Andrei Tchirkov

Publications (15)41.74 Total impact

• Article: NK cell immunotherapy for high-risk neuroblastoma relapse after haploidentical HSCT.

  Justyna Kanold, Catherine Paillard, Andrei Tchirkov, Peter Lang, Antony Kelly, Pascale Halle, Florentina Isfan, Etienne Merlin, Aurelien Marabelle, Emmanuelle Rochette, François Deméocq
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  ABSTRACT: Little is known on strategies to prevent or to treat relapses occurring after haploidentical stem cell transplantation (haplo-HSCT) performed for the high-risk neuroblastoma (NB). We describe a 6-year-old male with refractory NB who relapsed 22 months after haplo-HSCT. A complete remission was obtained with a combination of immuno-chemotherapy based on donor NK cells transplants, IL2 infusions and temozolomide/topotecan. This case is an incentive to explore both the immediate therapeutic effect of haplo-graft provided via haplo-NK cells and the immunogenic platform that haplo-HSCT offers for future treatment. Our post-relapse strategy shows that chemo- and bio-treatment should be viewed as complementary therapeutic options.
  Pediatric Blood & Cancer 12/2011; 59(4):739-42. · 1.89 Impact Factor
• Article: CD34+ immunoselection of autologous grafts for the treatment of high-risk neuroblastoma.

  Aurélien Marabelle, Etienne Merlin, Pascale Halle, Catherine Paillard, Marc Berger, Andrei Tchirkov, Raphaël Rousseau, Guy Leverger, Christophe Piguet, Jean-Louis Stephan, François Demeocq, Justyna Kanold
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  ABSTRACT: Graft contamination has been blamed for causing relapse in children with high-risk neuroblastoma (HRNB) after autologous hematopoietic stem cell transplantation (HSCT). We report the long-term results of hematopoietic reconstitution, post-transplant complications, and clinical outcome of 44 children with HRNB treated with busulfan/melphalan high-dose chemotherapy followed by transplantation of purged CD34+ immunoselected autologous peripheral HSCT. Minimal residual disease (MRD) of grafts was evaluated by anti-GD2 immunofluorescence or tyrosine hydroxylase reverse transcriptase-polymerase chain reaction (RT-PCR). Contaminating neuroblasts were found in 19/38 grafts (50%) before CD34+ positive selection, and none after (technique sensitivity of one cell in 10(5)). A median of 6.5 × 10(6) CD34+ cells/kg (range 0.8-23.7) were transplanted with only 2% of TRM. Neutrophils and platelet recovery occurred within a median of 12 days (range 9-47) and 44 days (range 12-259), respectively, without any secondary graft failure. Twenty-three percents of patients experienced a sepsis (10/44) and 14% a pyelonephritis (6/44). Recurrence of varicella zoster virus occurred in 21% of patients (9/44). Negative RT-PCR MRD within the leukapheresis product and cis-retinoic acid therapy were significantly and independently associated to a better survival (P < 0.05). Overall and event-free survivals at 5 years post-transplant were at 59.3% and 48.3% respectively. Besides high rates of manageable infections due to late immune recovery, transplantation with CD34+ immunoselected grafts in HRNB children was feasible and did not affect long-term hematopoiesis.
  Pediatric Blood & Cancer 11/2010; 56(1):134-42. · 1.89 Impact Factor
• Article: Increased expression of the oncogenic KLF6-SV1 transcript in human glioblastoma.

  Andrei Tchirkov, Vincent Sapin, Geoffroy Marceau, Emmanuel Chautard, Goutham Narla, Lauren Veronese, Scott Friedman, Toufic Khalil, Philippe Vago, Jean-Louis Kemeny, Pierre Verrelle
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  ABSTRACT: Gliomas constitute the vast majority of primary central nervous system tumors in adults. Glioblastoma multiforme (GBM) is the most aggressive form of these primary brain tumors. There is a need to define diagnostic and prognostic markers that may help to distinguish GBM from non-GBM tumors. The Krüppel-like factor 6 (KLF6) gene has recently emerged as a promising candidate. The goal of our study was to determine if there is a link between KLF6 splice variants expression and different grades of gliomas. Fifty-three primary gliomas tumor samples were analyzed using quantitative real-time PCR for the total KLF6, wild-type and alternatively spliced (SV1) KLF6 mRNA. Compared to the non-GBM group, the GBM group had a 2.2-fold increase in the mean level of total KLF6 mRNA expression. GBM showed a 2.1-fold increase in the KLF6 splicing ratio. In addition, KLF6-SV1 mRNA expression levels were also 2.2-fold higher in the GBM group, suggesting that the increase in the KLF6 splicing ratio was due to increased expression of the KLF6-SV1 oncogenic splice variant. Our study demonstrates that quantification of total and spliced forms of KLF6 may provide a new and useful supplementary molecular tool for grading glioma.
  Clinical Chemistry and Laboratory Medicine 08/2010; 48(8):1167-70. · 2.15 Impact Factor
• Article: Prenatal detection of cryptic rearrangements by multiplex ligation probe amplification in fetuses with ultrasound abnormalities.

