Cytosolic 5′-nucleotidase 1A autoimmunity in sporadic inclusion body myositis

Harry Larman, Mohammad Salajegheh, Remedios Nazareno, Theresa Lam, John Sauld, Hanno Steen, Sek Won Kong, Jack L. Pinkus, Anthony A. Amato, Stephen J. Elledge, Steven A. Greenberg

Research output: Contribution to journalArticle

Abstract

Objective We previously identified a circulating autoantibody against a 43 kDa muscle autoantigen in sporadic inclusion body myositis (IBM) and demonstrated the feasibility of an IBM diagnostic blood test. Here, we sought to identify the molecular target of this IBM autoantibody, understand the relationship between IBM autoimmunity and muscle degeneration, and develop an IBM blood test with high diagnostic accuracy. Methods IBM blood samples were screened using mass spectrometry and a synthetic human peptidome. Plasma and serum samples (N=200 patients) underwent immunoblotting assays, and results were correlated to clinical features. Muscle biopsy samples (n=30) were examined by immunohistochemistry and immunoblotting. Exome or whole genome sequencing was performed on DNA from 19 patients. Results Both mass spectrometry and screening of a 413,611 human peptide library spanning the entire human proteome identified cytosolic 5′-nucleotidase 1A (cN1A; NT5C1A) as the likely 43 kDa IBM autoantigen, which was then confirmed in dot blot and Western blot assays using recombinant cN1A protein. Moderate reactivity of anti-cN1A autoantibodies was 70% sensitive and 92% specific, and high reactivity was 34% sensitive and 98% specific for the diagnosis of IBM. One to 3 major cN1A immunodominant epitopes were identified. cN1A reactivity by immunohistochemistry accumulated in perinuclear regions and rimmed vacuoles in IBM muscle, localizing to areas of myonuclear degeneration. Interpretation Autoantibodies against cN1A are common in and highly specific to IBM among muscle diseases, and may provide a link between IBM's dual processes of autoimmunity and myodegeneration. Blood diagnostic testing is feasible and should improve early and reliable diagnosis of IBM. Ann Neurol 2013;73:408-418

Original languageEnglish (US)
Pages (from-to)408-418
Number of pages11
JournalAnnals of Neurology
Volume73
Issue number3
DOIs
StatePublished - Mar 2013
Externally publishedYes

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Inclusion Body Myositis
5'-Nucleotidase
Autoimmunity
Autoantibodies
Muscles
Autoantigens
Hematologic Tests
Immunoblotting
Mass Spectrometry
Immunohistochemistry
Exome
Peptide Library
Immunodominant Epitopes
Mass Screening
Proteome
Vacuoles
Recombinant Proteins
Routine Diagnostic Tests
Early Diagnosis

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology

Cite this

Larman, H., Salajegheh, M., Nazareno, R., Lam, T., Sauld, J., Steen, H., ... Greenberg, S. A. (2013). Cytosolic 5′-nucleotidase 1A autoimmunity in sporadic inclusion body myositis. Annals of Neurology, 73(3), 408-418. https://doi.org/10.1002/ana.23840

Cytosolic 5′-nucleotidase 1A autoimmunity in sporadic inclusion body myositis. / Larman, Harry; Salajegheh, Mohammad; Nazareno, Remedios; Lam, Theresa; Sauld, John; Steen, Hanno; Won Kong, Sek; Pinkus, Jack L.; Amato, Anthony A.; Elledge, Stephen J.; Greenberg, Steven A.

In: Annals of Neurology, Vol. 73, No. 3, 03.2013, p. 408-418.

Research output: Contribution to journalArticle

Larman, H, Salajegheh, M, Nazareno, R, Lam, T, Sauld, J, Steen, H, Won Kong, S, Pinkus, JL, Amato, AA, Elledge, SJ & Greenberg, SA 2013, 'Cytosolic 5′-nucleotidase 1A autoimmunity in sporadic inclusion body myositis', Annals of Neurology, vol. 73, no. 3, pp. 408-418. https://doi.org/10.1002/ana.23840
Larman, Harry ; Salajegheh, Mohammad ; Nazareno, Remedios ; Lam, Theresa ; Sauld, John ; Steen, Hanno ; Won Kong, Sek ; Pinkus, Jack L. ; Amato, Anthony A. ; Elledge, Stephen J. ; Greenberg, Steven A. / Cytosolic 5′-nucleotidase 1A autoimmunity in sporadic inclusion body myositis. In: Annals of Neurology. 2013 ; Vol. 73, No. 3. pp. 408-418.
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N2 - Objective We previously identified a circulating autoantibody against a 43 kDa muscle autoantigen in sporadic inclusion body myositis (IBM) and demonstrated the feasibility of an IBM diagnostic blood test. Here, we sought to identify the molecular target of this IBM autoantibody, understand the relationship between IBM autoimmunity and muscle degeneration, and develop an IBM blood test with high diagnostic accuracy. Methods IBM blood samples were screened using mass spectrometry and a synthetic human peptidome. Plasma and serum samples (N=200 patients) underwent immunoblotting assays, and results were correlated to clinical features. Muscle biopsy samples (n=30) were examined by immunohistochemistry and immunoblotting. Exome or whole genome sequencing was performed on DNA from 19 patients. Results Both mass spectrometry and screening of a 413,611 human peptide library spanning the entire human proteome identified cytosolic 5′-nucleotidase 1A (cN1A; NT5C1A) as the likely 43 kDa IBM autoantigen, which was then confirmed in dot blot and Western blot assays using recombinant cN1A protein. Moderate reactivity of anti-cN1A autoantibodies was 70% sensitive and 92% specific, and high reactivity was 34% sensitive and 98% specific for the diagnosis of IBM. One to 3 major cN1A immunodominant epitopes were identified. cN1A reactivity by immunohistochemistry accumulated in perinuclear regions and rimmed vacuoles in IBM muscle, localizing to areas of myonuclear degeneration. Interpretation Autoantibodies against cN1A are common in and highly specific to IBM among muscle diseases, and may provide a link between IBM's dual processes of autoimmunity and myodegeneration. Blood diagnostic testing is feasible and should improve early and reliable diagnosis of IBM. Ann Neurol 2013;73:408-418

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