scholarly journals Rapid Flow Cytometry–Based Structural Maintenance of Chromosomes 1 (SMC1) Phosphorylation Assay for Identification of Ataxia-Telangiectasia Homozygotes and Heterozygotes

2009 ◽  
Vol 55 (3) ◽  
pp. 463-472 ◽  
Author(s):  
Shareef A Nahas ◽  
Anthony W Butch ◽  
Liutao Du ◽  
Richard A Gatti
Keyword(s):  
Flow Cytometry ◽  
Dna Damage ◽  
Ataxia Telangiectasia ◽  
Mononuclear Cells ◽  
Geometric Mean ◽  
Kinase Assay ◽  
Increased Risk ◽  
Rapid Flow ◽  
Risk Of Cancer ◽  
Structural Maintenance Of Chromosomes

Abstract Background: No rapid reliable method exists for identifying ataxia-telangiectasia (A-T) homozygotes or heterozygotes. Heterozygotes are at an increased risk of cancer and are more sensitive to the effects of ionizing radiation (IR) than the general population. We report a rapid flow cytometry (FC)-based ataxia-telangiectasia mutated (ATM) kinase assay that measures ATM- dependent phosphorylation of structural maintenance of chromosomes 1 (SMC1) following DNA damage (FC-pSMC1 assay). Methods: After optimizing conditions with lymphoblastoid cell lines (LCLs), we studied peripheral blood mononuclear cells (PBMCs) isolated from 16 healthy donors (unknowns), 10 obligate A-T heterozygotes, and 6 unrelated A-T patients. One hour after DNA damage (by either IR or bleomycin), the cells were fixed and incubated with a primary antibody to SMC1pSer966. We analyzed the stained cells by FC to determine the difference in geometric mean fluorescence intensity (ΔGMFI) of untreated and treated cells; this difference was expressed as a percentage of daily experimental controls. Results: The FC-pSMC1 assay reliably distinguished ATM heterozygotes and homozygotes from controls. Average ΔGMFI percentages (SD) of daily controls were, for unknowns, 106.1 (37.6); for A-T heterozygotes, 37.0 (18.7); and for A-T homozygotes; −8.73 (16.2). Values for heterozygotes and homozygotes were significantly different from those of controls (P < 0.0001). Conclusions: The FC-pSMC1 assay shortens the turnaround time for diagnosing A-T homozygotes from approximately 3 months to approximately 3 h. It also identifies A-T heterozygotes and can be used for prenatal counseling or for screening individuals in large study cohorts for potential ATM heterozygosity, which can then be confirmed by sequencing.

High DNA damage levels to predict hematologic toxicity in testicular germ cell tumor (TGCT) patients treated with first-line chemotherapy.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e16055-e16055
Author(s):  
Michal Mego ◽  
Nikola Hapakova ◽  
Zuzana Sestakova ◽  
Andrea Holickova ◽  
Vera Miskovska ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Mononuclear Cells ◽  
Testicular Germ Cell Tumor ◽  
Hematologic Toxicity ◽  
Testicular Germ Cell ◽  
Peripheral Blood Mononuclear ◽  
Damage Level ◽  
Platinum Based Chemotherapy ◽  
Increased Risk

e16055 Background: TGCTs are an excellent example of chemosensitive disease. However, cisplatin-based chemotherapy has significant side effects, including myelosuppression. Previously, we found endogenous DNA damage level in peripheral blood mononuclear cells (PBMCs) to be an independent prognostic marker. We tested the hypothesis that patients with high endogenous DNA damage levels in PBMCs have an increased risk of developing hematologic toxicity. Methods: 120 chemotherapy-naïve patients with TGCTs treated in the National Cancer Institute and the St. Elisabeth Cancer Institute in Bratislava, Slovakia, from 2012 to 2018 were enrolled. All patients received platinum-based chemotherapy with G-CSF support. On the day of starting treatment, we measured the endogenous DNA damage levels in PBMCs using the Comet assay. We used a cut-off level of 5.25, a value previously found to stratify patients based on their prognosis. We monitored hematologic toxicity during the 1st cycle of chemotherapy. The mean and SEM were calculated for all variables. Results: Patients with high DNA damage levels ( > 5.25) had more significant hematologic toxicity with significantly lower nadir white blood cell count (6.0±1.1×109/L vs 9.8±1.0×109/L p = 0.001), absolute neutrophil count (4.1±1.0×109/L vs 7.0±0.9×109/L p = 0.013) and absolute lymphocyte count (ALC, 1.1±0.1×109/L vs 1.5±0.1×109/L p < 0.001). ALCs on day 0 (1.5±0.1×109/L vs 1.8±0.1×109/L p = 0.005) and day 22 (2.0±0.1×109/L vs 2.4±0.1×109/L p = 0.046) were also significantly lower in patients with high DNA damage levels. There were no significant differences in hemoglobin levels or platelet counts between the two groups. Neutrophil to lymphocyte ratio and systemic immune-inflammation index were lower at nadir in patients with high DNA damage levels, however, these differences were not statistically significant (p = 0.08 and p = 0.10, respectively). Conclusions: This study shows that higher endogenous DNA damage levels correlate with an increased risk of hematologic toxicity. The Comet assay data can be used to select patients for closer monitoring due to a higher risk of acute chemotherapy-related complications.

