Systematic functional validation of IKAROS variants from patients and laboratory-generated mutations
Issued Date
2026-03-10
Resource Type
ISSN
24739529
eISSN
24739537
Scopus ID
2-s2.0-105031456762
Pubmed ID
41380100
Journal Title
Blood Advances
Volume
10
Issue
5
Start Page
1522
End Page
1536
Rights Holder(s)
SCOPUS
Bibliographic Citation
Blood Advances Vol.10 No.5 (2026) , 1522-1536
Suggested Citation
Kuehn H.S., Gil Silva A.A., Klangkalya N., Esteve-Sole A., Goel S., Niemela J.E., Stoddard J.L., Chong H., Williams K., Ip W., Framil Seoane M., Loganathan S.K., Anantharachagan A., García-Soidán A., Shaffren S., Bindernagel C., Mendez P., Miller R.Z., Abolhassani H., Hoytema van Konijnenburg D.P., Geerlinks A.V., Cabanillas D., Zonneveld-Huijssoon E., Rutgers A., Tangye S., Ma C., Grey A., O'Young P., Aldave Becerra J.C., Lyons J.J., White A.A., Claiborne M., Doroudchi M.A., Modena B.D., Milner J.D., Solanich X., Farela Neves J., Platt C.D., Hammarström L., Routes J.M., Verbsky J., Butte M.J., Lesmana H., Griffiths W.J.H., Kumararatne D., Hauck F., Allende L.M., Rodríguez-Gallego C., Elfeky R., Rubin T., Boztug K., Fleisher T.A., Rosenzweig S.D. Systematic functional validation of IKAROS variants from patients and laboratory-generated mutations. Blood Advances Vol.10 No.5 (2026) , 1522-1536. 1536. doi:10.1182/bloodadvances.2025017243 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/115589
Title
Systematic functional validation of IKAROS variants from patients and laboratory-generated mutations
Author(s)
Kuehn H.S.
Gil Silva A.A.
Klangkalya N.
Esteve-Sole A.
Goel S.
Niemela J.E.
Stoddard J.L.
Chong H.
Williams K.
Ip W.
Framil Seoane M.
Loganathan S.K.
Anantharachagan A.
García-Soidán A.
Shaffren S.
Bindernagel C.
Mendez P.
Miller R.Z.
Abolhassani H.
Hoytema van Konijnenburg D.P.
Geerlinks A.V.
Cabanillas D.
Zonneveld-Huijssoon E.
Rutgers A.
Tangye S.
Ma C.
Grey A.
O'Young P.
Aldave Becerra J.C.
Lyons J.J.
White A.A.
Claiborne M.
Doroudchi M.A.
Modena B.D.
Milner J.D.
Solanich X.
Farela Neves J.
Platt C.D.
Hammarström L.
Routes J.M.
Verbsky J.
Butte M.J.
Lesmana H.
Griffiths W.J.H.
Kumararatne D.
Hauck F.
Allende L.M.
Rodríguez-Gallego C.
Elfeky R.
Rubin T.
Boztug K.
Fleisher T.A.
Rosenzweig S.D.
Gil Silva A.A.
Klangkalya N.
Esteve-Sole A.
Goel S.
Niemela J.E.
Stoddard J.L.
Chong H.
Williams K.
Ip W.
Framil Seoane M.
Loganathan S.K.
Anantharachagan A.
García-Soidán A.
Shaffren S.
Bindernagel C.
Mendez P.
Miller R.Z.
Abolhassani H.
Hoytema van Konijnenburg D.P.
Geerlinks A.V.
Cabanillas D.
Zonneveld-Huijssoon E.
Rutgers A.
Tangye S.
Ma C.
Grey A.
O'Young P.
Aldave Becerra J.C.
Lyons J.J.
White A.A.
Claiborne M.
Doroudchi M.A.
Modena B.D.
Milner J.D.
Solanich X.
Farela Neves J.
Platt C.D.
Hammarström L.
Routes J.M.
Verbsky J.
Butte M.J.
Lesmana H.
Griffiths W.J.H.
Kumararatne D.
Hauck F.
Allende L.M.
Rodríguez-Gallego C.
Elfeky R.
Rubin T.
Boztug K.
Fleisher T.A.
Rosenzweig S.D.
