NorMa: Normal mammary tissue and cancer development

­­NorMaNormal Mammary tissue

Main objective:

Molecular studies on breast tissue from healthy women, with varying risk of breast cancer development.

Aim of study:

The study is defined in two set of aims; short-term aims and long-term aims.

1. Short-term aims:

  • Reveal the differentiation route of epithelial cells
  • Explore the pattern of development of different breast cancer sub-types
  • Detect how the micro environment affect the differentiation of epithelial cells and cancer sub-type development

2. Long-term aims:

  • Reveal the molecular profiles of normal breast tissue associated with increased risk of breast cancer development
  • Achieve increased knowledge of breast cancer initiation and progression

Objects of investigation:

Three groups of women is investigated

  • Group 1. Low-risk: Reduction mammoplastics
  • Group 2. Increased-risk: Benign lesions
  • Group 3. High-risk: a) Breast cancer patients (Ipsilateral, contralateral) b) Profylactic mastectomies

 Collected materials:

  • Healthy breast tissue for isolation of viable cells/ organoids
  • Healthy breast tissue (fresh-frozen)
  • Tumor tissue (fresh-frozen)
  • Biopsy from healthy tissue (contralateral)
  • Blood samples (EDTA, plasma, serum)
  • Skin (Ipsilateral, contralateral)  

Methods:

Micro enviroment (ME) arrays: What micro environment will cause the progenitor cells to differentiate into luminal or myoepithelial cells? 

 

(A) Microarrays are typically fabricated on substrates such as poly-dimethylsiloxane (PDMS) or functionalized glass slides (step 1) to physically adsorb or covalently bind mixtures of growth factors and ECM components that are spotted using a robotic spotter or a printer from conventional multi-titer plate libraries (step 2). After passivation of the array to avoid nonspecific cell attachment, stem cells are cultured onto the arrays (step 3) and the effect of individual protein mixtures on proliferation or differentiation can be assessed using fluorescence stainings (step 4). Reprinted with permission from Reference 11. Copyright 2009 Royal society of Chemistry. (B) An example of primary human neural progenitors cultured on laminin (Ln) spots illustrating the strong influence of different growth factors combinations or concentrations on cell proliferation and the expression of neurogenic (TUJ1, in green) and glial (GFAP, in red) markers. Adapted with permission from Reference 10. Copyright 2005 Nature Publishing Group.

 

Micro environment array. A plate with 384-wells is coated with a hydrophobic layer to permit protein adhesion. The ME-array is printed with 192 highly specific combinations of different micro environments. Epithelial progenitor cells are grown to allow differentiation.

 

Project members:  Marie Fongaard, Grethe I. Grenaker Alnæs, Torben luders, Xavier Tekpli

 External colaborators: