 33049208 doi: 10.1016/j.ccell.2020.09.012
Asghar U, Witkiewicz AK, Turner NC, Knudsen ES. The history and future of targeting cyclin-dependent kinases in cancer therapy. Nat Rev Drug Disco. 2015;14:130–46.
doi: 10.1038/nrd4504
Rapisarda V, Borghesan M, Miguela V, Encheva V, Snijders AP, Lujambio A, et al. Integrin Beta 3 Regulates Cellular Senescence by Activating the TGF-beta Pathway. Cell Rep. 2017;18:2480–93.
pubmed: 28273461 pmcid: 5357738 doi: 10.1016/j.celrep.2017.02.012
Ruscetti M, Morris JPT, Mezzadra R, Russell J, Leibold J, Romesser PB, et al. Senescence-Induced Vascular Remodeling Creates Therapeutic Vulnerabilities in Pancreas Cancer. Cell. 2020;181:424–41.e421.
pubmed: 32234521 pmcid: 7278897 doi: 10.1016/j.cell.2020.03.008
Lee S, Schmitt CA. The dynamic nature of senescence in cancer. Nat Cell Biol. 2019;21:94–101.
pubmed: 30602768 doi: 10.1038/s41556-018-0249-2
Faget DV, Ren Q, Stewart SA. Unmasking senescence: context-dependent effects of SASP in cancer. Nat Rev Cancer. 2019;19:439–53.
pubmed: 31235879 doi: 10.1038/s41568-019-0156-2
Coppe JP, Desprez PY, Krtolica A, Campisi J. The senescence-associated secretory phenotype: the dark side of tumor suppression. Annu Rev Pathol. 2010;5:99–118.
pubmed: 20078217 pmcid: 4166495 doi: 10.1146/annurev-pathol-121808-102144
Fafian-Labora JA, O’Loghlen A. Classical and Nonclassical Intercellular Communication in Senescence and Ageing. Trends Cell Biol. 2020;30:628–39.
pubmed: 32505550 doi: 10.1016/j.tcb.2020.05.003
Borghesan M, Fafian-Labora J, Eleftheriadou O, Carpintero-Fernandez P, Paez-Ribes M, Vizcay-Barrena G, et al. Small Extracellular Vesicles Are Key Regulators of Non-cell Autonomous Intercellular Communication in Senescence via the Interferon Protein IFITM3. Cell Rep. 2019;27:3956–71.e3956.
pubmed: 31242426 pmcid: 6613042 doi: 10.1016/j.celrep.2019.05.095
Demaria M, O’Leary MN, Chang J, Shao L, Liu S, Alimirah F, et al. Cellular Senescence Promotes Adverse Effects of Chemotherapy and Cancer Relapse. Cancer Disco. 2017;7:165–76.
doi: 10.1158/2159-8290.CD-16-0241
Hamm CA, Moran D, Rao K, Trusk PB, Pry K, Sausen M, et al. Genomic and Immunological Tumor Profiling Identifies Targetable Pathways and Extensive CD8+/PDL1+ Immune Infiltration in Inflammatory Breast Cancer Tumors. Mol Cancer Ther. 2016;15:1746–56.
pubmed: 27196778 doi: 10.1158/1535-7163.MCT-15-0353
Finn RS, Dering J, Conklin D, Kalous O, Cohen DJ, Desai AJ, et al. PD 0332991, a selective cyclin D kinase 4/6 inhibitor, preferentially inhibits proliferation of luminal estrogen receptor-positive human breast cancer cell lines in vitro. Breast Cancer Res. 2009;11:R77.
pubmed: 19874578 pmcid: 2790859 doi: 10.1186/bcr2419
Pandey K, An HJ, Kim SK, Lee SA, Kim S, Lim SM, et al. Molecular mechanisms of resistance to CDK4/6 inhibitors in breast cancer: A review. Int J Cancer. 2019;145:1179–88.
pubmed: 30478914 pmcid: 6767051 doi: 10.1002/ijc.32020
Anders L, Ke N, Hydbring P, Choi YJ, Widlund HR, Chick JM, et al. A systematic screen for CDK4/6 substrates links FOXM1 phosphorylation to senescence suppression in cancer cells. Cancer Cell. 2011;20:620–34.
pubmed: 22094256 pmcid: 3237683 doi: 10.1016/j.ccr.2011.10.001
Alvarez-Fernandez M, Malumbres M. Mechanisms of Sensitivity and Resistance to CDK4/6 Inhibition. Cancer Cell. 2020;37:514–29.
