Association of Adjuvant Radiotherapy of Non-metastatic Breast Carcinoma With Immunomodulation and Circulating Tumor Cell Phenotype in Relation to Patient Age.

Study Overview
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Brief Summary
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In a prospective study, the influence of adjuvant radiotherapy in patients with non-metastatic breast carcinoma on the epithelial tumor cells (CETCs) circulating in the blood and their immunohistochemical characteristics depending on age will be investigated. In addition to a histopathological assignment of the CETCs as cells of the primary tumor, major trigger points of the immune system will be exploratively analyzed. For this purpose, blood samples are taken from the patients at different time points after tumor resection and during adjuvant radiotherapy. In addition to the detection, isolation and genetic characterization of the CETCs, the determination of immunological biomarkers by immunophenotyping, among other methods, is planned. Furthermore, analyses of tissue from the primary tumor with respect to immunohistochemical features as well as tumor-infiltrating lymphocytes (TILs) are planned. The results will be classified and correlated especially with regard to patient age. As there are insufficient data available for breast carcinoma regarding radiotherapeutic effects on the immune system depending on patient age, it is of great interest to better understand these molecular biological basics in order to identify potential prognostic biomarkers.

Detailed Description
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The primary aim of this prospective-explorative phase 0 study is to investigate the influence of adjuvant radiotherapy in patients with non-metastatic breast cancer on the circulating epithelial tumor cells (CETCs) in the peripheral blood and their immunohistochemical characteristics in younger high risk (premenopausal) versus older low risk (postmenopausal) patients. In addition, the immunophenotype will be determined using peripheral venous blood mononuclear cells (PBMCs) at different time points during the course of therapy and correlated with the results of CETC analysis. The aim of the study is to identify prognostic and predictive biomarkers for early stage breast cancer patients. No therapeutic consequences will be derived. For this purpose, the phenotype and molecular biological profile of CETC will be investigated and compared with the corresponding characteristics of the primary tumor in order to assign the tumor cells detected in the peripheral blood to the primary tumor. In this context, the number of tumor-infiltrating lymphocytes (TILs) will be correlated with circulating tumor cells and immunophenotype. Another goal is the characterization of individually isolated CETC by RT-PCR. The expression of selected marker genes involved in metastatic processes will be investigated during the course of therapy and compared with that of the primary tumor to identify therapy-associated changes. The in vitro cultivation of so-called tumor spheres and their characterization as a sign of a therapy-resistant phenotype is a secondary objective of the study.

Official Title
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Association of Adjuvant Radiotherapy of Non-metastatic Breast Carcinoma With Immunomodulation and Circulating Tumor Cell Phenotype in Relation to Patient Age.

Conditions
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Tumor Cell, Circulating

Intervention / Treatment
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* Radiation: Adjuvant radiotherapy
* Drug: neoadjuvant and adjuvant chemotherapy


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Primary diagnosis histologically confirmed invasive breast carcinoma pT1-4 pN0 or pN+ cM0 curative-intended breast-conserving therapy (BET) ± (neo-) adj. system therapy with subsequent adjuvant radiotherapy of the breast ± lymphatic drainage system planned informed consent of the patient Exclusion Criteria: DCIS, Inflammatory breast carcinoma Recurrence situation Presence of distant metastases Second malignancies < 10 years prior to diagnosis of breast carcinoma Prior exposure to radiotherapy

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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Female

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| premenopausal patients<br> | Radiation: Adjuvant radiotherapy<br>* Guideline-based therapy of primary non-metastatic early breast carcinoma<br>* Other names: breast-conserving therapy;Drug: neoadjuvant and adjuvant chemotherapy<br>* Guideline-based therapy of primary non-metastatic early breast carcinoma<br>|
| postmenopausal patients<br> | Radiation: Adjuvant radiotherapy<br>* Guideline-based therapy of primary non-metastatic early breast carcinoma<br>* Other names: breast-conserving therapy;Drug: neoadjuvant and adjuvant chemotherapy<br>* Guideline-based therapy of primary non-metastatic early breast carcinoma<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Age-dependent influence of radiotherapy on circulating epithelial tumor cell counts (CETC) and immunophenotype (PBMC). | Measurement of CETC count / µl blood in the course of therapy and correlation with patient age. | observation period 2 years | 
| Longitudinal quantification of CETCs as well as immunophenotype at specific time points during the course of therapy. | Measurement of CETC count / µl blood in the course of therapy and correlation with the cell count of detected T and B cells / µl blood. | observation period 2 years | 
| Comparison of immunohistochemical parameters of the primary tumor with the immunohistochemical and cytological parameters of CETC | Comparison of PD-L1 expression of primary tumor cells with CETC in percent. Comparison of hormone receptor status (estrogen, progesterone) values of primary tumor and CETC in percent. | observation period 2 years | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Recurrence-free survival (RFS) | Recurrences as a function of time (analysis using Kaplan-Meier curves, log-rank test, and Cox regression). | observation period 3 years follow up | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Early Stage Breast cancer, Circulating Tumor Cells, Tumor-infiltrating lymphocytes, Radiotherapy, Immunophenotyping