A systematic analysis of recombination activity and genotype-phenotype correlation in human recombination-activating gene 1 deficiency

Yu Nee Lee, Francesco Frugoni, Kerry Dobbs, Jolan E. Walter, Silvia Giliani, Andrew R. Gennery, Waleed Al-Herz, Elie Haddad, Francoise Ledeist, Jack H. Bleesing, Lauren A. Henderson, Sung Yun Pai, Robert P. Nelson, Dalia H. El-Ghoneimy, Reem A. El-Feky, Shereen M. Reda, Elham Hossny, Pere Soler-Palacin, Ramsay L Fuleihan, Niraj C. Patel & 9 others Michel J. Massaad, Raif S. Geha, Jennifer M. Puck, Paolo Palma, Caterina Cancrini, Karin Chen, Mauno Vihinen, Frederick W. Alt*, Luigi D. Notarangelo

*Corresponding author for this work

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

Background The recombination-activating gene (RAG) 1/2 proteins play a critical role in the development of T and B cells by initiating the VDJ recombination process that leads to generation of a broad T-cell receptor (TCR) and B-cell receptor repertoire. Pathogenic mutations in the RAG1/2 genes result in various forms of primary immunodeficiency, ranging from T - B - severe combined immune deficiency to delayed-onset disease with granuloma formation, autoimmunity, or both. It is not clear what contributes to such heterogeneity of phenotypes. Objective We sought to investigate the molecular basis for phenotypic diversity presented in patients with various RAG1 mutations. Methods We have developed a flow cytometry-based assay that allows analysis of RAG recombination activity based on green fluorescent protein expression and have assessed the induction of the Ighc locus rearrangements in mouse Rag1 -/- pro-B cells reconstituted with wild-type or mutant human RAG1 (hRAG1) using deep sequencing technology. Results Here we demonstrate correlation between defective recombination activity of hRAG1 mutant proteins and severity of the clinical and immunologic phenotype and provide insights on the molecular mechanisms accounting for such phenotypic diversity. Conclusions Using a sensitive assay to measure the RAG1 activity level of 79 mutations in a physiologic setting, we demonstrate correlation between recombination activity of RAG1 mutants and the severity of clinical presentation and show that RAG1 mutants can induce specific abnormalities of the VDJ recombination process.

Original languageEnglish (US)
JournalJournal of Allergy and Clinical Immunology
Volume133
Issue number4
DOIs
StatePublished - Jan 1 2014

Fingerprint

RAG-1 Genes
Genetic Association Studies
Genetic Recombination
V(D)J Recombination
Mutation
B-Lymphocytes
Phenotype
High-Throughput Nucleotide Sequencing
Severe Combined Immunodeficiency
B-Lymphoid Precursor Cells
Mutant Proteins
T-Cell Antigen Receptor
Green Fluorescent Proteins
Granuloma
Autoimmunity
Genes
Flow Cytometry
Technology
T-Lymphocytes

Keywords

  • Omenn syndrome
  • Recombination-activating gene 1
  • V(D)J recombination
  • autoimmunity
  • genotype-phenotype correlation
  • immune repertoire
  • severe combined immune deficiency

