Patient case: 35-year-old female with Breast cancer. Genetic variant: NRAS Amplification. Other conditions: None.

Clinical trial: Circulating miRNAs. ICORG 10-11, V2

Summary: To identify a panel of circulating miRNA markers which could help identify those breast cancer patients who are most likely to respond well to neoadjuvant and adjuvant chemotherapy, and indeed serve as an overall prognostic factor and stratify patients into risk categories which would further guide their management. Similarly, the investigators aim to identify a panel of circulating miRNA markers which could monitor patient's response to chemotherapy and hormonal therapies. Ideally a suitable panel of markers would show significant changes in expression level in good-responders whilst little or no change would be observed in miRNA expression in non-responders.

Detailed description: Primary Objectives:
  1. To identify a panel of miRNAs, detectable in the circulation, which are altered in breast cancer patients
  2. To identify specific combinations of miRNAs ('signatures') which associate with breast cancer intrinsic subtypes, and thereby could aid in prognostication and treatment planning on an individual patient basis.
  Secondary Objective:
  1. To determine if systemic miRNA analysis can be used as a biomarker for monitoring response to chemotherapy, in the neoadjuvant setting and in patients who present with breast cancer recurrence and are treated with upfront chemotherapy
  This is a prospective cohort studies, involving two study cohorts:
  Cohort 1: Newly diagnosed breast cancer patients, Cohort 2: Recurrent breast cancer patients
  Blood Sampling:
  -  1st (baseline) blood sample at presentation before commencing neoadjuvant (cohort 1) or systemic (cohort 2) treatment.
  -  2nd blood sample midway through their chemotherapy treatment (after 2nd cycle if they are enrolled in a 4 cycle regimen, or after 4th cycle if they are prescribed an 8 week regimen).
  -  3rd blood sample post-chemotherapy and before surgery (if applicable).
  -  4th blood sample 2-4 weeks after surgery, or after last blood sampling if surgery is not envisaged
  -  5th blood sample 12-18 month after surgery or after 3rd blood sampling if surgery is not envisaged.
  GUH only: Tissue samples will be taken at time of biopsy and/or at time of surgery.
  All samples will be processed for miRNA quantification - a panel of 9 cancer-specific miRNAs will be measured in each sample, and the change in each patient's miRNA expression levels monitored over the course of their treatment.
  Blood samples will be processed for miRNA analysis, which involves:
  1. Lysis using Trizol
  2. RNA isolation
  3. Assessing concentration and integrity of RNA using Nanodrop spectrophotometry
  4. cDNA synthesis (using miRNA specific stem loop primers)
  5. PCR amplification and relative quantification (using miRNA specific probes)

Eligibility: Inclusion Criteria:
  Patients must satisfy the following criteria:
  1. All patients with a new diagnosis of breast cancer, who are destined to undergo neoadjuvant chemotherapy.
  2. Patients with breast cancer recurrence or disease progression who will receive up-front chemotherapy.
  3. Patients must be aged 18 years or over.
  4. Patients must be able to give written informed consent.
  Exclusion Criteria:
  1. Patients who do not fulfil the inclusion criteria mentioned above

Relevance to patient case: not relevant