  Carole Goumy, Laetitia Gouas, Céline Pebrel-Richard, Lauren Véronèse, Eleonore Eymard-Pierre, Anne Debost-Legrand, Khadidja Haoud, Andrei Tchirkov, Philippe Vago
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  ABSTRACT: Congenital malformations are a major cause of morbidity and mortality in newborn infants, and genomic imbalances are a significant component of their etiology. The aim of this study was to evaluate the ability of prenatal multiplex ligation probe amplification screening to detect cryptic chromosomal imbalances in fetuses with ultrasound abnormalities of unknown etiology. Multiplex ligation probe amplification was performed with three separate sets of probes: two for subtelomeric regions and one for mental retardation syndrome loci. Sixty-one fetuses with significant ultrasound anomalies and normal karyotype at a minimum of 400-band resolution were tested between January 2007 and January 2009. We identified four unbalanced rearrangements: one del 18pter/amp 5pter, one del 9pter, one 15q11q13 microdeletion, and one 22q11 microdeletion with atypical presentation. After genetic counseling, two of the pregnancies were terminated. Multiplex ligation probe amplification analysis was able to identify clinically significant rearrangements in 6.5% of fetuses with prenatally identified sonographic abnormalities. This prospective study highlights that multiplex ligation probe amplification screening of fetuses with ultrasound abnormalities in the prenatal period is technically feasible and relevant for diagnosis and prognosis.
  Genetics in medicine: official journal of the American College of Medical Genetics 06/2010; 12(6):376-80. · 3.92 Impact Factor
• Article: Intraoperative imprint cytology examination of sentinel lymph nodes after neoadjuvant chemotherapy in breast cancer patients.

  P Gimbergues, M M Dauplat, X Durando, C Abrial, G Le Bouedec, M A Mouret-Reynier, F Cachin, F Kwiatkowski, Andrei Tchirkov, J Dauplat, F Penault-Llorca
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  ABSTRACT: Intraoperative imprint cytology (IC) is one of several accurate, proven methods to detect tumor cells in sentinel lymph nodes (SLN) from patients with operable breast cancer. In patients treated with neoadjuvant chemotherapy (NAC), studies have demonstrated the feasibility and accuracy of SLN biopsy procedure. We evaluated the validity of IC for SLN testing in patients after NAC. Patients with infiltrating breast carcinoma receiving NAC (n = 132) were studied prospectively. At surgery, SLN biopsy followed by axillary lymph node dissection was performed. SLN were evaluated using IC in 80 of 132 patients (60%). The results of IC in the adjuvant setting (100 patients) were used for comparison. SLN metastases were correctly identified using IC in 58 of 80 (72%) patients. False negative results were observed in 21 patients. The sensitivity of IC testing was 38.2% and specificity 97.8%. The positive and negative predictive values (PPV and NPV) were 92.9% and 68.2%, respectively. In univariate analysis and multivariate logistic regression analysis, patients with micrometastases or isolated tumor cells in SLN have 2.3 times higher risk of a false negative IC result than patients with macrometastases in SLN (P = .00021; relative risk [RR] = 2.3; 95% confidence interval, 1.37-3.85). The non-NAC group, which contained fewer micrometastatic cases, showed better sensitivity (47.4%) and NPV (88.9%). NAC does not seem to influence the accuracy and sensitivity of IC. Variations in sensitivity are related to the proportion of cases with micrometastases and ITC, as it was also shown in chemonaive patients.
  Annals of Surgical Oncology 02/2010; 17(8):2132-7. · 4.17 Impact Factor
• Article: Retinoid pathway and congenital diaphragmatic hernia: hypothesis from the analysis of chromosomal abnormalities.