Ataxia-Telangiectasia genes and breast cancer risk in a French family study

2005 ◽  
Vol 72 (S1) ◽  
pp. 73-80 ◽  
Author(s):  
Nadine Andrieu ◽  
Eve Cavaciuti ◽  
Anthony Laugé ◽  
Katia Ossian ◽  
Nicolas Janin ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Ataxia Telangiectasia ◽  
Screening Program ◽  
Neuronal Degeneration ◽  
Carrier Status ◽  
X Rays ◽  
Increased Risk ◽  
Exogenous Factors ◽  
Atm Protein ◽  
Risk Of Cancer

Ataxia-telangiectasia (AT) is a rare autosomal recessive early childhood disorder, characterized by progressive neuronal degeneration, immunological deficiency, radiosensitivity and an increased risk of cancer caused in most cases by mutations in the AT-mutated gene (ATM). Epidemiological studies on AT families have shown that AT heterozygous women have an increased risk of developing breast cancer (BC). The ATM protein plays a central role in the recognition and repair of DNA double-strand breaks and the subsequent activation of cell-cycle checkpoints. Whilst AT is a rare disease, 0·5–1% of the general population are estimated to be AT mutation carriers, thus any increases in the risks of cancer associated with ATM carrier status are of public health relevance. The main results of our published studies on the risk of BC in 34 French AT families according to heterozygote status, type of ATM mutation and exogenous factors are summarized here. The risk of BC was higher in ATM heterozygous (HetATM) women and did not differ significantly according to the type of ATM mutation (missense vs truncating) carried by the AT family members but appeared associated with the position of some truncating mutations in certain binding domains of the ATM protein. The effect of exogenous factors, such as reproductive life factors and exposure to ionizing radiation, on the risk of BC according to ATM heterozygote status was assessed. There was no evidence for interaction (except for age at first full-term pregnancy). These findings does not appear to justify a separate screening program from that already available to other women with a first-degree relative affected by BC, as their risks have similar amplitude. Chest X-rays did not appear to be a risk factor for BC in our study population. More powerful studies, using data sets pooled from international sources are being set up to confirm these observations.

scholarly journals Atypical Ataxia Presentation in Variant Ataxia Telangiectasia: Iranian Case-Series and Review of the Literature

2022 ◽  
Vol 12 ◽  
Author(s):  
Tannaz Moeini Shad ◽  
Reza Yazdani ◽  
Parisa Amirifar ◽  
Samaneh Delavari ◽  
Marzieh Heidarzadeh Arani ◽  
...  
Keyword(s):  
T Cells ◽  
Ataxia Telangiectasia ◽  
Late Onset ◽  
Case Series ◽  
Atypical Presentation ◽  
Missense Mutations ◽  
Multisystem Disorder ◽  
Atypical Form ◽  
Increased Risk ◽  
Risk Of Cancer

Ataxia-telangiectasia (AT) is a rare autosomal recessive neurodegenerative multisystem disorder. A minority of AT patients can present late-onset atypical presentations due to unknown mechanisms. The demographic, clinical, immunological and genetic data were collected by direct interview and examining the Iranian AT patients with late-onset manifestations. We also conducted a systematic literature review for reported atypical AT patients. We identified three Iranian AT patients (3/249, 1.2% of total registry) with later age at ataxia onset and slower neurologic progression despite elevated alpha-fetoprotein levels, history of respiratory infections, and immunological features of the syndrome. Of note, all patients developed autoimmunity in which a decrease of naïve T cells and regulatory T cells were observed. The literature searches also summarized data from 73 variant AT patients with atypical presentation indicating biallelic mild mutations mainly lead to an atypical phenotype with an increased risk of cancer. Variant AT patients present with milder phenotype or atypical form of classical symptoms causing under- or mis- diagnosis. Although missense mutations are more frequent, an atypical presentation can be associated with deleterious mutations due to unknown modifying factors.

scholarly journals A rapid flow cytometry test based on histone H2AX phosphorylation for the sensitive and specific diagnosis of ataxia telangiectasia

