Ex vivo gamma H2AX radiation sensitivity assay in prostate cancer: Inter-patient and intra-patient heterogeneity

Chiara De-Colle; Ala Yaromina; Joerg Hennenlotter; Howard Thames; Arndt-Christian Mueller; Tim Neumann; Arnulf Stenzl; Marcus Scharpf; Falko Fend; Umberto Ricardi; Michael Baumann; Daniel Zips; Apostolos Menegakis

doi:10.1016/j.radonc.2017.08.020

Ex vivo gamma H2AX radiation sensitivity assay in prostate cancer: Inter-patient and intra-patient heterogeneity

Chiara De-Colle, Ala Yaromina, Joerg Hennenlotter, Howard Thames, Arndt-Christian Mueller, Tim Neumann, Arnulf Stenzl, Marcus Scharpf, Falko Fend, Umberto Ricardi, Michael Baumann, Daniel Zips, Apostolos Menegakis^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Introduction: The aim of the study is to assess inter-patient and intralpatient heterogeneity in tumour cell radiosensitivity using the ex vivo gamma H2AX assay in prostate cancer specimens.

Methods: Excised specimens from untreated prostate cancer patients were cultivated 24 h in media, irradiated ex vivo and fixed after 24 h. Residual gamma H2AX foci were counted and the slope of the dose response was calculated. Intra-patient heterogeneity was studied from three to seven different biopsies.

Results: In pathology-confirmed tumour samples from 21 patients the slope of residual gamma H2AX foci and radiation dose showed a substantial heterogeneity ranging from 0.82 to 3.17 foci/Gy. No correlation was observed between the slope values and the Gleason score (p = 0.37), prostate specific antigen (p = 0.48) and tumour stage (p = 0.89). ANOVA indicated that only in 1 out of 9 patients, biopsies from different tumour locations yielded statistically significant differences. Variance component analysis indicated higher inter-patient than intra-patient variability. Bootstrap simulation study demonstrated that one biopsy is sufficient to estimate the mean value of residual gamma H2AX per dose level and account for intra-patient heterogeneity.

Conclusions: In prostate cancer inter-patient heterogeneity in tumour cell radiation sensitivity is pronounced and higher than intra-patient heterogeneity supporting the further development of the gamma H2AX ex vivo assay as a biomarker for individualized treatment. (C) 2017 Elsevier B.V. All rights reserved.

Original language	English
Pages (from-to)	386-394
Number of pages	9
Journal	Radiotherapy and Oncology
Volume	124
Issue number	3
DOIs	https://doi.org/10.1016/j.radonc.2017.08.020
Publication status	Published - Sept 2017
Event	15th International Wolfsberg Meeting on Molecular Radiation Biology/Oncology - SWITZERLAND, Switzerland Duration: 17 Jun 2017 → 19 Jun 2017

Keywords

Ex vivo
gamma H2AX foci
Intrinsic radiation sensitivity
Prostate cancer
Intra-patient variability
Inter-patient variability
Personalized radiation oncology
DOUBLE-STRAND BREAKS
TUMOR HETEROGENEITY
DNA-DAMAGE
CELL-SURVIVAL
HISTONE H2AX
DEPHOSPHORYLATES GAMMA-H2AX
INTRINSIC RADIOSENSITIVITY
RADICAL PROSTATECTOMY
CERVICAL-CARCINOMA
REPAIR

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10.1016/j.radonc.2017.08.020

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De-Colle, C., Yaromina, A., Hennenlotter, J., Thames, H., Mueller, A.-C., Neumann, T., Stenzl, A., Scharpf, M., Fend, F., Ricardi, U., Baumann, M., Zips, D., & Menegakis, A. (2017). Ex vivo gamma H2AX radiation sensitivity assay in prostate cancer: Inter-patient and intra-patient heterogeneity. Radiotherapy and Oncology, 124(3), 386-394. https://doi.org/10.1016/j.radonc.2017.08.020