  Carole Goumy, Laetitia Gouas, Geoffroy Marceau, Karen Coste, Lauren Veronese, Denis Gallot, Vincent Sapin, Philippe Vago, Andrei Tchirkov
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  ABSTRACT: Although there is strong evidence implicating genetic factors in congenital diaphragmatic hernia (CDH) pathogenesis, few causal genes have been identified. Many studies suggest that early disruption of the retinoid signaling pathway during gestation may contribute to CDH etiology. Chromosome abnormalities are detected in 10-20% of CDH cases. Chromosomal regions that are involved in balanced translocations or are recurrently deleted or duplicated in patients with CDH are of particular interest to researchers because they are more likely to harbor genes that cause or predispose one to the development of CDH. The aim of this review was to select chromosome loci which have been shown to be associated with CDH and to investigate if these loci contain candidate genes involved in the retinoic signaling pathway. We have re-examined the known CDH-critical chromosomal loci and searched in available databases, such as the UCSC Genome Browser and OMIM, to see whether candidate genes related to the retinoid pathway were present within these loci. Twelve retinoid-related genes have been proposed as potential candidates. Among them, COUP-TFII, FOG2 and GATA4 have already been well studied, especially in animal models. We propose other candidates such as STRA6, LRAT, CRBP1, CRBP2 and CRABP1 are directly implicated in retinoic acid metabolism. The identification of CDH-related genes and pathways affecting a normal diaphragm will contribute to the understanding of the pathophysiology of this severe embryopathy and might help to facilitate prenatal management and devise more individual treatment strategies. Further studies are necessary to screen large cohorts of patients with CDH for microimbalances or de novo mutations in these candidate genes. Moreover, functional analyses are needed to establish their exact role in CDH etiology.
  Fetal Diagnosis and Therapy 01/2010; 28(3):129-39. · 1.05 Impact Factor
• Source

  Available from: Sandrine Hayette

  Article: Detection of twelve nucleotides insertion in the BCR-ABL kinase domain in an imatinib-resistant but dasatinib-sensitive patient with bi-phenotypic acute leukemia.

  Sandrine Hayette, Kaddour Chabane, Andrei Tchirkov, Marc G Berger, Franck E Nicolini, Olivier Tournilhac
  Haematologica 10/2009; 94(9):1324-6. · 6.42 Impact Factor
• Article: Strong correlation between VEGF and MCL-1 mRNA expression levels in B-cell chronic lymphocytic leukemia.

  Lauren Véronèse, Olivier Tournilhac, Pierre Verrelle, Frédéric Davi, Guillaume Dighiero, Emmanuel Chautard, Richard Veyrat-Masson, Fabrice Kwiatkowski, Carole Goumy, Laetitia Gouas, Jacques-Olivier Bay, Philippe Vago, Andrei Tchirkov
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  ABSTRACT: Expression of the anti-apoptotic myeloid cell leukemia-1 (MCL-1) gene is a novel prognostic factor in B-cell chronic lymphocytic leukemia (B-CLL). Vascular and endothelial growth factor (VEGF) and interleukin-6 (IL-6) are able to upregulate MCL-1 via autocrine signaling loops. In 88 B-CLL patients, we found a strong correlation of MCL-1 gene expression with VEGF (P<10(-7)) but not with IL-6 mRNA levels. VEGF but not IL-6 expression influenced patient prognosis. VEGF may be a positive autocrine in vivo regulator of MCL-1 in B-CLL. Inhibition of VEGF and its signaling may prove to be useful in the treatment of B-CLL patients.
  Leukemia research 05/2009; 33(12):1623-6. · 2.36 Impact Factor
• Article: Cytogenetic characterization of seven human cancer cell lines by combining G- and R-banding, M-FISH, CGH and chromosome- and locus-specific FISH.