2008 ◽  
Vol 73A (6) ◽  
pp. 508-516 ◽  
Author(s):  
Paola Porcedda ◽  
Valentina Turinetto ◽  
Alfredo Brusco ◽  
Simona Cavalieri ◽  
Erica Lantelme ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Ataxia Telangiectasia ◽  
H2ax Phosphorylation ◽  
Specific Diagnosis ◽  
Histone H2ax ◽  
Rapid Flow ◽  
Histone H2ax Phosphorylation

Incidence, Presentation, and Prognosis of Malignancies in Ataxia-Telangiectasia: A Report From the French National Registry of Primary Immune Deficiencies

2015 ◽  
Vol 33 (2) ◽  
pp. 202-208 ◽  
Author(s):  
Felipe Suarez ◽  
Nizar Mahlaoui ◽  
Danielle Canioni ◽  
Chantal Andriamanga ◽  
Catherine Dubois d'Enghien ◽  
...  
Keyword(s):  
T Cell ◽  
B Cell ◽  
Hodgkin Lymphoma ◽  
Ataxia Telangiectasia ◽  
Lymphoblastic Leukemia ◽  
Response To Treatment ◽  
Acute Leukemias ◽  
Prolymphocytic Leukemia ◽  
Increased Risk ◽  
Risk Of Cancer

Purpose Biallelic mutations in ATM cause ataxia-telangiectasia (AT), a rare inherited disease with a high incidence of cancer. Precise estimates of the risk, presentation, and outcomes of cancer in patients with AT need to be addressed in large series. Patients and Methods In this large retrospective cohort, 69 patients with cancers (24.5%) were identified among 279 patients with AT. Centralized review was performed on 60% of the lymphomas. Incidence rates were compared with the French population, and risk factors were analyzed. Results Eight patients developed acute leukemias (including four T-cell acute lymphoblastic leukemias), 12 developed Hodgkin lymphoma (HL), 38 developed non-Hodgkin lymphoma (NHL), three developed T-cell prolymphocytic leukemia (T-PLL), and eight developed carcinoma at a median age of 8.3, 10.6, 9.7, 24.2, and 31.4 years, respectively (P < .001). The majority of NHLs were aggressive B-cell NHL. Epstein-Barr virus was associated with all of the HLs and 50% of the NHLs. Overall survival was shorter in patients with AT who developed cancer compared with those who did not develop cancer (15 v 24 years, respectively; P < .001). Survival was improved in patients who achieved a major response to treatment (3.46 v 0.87 years for major v minor responses, respectively; P = .011). Immunodeficiency was associated with increased risk of cancer. ATM mutation type was associated with a difference in survival in the entire cohort but not with cancer incidence or cancer survival. Conclusion B-cell NHL, HL, and acute lymphoblastic leukemia occur at a high rate and earlier age than carcinomas in AT. T-PLLs are rarer than initially reported. Prognosis is poor, but patients may benefit from treatment with an improved survival.

scholarly journals Cancer in Children With Fanconi Anemia and Ataxia-Telangiectasia—A Nationwide Register-Based Cohort Study in Germany

2021 ◽  
Author(s):  
Christina M. Dutzmann ◽  
Claudia Spix ◽  
Isabell Popp ◽  
Melanie Kaiser ◽  
Friederike Erdmann ◽  
...  
Keyword(s):  
Dna Repair ◽  
Childhood Cancer ◽  
Hodgkin Lymphoma ◽  
Fanconi Anemia ◽  
Ataxia Telangiectasia ◽  
Population Based ◽  
Cancer Data ◽  
Increased Risk ◽  
Risk Of Cancer ◽  
Cancer In Children

PURPOSE Fanconi anemia (FA) and ataxia-telangiectasia (AT) are rare inherited syndromes characterized by abnormal DNA damage response and caused by pathogenic variants in key DNA repair proteins that are also relevant in the pathogenesis of breast cancer and other cancer types. The risk of cancer in children with these diseases is poorly understood and has never been assessed in a population-based cohort before. METHODS We identified 421 patients with FA and 160 patients with AT diagnosed between 1973 and 2020 through German DNA repair disorder reference laboratories. We linked patients' laboratory data with childhood cancer data from the German Childhood Cancer Registry. RESULTS Among 421 patients with FA, we observed 33 cases of childhood cancer (15 cases of myelodysplastic syndrome; seven cases of acute myeloid leukemia; two cases of lymphoma, carcinoma, medulloblastoma, and nephroblastoma, respectively; and one case of rhabdomyosarcoma, acute lymphoblastic leukemia, and glioma, respectively) versus 0.74 expected (on the basis of population-based incidence rates in Germany). This corresponds to a 39-fold increased risk (standardized incidence ratio [SIR] = 39; 95% CI, 26 to 56). For all FA subgroups combined, the cancer-specific SIR for myeloid neoplasms was 445 (95% CI, 272 to 687). Among the 160 patients with AT, we observed 19 cases of childhood cancer (15 cases of lymphoma, three cases of leukemia, and one case of medulloblastoma) versus 0.32 expected. This corresponds to a 56-fold increased risk (SIR = 56; 95% CI, 33 to 88). The cancer-specific SIR for Hodgkin lymphoma was 215 (95% CI, 58 to 549) and for non-Hodgkin lymphoma 470 (95% CI, 225 to 865). CONCLUSION Approximately 11% of patients with FA and 14% of patients with AT develop cancer by age 18 years.