@article{0874447400de4453b2b786c0805e2db1,

title = "Ex vivo gamma H2AX radiation sensitivity assay in prostate cancer: Inter-patient and intra-patient heterogeneity",

abstract = "Introduction: The aim of the study is to assess inter-patient and intralpatient heterogeneity in tumour cell radiosensitivity using the ex vivo gamma H2AX assay in prostate cancer specimens.Methods: Excised specimens from untreated prostate cancer patients were cultivated 24 h in media, irradiated ex vivo and fixed after 24 h. Residual gamma H2AX foci were counted and the slope of the dose response was calculated. Intra-patient heterogeneity was studied from three to seven different biopsies.Results: In pathology-confirmed tumour samples from 21 patients the slope of residual gamma H2AX foci and radiation dose showed a substantial heterogeneity ranging from 0.82 to 3.17 foci/Gy. No correlation was observed between the slope values and the Gleason score (p = 0.37), prostate specific antigen (p = 0.48) and tumour stage (p = 0.89). ANOVA indicated that only in 1 out of 9 patients, biopsies from different tumour locations yielded statistically significant differences. Variance component analysis indicated higher inter-patient than intra-patient variability. Bootstrap simulation study demonstrated that one biopsy is sufficient to estimate the mean value of residual gamma H2AX per dose level and account for intra-patient heterogeneity.Conclusions: In prostate cancer inter-patient heterogeneity in tumour cell radiation sensitivity is pronounced and higher than intra-patient heterogeneity supporting the further development of the gamma H2AX ex vivo assay as a biomarker for individualized treatment. (C) 2017 Elsevier B.V. All rights reserved.",

keywords = "Ex vivo, gamma H2AX foci, Intrinsic radiation sensitivity, Prostate cancer, Intra-patient variability, Inter-patient variability, Personalized radiation oncology, DOUBLE-STRAND BREAKS, TUMOR HETEROGENEITY, DNA-DAMAGE, CELL-SURVIVAL, HISTONE H2AX, DEPHOSPHORYLATES GAMMA-H2AX, INTRINSIC RADIOSENSITIVITY, RADICAL PROSTATECTOMY, CERVICAL-CARCINOMA, REPAIR",

author = "Chiara De-Colle and Ala Yaromina and Joerg Hennenlotter and Howard Thames and Arndt-Christian Mueller and Tim Neumann and Arnulf Stenzl and Marcus Scharpf and Falko Fend and Umberto Ricardi and Michael Baumann and Daniel Zips and Apostolos Menegakis",

year = "2017",

month = sep,

doi = "10.1016/j.radonc.2017.08.020",

language = "English",

volume = "124",

pages = "386--394",

journal = "Radiotherapy and Oncology",

issn = "0167-8140",

publisher = "Elsevier Ireland Ltd",

number = "3",

note = "15th International Wolfsberg Meeting on Molecular Radiation Biology/Oncology ; Conference date: 17-06-2017 Through 19-06-2017",

}

De-Colle, C, Yaromina, A, Hennenlotter, J, Thames, H, Mueller, A-C, Neumann, T, Stenzl, A, Scharpf, M, Fend, F, Ricardi, U, Baumann, M, Zips, D & Menegakis, A 2017, 'Ex vivo gamma H2AX radiation sensitivity assay in prostate cancer: Inter-patient and intra-patient heterogeneity', Radiotherapy and Oncology, vol. 124, no. 3, pp. 386-394. https://doi.org/10.1016/j.radonc.2017.08.020

TY - JOUR

T1 - Ex vivo gamma H2AX radiation sensitivity assay in prostate cancer

T2 - 15th International Wolfsberg Meeting on Molecular Radiation Biology/Oncology

