RNA and Heat Shock Protein Biomarkers in Radiation-induced Fibrosis in Breast Cancer (SPLICI-Rad)

Study of RNA and Heat Shock Protein (HSP) Derived Biomarkers in Radiation-induced Fibrosis in Patients Treated for Breast Cancer.

The purpose of this study is to seeking a molecular signature of pathological radiation induced fibrosis based on the response of skin fibroblasts after irradiation, comparing two groups of patients distinguished by their individual radiosensitivity. The signature will integrate recent insights in terms of alternative splicing of mRNAs and level of expression of non-coding RNAs, particularly long non-coding RNAs, snRNAs, snoRNAs and microRNAs. In each group each expression patterns of candidate HSP proteins potentially predictive of pathological radiation induced fibrosis (HSP27, HSP70, αβ crystalline) in the serum and on cell culture will be characterized.

Study Overview

• Status: Completed
• Conditions: Fibrosis; Breast Carcinoma
• Intervention / Treatment: Other: skin biopsies; Other: blood samples

• Study Type: Interventional
• Enrollment (Actual): 20
• Phase: Not Applicable

Contacts and Locations

Study Locations

• France
  • Vandœuvre-lès-Nancy, France, 54519
    • Institut de Cancérologie de Lorraine

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 70 years (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: Female

Description

Inclusion Criteria:

• women
• age ≥ 18 and <70 years old
• non metastatic disease
• ECOG performance status 0 or 1
• chest size ≤ 110 cm et bra size <D
• absence of reconstructive breast surgery
• patient able to undergo blood samples (haematological conditions allowing blood sample)
• non-evolving carcinological disease
• absence of systemic inflammatory disease (other than scleroderma) or diabetes
• no inflammatory ou infectious flare on biopsy site at the time of inclusion
• invasive or in situ breast carcinoma
• ability to provide an informed written consent form
• affiliation to a social security system

Then stratification into two groups :

group 1 : radio-sensitive patients

• Post-operative radiotherapy completed at least 6 months ago AND
• radiation induced dermal and/or soft tissue toxicity (dermatitis, fibrosis, atrophy) rated > 2 (CTCAE v4.0 scale)

group 2 : radio-tolerant (control) patients

• Post-operative radiotherapy completed more than 4 years ago AND
• radiation induced dermal and/or soft tissue toxicity (dermatitis, fibrosis, atrophy) rated ≤1 (CTCAE v4.0 scale) .

Exclusion Criteria:

• age <18 or > 70 years old
• evolutive cancer / metastatic disease
• chest size > 110 cm et bra size ≥ D
• previous reconstructive breast surgery
• ECOG performance status > 1
• systemic inflammatory disease or diabetes
• inflammatory ou infectious flare on biopsy site at the time of inclusion, very significant ulceration in the treated breast
• anemic patients
• use of oral anticoagulants
• pregnant or likely to be in 6 months
• patients deprived of liberty or under supervision
• non-affiliation to a social security system

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Other
• Allocation: N/A
• Interventional Model: Single Group Assignment
• Masking: None (Open Label)

Number of Arms

1

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Biomarkers	Other: skin biopsies; Biopsies (12 G) will be performed : in non-irradiated breast skin; in irradiated breast skin; Other: blood samples; blood samples are collected: 10 ml in EDTA tube; 2,5 ml in PAXgene Blood RNA tube; 4 ml in EDTA tube

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Global mRNA alternative splicing and expression of non-coding RNAs profiles in healthy dermal fibroblasts	frequency of inclusion of individual exons within the set of mRNA isoforms (overall splicing profile) and variation in expression of non-coding RNAs	6 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Transcriptomic signature of pathological induced fibrosis when comparing the primary outcome between the two populations on cultured fibroblasts		6 months
Transcriptomic signature of pathological induced fibrosis when comparing the primary outcome between the two populations on serum		6 months
Individual radiosensitivity on healthy dermal fibroblasts	The micronuclei will be counted 24 hours after ex vivo irradiation with an indirect immunofluorescence assay (53BP1 + pATM antibodies)	6 months
Comparison of the overall mRNA splicing and non-coding RNA expression profiles between non irradiated and irradiated dermal fibroblasts in the same individual		6 months
Changes in cellular distribution of the main non-coding RNAs whose expression varies significantly within the pre-identified signature between the 2 groups of patients	The cellular distribution is defined as the compartment (nucleoplasm, nucleolus, intra-nuclear corpuscles, cytosol, RE, mitochondria ...) marked by the fluorescent probe labeled to the non-coding RNAs of interest (RNA-FISH)	6 months
seric HSP proteins potentially predictive of pathological induced fibrosis	HSP27, HSP70 and αB crystalline measured in serum with ELISA assay	6 months
Cellular distribution of specific HSP on fibroblast culture in each group of patients	immunolabeling of HSPs and spatial mapping and sub-nuclear distributions	6 months
Potential interactions between DNA damage response proteins and candidate HSP	Collocation of HSPs with pATM and 53-BP1 (confocal microscopy / FLIM)	6 months

Collaborators and Investigators

• Sponsor: Institut de Cancérologie de Lorraine
• Investigators: Principal Investigator: VOGIN GUILLAUME, MD, PhD, Institut de Cancérologie de Lorraine; Principal Investigator: BEHM-ANSMANT Isabelle, PhD, UMR 7365 CNRS-Université de Lorraine, IMoPA

Study record dates

Study Major Dates

• Study Start (Actual): May 10, 2017
• Primary Completion (Actual): April 18, 2018
• Study Completion (Actual): April 25, 2018

Study Registration Dates

• First Submitted: December 19, 2016
• First Submitted That Met QC Criteria: December 19, 2016
• First Posted (Estimate): December 22, 2016

Study Record Updates

• Last Update Posted (Actual): August 8, 2018
• Last Update Submitted That Met QC Criteria: August 7, 2018
• Last Verified: August 1, 2018

More Information

Terms related to this study

• Keywords: Radiotherapy; Gene expression; Skin biopsy; RNA biomarker; HSP biomarker; Exon junction array; Radiosensitivity prediction
• Additional Relevant MeSH Terms: Pathologic Processes; Skin Diseases; Neoplasms; Neoplasms by Site; Breast Diseases; Fibrosis; Breast Neoplasms
• Other Study ID Numbers: 2016-A00592-49

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No