scholarly journals Immunoassay to Measure Ataxia-Telangiectasia Mutated Protein in Cellular Lysates

2004 ◽  
Vol 50 (12) ◽  
pp. 2302-2308 ◽  
Author(s):  
Anthony W Butch ◽  
Helen H Chun ◽  
Shareef A Nahas ◽  
Richard A Gatti
Keyword(s):  
Ataxia Telangiectasia ◽  
Mononuclear Cells ◽  
Lymphoblastoid Cell Lines ◽  
Ataxia Telangiectasia Mutated ◽  
Peripheral Blood Mononuclear ◽  
Neurologic Disorder ◽  
Increased Risk ◽  
Atm Gene ◽  
Atm Protein ◽  
Ataxia Telangiectasia Mutated Protein

Abstract Background: Ataxia-telangiectasia (A-T) is a neurologic disorder caused by mutations in the ataxia-telangiectasia mutated (ATM) gene. A clinical diagnosis of A-T is confirmed by radiosensitivity testing and immunoblotting for ATM protein. Because both of these tests have long turnaround times (≥3 months), we developed a rapid immunoassay to measure ATM protein and determined its sensitivity and specificity for diagnosing A-T. Methods: Recombinant ATM protein was used for standardization. Lysates of lymphoblastoid cell lines (LCLs) and peripheral blood mononuclear cells (PBMCs) from A-T patients, controls, and A-T heterozygotes were tested for ATM protein by immunoassay. Results: Between-run imprecision (CV) was ≤13%. Nuclear lysates from control LCLs and PBMCs had ATM protein concentrations of 49–610 μg/L and 48–943 μg/L, respectively. ATM protein was not detectable in LCL nuclear lysates from 18 of 21 A-T patients. The three remaining A-T patients had trace amounts of ATM protein, which was confirmed on immuoblots. ATM protein was also detectable in whole-cell lysates from 4 × 106 cells at concentrations of 64–463 μg/L and 42–444 μg/L for control LCLs and PBMCs, respectively. A-T heterozygotes had ATM protein concentrations of 52–98 μg/L. ATM protein was stable in PBMCs stored for 1 month at −70 °C, but rapidly decreased after 1 day in unprocessed blood. Conclusions: This ATM protein immunoassay can be used to confirm a diagnosis of A-T in 2 days on small numbers of PBMCs and can potentially identify A-T carriers and individuals at increased risk for cancer.

scholarly journals ERS statement on the multidisciplinary respiratory management of ataxia telangiectasia

2015 ◽  
Vol 24 (138) ◽  
pp. 565-581 ◽  
Author(s):  
Jayesh M. Bhatt ◽  
Andrew Bush ◽  
Marjo van Gerven ◽  
Andreea Nissenkorn ◽  
Michael Renke ◽  
...  
Keyword(s):  
Lung Disease ◽  
Ataxia Telangiectasia ◽  
Causes Of Death ◽  
Respiratory Infections ◽  
Multisystem Disease ◽  
Respiratory Management ◽  
Increased Risk ◽  
Proactive Monitoring ◽  
Risk Of Cancer ◽  
Multidisciplinary Clinics

Ataxia telangiectasia (A-T) is a rare, progressive, multisystem disease that has a large number of complex and diverse manifestations which vary with age. Patients with A-T die prematurely with the leading causes of death being respiratory diseases and cancer. Respiratory manifestations include immune dysfunction leading to recurrent upper and lower respiratory infections; aspiration resulting from dysfunctional swallowing due to neurodegenerative deficits; inefficient cough; and interstitial lung disease/pulmonary fibrosis. Malnutrition is a significant comorbidity. The increased radiosensitivity and increased risk of cancer should be borne in mind when requesting radiological investigations. Aggressive proactive monitoring and treatment of these various aspects of lung disease under multidisciplinary expertise in the experience of national multidisciplinary clinics internationally forms the basis of this statement on the management of lung disease in A-T. Neurological management is outwith the scope of this document.

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