AU - De-Colle, Chiara

AU - Yaromina, Ala

AU - Hennenlotter, Joerg

AU - Thames, Howard

AU - Mueller, Arndt-Christian

AU - Neumann, Tim

AU - Stenzl, Arnulf

AU - Scharpf, Marcus

AU - Fend, Falko

AU - Ricardi, Umberto

AU - Baumann, Michael

AU - Zips, Daniel

AU - Menegakis, Apostolos

PY - 2017/9

Y1 - 2017/9

N2 - Introduction: The aim of the study is to assess inter-patient and intralpatient heterogeneity in tumour cell radiosensitivity using the ex vivo gamma H2AX assay in prostate cancer specimens.Methods: Excised specimens from untreated prostate cancer patients were cultivated 24 h in media, irradiated ex vivo and fixed after 24 h. Residual gamma H2AX foci were counted and the slope of the dose response was calculated. Intra-patient heterogeneity was studied from three to seven different biopsies.Results: In pathology-confirmed tumour samples from 21 patients the slope of residual gamma H2AX foci and radiation dose showed a substantial heterogeneity ranging from 0.82 to 3.17 foci/Gy. No correlation was observed between the slope values and the Gleason score (p = 0.37), prostate specific antigen (p = 0.48) and tumour stage (p = 0.89). ANOVA indicated that only in 1 out of 9 patients, biopsies from different tumour locations yielded statistically significant differences. Variance component analysis indicated higher inter-patient than intra-patient variability. Bootstrap simulation study demonstrated that one biopsy is sufficient to estimate the mean value of residual gamma H2AX per dose level and account for intra-patient heterogeneity.Conclusions: In prostate cancer inter-patient heterogeneity in tumour cell radiation sensitivity is pronounced and higher than intra-patient heterogeneity supporting the further development of the gamma H2AX ex vivo assay as a biomarker for individualized treatment. (C) 2017 Elsevier B.V. All rights reserved.

AB - Introduction: The aim of the study is to assess inter-patient and intralpatient heterogeneity in tumour cell radiosensitivity using the ex vivo gamma H2AX assay in prostate cancer specimens.Methods: Excised specimens from untreated prostate cancer patients were cultivated 24 h in media, irradiated ex vivo and fixed after 24 h. Residual gamma H2AX foci were counted and the slope of the dose response was calculated. Intra-patient heterogeneity was studied from three to seven different biopsies.Results: In pathology-confirmed tumour samples from 21 patients the slope of residual gamma H2AX foci and radiation dose showed a substantial heterogeneity ranging from 0.82 to 3.17 foci/Gy. No correlation was observed between the slope values and the Gleason score (p = 0.37), prostate specific antigen (p = 0.48) and tumour stage (p = 0.89). ANOVA indicated that only in 1 out of 9 patients, biopsies from different tumour locations yielded statistically significant differences. Variance component analysis indicated higher inter-patient than intra-patient variability. Bootstrap simulation study demonstrated that one biopsy is sufficient to estimate the mean value of residual gamma H2AX per dose level and account for intra-patient heterogeneity.Conclusions: In prostate cancer inter-patient heterogeneity in tumour cell radiation sensitivity is pronounced and higher than intra-patient heterogeneity supporting the further development of the gamma H2AX ex vivo assay as a biomarker for individualized treatment. (C) 2017 Elsevier B.V. All rights reserved.

KW - Ex vivo

KW - gamma H2AX foci

KW - Intrinsic radiation sensitivity

KW - Prostate cancer

KW - Intra-patient variability

KW - Inter-patient variability

KW - Personalized radiation oncology

KW - DOUBLE-STRAND BREAKS

KW - TUMOR HETEROGENEITY

KW - DNA-DAMAGE

KW - CELL-SURVIVAL

KW - HISTONE H2AX

KW - DEPHOSPHORYLATES GAMMA-H2AX

KW - INTRINSIC RADIOSENSITIVITY

KW - RADICAL PROSTATECTOMY

KW - CERVICAL-CARCINOMA

KW - REPAIR

U2 - 10.1016/j.radonc.2017.08.020

DO - 10.1016/j.radonc.2017.08.020

M3 - Article

C2 - 28919005

SN - 0167-8140

VL - 124

SP - 386

EP - 394

JO - Radiotherapy and Oncology

JF - Radiotherapy and Oncology

IS - 3

Y2 - 17 June 2017 through 19 June 2017

ER -