Abstract
MLKL is the essential effector of necroptosis, a form of programmed lytic cell death. We have isolated a mouse strain with a single missense mutation, MlklD139V, that alters the two-helix ‘brace’ that connects the killer four-helix bundle and regulatory pseudokinase domains. This confers constitutive, RIPK3 independent killing activity to MLKL. Homozygous mutant mice develop lethal postnatal inflammation of the salivary glands and mediastinum. The normal embryonic development of MlklD139V homozygotes until birth, and the absence of any overt phenotype in heterozygotes provides important in vivo precedent for the capacity of cells to clear activated MLKL. These observations offer an important insight into the potential disease-modulating roles of three common human MLKL polymorphisms that encode amino acid substitutions within or adjacent to the brace region. Compound heterozygosity of these variants is found at up to 12-fold the expected frequency in patients that suffer from a pediatric autoinflammatory disease, chronic recurrent multifocal osteomyelitis (CRMO).
| Original language | English |
|---|---|
| Article number | 3150 |
| Number of pages | 16 |
| Journal | Nature Communications |
| Volume | 11 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 1 Dec 2020 |
| Externally published | Yes |
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In: Nature Communications, Vol. 11, No. 1, 3150, 01.12.2020.
Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - A missense mutation in the MLKL brace region promotes lethal neonatal inflammation and hematopoietic dysfunction
AU - Hildebrand, Joanne M.
AU - Kauppi, Maria
AU - Majewski, Ian J.
AU - Liu, Zikou
AU - Cox, Allison J.
AU - Miyake, Sanae
AU - Petrie, Emma J.
AU - Silk, Michael A.
AU - Li, Zhixiu
AU - Tanzer, Maria C.
AU - Brumatti, Gabriela
AU - Young, Samuel N.
AU - Hall, Cathrine
AU - Garnish, Sarah E.
AU - Corbin, Jason
AU - Stutz, Michael D.
AU - Di Rago, Ladina
AU - Gangatirkar, Pradnya
AU - Josefsson, Emma C.
AU - Rigbye, Kristin
AU - Anderton, Holly
AU - Rickard, James A.
AU - Tripaydonis, Anne
AU - Sheridan, Julie
AU - Scerri, Thomas S.
AU - Jackson, Victoria E.
AU - Czabotar, Peter E.
AU - Zhang, Jian-Guo
AU - Varghese, Leila
AU - Allison, Cody C.
AU - Pellegrini, Marc
AU - Tannahill, Gillian M.
AU - Hatchell, Esme C.
AU - Willson, Tracy A.
AU - Stockwell, Dina
AU - de Graaf, Carolyn A.
AU - Collinge, Janelle
AU - Hilton, Adrienne
AU - Silke, Natasha
AU - Spall, Sukhdeep K.
AU - Chau, Diep
AU - Athanasopoulos, Vicki
AU - Metcalf, Donald
AU - Laxer, Ronald M.
AU - Bassuk, Alexander G.
AU - Darbro, Benjamin W.
AU - Fiatarone Singh, Maria A.
AU - Vlahovich, Nicole
AU - Hughes, David
AU - Kozlovskaia, Maria
AU - Ascher, David B.
AU - Warnatz, Klaus
AU - Venhoff, Nils
AU - Thiel, Jens
AU - Biben, Christine
AU - Blum, Stefan
AU - Reveille, John
AU - Hildebrand, Michael S.
AU - Vinuesa, Carola G.
AU - McCombe, Pamela
AU - Brown, Matthew A.
AU - Kile, Benjamin T.
AU - McLean, Catriona
AU - Bahlo, Melanie
AU - Masters, Seth L.
AU - Nakano, Hiroyasu
AU - Ferguson, Polly J.
AU - Murphy, James M.
AU - Alexander, Warren S.
