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RNA and Heat Shock Protein Biomarkers in Radiation-induced Fibrosis in Breast Cancer

Not Applicable
Completed
Conditions
Breast Carcinoma
Fibrosis
Interventions
Other: skin biopsies
Other: blood samples
Registration Number
NCT03000764
Lead Sponsor
Institut de Cancérologie de Lorraine
Brief Summary

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.

Detailed Description

Not available

Recruitment & Eligibility

Status
COMPLETED
Sex
Female
Target Recruitment
20
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 & Design

Study Type
INTERVENTIONAL
Study Design
SINGLE_GROUP
Arm && Interventions
GroupInterventionDescription
Biomarkersblood samples-
Biomarkersskin biopsies-
Primary Outcome Measures
NameTimeMethod
Global mRNA alternative splicing and expression of non-coding RNAs profiles in healthy dermal fibroblasts6 months

frequency of inclusion of individual exons within the set of mRNA isoforms (overall splicing profile) and variation in expression of non-coding RNAs

Secondary Outcome Measures
NameTimeMethod
Transcriptomic signature of pathological induced fibrosis when comparing the primary outcome between the two populations on cultured fibroblasts6 months
Transcriptomic signature of pathological induced fibrosis when comparing the primary outcome between the two populations on serum6 months
Individual radiosensitivity on healthy dermal fibroblasts6 months

The micronuclei will be counted 24 hours after ex vivo irradiation with an indirect immunofluorescence assay (53BP1 + pATM antibodies)

Comparison of the overall mRNA splicing and non-coding RNA expression profiles between non irradiated and irradiated dermal fibroblasts in the same individual6 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 patients6 months

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)

seric HSP proteins potentially predictive of pathological induced fibrosis6 months

HSP27, HSP70 and αB crystalline measured in serum with ELISA assay

Cellular distribution of specific HSP on fibroblast culture in each group of patients6 months

immunolabeling of HSPs and spatial mapping and sub-nuclear distributions

Potential interactions between DNA damage response proteins and candidate HSP6 months

Collocation of HSPs with pATM and 53-BP1 (confocal microscopy / FLIM)

Trial Locations

Locations (1)

Institut de Cancérologie de Lorraine

🇫🇷

Vandœuvre-lès-Nancy, France

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