PMID- 34917099
OWN - NLM
STAT- MEDLINE
DCOM- 20220228
LR  - 20220228
IS  - 1664-3224 (Electronic)
IS  - 1664-3224 (Linking)
VI  - 12
DP  - 2021
TI  - A Genome-Wide CRISPR/Cas9-Based Screen Identifies Heparan Sulfate Proteoglycans 
      as Ligands of Killer-Cell Immunoglobulin-Like Receptors.
PG  - 798235
LID - 10.3389/fimmu.2021.798235 [doi]
LID - 798235
AB  - 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.
CI  - Copyright (c) 2021 Klein, Holzemer, Wang, Kim, Dugan, Jost, Altfeld and 
      Garcia-Beltran.
FAU - Klein, Klara
AU  - Klein K
AD  - Broad Institute of MIT and Harvard, Cambridge, MA, United States.
AD  - Whitehead Institute for Biomedical Research, Cambridge, MA, United States.
AD  - Institute of Pharmacology and Toxicology, University of Veterinary Medicine 
      Vienna, Vienna, Austria.
FAU - Holzemer, Angelique
AU  - Holzemer A
AD  - Leibniz Institute for Experimental Virology, Hamburg, Germany.
AD  - First Department of Internal Medicine, Division of Infectious Diseases, 
      University Medical Centre Eppendorf, Hamburg, Germany.
AD  - German Center for Infection Research (DZIF), Site Hamburg-Lubeck-Borstel-Riems, 
      Hamburg, Germany.
FAU - Wang, Tim
AU  - Wang T
AD  - Broad Institute of MIT and Harvard, Cambridge, MA, United States.
AD  - Whitehead Institute for Biomedical Research, Cambridge, MA, United States.
AD  - Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 
      United States.
FAU - Kim, Tae-Eun
AU  - Kim TE
AD  - Ragon Institute of Massachusetts General Hospital (MGH), MIT, and Harvard, 
      Cambridge, MA, United States.
FAU - Dugan, Haley L
AU  - Dugan HL
AD  - Ragon Institute of Massachusetts General Hospital (MGH), MIT, and Harvard, 
      Cambridge, MA, United States.
AD  - Adimab, LLC, Lebanon, NH, United States.
FAU - Jost, Stephanie
AU  - Jost S
AD  - Ragon Institute of Massachusetts General Hospital (MGH), MIT, and Harvard, 
      Cambridge, MA, United States.
AD  - Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, 
      Boston, MA, United States.
FAU - Altfeld, Marcus
AU  - Altfeld M
AD  - Leibniz Institute for Experimental Virology, Hamburg, Germany.
FAU - Garcia-Beltran, Wilfredo F
AU  - Garcia-Beltran WF
AD  - Ragon Institute of Massachusetts General Hospital (MGH), MIT, and Harvard, 
      Cambridge, MA, United States.
AD  - Department of Pathology, Massachusetts General Hospital (MGH), Boston, MA, United 
      States.
LA  - eng
GR  - R01 AI067031/AI/NIAID NIH HHS/United States
GR  - P01 AI104715/AI/NIAID NIH HHS/United States
GR  - F31 AI116366/AI/NIAID NIH HHS/United States
GR  - T32 GM007753/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20211130
PL  - Switzerland
TA  - Front Immunol
JT  - Frontiers in immunology
JID - 101560960
RN  - 0 (Heparan Sulfate Proteoglycans)
RN  - 0 (KIR3DS1 protein, human)
RN  - 0 (Ligands)
RN  - 0 (Receptors, KIR)
RN  - 0 (Receptors, KIR3DS1)
RN  - EC 3.1.- (CRISPR-Associated Protein 9)
SB  - IM
MH  - CRISPR-Associated Protein 9/metabolism
MH  - CRISPR-Cas Systems
MH  - Genome-Wide Association Study
MH  - Heparan Sulfate Proteoglycans/*agonists
MH  - Humans
MH  - K562 Cells
MH  - Killer Cells, Natural/*immunology
MH  - Ligands
MH  - Receptors, KIR/*agonists
MH  - Receptors, KIR3DS1/*metabolism
MH  - Signal Transduction
PMC - PMC8669139
OTO - NOTNLM
OT  - CRISPR
OT  - KIR
OT  - NK cells
OT  - heparan sulfate
OT  - screen
COIS- Author HD is employed by Adimab, LLC. The remaining authors declare that the 
      research was conducted in the absence of any commercial or financial 
      relationships that could be construed as a potential conflict of interest.
EDAT- 2021/12/18 06:00
MHDA- 2022/03/01 06:00
PMCR- 2021/01/01
CRDT- 2021/12/17 06:53
PHST- 2021/10/19 00:00 [received]
PHST- 2021/11/11 00:00 [accepted]
PHST- 2021/12/17 06:53 [entrez]
PHST- 2021/12/18 06:00 [pubmed]
PHST- 2022/03/01 06:00 [medline]
PHST- 2021/01/01 00:00 [pmc-release]
AID - 10.3389/fimmu.2021.798235 [doi]
PST - epublish
SO  - Front Immunol. 2021 Nov 30;12:798235. doi: 10.3389/fimmu.2021.798235. eCollection 
      2021.