Author's Affiliation
University of California, Los Angeles
UNSW Sydney
Karolinska Institutet
Western University
Case Western Reserve University
University of Manitoba
Boston Children's Hospital
Klinikum der Universität München
Universitair Medisch Centrum Groningen
Columbia University Irving Medical Center
Medical College of Wisconsin
Hospital Clínic de Barcelona
College of Medicine
Addenbrooke's Hospital
Morsani College of Medicine
Department of Medicine
Universitätsklinikum Bonn
UCL Great Ormond Street Institute of Child Health
The Royal Free Hospital
Hospital Universitari de Bellvitge
Hospital Universitario 12 de Octubre
Royal Prince Alfred Hospital
Westmead Hospital
Universidad de Las Palmas de Gran Canaria
UPMC Children’s Hospital of Pittsburgh
VA San Diego Healthcare System
Christian Medical College, Vellore
Centro de Investigación Biomédica en Red de Enfermedades Respiratorias
Garvan Institute of Medical Research
NIH Clinical Center (CC)
Scripps Clinic
Faculty of Medicine Ramathibodi Hospital, Mahidol University
NOVA Medical School - Faculdade de Ciências Médicas, Universidade Nova de Lisboa
Johns Hopkins All Children's Hospital
Hospital de Gran Canaria Dr. Negrin
Lancashire Teaching Hospitals NHS Foundation Trust
Research Center for Immunodeficiencies
Hospital Nacional Edgardo Rebagliati Martins, EsSalud
Hospital de Dona Estefânia
Hospital de Ninos Sor Maria Ludovica
Universidad Fernando Pessoa Canarias
St. Anna Children’s Cancer Research Institute
Clinical Immunogenomics Research Consortium Australasia
Modena Health
UNSW Sydney
Karolinska Institutet
Western University
Case Western Reserve University
University of Manitoba
Boston Children's Hospital
Klinikum der Universität München
Universitair Medisch Centrum Groningen
Columbia University Irving Medical Center
Medical College of Wisconsin
Hospital Clínic de Barcelona
College of Medicine
Addenbrooke's Hospital
Morsani College of Medicine
Department of Medicine
Universitätsklinikum Bonn
UCL Great Ormond Street Institute of Child Health
The Royal Free Hospital
Hospital Universitari de Bellvitge
Hospital Universitario 12 de Octubre
Royal Prince Alfred Hospital
Westmead Hospital
Universidad de Las Palmas de Gran Canaria
UPMC Children’s Hospital of Pittsburgh
VA San Diego Healthcare System
Christian Medical College, Vellore
Centro de Investigación Biomédica en Red de Enfermedades Respiratorias
Garvan Institute of Medical Research
NIH Clinical Center (CC)
Scripps Clinic
Faculty of Medicine Ramathibodi Hospital, Mahidol University
NOVA Medical School - Faculdade de Ciências Médicas, Universidade Nova de Lisboa
Johns Hopkins All Children's Hospital
Hospital de Gran Canaria Dr. Negrin
Lancashire Teaching Hospitals NHS Foundation Trust
Research Center for Immunodeficiencies
Hospital Nacional Edgardo Rebagliati Martins, EsSalud
Hospital de Dona Estefânia
Hospital de Ninos Sor Maria Ludovica
Universidad Fernando Pessoa Canarias
St. Anna Children’s Cancer Research Institute
Clinical Immunogenomics Research Consortium Australasia
Modena Health
Corresponding Author(s)
Other Contributor(s)
Abstract
The transcription factor IKAROS plays an important role in lymphocyte development, differentiation, and as a tumor suppressor. To date, >70 IKAROS germ line heterozygous variants have been reported in patients with primary immunodeficiency (PID)/inborn errors of immunity (IEI) and leukemia, and this number continues to grow. Germ line IKAROS loss- and gain-of-function mutations have been linked to immunodeficiency, immune dysregulation, and hematologic malignancies, with a broad spectrum of clinical manifestations. Routine next-generation sequencing approaches in patients with PID/IEI have facilitated the identification of IKAROS variants, including several cases with variants of uncertain significance (VUS). To determine the VUS’ functional behavior, we systematically generated constructs recapitulating those changes and tested IKAROS functions in vitro. We also conducted an in-depth examination of the C-terminal dimerization domain using alanine-scanning mutagenesis to identify amino acids critical for dimerization and other functions. This work provides a comprehensive description of the biologic impact of 81 previously unreported and/or untested IKAROS variants, including 33 patient-detected germ line VUS and 48 laboratory-generated mutations in the dimerization domain. Among them, 15 of the patient-detected variants, primarily mapping to IKAROS DNA-binding or dimerization domains, and at least 21 of the laboratory-generated mutations, impaired IKAROS function and could explain or result in human disease. VUS located in between IKAROS DNA binding and dimerization domains were less likely to be functionally deleterious. Of note, both positive and negative functional data herein generated can be relevant for patients carrying these IKAROS variants, helping to establish a diagnosis and guide treatment decisions.