AU - Silke, John
N1 - Funding Information: MLKL variant quantification. 1000 Genomes: Vcf files from 1000 genomes were annotated and filtered as described previously79. Values for MLKL variants rs35589326 (S132P), rs34515646 (R146Q), and rs144526386 (G202V) as well as all MLKL coding variants were queried and tabulated for allele and genotype count for participants of all ancestry (n = 2504), and for those of European ancestry (n = 503). Compound heterozygous variants were evident due to the phasing of all variants in the 1000 genomes dataset. CRMO: Allele and genotype counts for all MLKL coding variants were tabulated in probands of European ancestry (n = 101) and for all probands (n = 128). Compound heterozygous variants were identified using parental sequence data. AS: DNA from all subjects in AS cohort were gen-otyped using the Illumina CoreExome chip following standard protocols at the Australian Translational Genomics Centre, Princess Alexandra Hospital, Brisbane. Bead intensity data was processed and normalized for each sample and genotypes called using the Illumina Genome Studio software. All the samples listed in the table have passed quality control process80. GB: Genotyping was performed in an ISO15189-accredited clinical genomics facility, Australian Translational Genomics Centre (ATGC), Queensland University of Technology. All samples were geno-typed by Illumina HumanOmniExpress (OmniExpress) BeadChip81. QUT controls: A collection of healthy control data of verified European ancestry from various cohort studies, complied by the Translational Genomics Group, QUT and typed on an Illumina CoreExome microarray. Includes data from The UK Household Longitudinal Study, led by the Institute for Social and Economic Research at the University of Essex and funded by the Economic and Social Research Council. The survey was conducted by NatCen and the genome-wide scan data were analysed and deposited by the Wellcome Trust Sanger Institute. University of Essex. Institute for Social and Economic Research, N. S. R., Kantar Public. Understanding Society: Waves 1–8, 2009–2017 and Harmonised BHPS: Waves 1–18, 1991–2009. [data collection]. 11th Edition. UK Data Service., (2018). Funding Information: We thank all the following people for their technical assistance; Jiami Han, Cynthia Liu, Jasmine McManus, Janelle Lochland and Emma Tovey (WEHI), Aira Nuguid and Tina Cardamone (APN histopathology – The University of Melbourne). Thomas Boudier (WEHI Centre for Dynamic Imaging). The WEHI Histology Service, WEHI Antibody Facility and WEHI Bioservices. Y. Uchiyama and S. Kakuta who advised the interpretation of the results of TEM and Annette Jacobsen (WEHI) for important insight and discussion. The generation of MlklD139V mice by CRISPR/Cas9 was performed by Andrew Kueh and Marco Herold (WEHI MAGEC laboratory) supported by the Australian Phenomics Network (APN) and the Australian Government through the National Collaborative Research Infrastructure Strategy (NCRIS) program. This work is supported by; Project grant (1105023) and Fellowships (0541951 and 1142669) from the Australian National Health and Medical Research Council (NHMRC) to J.M. H.; Fellowship (1107149) from the NHMRC to J.S.; Program grant (1113577) and Fellowship (1058344) from the NHMRC (W.S.A.); Project grant (1124735) and Fellowships (1105754, 1172929) from the NHMRC (J.M.M.); NIH training grants T32GM008629 and T32GM082729-01 (A.J.C.); R01AR059703 from the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) at the National Institutes of Health (P.J.F. and A.G.B.); the Marjorie K. Lamb Professorship (P.J.F.); Grants-in-Aid from Scientific Research (B) 17H04069 from Japan Society for the Promotion of Science, and Scientific Research on Innovative areas 26110003, the Japan Agency for Medical Research and Development (AMED) through AMED-CREST JP20gm1210002 and Private University Research Branding project from a MEXT (H. N.); Victorian International Research Scholarship (Z.L. and M.C.T.). Australia Post-graduate Award (C.A.D.); S.L.M. acknowledges funding from NHMRC grants (1144282, 1142354, and 1099262), The Sylvia and Charles