pubmed: 32289274 doi: 10.1016/j.ccell.2020.03.010
Goel S, DeCristo MJ, McAllister SS, Zhao JJ. CDK4/6 Inhibition in Cancer: Beyond Cell Cycle Arrest. Trends Cell Biol. 2018;28:911–25.
pubmed: 30061045 pmcid: 6689321 doi: 10.1016/j.tcb.2018.07.002
Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, et al. Genome-scale CRISPR-Cas9 knockout screening in human cells. Science. 2014;343:84–87.
pubmed: 24336571 doi: 10.1126/science.1247005
Wang T, Wei JJ, Sabatini DM, Lander ES. Genetic screens in human cells using the CRISPR-Cas9 system. Science. 2014;343:80–84.
pubmed: 24336569
Finn RS, Crown JP, Lang I, Boer K, Bondarenko IM, Kulyk SO, et al. The cyclin-dependent kinase 4/6 inhibitor palbociclib in combination with letrozole versus letrozole alone as first-line treatment of oestrogen receptor-positive, HER2-negative, advanced breast cancer (PALOMA-1/TRIO-18): a randomised phase 2 study. Lancet Oncol. 2015;16:25–35.
pubmed: 25524798 doi: 10.1016/S1470-2045(14)71159-3
Fafian-Labora JA, Rodriguez-Navarro JA, O’Loghlen A. Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage. Cell Metab. 2020;32:71–86.e75.
pubmed: 32574561 pmcid: 7342013 doi: 10.1016/j.cmet.2020.06.004
Ashida S, Orloff MS, Bebek G, Zhang L, Zheng P, Peehl DM, et al. Integrated analysis reveals critical genomic regions in prostate tumor microenvironment associated with clinicopathologic phenotypes. Clin Cancer Res. 2012;18:1578–87.
pubmed: 22275508 doi: 10.1158/1078-0432.CCR-11-2535
Finak G, Bertos N, Pepin F, Sadekova S, Souleimanova M, Zhao H, et al. Stromal gene expression predicts clinical outcome in breast cancer. Nat Med. 2008;14:518–27.
pubmed: 18438415 doi: 10.1038/nm1764
Nishida N, Nagahara M, Sato T, Mimori K, Sudo T, Tanaka F, et al. Microarray analysis of colorectal cancer stromal tissue reveals upregulation of two oncogenic miRNA clusters. Clin Cancer Res. 2012;18:3054–70.
pubmed: 22452939 doi: 10.1158/1078-0432.CCR-11-1078
Valencia T, Kim JY, Abu-Baker S, Moscat-Pardos J, Ahn CS, Reina-Campos M, et al. Metabolic reprogramming of stromal fibroblasts through p62-mTORC1 signaling promotes inflammation and tumorigenesis. Cancer Cell. 2014;26:121–35.
pubmed: 25002027 pmcid: 4101061 doi: 10.1016/j.ccr.2014.05.004
Acosta JC, O’Loghlen A, Banito A, Guijarro MV, Augert A, Raguz S, et al. Chemokine signaling via the CXCR2 receptor reinforces senescence. Cell. 2008;133:1006–18.
pubmed: 18555777 doi: 10.1016/j.cell.2008.03.038
Sheffer M, Bacolod MD, Zuk O, Giardina SF, Pincas H, Barany F, et al. Association of survival and disease progression with chromosomal instability: a genomic exploration of colorectal cancer. Proc Natl Acad Sci USA. 2009;106:7131–6.
pubmed: 19359472 pmcid: 2678450 doi: 10.1073/pnas.0902232106
Pawitan Y, Bjohle J, Amler L, Borg AL, Egyhazi S, Hall P, et al. Gene expression profiling spares early breast cancer patients from adjuvant therapy: derived and validated in two population-based cohorts. Breast Cancer Res. 2005;7:R953–964.
pubmed: 16280042 pmcid: 1410752 doi: 10.1186/bcr1325
Sanada F, Taniyama Y, Muratsu J, Otsu R, Shimizu H, Rakugi H, et al. Source of Chronic Inflammation in Aging. Front Cardiovasc Med. 2018;5:12.
pubmed: 29564335 pmcid: 5850851 doi: 10.3389/fcvm.2018.00012
