NEMO, a non-catalytically active component of the IkappaB alpha kinase (IKK) complex, is required for IKK enzymatic activity and localization. If NEMO is defective, NF-kB transcription factor family members do not translocate to the nucleus to regulate gene transcription, and normal development of the organism, including the immune system, does not occur. In addition to these developmental defects, the immune system does not function correctly if NEMO is defective. Individuals with specific mutations which result in NEMO immunodeficiency and inflammatory disease are investigated using molecular, cellular and biochemical assays/techniques. In addition to clinical and laboratory evaluation of human patients, these studies are conducted using patient-derived induced pluripotent stem cells, and a reconstitution system utilizing NEMO-deficient Jurkat T cells. Because NEMO syndrome is associated with defects in development of ectodermal structures in addition to secondary lymphoid structures in association with certain mutations, we seek to understand the function of NEMO in both of these roles in addition to its role in hematopoietically derived cells. We have generated human iPSC to study the effects of mutation on the development and function of these various cell types. NEMO is required for IKK activation, but also thought to serve as a scaffold to other signaling proteins. Its regulation is complex: splice isoforms exist, and it is post-translationally modified by serine phosphorylation, non-degradative ubiquitination, and sumoylation. Variability of clinical disease phenotypes and immune-function lab abnormalities make management of individuals with NEMO syndrome and inflammatory disease difficult. Because of our incomplete understanding, treatment paradigms such as the use of allogeneic hematopoietic stem cell transplantation cannot at this point be standardized and often result in poor outcomes. We are establishing protocols to evaluate patients at the NIH Clinical Center to be able to comprehensively characterize the spectrum of phenotypes, and are working to develop protocols to offer advanced treatment based on our clinical and laboratory findings. In a similar fashion, we are studying individuals with NEMO-Like Syndrome who have suspected monogenic diseases which phenocopy NEMO syndrome in some aspects. We hypothesize that these individuals will have defects in signaling proteins or other regulatory proteins that interact in the NF-kB signaling pathway. Our general approach to discover the genetic defects that cause disease is to use candidate gene sequencing based on the disease and clinical laboratory phenotype, or, if necessary, exome or whole genome sequencing. Candidate genes identified are evaluated using assays that interrogate different checkpoints along the NF-kB activation pathway. Insights into the role of proteins gained from study of patients with rare diseases will be applied to evaluation of the normal immune system to understand the function of these proteins in the normal immune response and more common inflammatory diseases.

Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2013
Total Cost
$502,097
Indirect Cost
Name
National Institute of Arthritis and Musculoskeletal and Skin Diseases
Department
Type
DUNS #
City
State
Country
Zip Code
Zhou, Qing; Wang, Hongying; Schwartz, Daniella M et al. (2016) Loss-of-function mutations in TNFAIP3 leading to A20 haploinsufficiency cause an early-onset autoinflammatory disease. Nat Genet 48:67-73
Zilberman-Rudenko, Jevgenia; Shawver, Linda Monaco; Wessel, Alex W et al. (2016) Recruitment of A20 by the C-terminal domain of NEMO suppresses NF-κB activation and autoinflammatory disease. Proc Natl Acad Sci U S A 113:1612-7
Wessel, Alex W; Hanson, Eric P (2015) A method for the quantitative analysis of stimulation-induced nuclear translocation of the p65 subunit of NF-κB from patient-derived dermal fibroblasts. Methods Mol Biol 1280:413-26
Rider, Nicholas L; Boisson, Bertrand; Jyonouchi, Soma et al. (2015) Corrigendum: Novel TTC37 Mutations in a Patient with Immunodeficiency without Diarrhea: Extending the Phenotype of Trichohepatoenteric Syndrome. Front Pediatr 3:28
Rider, Nicholas L; Boisson, Bertrand; Jyonouchi, Soma et al. (2015) Novel TTC37 Mutations in a Patient with Immunodeficiency without Diarrhea: Extending the Phenotype of Trichohepatoenteric Syndrome. Front Pediatr 3:2
Kotlarz, Daniel; Zietara, Natalia; Uzel, Gulbu et al. (2013) Loss-of-function mutations in the IL-21 receptor gene cause a primary immunodeficiency syndrome. J Exp Med 210:433-43
Keller, Michael D; Petersen, Maureen; Ong, Peck et al. (2011) Hypohidrotic ectodermal dysplasia and immunodeficiency with coincident NEMO and EDA mutations. Front Immunol 2:61
Karamchandani-Patel, Gital; Hanson, Eric P; Saltzman, Rushani et al. (2011) Congenital alterations of NEMO glutamic acid 223 result in hypohidrotic ectodermal dysplasia and immunodeficiency with normal serum IgG levels. Ann Allergy Asthma Immunol 107:50-6
Devora, Gene A; Sun, Lijun; Chen, Zhijian et al. (2010) A novel missense mutation in the nuclear factor-κB essential modulator (NEMO) gene resulting in impaired activation of the NF-κB pathway and a unique clinical phenotype presenting as MRSA subdural empyema. J Clin Immunol 30:881-5
Yatherajam, Gayatri; Banerjee, Pinaki P; McCorkell, Kelly A et al. (2010) Cutting edge: association with I kappa B kinase beta regulates the subcellular localization of Homer3. J Immunol 185:2665-9