Clinical measurement of cellular DNA damage hypersensitivity in patients with DNA repair defects
Abstract Background DNA repair deficiency disorders are rare inherited diseases arising from pathogenic (disease-causing) variants in genes involved in DNA repair. There are no standardized diagnostic assays for the investigation of pathological significance of unknown variants in DNA repair genes....
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-02-01
|
| Series: | Orphanet Journal of Rare Diseases |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13023-022-02199-8 |
| _version_ | 1818281538957082624 |
|---|---|
| author | Ola Hammarsten Anna Lyytikäinen Sofia Thunström Torben Ek Anders Fasth Olov Ekwall Sara Cajander Emilie Wahren Borgström C. I. Edvard Smith Pegah Johansson |
| author_facet | Ola Hammarsten Anna Lyytikäinen Sofia Thunström Torben Ek Anders Fasth Olov Ekwall Sara Cajander Emilie Wahren Borgström C. I. Edvard Smith Pegah Johansson |
| author_sort | Ola Hammarsten |
| collection | DOAJ |
| description | Abstract Background DNA repair deficiency disorders are rare inherited diseases arising from pathogenic (disease-causing) variants in genes involved in DNA repair. There are no standardized diagnostic assays for the investigation of pathological significance of unknown variants in DNA repair genes. We hypothesized that our assays for measuring in vitro patient blood cell hypersensitivity to DNA-damaging agents can be used to establish the pathological significance of unknown variants in DNA repair genes. Six patients with variants in the DNA repair genes PRKDC (two siblings), DCLRE1C (two siblings), NBN, and MSH6 were included. Here, we used the cell division assay (CDA) and the γ-H2AX assay, which were both developed and clinically validated by us, to measure patient cell hypersensitivity in response to ionizing radiation, mitomycin C, cytarabine and doxorubicin. Results Radiation hypersensitivity was detected in the two patients with variants in the PRKDC gene (p < 0.0001 for both at 3.5 Gy), and the two patients with DCLRE1C variants (p < 0.0001 at 3.5 Gy for sibling 1 and p < 0.0001 at 1 Gy for sibling 2). The cells from the patients with the PRKDC variant were also deficient in removing γ-H2AX (p < 0.001). The cells from the patient with variants in the NBN gene were hypersensitive to mitomycin C (p = 0.0008) and deficient in both induction and removal of γ-H2AX in response to radiation. Conclusions The combination of the CDA and the γ-H2AX assay is useful in investigating the significance of unknown variants in some DNA repair genes. |
| first_indexed | 2024-12-13T00:06:43Z |
| format | Article |
| id | doaj.art-75b1d9d18bce47b7b32a1fc70bdab469 |
| institution | Directory Open Access Journal |
| issn | 1750-1172 |
| language | English |
| last_indexed | 2024-12-13T00:06:43Z |
| publishDate | 2022-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Orphanet Journal of Rare Diseases |
| spelling | doaj.art-75b1d9d18bce47b7b32a1fc70bdab4692022-12-22T00:06:11ZengBMCOrphanet Journal of Rare Diseases1750-11722022-02-011711810.1186/s13023-022-02199-8Clinical measurement of cellular DNA damage hypersensitivity in patients with DNA repair defectsOla Hammarsten0Anna Lyytikäinen1Sofia Thunström2Torben Ek3Anders Fasth4Olov Ekwall5Sara Cajander6Emilie Wahren Borgström7C. I. Edvard Smith8Pegah Johansson9Laboratory of Clinical Chemistry, Sahlgrenska University HospitalLaboratory of Clinical Chemistry, Sahlgrenska University HospitalDepartment of Clinical Genetics, Sahlgrenska University HospitalChildren’s Cancer Centre, Queen Silvia Children’s HospitalDepartment of Pediatrics, , Institution of Clinical Sciences, Sahlgrenska Academy, University of GothenburgDepartment of Pediatrics, , Institution of Clinical Sciences, Sahlgrenska Academy, University of GothenburgDepartment of Infectious Diseases, Faculty of Medicine and Health, Örebro UniversityDepartment of Infectious Diseases, The Immunodeficiency Unit, Karolinska University