Boccaccio C, Sabatino G, Medico E, Girolami F, Follenzi A, Reato G, et al. The MET oncogene drives a genetic programme linking cancer to haemostasis. Nature. 2005;434:396–400.
pubmed: 15772665 doi: 10.1038/nature03357
Ashrani AA, Gullerud RE, Petterson TM, Marks RS, Bailey KR, Heit JA. Risk factors for incident venous thromboembolism in active cancer patients: A population based case-control study. Thromb Res. 2016;139:29–37.
pubmed: 26916293 pmcid: 4769375 doi: 10.1016/j.thromres.2016.01.002
Zagar TM, Cardinale DM, Marks LB. Breast cancer therapy-associated cardiovascular disease. Nat Rev Clin Oncol. 2016;13:172–84.
pubmed: 26598943 doi: 10.1038/nrclinonc.2015.171
Wiley CD, Liu S, Limbad C, Zawadzka AM, Beck J, Demaria M, et al. SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells. Cell Rep. 2019;28:3329–37.e3325.
pubmed: 31553904 pmcid: 6907691 doi: 10.1016/j.celrep.2019.08.049
Acosta JC, Banito A, Wuestefeld T, Georgilis A, Janich P, Morton JP, et al. A complex secretory program orchestrated by the inflammasome controls paracrine senescence. Nat Cell Biol. 2013;15:978–90.
pubmed: 23770676 pmcid: 3732483 doi: 10.1038/ncb2784
De Cecco M, Ito T, Petrashen AP, Elias AE, Skvir NJ, Criscione SW, et al. L1 drives IFN in senescent cells and promotes age-associated inflammation. Nature. 2019;566:73–78.
pubmed: 30728521 pmcid: 6519963 doi: 10.1038/s41586-018-0784-9
Wilkerson WR, Sane DC. Aging and thrombosis. Semin Thromb Hemost. 2002;28:555–68.
pubmed: 12536349 doi: 10.1055/s-2002-36700
Franchini M. Hemostasis and aging. Crit Rev Oncol Hematol. 2006;60:144–51.
pubmed: 16860994 doi: 10.1016/j.critrevonc.2006.06.004
Sathyan S, Ayers E, Gao T, Milman S, Barzilai N, Verghese J. Plasma proteomic profile of frailty. Aging Cell. 2020;19:e13193.
pubmed: 32762010 pmcid: 7511877
Engbers MJ, van Hylckama Vlieg A, Rosendaal FR. Venous thrombosis in the elderly: incidence, risk factors and risk groups. J Thromb Haemost. 2010;8:2105–12.
pubmed: 20629943 doi: 10.1111/j.1538-7836.2010.03986.x
Malehmir M, Pfister D, Gallage S, Szydlowska M, Inverso D, Kotsiliti E, et al. Platelet GPIbalpha is a mediator and potential interventional target for NASH and subsequent liver cancer. Nat Med. 2019;25:641–55.
pubmed: 30936549 doi: 10.1038/s41591-019-0379-5
Heit JA, Mohr DN, Silverstein MD, Petterson TM, O’Fallon WM, Melton LJ 3rd. Predictors of recurrence after deep vein thrombosis and pulmonary embolism: a population-based cohort study. Arch Intern Med. 2000;160:761–8.
pubmed: 10737275 doi: 10.1001/archinte.160.6.761
Mingozzi F, Liu YL, Dobrzynski E, Kaufhold A, Liu JH, Wang Y, et al. Induction of immune tolerance to coagulation factor IX antigen by in vivo hepatic gene transfer. J Clin Invest. 2003;111:1347–56.
pubmed: 12727926 pmcid: 154443 doi: 10.1172/JCI200316887
Fafian-Labora J, Carpintero-Fernandez P, Jordan SJD, Shikh-Bahaei T, Abdullah SM, Mahenthiran M, et al. FASN activity is important for the initial stages of the induction of senescence. Cell Death Dis. 2019;10:318.
pubmed: 30962418 pmcid: 6453932 doi: 10.1038/s41419-019-1550-0
Sanjana NE, Shalem O, Zhang F. Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014;11:783–4.
pubmed: 25075903 pmcid: 4486245 doi: 10.1038/nmeth.3047