  Michèle Cottier, Andrei Tchirkov, Bernard Perissel, Michel Giollant, Lydia Campos, Philippe Vago
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  ABSTRACT: Leukemia cell lines K562, KG1a, U937, HL60, Jurkat and solid tumor cell lines A549 and M4Beu are widely used in studies of cell cycle, apoptosis and adhesion mechanisms in cancer cells. Although the K562 and U937 cell lines were previously subjected to a detailed cytogenetic characterization, only a few molecular cytogenetic investigations have been performed on the other five cell lines. We combined several molecular cytogenetic techniques, such as fluorescence in situ hybridization (FISH), multicolor FISH (M-FISH), and comparative genomic hybridization (CGH) to demonstrate the precise genetic aberrations in tumor genomes of these seven cell lines. This information may be useful for multiple studies on these cell lines, providing a genetic basis for the interpretations of experimental findings.
  International Journal of Molecular Medicine 11/2004; 14(4):483-95. · 1.98 Impact Factor
• Article: Significance of molecular quantification of minimal residual disease in metastatic neuroblastoma.

  Andrei Tchirkov, Catherine Paillard, Pascale Halle, Frédéric Bernard, Pierre Bordigoni, Philippe Vago, François Deméocq, Justyna Kanold
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  ABSTRACT: Molecular detection of tumor cells is the most sensitive approach to study residual disease in bone marrow (BM), peripheral blood (PB), and peripheral blood stem cell (PBSC) autografts from children with metastatic neuroblastoma (NB). We have developed a real-time PCR assay that allows the quantification of tyrosine hydroxylase (TH) mRNA, a tissue-specific marker of neuroblasts. We investigated a total of 165 BM, PB, and PBSC samples from 30 children over 1 year of age with stage IV NB and correlated the findings with disease status and patient survival. The levels of TH mRNA agreed well with clinical status and were significantly different across the groups that included samples obtained from patients at diagnosis, after three cycles of chemotherapy, in complete or very good partial remission and at relapse. We found that overall survival was significantly worse for patients with >1000 TH copies in BM after initial chemotherapy (p=0.0075). In 57% of cases, autologous PBSC harvests were found to be contaminated by neuroblasts, the level of TH >500 copies being associated with a decreased survival (p=0.003). In addition, molecular quantification enabled an estimation of tumor depletion in contaminated autografts using CD34 selection (median, 3 logs). In conclusion, quantification of minimal residual disease in metastatic NB using real-time RT-PCR for TH mRNA appears to be of potential clinical value. Further studies are needed to ascertain prognostic implications of molecular analysis of residual disease.
  Journal of Hematotherapy &amp Stem Cell Research 08/2003; 12(4):435-42.
• Article: Simulation of neutron interactions at the single-cell level.

  Jean-Pierre Alard, Véronique Bodez, Andrei Tchirkov, Marie-Laure Nénot, Jack Arnold, Sylvain Crespin, Maryse Rapp, Pierre Verrelle, Claude Dionet
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  ABSTRACT: We recently reported that the exposure of cancer cells to 14 MeV neutrons at a very low dose rate (0.8 mGy min(-1)) produced a marked increase in cell killing at 5 cGy, followed by a plateau in survival and chromosomal damage. Simulation of the energy deposition events in irradiated cells may help to explain these unusual cell responses. We describe here a Monte Carlo simulation code, Energy Deposition in Cells Irradiated by Neutrons (EDCIN). The procedure considered the experimental setup and a hemispheric cell model. The simulation data fitted the dosimetric measurements performed using tissue-equivalent ionization chambers, Geiger-Müller counters, fission chambers, and silicon diodes. The simulation showed that 80% of the energy deposited in a single cell came from the interactions of neutrons outside the cell and only 20% came from neutron interactions inside the cell. Thus the "external" interactions that result in the production of recoil protons and secondary electrons may induce most of the biological damage, which may be repaired efficiently at low dose rate. The repair process may be triggered from a threshold level of damage, which would explain an initial increase cell death due to unrepaired sublethal damage, and then may compensate for induced damage, resulting in the plateaus.
  Radiation Research 12/2002; 158(5):650-6. · 2.68 Impact Factor
• Article: Interleukin-6-producing cells in a human glioblastoma cell line are not affected by ionizing radiation.