HospitalDepartment of Infectious Diseases, The Immunodeficiency Unit, Karolinska University HospitalLaboratory of Clinical Chemistry, Sahlgrenska University HospitalAbstract Background DNA repair deficiency disorders are rare inherited diseases arising from pathogenic (disease-causing) variants in genes involved in DNA repair. There are no standardized diagnostic assays for the investigation of pathological significance of unknown variants in DNA repair genes. We hypothesized that our assays for measuring in vitro patient blood cell hypersensitivity to DNA-damaging agents can be used to establish the pathological significance of unknown variants in DNA repair genes. Six patients with variants in the DNA repair genes PRKDC (two siblings), DCLRE1C (two siblings), NBN, and MSH6 were included. Here, we used the cell division assay (CDA) and the γ-H2AX assay, which were both developed and clinically validated by us, to measure patient cell hypersensitivity in response to ionizing radiation, mitomycin C, cytarabine and doxorubicin. Results Radiation hypersensitivity was detected in the two patients with variants in the PRKDC gene (p < 0.0001 for both at 3.5 Gy), and the two patients with DCLRE1C variants (p < 0.0001 at 3.5 Gy for sibling 1 and p < 0.0001 at 1 Gy for sibling 2). The cells from the patients with the PRKDC variant were also deficient in removing γ-H2AX (p < 0.001). The cells from the patient with variants in the NBN gene were hypersensitive to mitomycin C (p = 0.0008) and deficient in both induction and removal of γ-H2AX in response to radiation. Conclusions The combination of the CDA and the γ-H2AX assay is useful in investigating the significance of unknown variants in some DNA repair genes.https://doi.org/10.1186/s13023-022-02199-8Cell division assay (CDA)γ-H2AXDNA repair deficiency disordersIonizing radiation sensitivityMitomycin C sensitivityClinical diagnosis |
| spellingShingle | Ola Hammarsten Anna Lyytikäinen Sofia Thunström Torben Ek Anders Fasth Olov Ekwall Sara Cajander Emilie Wahren Borgström C. I. Edvard Smith Pegah Johansson Clinical measurement of cellular DNA damage hypersensitivity in patients with DNA repair defects Orphanet Journal of Rare Diseases Cell division assay (CDA) γ-H2AX DNA repair deficiency disorders Ionizing radiation sensitivity Mitomycin C sensitivity Clinical diagnosis |
| title | Clinical measurement of cellular DNA damage hypersensitivity in patients with DNA repair defects |
| title_full | Clinical measurement of cellular DNA damage hypersensitivity in patients with DNA repair defects |
| title_fullStr | Clinical measurement of cellular DNA damage hypersensitivity in patients with DNA repair defects |
| title_full_unstemmed | Clinical measurement of cellular DNA damage hypersensitivity in patients with DNA repair defects |
| title_short | Clinical measurement of cellular DNA damage hypersensitivity in patients with DNA repair defects |
| title_sort | clinical measurement of cellular dna damage hypersensitivity in patients with dna repair defects |
| topic | Cell division assay (CDA) γ-H2AX DNA repair deficiency disorders Ionizing radiation sensitivity Mitomycin C sensitivity Clinical diagnosis |
| url | https://doi.org/10.1186/s13023-022-02199-8 |
| work_keys_str_mv | AT olahammarsten clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT annalyytikainen clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT sofiathunstrom clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT torbenek clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT andersfasth clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT olovekwall clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT saracajander clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT emiliewahrenborgstrom clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT ciedvardsmith clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects AT pegahjohansson clinicalmeasurementofcellulardnadamagehypersensitivityinpatientswithdnarepairdefects |