Auteurs

Paula Carpintero-Fernández (P)

Epigenetics & Cellular Senescence Group; Blizard Institute; Barts and The London School of Medicine and Dentistry; Queen Mary University of London; 4 Newark Street, London, E1 2AT, United Kingdom.
CellCOM Research Group. Instituto de Investigación Biomédica de A Coruña (INIBIC). CH-Universitario A Coruña (XXIAC), Universidade da Coruña. Servizo Galego de Saúde (SERGAS) Xubias de Arriba, 84 15006A, Coruña, Spain.

Michela Borghesan (M)

Epigenetics & Cellular Senescence Group; Blizard Institute; Barts and The London School of Medicine and Dentistry; Queen Mary University of London; 4 Newark Street, London, E1 2AT, United Kingdom.

Olga Eleftheriadou (O)

Epigenetics & Cellular Senescence Group; Blizard Institute; Barts and The London School of Medicine and Dentistry; Queen Mary University of London; 4 Newark Street, London, E1 2AT, United Kingdom.

Belen Pan-Castillo (B)

Epigenetics & Cellular Senescence Group; Blizard Institute; Barts and The London School of Medicine and Dentistry; Queen Mary University of London; 4 Newark Street, London, E1 2AT, United Kingdom.
StarkAge Therapeutics, Campus de l'Institut Pasteur de Lille, 1 rue du Professeur Calmette, 59800, Lille, France.

Juan Antonio Fafián-Labora (JA)

Epigenetics & Cellular Senescence Group; Blizard Institute; Barts and The London School of Medicine and Dentistry; Queen Mary University of London; 4 Newark Street, London, E1 2AT, United Kingdom.
Grupo de investigación en Terapia Celular y Medicina Regenerativa, Departamento de Fisioterapia, Medicina y Ciencias Biomédicas, Facultad de Ciencias de la Salud, Universidade da Coruña, INIBIC-Complejo Hospitalario Universitario A Coruña (CHUAC), Agrupación estratégica CICA-INIBIC, As Xubías, 15006, A Coruña, Spain.

Tom P Mitchell (TP)

Centre for Microvascular Research. The William Harvey Research Institute. Charterhouse Square Barts and the London School of Medicine and Dentistry Queen Mary University of London, EC1M 6BQ, London, UK.

Alejandro Yuste (A)

CellCOM Research Group. Instituto de Investigación Biomédica de A Coruña (INIBIC). CH-Universitario A Coruña (XXIAC). Universidade da Coruña. Servizo Galego de Saúde (SERGAS). Xubias de Arriba, 84 15006, A Coruña, Spain.

Muge Ogrunc (M)

StarkAge Therapeutics, Campus de l'Institut Pasteur de Lille, 1 rue du Professeur Calmette, 59800, Lille, France.
SynLab Hauts-de-France, Campus de l'Institut Pasteur de Lille, 1 rue du Professeur Calmette, 59800, Lille, France.

Thomas D Nightingale (TD)

Centre for Microvascular Research. The William Harvey Research Institute. Charterhouse Square Barts and the London School of Medicine and Dentistry Queen Mary University of London, EC1M 6BQ, London, UK.

Maria Mayan (M)

CellCOM Research Group. Instituto de Investigación Biomédica de A Coruña (INIBIC). CH-Universitario A Coruña (XXIAC). Universidade da Coruña. Servizo Galego de Saúde (SERGAS). Xubias de Arriba, 84 15006, A Coruña, Spain.

Ana O'Loghlen (A)

Epigenetics & Cellular Senescence Group; Blizard Institute; Barts and The London School of Medicine and Dentistry; Queen Mary University of London; 4 Newark Street, London, E1 2AT, United Kingdom. a.ologhlen@qmul.ac.uk.

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