A Genome-Wide CRISPR/Cas9-Based Screen Identifies Heparan Sulfate Proteoglycans as Ligands of Killer-Cell Immunoglobulin-Like Receptors

被引:3
|
作者
Klein, Klara [1 ,2 ,3 ]
Hoelzemer, Angelique [4 ,5 ,6 ]
Wang, Tim [1 ,2 ,7 ]
Kim, Tae-Eun [8 ]
Dugan, Haley L. [8 ,9 ]
Jost, Stephanie [8 ,10 ]
Altfeld, Marcus [4 ]
Garcia-Beltran, Wilfredo F. [8 ,11 ]
机构
[1] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA
[2] Whitehead Inst Biomed Res, 9 Cambridge Ctr, Cambridge, MA 02142 USA
[3] Univ Vet Med Vienna, Inst Pharmacol & Toxicol, Vienna, Austria
[4] Leibniz Inst Expt Virol, Hamburg, Germany
[5] Univ Med Ctr Eppendorf, Dept Internal Med 1, Div Infect Dis, Hamburg, Germany
[6] German Ctr Infect Res DZIF, Site Hamburg Lubeck Borstel Riems, Hamburg, Germany
[7] MIT, Dept Biol, Cambridge, MA USA
[8] Ragon Inst Massachusetts Gen Hosp MGH MIT & Harva, Cambridge, MA 02138 USA
[9] Adimab LLC, Lebanon, NH USA
[10] Beth Israel Deaconess Med Ctr, Ctr Virol & Vaccine Res, Boston, MA 02215 USA
[11] Massachusetts Gen Hosp MGH, Dept Pathol, Boston, MA 02114 USA
来源
FRONTIERS IN IMMUNOLOGY | 2021年 / 12卷
基金
美国国家卫生研究院;
关键词
CRISPR; screen; KIR; heparan sulfate; NK cells; HLA-F; BINDING; PROTEIN; MICE;
D O I
10.3389/fimmu.2021.798235
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
While human leukocyte antigen (HLA) and HLA-like proteins comprise an overwhelming majority of known ligands for NK-cell receptors, the interactions of NK-cell receptors with non-conventional ligands, particularly carbohydrate antigens, is less well described. We previously found through a bead-based HLA screen that KIR3DS1, a formerly orphan member of the killer-cell immunoglobulin-like receptor (KIR) family, binds to HLA-F. In this study, we assessed the ligand binding profile of KIR3DS1 to cell lines using Fc fusion constructs, and discovered that KIR3DS1-Fc exhibited binding to several human cell lines including ones devoid of HLA. To identify these non-HLA ligands, we developed a magnetic enrichment-based genome-wide CRISPR/Cas9 knock-out screen approach, and identified enzymes involved in the biosynthesis of heparan sulfate as crucial for the binding of KIR3DS1-Fc to K562 cells. This interaction between KIR3DS1 and heparan sulfate was confirmed via surface plasmon resonance, and removal of heparan sulfate proteoglycans from cell surfaces abolished KIR3DS1-Fc binding. Testing of additional KIR-Fc constructs demonstrated that KIR family members containing a D0 domain (KIR3DS1, KIR3DL1, KIR3DL2, KIR2DL4, and KIR2DL5) bound to heparan sulfate, while those without a D0 domain (KIR2DL1, KIR2DL2, KIR2DL3, and KIR2DS4) did not. Overall, this study demonstrates the use of a genome-wide CRISPR/Cas9 knock-out strategy to unbiasedly identify unconventional ligands of NK-cell receptors. Furthermore, we uncover a previously underrecognized binding of various activating and inhibitory KIRs to heparan sulfate proteoglycans that may play a role in NK-cell receptor signaling and target-cell recognition.
引用
收藏
页数:10
相关论文
共 50 条
  • [21] A genome-wide CRISPR screen identifies novel ligands for the Siglec family of glyco-immune checkpoint receptors.