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Lee, Yu Nee ; Frugoni, Francesco ; Dobbs, Kerry ; Walter, Jolan E. ; Giliani, Silvia ; Gennery, Andrew R. ; Al-Herz, Waleed ; Haddad, Elie ; Ledeist, Francoise ; Bleesing, Jack H. ; Henderson, Lauren A. ; Pai, Sung Yun ; Nelson, Robert P. ; El-Ghoneimy, Dalia H. ; El-Feky, Reem A. ; Reda, Shereen M. ; Hossny, Elham ; Soler-Palacin, Pere ; Fuleihan, Ramsay L ; Patel, Niraj C. ; Massaad, Michel J. ; Geha, Raif S. ; Puck, Jennifer M. ; Palma, Paolo ; Cancrini, Caterina ; Chen, Karin ; Vihinen, Mauno ; Alt, Frederick W. ; Notarangelo, Luigi D. / A systematic analysis of recombination activity and genotype-phenotype correlation in human recombination-activating gene 1 deficiency. In: Journal of Allergy and Clinical Immunology. 2014 ; Vol. 133, No. 4.
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title = "A systematic analysis of recombination activity and genotype-phenotype correlation in human recombination-activating gene 1 deficiency",
abstract = "Background The recombination-activating gene (RAG) 1/2 proteins play a critical role in the development of T and B cells by initiating the VDJ recombination process that leads to generation of a broad T-cell receptor (TCR) and B-cell receptor repertoire. Pathogenic mutations in the RAG1/2 genes result in various forms of primary immunodeficiency, ranging from T - B - severe combined immune deficiency to delayed-onset disease with granuloma formation, autoimmunity, or both. It is not clear what contributes to such heterogeneity of phenotypes. Objective We sought to investigate the molecular basis for phenotypic diversity presented in patients with various RAG1 mutations. Methods We have developed a flow cytometry-based assay that allows analysis of RAG recombination activity based on green fluorescent protein expression and have assessed the induction of the Ighc locus rearrangements in mouse Rag1 -/- pro-B cells reconstituted with wild-type or mutant human RAG1 (hRAG1) using deep sequencing technology. Results Here we demonstrate correlation between defective recombination activity of hRAG1 mutant proteins and severity of the clinical and immunologic phenotype and provide insights on the molecular mechanisms accounting for such phenotypic diversity. Conclusions Using a sensitive assay to measure the RAG1 activity level of 79 mutations in a physiologic setting, we demonstrate correlation between recombination activity of RAG1 mutants and the severity of clinical presentation and show that RAG1 mutants can induce specific abnormalities of the VDJ recombination process.",
keywords = "Omenn syndrome, Recombination-activating gene 1, V(D)J recombination, autoimmunity, genotype-phenotype correlation, immune repertoire, severe combined immune deficiency",
author = "Lee, {Yu Nee} and Francesco Frugoni and Kerry Dobbs and Walter, {Jolan E.} and Silvia Giliani and Gennery, {Andrew R.} and Waleed Al-Herz and Elie Haddad and Francoise Ledeist and Bleesing, {Jack H.} and Henderson, {Lauren A.} and Pai, {Sung Yun} and Nelson, {Robert P.} and El-Ghoneimy, {Dalia H.} and El-Feky, {Reem A.} and Reda, {Shereen M.} and Elham Hossny and Pere Soler-Palacin and Fuleihan, {Ramsay L} and Patel, {Niraj C.} and Massaad, {Michel J.} and Geha, {Raif S.} and Puck, {Jennifer M.} and Paolo Palma and Caterina Cancrini and Karin Chen and Mauno Vihinen and Alt, {Frederick W.} and Notarangelo, {Luigi D.}",
year = "2014",
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journal = "Journal of Allergy and Clinical Immunology",
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Lee, YN, Frugoni, F, Dobbs, K, Walter, JE, Giliani, S, Gennery, AR, Al-Herz, W, Haddad, E, Ledeist, F, Bleesing, JH, Henderson, LA, Pai, SY, Nelson, RP, El-Ghoneimy, DH, El-Feky, RA, Reda, SM, Hossny, E, Soler-Palacin, P, Fuleihan, RL, Patel, NC, Massaad, MJ, Geha, RS, Puck, JM, Palma, P, Cancrini, C, Chen, K, Vihinen, M, Alt, FW & Notarangelo, LD 2014, 'A systematic analysis of recombination activity and genotype-phenotype correlation in human recombination-activating gene 1 deficiency', Journal of Allergy and Clinical Immunology, vol. 133, no. 4. https://doi.org/10.1016/j.jaci.2013.10.007

A systematic analysis of recombination activity and genotype-phenotype correlation in human recombination-activating gene 1 deficiency. / Lee, Yu Nee; Frugoni, Francesco; Dobbs, Kerry; Walter, Jolan E.; Giliani, Silvia; Gennery, Andrew R.; Al-Herz, Waleed; Haddad, Elie; Ledeist, Francoise; Bleesing, Jack H.; Henderson, Lauren A.; Pai, Sung Yun; Nelson, Robert P.; El-Ghoneimy, Dalia H.; El-Feky, Reem A.; Reda, Shereen M.; Hossny, Elham; Soler-Palacin, Pere; Fuleihan, Ramsay L; Patel, Niraj C.; Massaad, Michel J.; Geha, Raif S.; Puck, Jennifer M.; Palma, Paolo; Cancrini, Caterina; Chen, Karin; Vihinen, Mauno; Alt, Frederick W.; Notarangelo, Luigi D.

In: Journal of Allergy and Clinical Immunology, Vol. 133, No. 4, 01.01.2014.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A systematic analysis of recombination activity and genotype-phenotype correlation in human recombination-activating gene 1 deficiency

AU - Lee, Yu Nee

AU - Frugoni, Francesco

AU - Dobbs, Kerry

AU - Walter, Jolan E.

