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GENOMICS AND GENE THERAPY

Implication of PIGA genotype on erythrocytes phenotype in Paroxysmal Nocturnal Hemoglobinuria

Leukemia volume 35, pages 2431–2434 (2021)Cite this article

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Somatic mutations of the X-linked phosphatidylinositol N-acetylglucosaminyltransferase subunit A (PIGA) gene in hematopoietic stem cells are the key molecular events in the pathogenesis of paroxysmal nocturnal hemoglobinuria (PNH) [1]. PIGA mutations lead to impaired biosynthesis of glycosylphosphatidylinositol (GPI)-anchor [2]. Resultant lack of GPI-anchored proteins (GPI-AP), primarily CD55 and CD59, generates clonal selection of PNH clone(s) and increases sensitivity of PNH red cells (RBCs) to complement-mediated hemolysis [3]. Based on immunofluorescent staining, the grade of GPI-APs deficiency on the RBCs can distinguish: PNH type-1 (TI) with normal expression; PNH type-2 (TII) with partial reduction; and PNH type-3 (TIII) with total absence of two common GPI-APs (CD55 and CD59) [4]. Fractions of these RBC types are believed to be a result of the molecular spectrum of PIGA mutations, ranging from missense in various amino acids positions to truncations and locus deletions, which can be concurrent in some patients [5, 6]. Consequently, the presence of TII and TIII cells may indicate the overall severity of PIGA defect, and thus, individual phenotypic subtypes can be defined as predominantly TII or TIII according to the dominant clonal defect [7, 8].

We hypothesize that PIGA genotype may shape the PNH phenotype trajectory with regard to the severity of GPI-AP deficiency and clinical course. For instance, incomplete TII RBCs PNH phenotype should result in slower expansion due to incomplete selection advantage as compared to complete loss of PIGA function indicated by TIII RBCs, typical of hemolytic PNH. However, the process of “GPI-AP painting”, a passive transfer from normal to TIII RBC could result in the presence of pseudo-TII RBCs with an intermediate phenotype [9]. Here, we first used modern deep targeted DNA sequencing to characterize the molecular landscape of patients with PNH and then correlated the type of PIGA mutations with flow-cytometric phenotypes. For improved resolution, our study focused on patients with large clones and hemolytic PNH.

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Fig. 1: PIGA mutational landscape of hemolytic PNH.

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Acknowledgements

We thank Edward P. Evans Foundation, the HENRY & MARILYN TAUB FOUNDATION, grants R01HL118281, R01HL123904, R01HL132071, R35HL135795 (all to J.P.M), AA&MDSIF, VeloSano Pilot Award, and Vera and Joseph Dresner Foundation–MDS (to VV), and Lilli Foundation (to CG). Supported by the American-Italian Cancer Foundation Post-Doctoral Research Fellowship (to CG).

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Authors and Affiliations

  1. Translational Hematology and Oncology Research Department, Taussig Cancer Center, Cleveland Clinic, Cleveland, OH, USA

    Carmelo Gurnari, Simona Pagliuca, Bhumika J. Patel, Hassan Awada, Sunisa Kongkiatkamon, Laila Terkawi, Misam Zawit, Seth Corey, Alan E. Lichtin, Hetty E. Carraway, Valeria Visconte & Jaroslaw P. Maciejewski

  2. Department of Biomedicine and Prevention, PhD in Immunology, Molecular Medicine and Applied Biotechnology, University of Tor Vergata, Rome, Italy

    Carmelo Gurnari

  3. Université de Paris, Paris, France

    Simona Pagliuca

  4. Leukemia Program, Department of Hematology and Medical Oncology, Taussig Cancer Institute, Cleveland Clinic, Cleveland, OH, USA

    Bhumika J. Patel

  5. Torsten-Haferlach Leukemia Diagnostics Foundation, Munich, Germany

    Adam Wahida

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  1. Carmelo Gurnari
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Contributions

CG and JPM generated and conceived the study design, figures, tables and manuscript. JPM and BJP collected and analyzed experimental patients’ data. CG and SP performed statistical analyses. VV, SP, BJP, HA, SK, LT, MZ, SC, AEL, HEC, and AW kindly reviewed the clinical data, took a part in samples selection and helped edited the manuscript. All authors participated in data interpretation and critical review of the final paper and submission.

Corresponding author

Correspondence to Jaroslaw P. Maciejewski.

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The authors declare that they have no conflict of interest.

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Written informed consent was obtained from all patients.

Ethics approval and patients’ consent to participate

Ethics approval and patients’ consent to participate to the study was approved by The Institutional Review Board of the Cleveland Clinic Foundation. All procedures were carried out in accordance with guidelines set forth by the Declaration of Helsinki.

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Gurnari, C., Pagliuca, S., Patel, B.J. et al. Implication of PIGA genotype on erythrocytes phenotype in Paroxysmal Nocturnal Hemoglobinuria. Leukemia 35, 2431–2434 (2021). https://doi.org/10.1038/s41375-020-01113-0

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