Nature | Vol 577 | 2 January 2020 | 109Article
A dominant autoinflammatory disease
caused by non-cleavable variants of RIPK1
Panfeng Tao1,1 6, Jinqiao Sun2 ,1 6, Zheming Wu3,1 6, Shihao Wang1,1 6, Jun Wang1,1 6, Wanjin Li4 ,1 6,
Heling Pan^3 , Renkui Bai^5 , Jiahui Zhang^1 , Ying Wang^2 , Pui Y. Lee^6 , Wenjing Ying^2 , Qinhua Zhou^2 ,
Jia Hou^2 , Wenjie Wang^2 , Bijun Sun^2 , Mi Yang^2 , Danru Liu^2 , Ran Fang^1 , Huan Han^1 , Zhaohui Yang^1 ,
Xin Huang^3 , Haibo Li^7 , Natalie Deuitch^8 , Yuan Zhang^9 , Dilan Dissanayake^10 , Katrina Haude^5 ,
Kirsty McWalter^5 , Chelsea Roadhouse^11 , Jennifer J. MacKenzie1 1,1 2, Ronald M. Laxer^13 ,
Ivona Aksentijevich^14 , Xiaomin Yu1,1 7*, Xiaochuan Wang2 ,1 7*, Junying Yuan4 ,1 7* & Qing Zhou1,1 5,1 7*Activation of RIPK1 controls TNF-mediated apoptosis, necroptosis and inflammatory
pathways^1. Cleavage of human and mouse RIPK1 after residues D324 and D325,
respectively, by caspase-8 separates the RIPK1 kinase domain from the intermediate
and death domains. The D325A mutation in mouse RIPK1 leads to embryonic lethality
during mouse development^2 ,^3. However, the functional importance of blocking
caspase-8-mediated cleavage of RIPK1 on RIPK1 activation in humans is unknown.
Here we identify two families with variants in RIPK1 (D324V and D324H) that lead to
distinct symptoms of recurrent fevers and lymphadenopathy in an autosomal-
dominant manner. Impaired cleavage of RIPK1 D324 variants by caspase-8 sensitized
patients’ peripheral blood mononuclear cells to RIPK1 activation, apoptosis and
necroptosis induced by TNF. The patients showed strong RIPK1-dependent activation
of inflammatory signalling pathways and overproduction of inflammatory cytokines
and chemokines compared with unaffected controls. Furthermore, we show that
expression of the RIPK1 mutants D325V or D325H in mouse embryonic fibroblasts
confers not only increased sensitivity to RIPK1 activation-mediated apoptosis and
necroptosis, but also induction of pro-inflammatory cytokines such as IL-6 and TNF.
By contrast, patient-derived fibroblasts showed reduced expression of RIPK1 and
downregulated production of reactive oxygen species, resulting in resistance to
necroptosis and ferroptosis. Together, these data suggest that human non-cleavable
RIPK1 variants promote activation of RIPK1, and lead to an autoinflammatory disease
characterized by hypersensitivity to apoptosis and necroptosis and increased
inflammatory response in peripheral blood mononuclear cells, as well as a
compensatory mechanism to protect against several pro-death stimuli in fibroblasts.RIPK1 is a key mediator of apoptotic and necrotic cell death as well as
inflammatory pathways^1. Activation of RIPK1 promotes several cell
death responses, including apoptosis and necroptosis, downstream
of TNFR1. Caspase-8-mediated cleavage after Asp324 in human RIPK1
(or Asp325 in mouse RIPK1) separates the kinase domain in the N-termi-
nal part of RIPK1 from its intermediate and death domains. The death
domain is involved in mediating the activation of the N-terminal kinase
by dimerization^1 ,^4 ,^5. The D324A variant in human RIPK1 blocks cleavage
by caspase-8^6. Homozygous D325A mutation in mouse RIPK1 sensitizes
cells to both apoptosis and necroptosis induced by TNF and leads to
embryonic lethality. The early demise of Ripk1D325A/D325A mice can be
rescued by simultaneous deletion of Ripk3 and Fadd^2 , or Mlkl and Fadd^3 ,
but not of either gene alone. However, the functional importance of
caspase-8-mediated cleavage of RIPK1 in humans is unknown. Here, we
identified a human autoinflammatory disease caused by non-cleavable
RIPK1 variants with mutations at D324. We show that disrupted cleav-
age of RIPK1 by caspase-8 in humans leads to a dominantly inherited
condition by promoting the activation of RIPK1.https://doi.org/10.1038/s41586-019-1830-y
Received: 24 May 2019
Accepted: 21 October 2019
Published online: 11 December 2019
(^1) The MOE Key Laboratory of Biosystems Homeostasis & Protection, Life Sciences Institute, Zhejiang University, Hangzhou, China. (^2) Department of Clinical Immunology, Children’s Hospital of
Fudan University, Shanghai, China.^3 Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.
(^4) Department of Cell Biology, Harvard Medical School, Boston, MA, USA. (^5) GeneDx, Gaithersburg, MD, USA. (^6) Division of Immunology, Boston Children’s Hospital, Boston, MA, USA. (^7) Ningbo
Women and Children’s Hospital, Ningbo, China.^8 Department of Human Genetics, Stanford University School of Medicine, Stanford, CA, USA.^9 Laboratory of Allergic Diseases, National Institute
of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.^10 Division of Rheumatology, Department of Paediatrics, The Hospital for Sick Children and the University of
Toronto, Toronto, Ontario, Canada.^11 Department of Pediatrics, McMaster Children’s Hospital, Hamilton, Ontario, Canada.^12 McMaster University, Hamilton, Ontario, Canada.^13 Division of
Rheumatology, Departments of Paediatrics and Medicine, The Hospital for Sick Children and the University of Toronto, Toronto, Ontario, Canada.^14 Inflammatory Disease Section, National
Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA.^15 Women’s Hospital, Zhejiang University School of Medicine, Hangzhou, China.^16 These authors
contributed equally: Panfeng Tao, Jinqiao Sun, Zheming Wu, Shihao Wang, Jun Wang, Wanjin Li.^17 These authors jointly supervised this work: Xiaomin Yu, Xiaochuan Wang, Junying Yuan, Qing
Zhou. *e-mail: [email protected]; [email protected]; [email protected]; [email protected]