PMC1215484_01
Pedigree of the patient (indicated by arrow). She, one maternal aunt and one maternal cousin had breast cancer diagnosed at 36, 44 and 41 years old, respectively, as indicated by the half-filled symbols, and her aunt died of the disease. Her cousin underwent lumpectomy followed by chemotherapy, radiotherapy and is presently on tamoxifen. Her mother had breast cancer in both breasts, diagnosed at ages 41 and 42, as indicated by the completely filled symbol. She underwent bilateral mastectomy and hysterectomy followed by chemotherapy and radiotherapy and died of the disease at age 44. A second maternal aunt was diagnosed with colon cancer at age 52 (light half-filled symbol) and breast cancer at age 55 (dark half-filled symbol). Based on this pattern of familial cancer the patient was considered to be at high risk of developing breast cancer and was entered into the low power MRI screening validation and feasibility study. Following her diagnosis, she was confirmed as carrying a Q1200X mutation in the BRCA1 gene.
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F1
The patient was a 35.7 year old woman who presented with a very strong family history of breast cancer as depicted in Figure 1, and negative physical and mammographic examination.
Ultrasound of the MRI-detected lesion. Following MRI, the patient was scheduled for ultrasound to identify the questionable lesions seen on MRI for possible core biopsy. Under ultrasound the lesion of concern was identified and biopsied at the 1:00 location in the left breast. Additionally, one lesion seen by MRI in the right breast at the 4:00 location was identified and biopsied.
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F2
Under ultrasound, the lesion of concern in the left breast was identified and biopsied, as well as one lesion in the right breast (Figure 2).
Micrographs of the non-diseased primary human mammary epithelial cultures (HMEC) from the BRCA1 mutation carrier. A) Contralateral breast - A cluster of epithelial cells called a mammosphere is shown on the left center of the image sitting on a field of fibroblasts. B) Ipsilateral breast - The original fresh tissue block from which this culture was derived was located 4 cm from the infiltrating ductal carcinoma. The structure shown is a cluster of rounded epithelial cells manifesting a column configuration called "pre-ductal linearization". Both images were captured under Differential Interference Contrast (DIC) optics on a Zeiss Axiovert 100 microscope at a total of 140x magnification.
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F3
After several days in culture without passaging, the epithelial cells began to self-organize, initially forming three-dimensional mammospheres (Figure 3A), and, after 2 weeks in culture, more complex pre-ductal linear columns of epithelial cells (Figure 3B). The tissue explants from both breasts showed similar patterns of behavior (Figure 3).
Comparison of the NER capacity of a PBL sample from our BRCA1 mutation carrier patient with those of a population of disease-free controls. The dark horizontal line indicates the average for the normal population, while the dotted lines indicate upper limits for residual NER activity in patients with the hereditary NER deficiency disease XP (0.50) and the cut-off established in our breast tissue study that identified tumors with high sensitivity and specificity (0.70).
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F4
Analysis of cultured blood lymphocytes from the patient established that they had normal NER capacity (99.6% of the average of our 33 normal samples) (Figure 4).
Comparison of the NER capacities of two samples of normal breast epithelium from our BRCA1 mutation carrier patient with those of a population of disease-free controls who underwent breast reduction mammoplasty. The dark horizontal line indicates the average for the normal population of breast reduction epithelium (BRE), while the dotted lines indicate upper limits for residual NER activity in patients with the hereditary NER deficiency disease XP (0.50) and the cut-off established in our breast tissue study that identified tumors with high sensitivity and specificity (0.70). The patient sample on the left was derived from the ipsilateral (left) breast, while the sample on the right was from the contralateral (right) breast.
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F5
The NER capacity of the ipsilateral breast epithelial sample was 1.05 times the average of our population of BRE controls, clearly exhibiting no overt DNA repair deficiency (Figure 5).