AU - Giliani, Silvia

AU - Gennery, Andrew R.

AU - Al-Herz, Waleed

AU - Haddad, Elie

AU - Ledeist, Francoise

AU - Bleesing, Jack H.

AU - Henderson, Lauren A.

AU - Pai, Sung Yun

AU - Nelson, Robert P.

AU - El-Ghoneimy, Dalia H.

AU - El-Feky, Reem A.

AU - Reda, Shereen M.

AU - Hossny, Elham

AU - Soler-Palacin, Pere

AU - Fuleihan, Ramsay L

AU - Patel, Niraj C.

AU - Massaad, Michel J.

AU - Geha, Raif S.

AU - Puck, Jennifer M.

AU - Palma, Paolo

AU - Cancrini, Caterina

AU - Chen, Karin

AU - Vihinen, Mauno

AU - Alt, Frederick W.

AU - Notarangelo, Luigi D.

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Background The recombination-activating gene (RAG) 1/2 proteins play a critical role in the development of T and B cells by initiating the VDJ recombination process that leads to generation of a broad T-cell receptor (TCR) and B-cell receptor repertoire. Pathogenic mutations in the RAG1/2 genes result in various forms of primary immunodeficiency, ranging from T - B - severe combined immune deficiency to delayed-onset disease with granuloma formation, autoimmunity, or both. It is not clear what contributes to such heterogeneity of phenotypes. Objective We sought to investigate the molecular basis for phenotypic diversity presented in patients with various RAG1 mutations. Methods We have developed a flow cytometry-based assay that allows analysis of RAG recombination activity based on green fluorescent protein expression and have assessed the induction of the Ighc locus rearrangements in mouse Rag1 -/- pro-B cells reconstituted with wild-type or mutant human RAG1 (hRAG1) using deep sequencing technology. Results Here we demonstrate correlation between defective recombination activity of hRAG1 mutant proteins and severity of the clinical and immunologic phenotype and provide insights on the molecular mechanisms accounting for such phenotypic diversity. Conclusions Using a sensitive assay to measure the RAG1 activity level of 79 mutations in a physiologic setting, we demonstrate correlation between recombination activity of RAG1 mutants and the severity of clinical presentation and show that RAG1 mutants can induce specific abnormalities of the VDJ recombination process.

AB - Background The recombination-activating gene (RAG) 1/2 proteins play a critical role in the development of T and B cells by initiating the VDJ recombination process that leads to generation of a broad T-cell receptor (TCR) and B-cell receptor repertoire. Pathogenic mutations in the RAG1/2 genes result in various forms of primary immunodeficiency, ranging from T - B - severe combined immune deficiency to delayed-onset disease with granuloma formation, autoimmunity, or both. It is not clear what contributes to such heterogeneity of phenotypes. Objective We sought to investigate the molecular basis for phenotypic diversity presented in patients with various RAG1 mutations. Methods We have developed a flow cytometry-based assay that allows analysis of RAG recombination activity based on green fluorescent protein expression and have assessed the induction of the Ighc locus rearrangements in mouse Rag1 -/- pro-B cells reconstituted with wild-type or mutant human RAG1 (hRAG1) using deep sequencing technology. Results Here we demonstrate correlation between defective recombination activity of hRAG1 mutant proteins and severity of the clinical and immunologic phenotype and provide insights on the molecular mechanisms accounting for such phenotypic diversity. Conclusions Using a sensitive assay to measure the RAG1 activity level of 79 mutations in a physiologic setting, we demonstrate correlation between recombination activity of RAG1 mutants and the severity of clinical presentation and show that RAG1 mutants can induce specific abnormalities of the VDJ recombination process.

KW - Omenn syndrome

KW - Recombination-activating gene 1

KW - V(D)J recombination

KW - autoimmunity

KW - genotype-phenotype correlation

KW - immune repertoire

KW - severe combined immune deficiency

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UR - http://www.scopus.com/inward/citedby.url?scp=84897389616&partnerID=8YFLogxK

U2 - 10.1016/j.jaci.2013.10.007

DO - 10.1016/j.jaci.2013.10.007

M3 - Article

VL - 133

JO - Journal of Allergy and Clinical Immunology

JF - Journal of Allergy and Clinical Immunology

SN - 0091-6749

IS - 4

ER -