    Wisnovsky, Simon
    Bertozzi, Carolyn
    CANCER IMMUNOLOGY RESEARCH, 2020, 8 (04) : 80 - 81
  • [22] A GENOME-WIDE CRISPR-Cas9 SCREEN FOR GENES REGULATING QUIESCENT-LIKE STATES IN GLIOBLASTOMA
    Feldman, Heather
    Mihalas, Anca
    Plaisier, Christopher
    Patel, Anoop
    Paddison, Patrick
    NEURO-ONCOLOGY, 2019, 21 : 42 - 42
  • [23] Genome-wide CRISPR/Cas9 deletion screen defines mitochondrial gene essentiality and identifies routes for tumour cell viability in hypoxia
    Luke W. Thomas
    Cinzia Esposito
    Rachel E. Morgan
    Stacey Price
    Jamie Young
    Steven P. Williams
    Lucas A. Maddalena
    Ultan McDermott
    Margaret Ashcroft
    Communications Biology, 4
  • [24] Genome-Wide CRISPR/Cas9-Based Screening for Deubiquitinase Subfamily Identifies Ubiquitin-Specific Protease 11 as a Novel Regulator of Osteogenic Differentiation
    Kaushal, Kamini
    Tyagi, Apoorvi
    Karapurkar, Janardhan Keshav
    Kim, Eun-Jung
    Tanguturi, Parthasaradhireddy
    Kim, Kye-Seong
    Jung, Han-Sung
    Ramakrishna, Suresh
    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2022, 23 (02)
  • [25] Genome-wide CRISPR/Cas9 deletion screen defines mitochondrial gene essentiality and identifies routes for tumour cell viability in hypoxia
    Thomas, Luke W.
    Esposito, Cinzia
    Morgan, Rachel E.
    Price, Stacey
    Young, Jamie
    Williams, Steven P.
    Maddalena, Lucas A.
    McDermott, Ultan
    Ashcroft, Margaret
    COMMUNICATIONS BIOLOGY, 2021, 4 (01)
  • [26] A Genome-Wide CRISPR/Cas9 Screen Identifies Druggable Targets that Synergize with TKIs for Potential Combination Treatment of Leukemia
    Herrera, Maria
    Andersen, Aram
    Piechaczyk, Laure
    Floisand, Yngvar
    Enserink, Jorrit
    CLINICAL LYMPHOMA MYELOMA & LEUKEMIA, 2019, 19 : S183 - S183
  • [27] A Genome-Wide CRISPR/Cas9 Screen Identifies an Essential Role of SLC35A1 in rAAV Transduction
    Zhang, Xiujuan
    Ning, Kang
    Hao, Siyuan
    Feng, Zehua
    Kuz, Cagla Aksu
    McFarlin, Shane
    Richart, Donovan G.
    Cheng, Fang
    Yan, Ziying
    Qiu, Jianming
    MOLECULAR THERAPY, 2024, 32 (04) : 37 - 37
  • [28] A Genome-Wide CRISPR/Cas9 Screen Identifies a Role for Early Endosomes in Hepatitis E Virus RNA Replication
    Oechslin, Noemie
    Da Silva, Nathalie
    Ankavay, Maliki
    Pollan, Angela
    Moradpour, Darius
    Gouttenoire, Jerome
    SWISS MEDICAL WEEKLY, 2022, 152 : 8S - 8S
  • [29] A genome-wide CRISPR-Cas9 knockout screen identifies novel PARP inhibitor resistance genes in prostate cancer
    Malene Blond Ipsen
    Ea Marie Givskov Sørensen
    Emil Aagaard Thomsen
    Simone Weiss
    Jakob Haldrup
    Anders Dalby
    Johan Palmfeldt
    Peter Bross
    Martin Rasmussen
    Jacob Fredsøe
    Søren Klingenberg
    Mads R. Jochumsen
    Kirsten Bouchelouche
    Benedicte Parm Ulhøi
    Michael Borre
    Jacob Giehm Mikkelsen
    Karina Dalsgaard Sørensen
    Oncogene, 2022, 41 : 4271 - 4281
  • [30] A Crispr/Cas9 Drop-out Screen Identifies Genome-Wide Genetic Valnerubilities in Acute Myeloid Leukaemia
    Tzelepis, Konstantinos
    Koike-Yusa, Hiroko
    De Braekeleer, Etienne
    Li, Yilong
    Metzakopian, Emmanouil
    Dovey, Oliver M.
    Mupo, Annalisa
    Grinkevich, Vera
    Mazan, Milena M.
    Gozdecka, Malgorzata
    Cooper, Jonathan L.
    Patel, Miten
    McKerrell, Thomas David Hardman
    Chen, Bin
    Ponstingl, Hannes
    Huntly, Brian J. P.
    McDermott, Ultan
    Saez-Rodriguez, Julio
    Iorio, Francesco
    Yusa, Kosuke
    Vassiliou, George S.
    BLOOD, 2015, 126 (23)