Viertel Foundation, HHMI-Wellcome International Research Scholarship and Glaxosmithkline; Fellowship from the Lorenzo and Pamela Galli Charitable Trust (E.C.J.), NHMRC grants 1107425 and 1045549 and The Sylvia & Charles Viertel Senior Medical Research Fellowship (M.P.); D.B.A. was supported by the Jack Brockhoff Foundation (JBF 4186, 2016) and NHMRC Fellowship (APP1072476); NHMRC IRIISS 9000587 and Victorian Government Operational Infrastructure Support schemes; NHMRC Project and Targeted Research grants 1006769, 512672, and 512381 (M.F.S.); The Department of Industry, Innovation, Science, Research and Tertiary Education Collaborative Research Network and Diabetes Australia (MAB); I.J.M. was supported by the Victorian Cancer Agency, and by generous support from the Felton Bequest; Cancer Australia and Leukemia Foundation Australia priority grant (PdCCRS 1162023) and a Victoria Cancer Agency (VCA) mid-career fellowship (MCRF 15027) to G.B. We gratefully acknowledge the contribution of genotype data by Dr Yorgi Mavros (University of Sydney), Professor Nick Martin (QIMR), Professor Jim Rosenbaum (Oregon Health and Science University), and Professor Maxime Breban and the Groupe Française d’Etude Génétique des Spondylarthrites (GFEGS). We are grateful to Professor BP Wordsworth of the University of Oxford, UK for access to genotype data on ankylosing spondylitis cases collected in studies funded, in part, by Arthritis Research UK (grants 19536 and 18797), by the Wellcome Trust (grant 076113) and by the Oxford Comprehensive Biomedical Research Centre ankylosing spondylitis chronic disease cohort (theme A91202). Publisher Copyright: © 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - MLKL is the essential effector of necroptosis, a form of programmed lytic cell death. We have isolated a mouse strain with a single missense mutation, MlklD139V, that alters the two-helix ‘brace’ that connects the killer four-helix bundle and regulatory pseudokinase domains. This confers constitutive, RIPK3 independent killing activity to MLKL. Homozygous mutant mice develop lethal postnatal inflammation of the salivary glands and mediastinum. The normal embryonic development of MlklD139V homozygotes until birth, and the absence of any overt phenotype in heterozygotes provides important in vivo precedent for the capacity of cells to clear activated MLKL. These observations offer an important insight into the potential disease-modulating roles of three common human MLKL polymorphisms that encode amino acid substitutions within or adjacent to the brace region. Compound heterozygosity of these variants is found at up to 12-fold the expected frequency in patients that suffer from a pediatric autoinflammatory disease, chronic recurrent multifocal osteomyelitis (CRMO).
AB - MLKL is the essential effector of necroptosis, a form of programmed lytic cell death. We have isolated a mouse strain with a single missense mutation, MlklD139V, that alters the two-helix ‘brace’ that connects the killer four-helix bundle and regulatory pseudokinase domains. This confers constitutive, RIPK3 independent killing activity to MLKL. Homozygous mutant mice develop lethal postnatal inflammation of the salivary glands and mediastinum. The normal embryonic development of MlklD139V homozygotes until birth, and the absence of any overt phenotype in heterozygotes provides important in vivo precedent for the capacity of cells to clear activated MLKL. These observations offer an important insight into the potential disease-modulating roles of three common human MLKL polymorphisms that encode amino acid substitutions within or adjacent to the brace region. Compound heterozygosity of these variants is found at up to 12-fold the expected frequency in patients that suffer from a pediatric autoinflammatory disease, chronic recurrent multifocal osteomyelitis (CRMO).
UR - http://www.scopus.com/inward/record.url?scp=85086677563&partnerID=8YFLogxK
U2 - 10.1038/s41467-020-16819-z
DO - 10.1038/s41467-020-16819-z
M3 - Article
C2 - 32561755
AN - SCOPUS:85086677563
SN - 2041-1723
VL - 11
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 3150
ER -