  Jean-Jacques Dubost, Christine Rolhion, Andrei Tchirkov, Suzanne Bertrand, Jacques Chassagne, Annie Dosgilbert, Pierre Verrelle
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  ABSTRACT: We investigated the production of interleukin 6 (IL-6) by a radioresistant human glioblastoma cell line G5 after single radiation events of 3, 6 and 9 Gy. The total cell number and IL-6 concentration in culture supernatant were assessed 24-96 h after irradiation. The radiation impeded or stopped G5 cell growth in a dose-dependent manner, but unexpectedly did not affect the IL-6 concentration in cell culture media that increased in the same range as in non-irradiated cultures. Furthermore, using flow cytometry, we found that the IL-6 positive cells expansion was unaffected by radiation. These findings suggested that this small (about 1%) fraction of G5 cells, constitutively producing IL-6, is highly radioresistant.
  Journal of Neuro-Oncology 02/2002; 56(1):29-34. · 3.21 Impact Factor
• Article: Interphase cytogenetics and competitive RT‐PCR for residual disease monitoring in patients with chronic myeloid leukaemia during interferon‐α therapy

  Andrei Tchirkov, Michel Giollant, FranÇoise Tavernier, Georges BrianÇon, Olivier Tournilhac, Fabrice Kwiatkowski, Pierre Philippe, Bachra Choufi, FranÇois DemÉocq, Philippe Travade, Paul Malet
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  ABSTRACT: There is a need for fast and sensitive methods to evaluate the response of patients with chronic myeloid leukaemia (CML) to interferon-α (IFN-α) therapy to complement cytogenetic analysis of Philadelphia (Ph) chromosome-positive metaphases. We have used interphase FISH (fluorescence in situ hybridization) and competitive RT-PCR (reverse transcriptase-polymerase chain reaction) techniques for detection of BCR-ABL-positive cells to measure suppression of leukaemic clone in a series of 51 follow-up samples from 24 CML patients undergoing IFN-α treatment. Interphase FISH analysis of the malignant clone in bone marrow using BCR and ABL probes was found to be highly correlated to conventional G-banding metaphase examination (r = 0.98). RT-PCR quantification of BCR-ABL mRNA transcripts in blood also showed a high degree of concordance with the proportion of Ph-positive metaphases (r = 0.93). In addition, the degree of cytogenetic response did not influence the equivalence between karyotype analysis and molecular methods. We concluded that interphase FISH and competitive RT-PCR provide reliable information on residual tumour burden and response to IFN-α in CML patients. These molecular methods may significantly improve the efficiency of residual disease monitoring during IFN-α therapy of CML.
  British Journal of Haematology 12/2001; 101(3):552 - 557. · 4.94 Impact Factor
• Source

  Available from: riedlab.nci.nih.gov

  Article: Characterization of double minute chromosomes’ DNA content in a human high grade astrocytoma cell line by using comparative genomic hybridization and fluorescence in situ hybridization

  Michel Giollant, Suzanne Bertrand, Pierre Verrelle, Andrei Tchirkov, Stanislas du Manoir, Thomas Ried, Françoise Mornex, Jean-François Doré, Thomas Cremer, P. Malet, A Tchirkov
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  ABSTRACT: The presence of double minute chromosomes (dmin) in cancer cells is known to be correlated with gene amplifications. In human high grade astrocytomas or glioblastomas, about 50% of cytogenetically characterized cases display dmin. G5 is a cell line which has been established from a human glioblastoma containing multiple dmin. In order to identify the DNA content of these dmin, three techniques were successively used: conventional cytogenetic analysis, comparative genomic hybridization (CGH), and fluorescent in situ hybridization (FISH). The karyotype of G5 cells showed numerical chromosome changes (hypertriploidy), several marker chromosomes, and multiple dmin. CGH experiments detected two strong DNA amplification areas located in 9p21-22 and 9p24, as well as an underrepresentation of chromosomes 6, 10, 11, 13, 14, and 18q. By using FISH with a chromosome 9-specific painting probe to metaphase chromosomes of the G5 cell line, dmin were shown to contain DNA sequences originating from chromosome 9. This study demonstrates the usefulness of a combination of classical karyotyping, CGH, and FISH to identify the chromosomal origin of amplified DNA sequences in dmin.
  Human Genetics 06/1996; 98(3):265-270. · 5.07 Impact Factor
• Article: Major molecular response to imatinib in a patient with chronic myeloid leukemia expressing a novel form of e8a2 BCR-ABL transcript.

  Andrei Tchirkov, Jean-Louis Couderc, Bernard Périssel, Carole Goumy, Aline Regnier, Nancy Uhrhammer, Pierre Verrelle, M. Berger
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  ABSTRACT: N.A